They can only detect previously identified mutations, and these methods would need adaptation to detect mutants that confer resistance to a growing list of nucleos(t)ide reverse transcriptase inhibitors [110]. 4.2.2.4 HBV genotyping. Currently, there is no indication for performing this as standard of care, except possibly in patients being considered for interferon therapy. It may be more relevant in the future as information on the differences among genotypes emerges. HBV genotypes have been reported to

correlate with Bortezomib spontaneous and interferon-induced HBeAg seroconversion, activity of liver disease, and progression to cirrhosis and HCC [101,111,112]. For example, HBV genotypes C and D are more difficult to treat than genotypes A and B [113,114]. There is also some evidence suggesting an increased pathogenicity of genotype C over B, with a greater likelihood of developing HCC [115,116]. Much of the current data examining the clinical relevance of HBV genotype should be viewed with caution. Many studies were small and cross-sectional in design, comparing two of the major genotypes with each other, and may be affected by referral bias. The predictive values of genotype in prognosis and treatment response have not been evaluated in prospective Luminespib mw trials, and, currently, most clinicians do not base their management

decisions selleck kinase inhibitor on the viral genotype. However, this approach is likely to change as more data become available. Further studies are still needed in this area [117]. HBV is directly carcinogenic and may promote the development of HCC in the absence of cirrhosis, especially in populations where HBV may

have been acquired at birth or in early childhood [53]. High HBV viral loads and low CD4 cell counts may be linked to the development of HCC [54,55]. Screening programmes utilizing serum AFP measurements together with 6-monthly USSs have been demonstrated to improve survival in non-HIV-infected patients [57]. Treatment decisions should be guided by the algorithms in Figures 1 and 2. Central to optimal management is the need for adequate initial assessment of both HBV and HIV status to inform the decision as to whether neither, HBV alone or both viruses require treatment [118]. This includes consideration of the severity of liver disease [119]. In HBV monoinfection, the decision on who to treat is based primarily on the ALT level, liver histology, HBeAg status and HBV DNA level [118–123]. ALT normality should not be used to assume that treatment is not necessary, although raised ALT often reflects HBV-induced inflammation and the need for treatment. As significant liver damage may be present without raised liver enzymes, assessment of liver fibrosis by transient elastometry (e.g.

The resulting PCR amplicons consisted of two types, differing according to size. Comparative sequence analysis and structural prediction of the flagellin amino acid sequences revealed the presence of numerous large gaps in the D2/D3

domains, which located in flagellum surface. Phylogenetic analysis using partial Anti-infection Compound high throughput screening N-terminal flagellin sequences revealed that the Actinoplanes species grouped into three subclusters. The diversity of flagellin gene provides us useful information to discuss the evolution of motile actinomycetes. This study was supported in part by a research grant from the Institute for Fermentation, Osaka (IFO). “
“Saccharomyces cerevisiae was engineered for assembly of minicellulosomes by heterologous expression of a recombinant scaffolding protein from Clostridium cellulovorans and a chimeric endoglucanase E from Clostridium thermocellum. The chimeric endoglucanase E fused with the dockerin domain of endoglucanase B from C. cellulovorans

was assembled with the recombinant scaffolding protein. The resulting strain was able to ferment amorphous cellulose [carboxymethyl-cellulose (CMC)] into ethanol with the aid of β-glucosidase 1 produced from Saccharomycopsis fibuligera. The minicellulosomes assembled in vivo retained the synergistic effect for cellulose hydrolysis. The minicellulosomes containing the cellulose-binding domain were purified by crystalline cellulose affinity in a single Akt inhibitor step. In the fermentation test at 10 g L−1 initial CMC, approximately 3.45 g L−1 ethanol was produced after 16 h. The yield (in grams of ethanol produced per substrate) was 0.34 g g−1 from CMC. This result indicates that a one-step processing of cellulosic biomass in a consolidated bioprocessing configuration is technically feasible by recombinant yeast cells expressing functional

minicellulosomes. Bioethanol is currently one of the most promising alternatives to conventional transport fuels because of its desirable characteristics, Lck such as high octane value and good combustion efficiency (Madhavan et al., 2009). Cellulosic materials of plant origin as a source of bioethanol production are the most abundant utilizable biomass resource. However, as alcohol production from cellulosic materials remains unfeasible economically, the development of a more effective and high-yield ethanol fermentation process is required to bring about a necessary dramatic reduction of production costs (Kondo et al., 2002). One-step conversion of lignocellulose to ethanol with an organism capable of cellulose degradation and efficient fermentation [consolidated bioprocessing (CBP)] would greatly enhance the cost effectiveness of bioethanol production (Lynd et al., 2005).

