In addition, vital signs, physical examination and laboratory res

In addition, vital signs, physical examination and laboratory results should not have exhibited any evidence of diseases such as psychiatric or cognitive disturbance, cirrhosis or advanced liver disease. Patients agreed not to take any herbal medicine or medication that was contraindicated with VPA for the duration of the study. VPA (valproate sodium; Epival®, Abbott Laboratories, Quebec, Canada) was administered at an initial dose of 500 mg on the first evening and increased to 500 mg twice a day (bid) per os over 1–7 days according to clinical tolerance. VPA serum concentration was assessed in all participants 12 h after the last dose at CHIR-99021 purchase week 1

and every 4 weeks thereafter. If the participant had not reached the therapeutic VPA concentration at the end of the first week, an unscheduled visit was arranged and the drug level retested. The dose was adjusted to the therapeutic range (50–100 μg/mL), which is used in patients with seizures. Venous blood samples were drawn into ethylenediaminetetraacetic acid (EDTA) tubes and processed within 1 h of collection as previously described [16]. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll-Hypaque density centrifugation, washed and re-suspended in phosphate-buffered saline containing heat-inactivated fetal calf serum and then stored in liquid nitrogen until used. CD4 and CD8 T cells were enumerated by flow

cytometry, and plasma viral load was measured using the Roche Amplicor Assay HKI-272 manufacturer (Roche Diagnostics,

Mississauga, Canada) as previously described [16]. A quantitative limiting-dilution culture assay was used to determine the frequency of cells harbouring replication-competent virus as previously described [17]. In brief, PBMC were resuspended at a concentration of 106 cells/mL in RPMI-1640 medium (Sigma, St Louis, MO) supplemented with 10% heat-inactivated fetal calf serum, penicillin (50 U/mL), streptomycin (50 mg/mL), L-glutamine (2 mM), HEPES buffer (10 mM), and recombinant human interleukin-2 (Hoffmann-La Roche, Nutley, NJ) (100 U/mL). Six fivefold dilutions of PBMC were cultured starting at a concentration of 25 × 106 PBMC. A CD3/CD8-bispecific monoclonal antibody, which selectively depletes CD8 T cells while activating CD4 T cells, was added at a final concentration of 1 mg/mL. Cell cultures were Urease incubated at 37°C in a humidified 5% CO2 atmosphere and maintained for a 21-day period with medium changes twice a week. Supernatants were collected weekly prior to the medium change, for the measurement of HIV-1 p24 antigen using an enzyme-linked immunosorbent assay (Vironostika, Bio Mérieux, France). The number of infectious units per log10 billion (IUPB) PBMC was calculated from the pattern of positive wells using the method of maximum likelihood. IUPB were assessed at baseline and at weeks 16 and 48. Quantitative data were summarized using the mean, median, the standard deviation and the range.

Enteritidis 11 (SE11) strain After selecting for the ApR marker

Enteritidis 11 (SE11) strain. After selecting for the ApR marker of the plasmid, the presence of pFOL1111 and the expression of IS30–FljA fusion transposase were confirmed. Subsequently, the insertion donor pFOL1069 from E. coli S17-1 λpir bacteria was conjugated to SE11(pFOL1111)ApR and the transconjugant bacteria were selected for CmR of pFOL1069 and the auxotrophy of the wt S. Enteritidis strain (Fig. 2). In the control experiment, the wt IS30 transposase producer plasmid pJKI132 was used instead of pFOL1111, where only the IS30 transposase was expressed without the FljA domain. In this case, the insertion pattern of

Selleckchem Talazoparib wt IS30 was expected due to the lack of the FljA-specific DNA-binding ability. Performing the transposon mutagenesis on the wt SE11 strain using both the IS30–FljA fusion or the wt

IS30 transposase, the results of three independent experiments (Supporting Information, Table S1) showed that the transpositional frequency mediated by the IS30–FljA fusion transposase (1.78E-04–1.62E-04) was as high as that of the wt IS30 transposase (1.45E-04–8.35E-05). The Doxorubicin data indicated that the fusion transposase maintained full activity compared with the wild type. The CmR transposon mutant Salmonella bacteria carrying pFOL1069 insertion in their genome were selected and tested for motility. As a result of the mutagenesis experiments, altogether 1200 randomly selected not ApRCmR SE11 transposon mutants were isolated and investigated: 600 were generated by the IS30–FljA fusion transposase and 600 by the wt IS30 transposase, respectively. The motility of the mutants was tested individually using the motility agar tube test. Four out of 600 mutants (0.67%) generated by the site-directed system proved to be completely nonmotile. In contrast, no nonmotile mutants were detected among the 600 mutants (<0.16%) generated by the wt IS30

