Only 10 patients (20%) had taken any prophylaxis to prevent malaria. Five of these took a drug that was inappropriate for the country to which they traveled. P. falciparum was most common (74%). P. vivax, P. ovale, and P. malariae were present in five, three, and one case, respectively.

In four cases, definitive species identification was not possible due to the low percent parasitemia, with just a few ring forms present. No coinfections were seen. The majority of patients (52%) had parasitemia <1%; only seven patients had hyperparasitemia (>5%). The maximum parasitemia was 28.6%. All cases with >5% parasitemia were P. falciparum. Nonfalciparum forms made up 42% of patients with ≤1% parasitemia GSK2118436 in vitro and 12% of those with 1% to 5% parasitemia. Gametocytes were rarely identified. Laboratory results are presented in Table 2. Thrombocytopenia and anemia were the most commonly observed laboratory abnormalities. Mild hyponatremia was also relatively common (36% had sodium ≤135 mEq/L and 12% had sodium ≤130 mEq/L). G6PD levels were measured in 10 children; only one was G6PD deficient. Six patients were tested for sickle cell disease; all were negative. Two patients had known sickle trait. Thirty-four children (68%) were hospitalized for treatment of malaria, with a maximum stay of 9 days. Among those with P. falciparum malaria, 75% were hospitalized;

17% stayed for only 1 day. Documentation of treatment available in 41 children: 18 patients (44%) received quinine and doxycycline, eight (19%)

quinine/quinidine and clindamycin, four (9.7%) received atovaquone–proguanil, six (15%) received only one drug (quinine, choloroquine, Cyclin-dependent kinase 3 or primaquine), and the rest received other Z-VAD-FMK cost combinations. Several children received antibiotic therapy due to concern for additional diagnoses. Sixteen patients had received antimalarial therapy previously, although in some cases this was several months prior. One patient received a blood transfusion for anemia (hemoglobin 5.4 mg/dL). No exchange transfusions were performed. One patient received platelet transfusion for a platelet count of 32,000/ul. All of the patients recovered without serious complications. This case series demonstrates the wide spectrum of possible clinical presentations which may be seen with malaria including vomiting, diarrhea, headache, abdominal pain, etc. Gastrointestinal symptoms can be so severe that an intestinal infection may be suspected. Hepatosplenomegaly may be seen; this was less common in our series than in other reports.14,15 In contrast to the report by Viani and Bromberg,14 hyponatremia was not a common finding. One almost universal symptom is fever, either by history or at presentation. Because malaria may present with a wide variety of clinical symptoms, a high index of suspicion is required to ensure prompt diagnosis. Primary care providers seeing patients with a history of fever should always ask about a history of travel and request the appropriate diagnostic tests.

, 2001; Werling & Jungi, 2003; Doherty & Arditi, 2004). In our study, the expressions of TLR2 and TLR4 were increased in both tuberculosis and healthy control groups 3-h poststimulation; however, only TLR2 expression showed a significant difference (P≤0.05) between the two PLX3397 solubility dmso groups. The increase in the expression of the TLR2 gene is more significant in healthy cattle (6.54-fold)

response to stimulation than the tuberculosis group (2.64-fold). This demonstrates that the TLR2 signal pathway may play a larger role in healthy control cattle than the tuberculosis group, possibly resulting in a more efficient proinflammatory gene activation. An important anti-inflammatory cytokine, IL10, was also examined in our study. IL10 downregulates the Th1 type immune response and upregulates the Th2 type in pathogen–host interaction (Jacobs et al., 2000). It has been shown in previous studies, in both humans and mice, that increased

expression of IL10 is associated with the decreased ability of macrophages to restrict the growth of intracellular Mycobacterium (Jamil et al., 2007; Bilenki et al., 2010). Our results show that IL10 expression is more increased in tuberculosis cattle (8.74-fold) than the healthy group (2.90-fold) relative to 3-h poststimulation compared with nonstimulated cells. The differential regulation of IL10 in tuberculosis cattle may reflect vulnerability in the defense of macrophages against M. bovis. The development trend of gene expressions in this study is consistent with that seen by Meade Ku-0059436 price and colleagues, while the different levels of gene expression seen could be attributed to cell populations (PBMCs vs. MDMs), stimulator (bovine tuberculin vs. M. bovis) or comparison of different clinical phenotypes [active BTB infection vs. Latent TB (LTB)]. Our study provides evidence of differences in gene expression between tuberculosis and healthy cattle, which confirms that the innate immune response, TLRs signal pathway