26; 95% CI 0.07–1.01). There was also a trend to lower HCV viral load in this group, which may go some way to explaining this. Also, in a small French cohort of co-infected women (29% on cART), rate of transmission Ruxolitinib molecular weight did not differ significantly between children

born by vaginal delivery or CS [227]. cART should be given to all HCV/HIV co-infected pregnant women, regardless of CD4 cell count or HIV viral load because of the evidence of increased HIV transmission in co-infected mothers. 6.2.7 Where the CD4 cell count is < 500 cells/μL, cART should be continued if HCV viraemia exists because of the increased risk of progressive HCV-related liver disease. Grading: 1B 6.2.8 Where the pre-cART CD4 cell count was > 500 cells/μL and there is no HCV viraemia or fibrosis, cART should be discontinued. Grading: 2C 6.2.9 Where the CD4 cell count is > 500 cells/μL and there is HCV viraemia and evidence of liver inflammation or fibrosis, continuing cART is preferable because of a benefit on fibrosis progression.

Grading: 2B 6.2.10 Where the CD4 cell count is between 350 and 500 cells/μL and there is no evidence of viraemia, inflammation or fibrosis, continuing cART is recommended. Grading: 1C The decision to continue ARV or not postpartum depends on both HIV and HCV factors. There is consensus amongst guidelines that all persons with active (HCV-viraemic) co-infection should receive cART if their CD4 cell count is < 500 cells/μL [175, 176, 228]. In those women with CD4 cell counts of 350–500 cells/μL

Sorafenib concentration who have cleared infection either spontaneously (around 25%) or after treatment and with a sustained virological response (SVR) and who have normal liver histology as judged by biopsy or hepatic elastometry, consideration should be given to continuing cART where the patient expresses a preference to do so. This is because until the completion of the randomized PROMISE trial, which addresses the question of whether to continue cART postnatally in mothers with CD4 cell counts > 400 cells/μL, there is equipoise as to correct management. In those with CD4 cell counts over 500 cells/μL, who received Methane monooxygenase cART to prevent MTCT, and who are not HCV-viraemic and have no evidence of established liver disease, ARVs can be discontinued. Without additional risk factors (such as alcohol, steatosis) and assuming they do not get re-infected, these women should have no further histological progression of their liver. In women with CD4 cell counts over 500 cells/μL who have established liver disease (inflammation or fibrosis), therapy should be continued. Interruption of ART in the SMART study was shown to lead to a greater risk of non-opportunistic disease-related death, particularly among those with HIV/HCV co-infection.

Prognosis is severe in children, especially in those with age less than 1 year or severe malnutrition.[1] Adult mortality rates are also high in immunocompromised patients.[3, 6] Conversely, elderly patients without underlying disease and young immune-competent

adults are much more likely to have a favorable outcome,[4, 5, 7] as observed in our two patients. Shigellosis, because of its severity, should always be treated, whether bacteremia is found or not. But the global increase in antibiotic resistance limits the choice of drugs.[1] Among www.selleckchem.com/products/MK-2206.html recommended treatments, fluoroquinolones or third-generation cephalosporins are the best choices for Shigella bacteremias, but sensitivity must be confirmed. Due to an absence of controlled studies, there is no consensus on treatment duration. Courses generally range between 5 and 14 days, depending on the severity and duration of symptoms.[3-5] Shigella bacteremia is fortunately uncommon in healthy travelers. When an underlying disease is absent, it should alert the physician to the possibility

of a transient co-morbid condition. These case reports underline the importance for travelers to seek pre-travel advice and be prepared to prompt self-treatment of diarrhea with an antibiotic-containing regimen. The authors thank Dr M. Nesemann for linguistic assistance. The authors state they have no conflicts of interest to declare. “
“Clinical and laboratory findings are described from 77 persons from Nairobi, Kenya, of whom 66 Crizotinib ic50 G protein-coupled receptor kinase were diagnosed with acute Schistosoma mansoni infection following a trip to Mwanza, Tanzania. Unusual ocular symptoms were observed as a rare manifestation of acute schistosomiasis. The outbreak highlights