transposase. At least three of the four nonmotile insertional mutants could be considered as independent mutants, originating from three independent experiments (Fig. 3b, column 3). These insertional mutants were confirmed as nonflagellated phenotypes using S. Enteritidis-specific Hg,m antiserum. At the same time, all of the four investigated mutants retained their agglutinability in group D antiserum. Thus, they were confirmed as flagella-free derivatives of SE11. In order to determine the target specificity of the IS30–FljA fusion transposase, altogether 40 different pFOL1069 insertions were cloned (see Materials and methods) and the integration sequences were identified. On analysing the target sequences (Table 1a), it was found that the IS30–FljA fusion transposase show pronounced target specificity. The consensus sequence derived from 24 insertion sites (Table 1b) showed high similarity to the previously determined CIG consensus of insertions of the wt IS30 in the genome of E. coli.

Self-reported symptoms of infection were common in travelers depa

Self-reported symptoms of infection were common in travelers departing Australia and Thailand with a total of 200/843 (23.7%) reporting at least one of the five symptoms in the two weeks prior to departure and 46 (5.5%) reporting two or more of these symptoms. Overall, 3.4% of respondents reported fever, 14.8% reported sore throat, 5.6% reported myalgia, 4.3% reported diarrhea, and 2.1% reported rash. The reporting of fever, sore throat, and myalgia were not significantly different between sites;

however, significant differences were reported for diarrhea (Sydney 3.0%, Bangkok 12.3%, p < 0.001) and rash (Sydney GSI-IX mouse 1.6%, Bangkok 5.3%, p = 0.03). Respondents departing Bangkok reported higher rates of any symptom (32.5%; p = 0.02) and two or more symptoms (12.3%; p = 0.001) compared to respondents departing Sydney (22.4 and 4.4%, respectively). Respondents who were departing from their country of residence were less likely to report any symptom of infection compared to departing visitors (p = 0.04). However, departure country residence was not significantly associated with the reporting of two

or more symptoms of infection (4.5% residents, 6.1% visitors, p = 0.3). Compared to departing visitors, departing residents reported lower rates of diarrhea (residents 1.5%, visitors 6.1%, p = 0.001) and rash (residents 0.9%, visitors 3.0%, p = 0.04) but not other symptoms. Female respondents were GSK3235025 more likely to report sore throat (females 17.8%, males 12.3%, p = 0.03), myalgia (females 7.1%, males 4.0%, p = 0.05), and diarrhea (females 6.1%, males 2.7%, p = 0.02)

than male respondents. A higher proportion GNE-0877 of holiday travelers reported diarrheal symptoms (23/357, 6.4%) compared to other travelers (13/486, 2.7%, p = 0.008). Contact in the 2 weeks prior to departure with a person the respondent perceived as having a fever was reported by 78/843 (9.2%) respondents and was not significantly associated with country of departure (p = 0.8). A significant association was seen in reporting febrile contacts by those departing from their country of residence (13.1%) compared to departing visitors (6.5%, p = 0.001). Of the 78 respondents who reported contact with a febrile person, the majority reported that contact to be a household family member (35.9%), followed by a work colleague (26.9%) and a non-household family member or friend (23.1%). Other contacts included hotel guests and the patients of health care workers. On multivariate logistic regression analysis, variables that were found to be independent predictors of reporting one or more symptoms were found to differ between Australian residents departing Australia, visitors departing Australia, and visitors departing Bangkok, and independent predictors identified from separate models are shown in Table 3.