and Th1/Th2 bias are important in BTB infection. The current techniques cannot predict the risk of an individual LTB animal developing into the active disease, and genes implicated in susceptibility and ZD1839 concentration resistance of tuberculosis in cattle cannot point to clear solutions. Building on the differences in gene expression regulation demonstrated in this study, it may provide insights into the diagnosis and treatment of tuberculosis cattle and lead to diagnostics that may characterize the immune response prognostic information in BTB infection. This work was supported by the key project of Ministry of Agriculture, China (Project no. 2009ZX08009-183B), the Beijing Science Foundation of China (Project no. 6101002) and the Natural Science Foundation of China (Project no. 30972164). Y.W. and X.Z. contributed equally to this work. Table S1. Data of IFN-γ ELISA assays (shown as values of optical density at 450 nm, OD 450 nm). Table S2.

, 2001), a characteristic that was confirmed by the sequencing of other S. Typhi strains (Deng et al., 2003; Holt et al., 2009). We will point out the different pseudogenes in each of the following sections. Surprisingly, most of the pseudogenes in S. Typhi are intact and fully functional in S. Typhimurium (McClelland et al., 2001) and could explain in part the loss of host range for serovar S. Typhi. Interestingly, many pseudogenes from S. Typhi are also LDE225 cost conserved in Paratyphi A, a serovar that has the ability to cause enteric fever that afflicts only humans (McClelland et al., 2004; Holt et al., 2009). Most S. Typhimurium strains contain a self-transmissible

virulence plasmid (pSLT) of about 90 kb harbouring virulence genes such as the spv operon, involved in intramacrophage survival, and the plasmid-encoded fimbriae (pef) fimbrial operon (Gulig & Doyle, 1993; Ahmer et al.,

1999; Rotger & Casadesús, 1999). When S. Typhimurium is cured selleck of the plasmid, virulence in the mouse is decreased (Jones et al., 1982) and can be complemented by the sole addition of the spv operon (Gulig et al., 1992) encoding the SpvB toxin (Lesnick et al., 2001). Additionally, S. Typhimurium can also carry multidrug-resistance plasmids of high molecular weight (up to 200 kb) and much smaller plasmids (<20 kb) with unknown functions (Rychlik et al., 2006). The pSLT virulence plasmid is absent in S. Typhi strains. In S. Typhi, incHI plasmids involved in multiple-drug resistance are commonly found (Maher & Taylor, 1993; Fica et al., 1997; Wain et al., 2003). Salmonella enterica serovar Typhi strain CT18 harbours plasmid pHCM1, an incHI1 plasmid of about 218 kb with genes for resistance to antibiotics PAK5 and heavy metals (Parkhill et al., 2001). Salmonella enterica serovar Typhi can also carry cryptic plasmids. Salmonella enterica serovar Typhi strain CT18 harbours the cryptic plasmid pHCM2 of

about 106 kb whose function is unknown, but it is rarely present in other strains (Parkhill et al., 2001; Kidgell et al., 2002a, b). Additionally, a 27-kb linear plasmid was recently isolated in S. Typhi strains originating from Indonesia. This plasmid carries the fljBz66 gene, encoding a flagellin antigen known as H:z66 (Baker et al., 2007b). However, no plasmid has been identified yet in S. Typhi that has been associated with virulence. Integrated bacteriophages represent major loci of genetic diversity in bacterial genomes (Brüssow et al., 2004). Salmonella genomes contain several prophages or prophage remnants with similarity to the lambda, Mu, P2 and P4 families (Thomson et al., 2004; Bossi & Figueroa-Bossi, 2005). The contribution of prophages to S. enterica virulence has been recognized only recently. Some prophages carry nonessential ‘cargo’ genes involved in fitness and/or virulence, including several type three secreted effectors (Ehrbar & Hardt, 2005). Each strain of S.