the risk of swimming in Lake Victoria. In August 2008, the Seventh Day Adventist (SDA) group of churches in East Africa organized a family retreat to Mwanza in Tanzania, located on the shores of Lake Victoria. They were there for several days, during which most of them swam in the lake, having been assured that the water was “safe.” Once the retreat was over, the families returned to their homes all over the East African region and beyond. Approximately 8 weeks after exposure to the lake water, on October 28, 2008, a 10-year-old girl was referred to the Centre for Tropical and Travel Medicine (CTTM) laboratory for malaria and hemoglobin testing. The child presented with general malaise which her mother thought was malaria, but she also had periorbital edema. On examination of the Giemsa-stained blood slide, marked eosinophilia was noted in the absence of malaria. Given the history of recent swimming in Lake Victoria during the church retreat to Mwanza, a blood test for bilharzia antibody was requested, which was positive at a titer of 1 : 1024. The child was put on treatment with praziquantel at a dose of 40 mg/kg daily for 4 days, and her symptoms subsided rapidly.

, 2001) (Fig. 1). The performance of these genetic tools for tagging various Gram-negative bacteria was compared. The three different vectors were chosen for their difference selleck inhibitor in antibiotic selection gene (gentamycin, tetracyclin and kanamycin, respectively) and the opportunities for maintenance as a plasmid (pBBRMCS-5 and pME6031) or integration into the chromosome (pBK-miniTn7). In addition, pBBRMCS-5 (a derivative of the general cloning vector pBBR) is assumed to have a higher copy number than pME6031 (containing the pVS1 replicon). pME6031 was described as being maintainable without the selective

pressure of tetracyclin (Heeb et al., 2000). All vectors were reported to have a broad

host range in Gram-negative bacteria. Pseudomonas putida strain PCL1445, which is an excellent root colonizer and is able to form biofilms on abiotic surfaces such as polyvinylchloride (Kuiper et al., 2004a), was selected to examine the new constructs containing mcherry. Growth curves of the transformed strains did not show an effect of the constructs 3-Methyladenine manufacturer and mcherry expression on growth (data not shown). However, care should be taken when using these plasmids under other growth conditions. As expected, the pME6031-derived plasmid pMP7604 was maintained without antibiotic pressure (no loss was observed), whereas the pBBRMCS-5-derived plasmid pMP7607 showed a loss of 3% in cells of the population after 3 days of subculturing without antibiotic pressure. Qualitative and quantitative analyses showed that all constructs can be used for visualization at the single-cell level and that the intensity of fluorescence resulting from the use of the different selleck chemical genetic constructs correlates with the copy number of the different plasmids and the transposon used (Fig. 2). The mcherry constructs created were shown to be functional in different Pseudomonas spp. (i.e. P. putida PCL1445, P. fluorescens WCS365 and P. aeruginosa PAO1) and the fish pathogen E. tarda, with comparable mCherry production

levels (Fig. 3). In addition, fluorescence was observed during cloning in E. coli. Labeled strains under in vitro (biofilm formation on glass) and in vivo (tomato root colonization) conditions showed that the constructs are well suited for the visualization at the single-cell level (Figs 4 and 5). In addition, tagging with the mcherry plasmid constructs was shown to be useful for the simultaneous visualization with the eGFP-tagged strain of P. putida PCL1445 as shown for biofilms formed on glass and tomato roots (Fig. 5). Also, single strains tagged with eGFP and mCherry were recently shown to be useful for bioreporter studies (Tecon et al., 2009). The vectors constructed in this study could function as markers to locate bacteria in such studies.

A double-strand break is recognized by the sensor protein complex MRN (MRE11-RAD50-NBS1). The sensor recruits ATM, which further activates its targets CHK1/CHK2. A single-strand DNA is sensed by ATRIP (ATR interacting protein) and recruits ATR. ATR also activates CHK1/CHK2. It has been found that acute severe hypoxia (<0.02% O2 for less than 24 h) activates both ATR and ATM without DNA damage.53 It is assumed that the activation of ATR is not transducing DNA damage but directed toward maintaining replication folk stability during severe hypoxia by phosphorylating the replisome components, MCM2 and MCM3.54 However, when cells are re-exposed to oxygen, reactive oxygen species (ROS) are very quickly

generated and damage cellular DNA. In response to the damage, ATM is activated and phosphorylates a downstream protein, CHK2.55,56 The activated CHK2 causes Sorafenib clinical trial G2 cell cycle arrest through phosphorylation of Cdc25C and Cdc2.56 There is a possibility that cancer cells may propagate new genetic alterations caused by reoxygenation-induced ROS if the cells