, 1998; Holo et al, 2001; Maldonado et al, 2003; Diep et al, 2

, 1998; Holo et al., 2001; Maldonado et al., 2003; Diep et al., 2009). Strain-related differences in bactericidal activity affect the susceptibility of other microorganisms to plantaricins and organic acids (Ehrmann et al., 2000; Omar et al., 2006; Nielsen et al., 2010). None of the strains had genes for plantaricins NC8, S, or W (Table 1). With the methodology used, plantaricin A-, EF-, JK-, and N-related genes were detectable in all strains except for TO1001 (Table 1). Similar to the case of TO1001, L. plantarum strain 3.9.1, isolated from an African fermented

food, does not have any of these plantaricin genes (Omar et al., 2006). Certain L. plantarum strains show the following different types of plantaricin-related gene combinations: (1)

plnEF and plnW; (2) plnD, plnEF, plnI, and plnG; (3) plnD, plnJ, plnK, and plnG; (4) PI3K inhibitor plnD, plnEF, plnI, plnK, and plnG; (5) plnA, plnC, plnD, plnEF, plnI, plnJ, plnK, and plnN (Omar et al., 2006; Moghadam et al., BGJ398 2010). Thus, the characteristics of the gene combinations carried for the production of plantaricins in TO1000, TO1002, and TO1003 are unique among the known L. plantarum strains isolated from fermented products. The synthesis of plantaricin A is observed from early exponential to early stationary phase. During stationary phase, the amount of plantaricin A strikingly declines (Diep et al., 1994). The addition of sucrose to the medium enhances production of nisin, another bacteriocin produced by Lactococcus lactis, (Devuyst & Vandamme, 1992). Thus, bacterial growth rate and available nutrients are associated with antimicrobial activity. In fact, the rates of fermentation differed among the four strains at 30 and 60 days of storage (Tables 3 and 4), suggesting that, in addition to the divergence in the available carbohydrates, the capacity for production of organic acids, and

the pH and temperature preferences for growth, antimicrobial activity may also be an important factor in the regulation of silage fermentation quality. Further selleck products studies are needed both to elucidate the production of plantaricins by the TO strains inoculated in silage and to understand their roles in the improvement of silage quality. In conclusion, phenotypic and genotypic differences were present among LAB strains in spite of their belonging to the same species and subspecies, and the fermentation quality of silage inoculated with different conspecific strains differed significantly, supporting the idea that suitable LAB inoculants should be selected on a strain basis. Because TO1002 most effectively improved the fermentation quality in terms of pH decrease, regulation of undesirable microorganisms, and high DM recovery, this strain should be the most suitable inoculant for longer storage of paddy rice silage. The selected L. plantarum subsp.

[9] For example, in Taiwan, 18 years after universal HBV vaccinat

[9] For example, in Taiwan, 18 years after universal HBV vaccination of children began,

the prevalence of chronic HBV infection (HBsAg+ve) in university students has decreased from 14.5% to 1.9%.[6, 10] However, some low-prevalence countries (eg, UK) have not implemented a universal vaccination policy.[11] Thus, many adult travelers born before the implementation of childhood immunization programs (or from countries where such programs do not exist) remain susceptible to HBV infection.[12] Transmission of HBV is through percutaneous or mucosal exposure to HBV-infected blood or bodily fluids including saliva or semen. It may also occur from mother to infant (perinatal), between children (horizontal), via sexual contact, contaminated blood products, contaminated medical equipment, and via sharing needles and injecting apparatus.[13, 14] The incubation period for HBV Hydroxychloroquine clinical trial may be up to 180 days.[14] Acute HBV infection results in symptomatic illness in approximately 30% to 80% of adults (1% fulminant hepatitis),[4] whereas children under 1 year are usually asymptomatic. Symptoms include malaise, fever, jaundice, dark urine, pale stools, right upper quadrant pain, anorexia, and nausea.[14] The risk of chronic disease after HBV infection depends on the age of acquisition.

About Selleckchem CHIR-99021 90% of infected neonates,[8] 30% to 50% of children aged 1 to 4 years, and 1% to 10% of acutely infected adults develop persistent infection.[14, 15] Approximately 15% to 40% with persistent infection develop advanced liver disease, cirrhosis, and/or HCC.[3] Apart from hepatitis A and influenza, HBV infection is among the commonest vaccine-preventable infections in travelers.[16-18] HBV acquisition during travel is associated with travel duration, the immune status of the traveler, and the prevalence of HBV in the destination country.[16] Additionally, specific populations of travelers may be

at greater risk including expatriates, those visiting friends and relatives, and travelers engaging in casual sex, dental surgery, and medical procedures.[16, Digestive enzyme 19-23] Emerging data suggest that travelers seeking urgent, unforeseen medical or dental care are common,[24] which places travelers at risk of HBV infection. The unpredictable nature of emergency care makes it difficult to target advice according to traveler characteristics. While there is little evidence to quantify the risk, travelers may also be exposed to HBV via activities including tattoos, piercings, and acupuncture.[20] HBV infection has been associated with travel. Nine percent of all HBV cases reported in the Netherlands between 1992 and 2003 were travel-related with an estimated incidence of HBV infection of 4.5 per 100,000 travelers.