A single, double dose of tenofovir administered shortly before delivery resulted in plasma concentrations similar to those observed in non-pregnant adults following a standard 300 mg dose and adequate levels in the neonate [66] (see

Section 8: Neonatal management). New data on emtricitabine show that while third-trimester concentrations are lower Alectinib mw than postpartum the absolute concentrations achieved during pregnancy are adequate and dose adjustment is not required [67]. Among the NNRTIs, nevirapine has been extensively studied in pregnancy and plasma concentrations are similar to those in non-pregnant adults [[27],[29]]. No dose adjustment is required when using licensed doses. There are no data on the prolonged release formulation of nevirapine in pregnant women. Efavirenz 600 mg daily has been reported in Veliparib chemical structure one study of 25 pregnant women to result in third-trimester plasma concentrations that were similar to 6–12-week postpartum concentrations in the same women. Cord blood to maternal blood ratio was 0.49 resulting in transplacental concentrations in the therapeutic range [68]. There are currently no data on the pharmacokinetics of etravirine and rilpivirine in pregnant women. PIs are highly protein-bound and placental transfer in humans appears to be limited. During the third trimester of pregnancy,

small reductions in protein binding can significantly increase free drug levels. For example, the protein binding of lopinavir reduces marginally to 99.04%, which results in 17% more unbound lopinavir [69]. It is therefore difficult to interpret the significance of studies that show reduced total plasma levels, with an increased likelihood of trough levels below the target during pregnancy. Compared with postpartum concentrations, third-trimester concentrations of lopinavir (lopinavir 400 mg/ritonavir 100 mg) are reduced by 28%. The protein-free fraction is moderately increased (17%) and, at the standard dose, lopinavir appears to be clinically effective with a wide

variation in individual plasma trough concentrations. A study using the tablet formulation concluded that women Etofibrate taking three tablets bd (lopinavir 600 mg/ritonavir 150 mg) achieved similar area under the curve (AUC) levels to non-pregnant adults taking the standard dose of two tablets bd [70]. The improved bioavailability of the tablet formulation is also found in pregnant women and this, together with the impact of pregnancy on changes in protein binding, increases the protein-free fraction in the third trimester [71]. Cohort studies have suggested that the majority of mothers taking the standard adult dose, even with the capsule formulation, have adequate trough concentrations and achieve an effective virological response [72]. The plasma concentrations of saquinavir achieved with the tablet formulation when boosted by ritonavir appear to be generally therapeutic and no dose adjustment is routinely required.

Amplification reactions generated a fragment of the HIV genome from nucleotide 2057 to 3623, numbering according to the HXB2 (K03455) genome. The resultant PCR products were diluted 1:20 using nuclease-free water. Allele-specific PCR (AS-PCR) reactions for drug-resistant point mutations K103N, Y181C and M184V were performed, using previously published oligonucleotides [8]. Real-time PCR conditions were modified to accommodate use of the Taqman Fast Universal Master Mix (Applied Biosystems, Warrington, UK). For the M184V minority assay, we used a modified protocol learn more with 55% M184V-F1, 30% M184V-F2 and 15% M184V-F3 type-specific

primers. Reactions were cycled using an ABI 7500 Fast Taqman instrument (Applied Biosystems), with an extension time of 35 s, and a reaction volume of 20 μL. Control reactions containing 1% mutant were included with each minority PCR run to provide a sensitivity cut-off point. Control reactions were generated by mixing plasmid DNA containing a subtype B reference sequence, with a second plasmid containing the same sequence with resistance point mutations. These plasmid mixtures were used to generate PCR fragments, akin to targets in

patients’ specimens, and were run alongside Selleck NVP-BEZ235 study specimens. All assay control reagents were identical to those used by Johnson et al. for their original technical validation investigations [8]. However, in contrast to the published methods, we included a 1% mutant control in each minority assay run. The ΔCt of these

reactions provided the sensitivity cut-off experimentally determined for each assay run. Patient-derived material with a ΔCt equal to or less than that recorded for the 1% control was scored as having minority drug resistance. All three assays underwent technical validation in house by triplicate testing of samples for reproducibility and precision, linearity of minority titrations and by testing of samples with known mixed nucleotides at relevant drug resistance codons. We also performed DNA sequencing on all patient-derived pol gene sequences. TDR was defined using mutations according ADP ribosylation factor to a published World Health Organization (WHO) list [13]. Statistical analyses were performed using McNemar’s test for paired data to compare AS-PCR methods vs. standard genotyping. Using HIV genotyping by population DNA sequencing, the K103N mutation was detected in 10 of 165 samples [6.1%; 95% confidence interval (CI) 2.9–10.9%]. Using the minority species assays, we found K103N in 12 of 165 samples (7.3%; 95% CI 3.8–12.4%). Thus, the minority-specific method increased the rate of detection of K103N by 20%; however, this was not statistically significant (P=0.5; 95% CI 0–54%).