are insensitive to the G2 arrest.54 The concept of ‘genetic instability’ was introduced to define the cancer cells’ property of new mutations with Ulixertinib mw each cell division. Using tissue cultured cancer cells, Lengauer and Vogelstein first demonstrated that some, but not all, cancer cells continuously change their chromosome numbers with each cell division.57 They termed this type of genetic instability as chromosome instability (CIN). Later, CIN was extended to characterize persistent changes, not only in the number of whole or part chromosomes (whole chromosome instability, W-CIN), but also changes in the structure of chromosomes (amplification, deletion and translocations: segmental chromosome instability, S-CIN) during the lifetime of cancer cells. Based on CIN observed in tissue cultures, it is assumed that the frequent occurrence of the chromosomal abbreviations observed in human tumor tissues is caused by CIN mechanisms. Great Florfenicol progress in understanding the molecular basis of CIN has been made through the

use of experimental in vitro and animal models.58 These studies have shown that W-CIN is caused by failures in the correct transmission of chromosomes into daughter cells or the spindle mitotic checkpoint.57 On the other hand, some inherited conditions, such as ataxia telangiectasis, Bloom syndrome, Fanconi anemia and Nijmegen breakage syndrome, are called chromosome instability syndromes and associated with S-CIN and a predisposition to certain types of cancer. Through identification of the genes responsible for these conditions, it is known that S-CIN is caused by mutations of the genes involved in replication, repair and S-phase checkpoints.59 Before CIN was fully understand, another type of genetic instability, microsatellite instability (MSI or MIN), had been recognized in a small fraction of cancers.

, 2005). Both genomes also encode proteins (GI:289669426 and GI:289663837) sharing FK228 purchase 30% amino acid sequence identity with the putative T3SS effector RipT (RSc3212), a YopT-like cysteine protease from the betaproteobacterium Ralstonia solanacearum GMI1000 (Poueymiro & Genin, 2009). Close homologues are not found in any other Xanthomonas genomes, but a protein (GI:270492983) from another plant-pathogenic betaproteobacterium, Acidovorax avenae ssp. avenae ATCC 19860, shares 48% sequence identity with the Xvv and Xcm RipT-like proteins. There are some differences between Xcm 4381 and Xvv 702 with respect to their complements of effectors that might contribute to their different host ranges. Xcm 4381 encodes two predicted

YopJ-like C55 cysteine proteases (GI:289670655 and GI:289671144) that are absent from Xvv 702. On the other hand, Xvv 702 encodes a protein (GI:289661936) sharing 87% amino acid sequence identity with Xanthomonas euvesicatoria XopAF (also known as AvrXv3) (Astua-Monge et al., 2000). This gene is absent from Xcm 4381, but shares 35% identity (at the amino acid level) with the HopAF1-like genes found at the integron locus in both Xcm and Xvv. Such differences in effector repertoires

have previously been shown to be significant for host adaptation (Wei et al., 2007; Kvitko et al., 2009; DAPT supplier Lindeberg et al., 2009). For example, HopQ1-1 is present in P. syringae pathovar phaseolicola, where it suppresses immunity in beans, but is absent from P. syringae pathovar tabaci, and triggers defences in tobacco (Ferrante et al., 2009). It is possible O-methylated flavonoid that the differences in effector repertoires of Xcm 4381 and Xvv 702 are significant

for the adaptation of Xcm 4381 to a new host (i.e. banana). It remains to be tested whether the two Xcm 4381 YopJ- and HopR-like proteins suppress defences and whether the Xvv 702 AvrXv3 confers avirulence in banana. The outer membranes of Gram-negative bacteria are covered with lipopolysaccharides (Lerouge & Vanderleyden, 2002). Among different strains of X. campestris pathovar campestris and X. oryzae pathovar oryzae, the lipopolysaccharide biosynthesis locus shows hypervariability arising from horizontal transfer (Patil & Sonti, 2004; Patil et al., 2007). The lipopolysaccharide locus in Xcm 4381 (GenBank: ACHT01000245.1) most closely matches that of Xanthomonas axonopodis pathovar citri 306 (93% nucleotide sequence identity). The lipopolysaccharide locus in Xvv 702 (GenBank: ACHS01000380.1) shows no significant sequence similarity to that of Xcm 4381. It does, however, share 86% nucleotide sequence identity with Xanthomonas albilineans strain GPE PC73 (Pieretti et al., 2009). This is incongruent with the close phylogenetic relationship between Xcm 4381 and Xvv 702 and indicates recent horizontal transfer in one or both strains from independent sources. Any significance of this variation between Xcm 4381 and Xvv 702 for virulence and host specificity remains unclear.