[9] For example, in Taiwan, 18 years after universal HBV vaccinat

[9] For example, in Taiwan, 18 years after universal HBV vaccination of children began,

the prevalence of chronic HBV infection (HBsAg+ve) in university students has decreased from 14.5% to 1.9%.[6, 10] However, some low-prevalence countries (eg, UK) have not implemented a universal vaccination policy.[11] Thus, many adult travelers born before the implementation of childhood immunization programs (or from countries where such programs do not exist) remain susceptible to HBV infection.[12] Transmission of HBV is through percutaneous or mucosal exposure to HBV-infected blood or bodily fluids including saliva or semen. It may also occur from mother to infant (perinatal), between children (horizontal), via sexual contact, contaminated blood products, contaminated medical equipment, and via sharing needles and injecting apparatus.[13, 14] The incubation period for HBV see more may be up to 180 days.[14] Acute HBV infection results in symptomatic illness in approximately 30% to 80% of adults (1% fulminant hepatitis),[4] whereas children under 1 year are usually asymptomatic. Symptoms include malaise, fever, jaundice, dark urine, pale stools, right upper quadrant pain, anorexia, and nausea.[14] The risk of chronic disease after HBV infection depends on the age of acquisition.

About Cabozantinib clinical trial 90% of infected neonates,[8] 30% to 50% of children aged 1 to 4 years, and 1% to 10% of acutely infected adults develop persistent infection.[14, 15] Approximately 15% to 40% with persistent infection develop advanced liver disease, cirrhosis, and/or HCC.[3] Apart from hepatitis A and influenza, HBV infection is among the commonest vaccine-preventable infections in travelers.[16-18] HBV acquisition during travel is associated with travel duration, the immune status of the traveler, and the prevalence of HBV in the destination country.[16] Additionally, specific populations of travelers may be

at greater risk including expatriates, those visiting friends and relatives, and travelers engaging in casual sex, dental surgery, and medical procedures.[16, Bacterial neuraminidase 19-23] Emerging data suggest that travelers seeking urgent, unforeseen medical or dental care are common,[24] which places travelers at risk of HBV infection. The unpredictable nature of emergency care makes it difficult to target advice according to traveler characteristics. While there is little evidence to quantify the risk, travelers may also be exposed to HBV via activities including tattoos, piercings, and acupuncture.[20] HBV infection has been associated with travel. Nine percent of all HBV cases reported in the Netherlands between 1992 and 2003 were travel-related with an estimated incidence of HBV infection of 4.5 per 100,000 travelers.

A limitation of the study is that the patient population consiste

A limitation of the study is that the patient population consisted of young college students and may not represent the general population. However, their destinations and itineraries mirror populations in other reports.1,2 Additionally, appropriate use of vaccines and medications could only be determined by the amount of information provided in the progress note; therefore, if a recommendation was not documented it was assumed that it did not occur. Lastly, due to the retrospective nature of the study, differences in postgraduate AZD1208 training of the PCPs and the volume of patients they saw could not be controlled. A pharmacist-run

pretravel health clinic can provide more consistent evidence-based care compared to primary care practitioners not specifically trained in travel medicine and may improve patient compliance

with recommendations. Pretravel health is a dynamic and specialized field that requires adequate time, resources, and expertise to Selleck AZD1152 HQPA deliver the best possible care. J. A. G. has received honoraria from speaking for Merck and Sanofi Pasteur. The other authors state that they have no conflicts of interest to declare. “
“Background. Older individuals represent a substantial proportion of international travelers. Because of physiological changes and the increased probability of underlying medical conditions, older travelers might be at higher risk for at least some travel-associated diseases. Methods. With the aim of describing the epidemiology of travel-associated diseases in older adults, medical data were prospectively collected on ill international travelers presenting to GeoSentinel sites from 1997 to 2009. Seven thousand thirty-four patients aged 60 years and over

were identified as older travelers and were compared to 56,042 patients aged 18–45 years, who were used as the young adult reference population. Results. The proportionate morbidity GNA12 of several etiological diagnoses was higher in older ill travelers compared to younger ill, including notably lower respiratory tract infections, high-altitude pulmonary edema, phlebitis and pulmonary embolism, arthropod bites, severe malaria, rickettsiosis, gastritis, peptic ulcers, esophagitis and gastroesophageal reflux disease, trauma and injuries, urinary tract infections, heart disease, and death. In contrast, acute diarrhea, upper respiratory tract infections, flu and flu-like illnesses, malaria, dengue, genital infections, sexually transmitted diseases, and schistosomiasis proportionate morbidities were lower among the older group. Conclusion.

