42.4% of children with enamel defects were born prematurely (<37 weeks) where as only 23.2% of them were born at normal gestational

age. No statistically significant difference in the prevalence of enamel defects was found in relation to birth weight (P > 0.05). Conclusions.  A high prevalence of developmental enamel defects was found among the children with CP. The prevalence of defects varied with the tooth type and was associated with gestational age of the children. “
“Estimating fluoride intake (FI) using the ‘duplicate plate’ method is difficult and can raise ethical dilemmas. To apply a semiquantitative food frequency questionnaire (FFQ) to 2- to 6-year-old Brazilian children in a non-fluoridated area (i) to estimate their FI and (ii) to provide additional validity to the questionnaire by comparing the results obtained with those found previously in a fluoridated municipality. Dapagliflozin cost The FFQ was administered to parents of 398 children residing in a non-fluoridated community. Constituents of the diet were divided into solids, water and other beverages and their fluoride content was analysed with the electrode. Data were analysed using unpaired t-test. The mean (±SD) FIs from

solids, water and other beverages were 0.009 ± 0.004, 0.001 ± 0.001 and 0.007 ± 0.007 mg F/kg body weight/day, respectively, totalling 0.017 ± 0.009 mg F/kg body weight/day. Total FI from food/beverage items ingested in the non-fluoridated area was significantly lower than that observed in a study previously Staurosporine conducted in a fluoridated Lck area (P < 0.0001). This result reinforces the use of the FFQ as a promising alternative to duplicate diet in order to estimate FI in children in this age range, with potential application in broad epidemiological surveys. "
“International Journal of Paediatric Dentistry 2011; 21: 35–42 Background.  Recent research has been focused on those attributes that appear to buffer a person against the stresses and strains of living with a visible difference. Aim.  To provide some insight on how young adults with Crouzon syndrome handle their life. Design.  Telephone

interviews were carried out with eight Crouzon syndrome individuals (six males, two females, mean age 25.4 years) and data were analysed according to the qualitative method of grounded theory. Results.  The informants’ main concern was to make the best of their situation, showing that even in adverse conditions, as in Crouzon syndrome, several individuals do find ways to live with their difference and to succeed in various aspects of life, using strategies they construct. Such strategies, as identified from the present investigation, were labelled: committed to an engaging activity, avoiding exposed situations, actively launching oneself, struggling with normalizing facial appearance, and lowering the expectations of finding a love partner. Conclusions.

Little variation was observed during the replicate experiments. The standard deviation for

the antirestriction results is 25% or less. Data on antirestriction activity of the recombinant plasmid pKLH53.1, containing Tn5053, are given in Table 2. The factor of restriction relief (R) is about 100. We suspected that the nucleotide sequence of the mercury-resistance transposon Tn5053 contains a fragment encoding an antirestriction protein. We used both insertion and deletion mutants of Tn5053 for all transposition genes (tni) as well as plasmid constructs containing various fragments of the Tn5053 DNA, while searching for the locus responsible for the antirestriction activity (Fig. 1). The results of searches http://www.selleckchem.com/products/pf-562271.html for the determinant of antirestriction activity Selleckchem Ku 0059436 within Tn5053 are shown in Table 2. It is evident that neither insertion (plasmids pKLH53.1tniA, pKLH53.1tniB2) or deletion (plasmids pKLH53.1tniQ2 and pKLH53.1tniQ1) mutations of the tni genes have any effect on antirestriction activity: about 100-fold decrease in EcoKI restriction level is preserved. Deletion of the major part of the mer operon (plasmid

pTLΔHindIII) completely removed the effect of antirestriction (Table 2). We assumed that the location of the gene coding for an antirestriction protein is within the mer operon. However, the recombinant plasmids pTLHindIII-ClaI and pTL2.5 with fragments HindIII-ClaI and HindIII from the mer operon (without the merR gene) in vector pUC19 show no antirestriction effect (Table 2). No antirestriction effect was also observed for the hybrid plasmid pKLH53.2, containing all the genes tni Tn5053 under its own promoter (in vector pACYC184; Fig. 1, Table 2). A paradox appeared: the mer operon together with the transposition genes (tni) of Tn5053 produce an antirestriction effect, while the plasmids with separately cloned mer operon or tni genes show no antirestriction effect.