Visualization of the antibody–antigen interaction was achieved using the enhanced chemiluminescent reaction (Agilent Technologies). The affinity-purified antibodies showed cross-reactions with other as yet unidentified polypeptides in E. coli extracts. Blue-native (BN)-PAGE was performed with 5–15% gradient gels according to Schägger & von Jagow (1991). Far-UV CD spectra were recorded on a Jasco J710 spectropolarimeter. Spectra of purified FocA (0.3 mg mL−1) were recorded in 20 mM Tris-HCl, 150 mM NaCl, 0.2 mM EDTA,

pH 8, at 20 °C in a 0.5-cm cuvette. Sirolimus research buy Before recording spectra, FocA was centrifuged at 100 000 g for 1 h. The α-helical content of FocA based on the CD spectrum was determined using the program cdnn (Böhm et al., 1992). The β-galactosidase see more activity was determined and calculated according to Miller (1972). Each experiment was performed three times independently, and the activities for each sample were determined in triplicate. The activities are presented with SDs. Plasmids for the overproduction of N- (FocAStrep–N) and C-terminally (FocAStrep–C) Strep-tagged FocA protein were constructed as described (see Materials and methods).

In order to assess whether FocAStrep–N and FocAStrep–C were functional in vivo, plasmids pASK-IBA5focA and pASK-IBA3focA were introduced into E. coli strain RM201 (ΔfocA-pflB) containing a single-copy fdhF∷lacZ transcriptional fusion to monitor alterations in the intracellular formate concentration. RM201 cannot generate formate endogenously (Sawers & Böck, 1989) and therefore the fdhF promoter cannot be activated by formate-dependent FhlA unless formate is supplied exogenously (Rossmann et al., 1991). When grown anaerobically with glucose, but

in the absence of exogenous formate, RM201 λfdhF∷lacZ showed a basal β-galactosidase activity of approximately ID-8 30–35 U, regardless of which plasmid was transformed into the strain (Table 2). Inclusion of 50 mM formate in the anaerobic growth medium resulted in a β-galactosidase activity of approximately 1000 U for the strain transformed with the empty vectors pASK-IBA5 and pASK-IBA3 (Table 2). This high β-galactosidase enzyme activity indicates that formate was transported into the cells in the absence of FocA, representing the transport of formate by an as yet unidentified system(s) and diffusion of undissociated formic acid (see also Suppmann & Sawers, 1994). Introduction of the tagged FocA derivatives into the RM201 λfdhF∷lacZ increased β-galactosidase activity roughly 2–2.5-fold (Table 2). This increase indicates that the intracellular formate levels increased in the presence of both FocAStrep–N and FocAStrep–C, and demonstrates that both proteins were active in importing exogenous formate into anaerobic E. coli cells.

IFN-γ inhibits EC growth as well as the expression of MMP-2 and MMP-9.[41] It can also induce expression of anti-angiogenic chemokines, such as CXCL10 and CXCL11 and down-regulate expression of pro-angiogenic CXCL12 chemokine.[1] In RA, other chemokines, such as CCL21, fractalkine and MIF mediate the synovial angiogenesis and migration of inflammatory leukocytes into the synovium.[86, 87] MIF has drawn

significant attention selleckchem recently. This chemokine is involved in macrophage activation and cytokine production.[73] MIF is primarily produced by synovial macrophages and is involved in macrophage-derived synovial angiogenesis.[73, 88] MIF acts via the induction of VEGF and IL-8/CXCL8 release by RA synovial fibroblasts.[89] Moreover, IL-8 is an angiogenic factor. This cytokine seems to be an important factor in which synovial tissue macrophages derive chemotactic activity in ECs, so that angiogenesis could be significantly decreased if IL-8 is immunodepleted.[90] A disintegrin and metalloproteinases (ADAMs) comprise a new family of metalloproteinases, responsible for liberating a variety of cell surface expressed proteins. ADAMs has been implicated in several inflammatory reactions as RA.[91]