We conclude that glucose self-monitoring in the weeks prior to outpatient CF clinic review could become the preferred tool for screening and monitoring of dysglycaemia in adult CF. Copyright © 2011 John Wiley & Sons. “
“In the UK, optometrists examine 17 million people yearly, many of whom will not have consulted a doctor and may have undiagnosed diabetes. Selective testing in optometry practices presents a new detection strategy. The purpose of this research was to ascertain optometrists’ perceptions, attitudes and beliefs towards diabetes and screening, prior to evaluating selleck kinase inhibitor a

pilot service. Focus groups and interviews were conducted with 21 optometrists in Northern England. Analysis was based on grounded theory. Four themes emerged: varying awareness of diabetes and

its early diagnosis, a reluctance in accepting a screening role, organisational barriers in implementing such a service, and controversies around the changing roles of optometrists. Although optometrists’ awareness of diabetes was varied, all had seen patients they suspected of having diabetes and felt that the public under-estimated risks of diabetes. Some felt www.selleckchem.com/products/ly2157299.html that diagnosis of asymptomatic diabetes was unnecessary, although most felt that early diagnosis would be beneficial. Optometrists believed that the public and doctors had mixed attitudes to their possible involvement in screening. Specific barriers included additional Bay 11-7085 cost, time, remuneration and litigation fears. However, optometrists felt that their professional role has evolved and that a greater, extended clinical involvement would be positive. In conclusion, optometrists are willing to carry out capillary blood glucose tests, provided that the scheme is simple, is supported by other health care professionals and is properly funded.

There is a clear advantage in identifying undiagnosed diabetes in people attending optometry practices who are not accessing other health care providers. Copyright © 2010 John Wiley & Sons. “
“There are four major hypertensive disorders in pregnancy: chronic hypertension, gestational hypertension, pre-eclampsia and chronic hypertension with superimposed pre-eclampsia. The indications for and efficacy of antihypertensive treatment in the different hypertensive disorders are evaluated. Advantages and disadvantages of different classes of antihypertensive drugs during pregnancy and lactation are described. “
“Detection of ketonaemia is a key factor in diagnosing diabetic ketoacidosis (DKA). Measurement of urinary ketones via the nitroprusside reaction is the most commonly employed diagnostic test; however, near patient testing of blood ketones is now widely available. In the clinical setting we wished to compare the utility of urine and blood ketone measurements to predict acid base balance and need for admission in patients with type 1 diabetes.

We therefore used the following method to determine the coordinate for fixation. If, within one set of eight blocks with the same calibration, the difference between median eye position in the different blocks was < 1°, we used the median x-values and y-values Gemcitabine concentration across all blocks as the fixation point coordinates. Otherwise, the eye-tracking data were analysed without this correction. On this filtered data, we removed all trials in which the subjects’ eyes moved by more than 1.75° from the fixation point. Two participants were excluded

because of excessive eye movements. All EEG data analyses were performed in matlab with the fieldtrip toolbox (Oostenveld et al., 2011) as well as custom scripts. The EEG recording was high-pass-filtered with a low cut-off of 0.5 Hz, by the use of fourth-order Chebyshev filters with zero phase-shift. This filter has the advantage selleckchem of very high attenuation in the stop band with minimal attenuation in the pass-band (< 0.1 dB). After filtering, bad channels were determined from the statistics of neighboring channels, and interpolated by the use of linear, distance-weighted interpolation. The EEG data were

then referenced to the average. In addition to the deletion of trials on the basis of eye movements, there was also an EEG threshold of ±125 μV. If more than six channels or any of the occipital electrodes of interest exceeded this threshold, the trial was discarded. Otherwise, high-amplitude channels were interpolated by the use of linear, distance-weighted interpolation. Three participants were excluded because of large numbers of trials with EEG artefacts, bringing the total number of participants used in further analysis selleck inhibitor to 14. After removal of artefact trials, an average of 117 trials per condition and participant

remained. Temporal second-order kernels (e.g. Sutter, 2000) representing evoked cortical responses were extracted for each electrode and each of the four stimulus locations, by reverse-correlating the EEG response with the known sequence of pattern reversals. The second-order response takes into account the history of visual stimulation, i.e. whether the current pattern is the same as the one presented one monitor refresh before. Given the findings of previous studies on spatial attention (e.g. Lalor et al., 2007; Kelly et al., 2008; Frey et al., 2010), we expect attentional modulation of the evoked responses during early cortical processing, as represented by responses in the C1 and P1 time-frame. As evoked response kernels represent activity in the early retinotopic cortex, which is very variable across participants (Ales et al., 2010a), the topographical distribution of peak activity was inconsistent across participants. For each stimulus location, we therefore selected two electrodes for each participant by determining mean activity across all four experimental conditions and selecting the two electrodes on the peak of the C1 and P1 topography, respectively.