Where

possible, amniocentesis should be deferred until th

Where

possible, amniocentesis should be deferred until the viral load is < 50 HIV RNA copies/mL. The fetal medicine team should discuss management with an HIV physician if the woman is HIV positive and has a detectable viral load. 7.1.4 If not on treatment and the invasive diagnostic test procedure cannot be delayed until viral suppression is complete, it is recommended that women should commence cART to include raltegravir and be given a single dose of nevirapine 2–4 hours prior to the procedure. Grading: 1D The French Pediatric HIV Infection Study Group observed a relative risk of HIV transmission of 1.9 (95% CI 1.3–2.7; P = 0.003) with ‘antenatal procedures’ that Sotrastaurin purchase included amniocentesis, cerclage, laser therapy and amnioscopy [241]. This study was conducted between 1985 and 1993 and, of the 1632 mother–infant

pairs (overall transmission 19%), only 100 mothers had received zidovudine, mostly for advanced HIV infection. There are few studies on the safety of invasive testing in the cART era. A study of 9302 pregnancies in France in 2009 (of which 166 had an amniocentesis) showed that the risk of MTCT in the untreated rose from 16% to 25% in those who had an amniocentesis, in those on zidovudine alone the risk rose from 3.3% to 6.1% and in those on cART there were no transmissions in 81 mothers Alectinib who underwent amniocentesis [242]. Viral load data were not reported, but in other settings suppression of viral load reduces transmission. A further study of nine women in France on cART in 2008 [243] and 17 women on cART in Portugal (1996–2009) showed no transmissions, while transmission occurred in one of six women either not diagnosed with HIV prior to amniocentesis, or not treated prior to the procedure. There are no studies and few case reports in the cART era reporting on chorionic villus sampling (CVS) or cordocentesis [244]. For evidence relating to choice of antiretroviral therapy to reduce transmission risk

associated with amniocentesis, see Section 5.4: Late-presenting woman not on treatment. 7.1.5 External cephalic medroxyprogesterone version (ECV) can be performed in women with HIV. Grading: 2D ECV should be offered to women with a viral load < 50copies/mL and a breech presentation at > 36 + 0 in the absence of obstetric contraindications There is less obstetric risk to the baby and mother when the fetus is head-down at the time of birth. External cephalic version (ECV) is a procedure by which the fetus, which is lying bottom first, is manipulated through the mother’s abdominal wall to the head-down position. If the fetus is not head down by about 36 weeks of pregnancy, ECV reduces the chance that the fetus will present as breech at the time of birth, and thus reduces the chance of Caesarean section. There is no published evidence that helps decision-making regarding ECV in the HIV-positive pregnant woman. For the general maternity population, ECV is recommended [233].

Where

possible, amniocentesis should be deferred until th

Where

possible, amniocentesis should be deferred until the viral load is < 50 HIV RNA copies/mL. The fetal medicine team should discuss management with an HIV physician if the woman is HIV positive and has a detectable viral load. 7.1.4 If not on treatment and the invasive diagnostic test procedure cannot be delayed until viral suppression is complete, it is recommended that women should commence cART to include raltegravir and be given a single dose of nevirapine 2–4 hours prior to the procedure. Grading: 1D The French Pediatric HIV Infection Study Group observed a relative risk of HIV transmission of 1.9 (95% CI 1.3–2.7; P = 0.003) with ‘antenatal procedures’ that Selleck Gefitinib included amniocentesis, cerclage, laser therapy and amnioscopy [241]. This study was conducted between 1985 and 1993 and, of the 1632 mother–infant