Several recent studies have demonstrated the effect of cytokines, such as IL-1β, LDK378 nmr TNF-α and TGF-β, on the expression of ADAMs with thrombospondin motifs 4 (ADAMTS-4) and ADAMTS-5 in FLS. Mimata and colleagues suggest that IL-6 may participate in cartilage destruction in RA as an inducer of ADAMTS-4 expression from FLS.[92] Furthermore, ADAMTS-12 has been observed in the cartilage, synovial fluid and serum of arthritic patients, which may

play an important role in the pathogenesis of arthritis. Nah et al. suggest that ADAMTS12 may be a susceptible gene for RA development.[93] In particular, ADAM10 Adenosine has been shown to cleave various inflammatory and angiogenic mediators from the cell surface, including CXCL16 and CX3CL1. Soluble CXCL16 plays an important role in T cell accumulation and stimulation in RA synovium, and ADAM10 was identified as a major protease responsible for the conversion of CXCL16 from a membrane-bound scavenger receptor to a soluble chemoattractant for T cells.[94] Also, ADAM10 is involved in the constitutive cleavage of CX3CL1 and thereby may regulate the recruitment of monocytic cells to CX3CL1-expressing cell layers.[95] Recently, Isozaki et al. show that ADAM10 is overexpressed in RA and suggest that ADAM10 may play a role in RA angiogenesis.[96] Moreover, other studies have shown that ADAM15 and ADAM17 are active in RA.[97] Komiya et al. in 2005 indicated that among the ADAMs, ADAM15 mRNA was more frequently expressed in the RA patients and also it was expressed in the synovial lining cells, ECs of blood vessels and macrophage-like cells in the sublining layer of RA synovium.

During the exponential growth phase, high PPi levels (approximately

4 ± 2 mM) and relatively low ATP levels (0.43 ± 0.07 mM) were found, and the PPi/ATP ratio decreased 13-fold when the cells entered the stationary phase. Pyruvate kinase activity appeared to be allosterically affected by PPi. Altogether, these findings suggest an important role for PPi in the central energy metabolism of C. saccharolyticus. The extremely thermophilic and strictly anaerobic bacterium Caldicellulosiruptor saccharolyticus belongs to the class of the Clostridia. This bacterium has potential for industrial applications because R788 clinical trial of its ability (1) to produce high hydrogen levels (de Vrije et al., 2007), (2) to grow on complex lignocellulosic material (Ivanova et al.,

2008; de Vrije et al., 2009) and (3) to cometabolize a number of monosaccharides without revealing any form of carbon catabolite repression (van de Werken et al., 2008; VanFossen et al., 2009). For these reasons, C. saccharolyticus recently became the subject of various research projects focusing on renewable energy production (van Niel et al., 2002; Ivanova et al., 2008; de Vrije et al., 2009). The classical Embden–Meyerhof (EM) pathway is the main route of glycolysis in this organism (de Vrije et al., 2007), and analysis of the C. saccharolyticus genome sequence has revealed the presence of all the EM-pathway enzymes (van de Werken et al., 2008). However, the authors of this study indicated further that the C. saccharolyticus genome contains genes coding for an inorganic Ruxolitinib ic50 pyrophosphate (PPi)-dependent pyruvate phosphate dikinase (PPDK) in addition to the pyruvate kinase (PK). Genes coding for typical gluconeogenic enzymes such as pyruvate water dikinase (or PEP synthase) and fructose bisphosphatase next are absent (van de Werken et al., 2008). Interestingly, recent studies on the acetate–lactate metabolic shift in C. saccharolyticus revealed that PPi is a strong modulator of the lactate dehydrogenase (LDH) (Willquist & van Niel, 2010). These observations motivated us to investigate

the role of PPi in the energy metabolism of C. saccharolyticus. PPi-dependent reactions have regularly been described for plants and primitive eukaryotes (Heinonen, 2001). However, little is known about PPi dependency in heterotrophic prokaryotes. Caldicellulosiruptor saccharolyticus DSM 8903 (Rainey et al., 1994) was purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen. For the enzyme and nucleotide measurements, cell extracts (CEs) were prepared from C. saccharolyticus cells, which were cultured batchwise in pH-controlled reactors and in a medium as described previously (van de Werken et al., 2008; Willquist et al., 2009), using glucose as a carbon source (4 g L−1 for the determination of enzyme levels and 10 g L−1 for the determination of nucleotide levels). For the determination of nucleotide levels, the working volume was 1.7 L to minimize the effect of sampling on the culture.