We therefore used the following method to determine the coordinate for fixation. If, within one set of eight blocks with the same calibration, the difference between median eye position in the different blocks was < 1°, we used the median x-values and y-values HSP signaling pathway across all blocks as the fixation point coordinates. Otherwise, the eye-tracking data were analysed without this correction. On this filtered data, we removed all trials in which the subjects’ eyes moved by more than 1.75° from the fixation point. Two participants were excluded

because of excessive eye movements. All EEG data analyses were performed in matlab with the fieldtrip toolbox (Oostenveld et al., 2011) as well as custom scripts. The EEG recording was high-pass-filtered with a low cut-off of 0.5 Hz, by the use of fourth-order Chebyshev filters with zero phase-shift. This filter has the advantage Ku-0059436 molecular weight of very high attenuation in the stop band with minimal attenuation in the pass-band (< 0.1 dB). After filtering, bad channels were determined from the statistics of neighboring channels, and interpolated by the use of linear, distance-weighted interpolation. The EEG data were

then referenced to the average. In addition to the deletion of trials on the basis of eye movements, there was also an EEG threshold of ±125 μV. If more than six channels or any of the occipital electrodes of interest exceeded this threshold, the trial was discarded. Otherwise, high-amplitude channels were interpolated by the use of linear, distance-weighted interpolation. Three participants were excluded because of large numbers of trials with EEG artefacts, bringing the total number of participants used in further analysis out to 14. After removal of artefact trials, an average of 117 trials per condition and participant

remained. Temporal second-order kernels (e.g. Sutter, 2000) representing evoked cortical responses were extracted for each electrode and each of the four stimulus locations, by reverse-correlating the EEG response with the known sequence of pattern reversals. The second-order response takes into account the history of visual stimulation, i.e. whether the current pattern is the same as the one presented one monitor refresh before. Given the findings of previous studies on spatial attention (e.g. Lalor et al., 2007; Kelly et al., 2008; Frey et al., 2010), we expect attentional modulation of the evoked responses during early cortical processing, as represented by responses in the C1 and P1 time-frame. As evoked response kernels represent activity in the early retinotopic cortex, which is very variable across participants (Ales et al., 2010a), the topographical distribution of peak activity was inconsistent across participants. For each stimulus location, we therefore selected two electrodes for each participant by determining mean activity across all four experimental conditions and selecting the two electrodes on the peak of the C1 and P1 topography, respectively.

In addition, all sequenced strains have the gene encoding archaeal form III ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), leaving the question as to whether only one or multiple pathways are functioning R428 in these species (Berg et al., 2007). The possibility of the

functioning of two different pathways of autotrophic CO2 fixation has been shown recently for an uncultured endosymbiont of a deep-sea tube worm (Markert et al., 2007). The goal of our work was to study the presence of the enzymes of the dicarboxylate/hydroxybutyrate and hydroxypropionate/hydroxybutyrate cycles in A. lithotrophicus. This species was chosen for the study as it is the only strictly autotrophic representative of this group known so far. Also, the possible function of Rubisco in this species was addressed. Materials were as described previously (Berg et al., 2010b). Acetyl-CoA, propionyl-CoA, succinyl-CoA and crotonyl-CoA were synthesized from the respective anhydrides, and acetoacetyl-CoA from diketene using the method of Simon & Shemin (1953). The dry powders of the CoA-esters were stored at −20 °C. (R)- and (S)-3-hydroxybutyryl-CoA were synthesized using the mixed anhydride method (Stadtman, 1957). Archaeoglobus lithotrophicus’ strain TF2 was obtained from the culture collection of the Lehrstuhl für Mikrobiologie, University of Regensburg. Cells were grown

autotrophically under anoxic conditions Epacadostat price in MGG medium (Huber et al., 1982) at 80 °C and pH 6.0 using sulfate (2 g L−1) as an electron acceptor. In the 300-L fermentor, a gassing rate of 1 L min−1 was applied (using a gas mixture of 80% H2 and 20% CO2, v/v). The cells were harvested by centrifugation in the late exponential growth phase and stored at −70 °C until use. Metallosphaera Glycogen branching enzyme sedula TH2T (DSMZ 5348) was grown autotrophically as reported before (Alber et al., 2006). Archaeoglobus fulgidus VC16T