pairs (overall transmission 19%), only 100 mothers had received zidovudine, mostly for advanced HIV infection. There are few studies on the safety of invasive testing in the cART era. A study of 9302 pregnancies in France in 2009 (of which 166 had an amniocentesis) showed that the risk of MTCT in the untreated rose from 16% to 25% in those who had an amniocentesis, in those on zidovudine alone the risk rose from 3.3% to 6.1% and in those on cART there were no transmissions in 81 mothers Cabozantinib chemical structure who underwent amniocentesis [242]. Viral load data were not reported, but in other settings suppression of viral load reduces transmission. A further study of nine women in France on cART in 2008 [243] and 17 women on cART in Portugal (1996–2009) showed no transmissions, while transmission occurred in one of six women either not diagnosed with HIV prior to amniocentesis, or not treated prior to the procedure. There are no studies and few case reports in the cART era reporting on chorionic villus sampling (CVS) or cordocentesis [244]. For evidence relating to choice of antiretroviral therapy to reduce transmission risk

associated with amniocentesis, see Section 5.4: Late-presenting woman not on treatment. 7.1.5 External cephalic Bacterial neuraminidase version (ECV) can be performed in women with HIV. Grading: 2D ECV should be offered to women with a viral load < 50copies/mL and a breech presentation at > 36 + 0 in the absence of obstetric contraindications There is less obstetric risk to the baby and mother when the fetus is head-down at the time of birth. External cephalic version (ECV) is a procedure by which the fetus, which is lying bottom first, is manipulated through the mother’s abdominal wall to the head-down position. If the fetus is not head down by about 36 weeks of pregnancy, ECV reduces the chance that the fetus will present as breech at the time of birth, and thus reduces the chance of Caesarean section. There is no published evidence that helps decision-making regarding ECV in the HIV-positive pregnant woman. For the general maternity population, ECV is recommended [233].

saccharolyticus (van de Werken et al, 2008), including two genes

saccharolyticus (van de Werken et al., 2008), including two genes that were annotated to code for a PFK (Csac_1830 and Csac_2366). A sequence alignment of these kinases against PFK-A family members (data not shown) and a more detailed analysis of their amino acid learn more sequences (Fig. 1), revealed that Csac_1830 belongs to the B1 group and contains the typical ATP-dependent PFK amino acid combination G104 and G124 (Chi & Kemp, 2000; Bapteste et al., 2003). On the

other hand, Csac_2366 belongs to the B2 group and contains the typical PPi-dependent PFK amino acid residues D104 and K124 (Chi & Kemp, 2000; Moore et al., 2002). These results strongly suggest that Csac_1830 is an ATP-dependent PFK and that Csac_2366 is a PPi-dependent PFK. The genome also contains the genes encoding a PK (Csac_1831) and a PPDK (Csac_1955), which are both able to perform the catabolic conversion of PEP to pyruvate (van de Werken et al., 2008). PPDK catalyzes a PPi-dependent reaction, whereas PK requires ADP (Fig. 2a). The anticipated PPi dependence of the specified glycolytic steps prompted us to seek other PPi-dependent reactions. The C. saccharolyticus genome lacks any homolog to cytosolic pyrophosphatases (COG0221, COG1227) (Bacillus type; Shintani et al., 1998). Instead, BAY 73-4506 concentration it was found to contain a gene coding

for a V-type proton-pumping membrane-bound pyrophosphatase (H+-PPase, COG3808, Csac_0905). The anticipated H+-PPase has 14 predicted membrane

helices and is expected to be an integral membrane protein. Sequence-based phylogenetic analysis of the H+-PPase revealed Endonuclease it as a member of the K+-insensitive group of the H+-PPase protein family (data not shown; Serrano et al., 2004). To confirm the genome sequence-based predictions, we performed enzyme assays on crude CEs. The activities of PK, PPDK, PPi-PFK, ATP-PFK and membrane-bound PPase could all be detected in CEs of C. saccharolyticus (Table 1). Under the specified assay conditions, the average PPDK activity (0.4 U mg−1) was twice the PK activity. For the ATP- and PPi-dependent PFKs, no significant difference was observed in the activity levels (∼0.05 U mg−1). In line with the presence of a membrane-associated pyrophosphatase, high levels of PPase activity (∼0.15 U mg−1) could be demonstrated in the membrane fraction, while the membrane-free CE barely showed PPase activity. Whether the membrane-bound PPase in C. saccharolyticus couples pyrophosphatase activity to the translocation of protons across the membrane remains to be investigated. The presence of PPi-dependent enzymes in the central metabolic pathway suggested the involvement of PPi as an energy carrier in the metabolism of C. saccharolyticus. Therefore, the levels of both ATP and PPi were determined during growth (Fig. 3). The PPi levels were relatively high (approximately 4 ± 2 mM), while the ATP levels were remarkably low (0.43 ± 0.07 mM).