(DSMZ 4304) was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and grown according to the recommendations of DSMZ. Cell extracts were prepared under anoxic conditions using a French pressure cell as described previously (Berg et al., 2010b). Spectrophotometric enzyme assays (0.5 mL assay mixture) were performed in 0.5-mL cuvettes at 65 °C. Radiochemical enzyme assays were performed at 80 °C. Anoxic assays were performed with N2 as the headspace. For the wavelengths and extinction coefficients used in spectrophotometric assays, see Berg et al. (2010b). Pyruvate and 2-oxoglutarate:acceptor oxidoreductase were measured anoxically as a pyruvate- or 2-oxoglutarate-dependent reduction of methyl viologen and as a 14CO2 exchange reaction with the carboxyl group of pyruvate or 2-oxoglutarate (Ramos-Vera et al., 2009). Phosphoenolpyruvate (PEP) carboxylase was measured radiochemically as PEP-dependent fixation of 14CO2 (Ramos-Vera et al., 2009).

In addition, all sequenced strains have the gene encoding archaeal form III ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), leaving the question as to whether only one or multiple pathways are functioning U0126 research buy in these species (Berg et al., 2007). The possibility of the

functioning of two different pathways of autotrophic CO2 fixation has been shown recently for an uncultured endosymbiont of a deep-sea tube worm (Markert et al., 2007). The goal of our work was to study the presence of the enzymes of the dicarboxylate/hydroxybutyrate and hydroxypropionate/hydroxybutyrate cycles in A. lithotrophicus. This species was chosen for the study as it is the only strictly autotrophic representative of this group known so far. Also, the possible function of Rubisco in this species was addressed. Materials were as described previously (Berg et al., 2010b). Acetyl-CoA, propionyl-CoA, succinyl-CoA and crotonyl-CoA were synthesized from the respective anhydrides, and acetoacetyl-CoA from diketene using the method of Simon & Shemin (1953). The dry powders of the CoA-esters were stored at −20 °C. (R)- and (S)-3-hydroxybutyryl-CoA were synthesized using the mixed anhydride method (Stadtman, 1957). Archaeoglobus lithotrophicus’ strain TF2 was obtained from the culture collection of the Lehrstuhl für Mikrobiologie, University of Regensburg. Cells were grown

autotrophically under anoxic conditions Enzalutamide in MGG medium (Huber et al., 1982) at 80 °C and pH 6.0 using sulfate (2 g L−1) as an electron acceptor. In the 300-L fermentor, a gassing rate of 1 L min−1 was applied (using a gas mixture of 80% H2 and 20% CO2, v/v). The cells were harvested by centrifugation in the late exponential growth phase and stored at −70 °C until use. Metallosphaera PRKACG sedula TH2T (DSMZ 5348) was grown autotrophically as reported before (Alber et al., 2006). Archaeoglobus fulgidus VC16T

(DSMZ 4304) was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and grown according to the recommendations of DSMZ. Cell extracts were prepared under anoxic conditions using a French pressure cell as described previously (Berg et al., 2010b). Spectrophotometric enzyme assays (0.5 mL assay mixture) were performed in 0.5-mL cuvettes at 65 °C. Radiochemical enzyme assays were performed at 80 °C. Anoxic assays were performed with N2 as the headspace. For the wavelengths and extinction coefficients used in spectrophotometric assays, see Berg et al. (2010b). Pyruvate and 2-oxoglutarate:acceptor oxidoreductase were measured anoxically as a pyruvate- or 2-oxoglutarate-dependent reduction of methyl viologen and as a 14CO2 exchange reaction with the carboxyl group of pyruvate or 2-oxoglutarate (Ramos-Vera et al., 2009). Phosphoenolpyruvate (PEP) carboxylase was measured radiochemically as PEP-dependent fixation of 14CO2 (Ramos-Vera et al., 2009).