We considered that the nucleotide sequence coding for the ORF with antirestriction activity is located within the region of the tni genes, but orientated in reverse to the direction of transcription of the tni genes. Consequently, the coding strand for this ORF is the same as for the mer operon. If so, transcription of this DNA fragment passes Meloxicam from the side of the mer operon. We analysed the DNA sequence from the region of the tni genes of Tn5053 in reverse direction, and found several orfs. Of main interest was orf-5, encoding a negatively charged protein with a motif close to the antirestriction motif of the proteins Ard (Fig. 2). The protein ORF-5 contains 147 amino acid residues of summary charge −1. It is encoded by orf-5 at positions 7511–7954 on the complementary strand of the tniA gene (positions numbered according to the nucleotide sequence of Tn5053, deposited in DBJ/EMBL/GenBank under accession number L40585).

[9] At 4559 m, inhalation of NO led to a marked decrease in PAP and an increase in arterial oxygen saturation especially in subjects susceptible to HAPE.[10] In addition, decreased pulmonary NO production during acute hypoxia was suggested to contribute among other Cobimetinib order factors to the enhanced hypoxic pulmonary vascular response in HAPE-susceptible subjects[11] and therefore might contribute to exaggerated hypoxic pulmonary vasoconstriction and in turn to pulmonary edema.[12] As

it is an NOS inhibitor, ADMA should cause an increase in PAP and raise the risk of developing altitude sickness and HAPE. By measuring ADMA serum levels during standardized altitude exposure, we were able to assess this approach both from a principal therapeutic perspective as described in the aforementioned studies and from a diagnostic perspective. This prospective comparative study was conducted to test the hypothesis that there is a relationship between Δ-ADMA in blood and a hypoxia-induced increase in PAP and AMS and that ADMA could be a predictive value for the development of AMS or a PAP > 40 mmHg. The tests

were performed in the altitude and climate chamber of the German Air Force Institute of Aviation Medicine in Koenigsbrueck, Germany (134 m). This hypobaric SB203580 datasheet chamber has a capacity of six individuals for an overnight stay. Two tests were performed and 12 subjects could be investigated. Each trial consisted of two overnight stays in the chamber. The subjects were allowed to sleep. For intraindividual comparison, both nights followed the same protocol. Altitude conditions, however, were simulated only during the second night, when the

subjects were decompressed over a period of 53 minutes to a pressure equivalent to an altitude Idoxuridine of 4000 m. The subjects spent 12 hours in the chamber under these altitude conditions. At all time points, the subjects could have been rapidly recompressed or could have left the chamber through an airlock. An emergency physician with expertise in altitude medicine was continuously present. The study design had been approved by the ethics committee of the Society of Physicians of the state of Baden-Wuerttemberg, Stuttgart, Germany. All participants had given their written informed consent to take part in the study. Twelve male subjects (median age: 23 years, range: 18–33 years; median height: 182.5 cm, range: 169–194 cm; median weight: 76 kg, range: 55–100 kg; median body mass index: 22.5 kg/m2, range: 19–29 kg/m2) without altitude exposure higher than 1500 m in the last month prior to this study showed a minor tricuspid valve insufficiency found incidentally in the context of this study and were otherwise healthy. Prior to the tests, the subjects received an echocardiogram (ECG). Blood tests (HBG, HCT, RBC, MCH, MCV, PLT) and a 12-channel ECG were performed immediately before the trial. All results were unremarkable.

14 Second, occult strongyloidiasis cannot be ruled Trametinib in vitro out easily and a presumptive

treatment against strongyloidiasis should also be considered.15 Finally, the timing of corticosteroids use is unclear. In one of these outbreaks its use was postponed in case of worsening evolution.1 According to our experience and as previously underlined we believe that corticosteroids use should be restricted to patients with severe forms (neurovasculitis, myocarditis, etc.) and PZQ only initiated when ova are detected in stools or urines according to the culprit species or when there is no more symptoms of AS.4,16 An effective and well-tolerated treatment for the management of patients with AS is still needed. A promising treatment could be artemisinin derivatives as they showed some activity against young schistosomulae.17,18 Therefore, a prospective study should be implemented to evaluate

the use of artemisin derivatives in AS. In conclusion, AS is difficult to diagnose and treat. The current diagnostic tools lack in sensitivity, the current treatment lack in efficacy and could cause complications. Therefore, research in diagnosis and Selleckchem Z-VAD-FMK treatment is needed. The authors state they have no conflicts of interest to declare. “
“We report an outbreak of Manchineel dermatitis and ophthalmitis in four students from North America who visited the island of Bequia, West Indies. The exposure resulted from taking shelter during a rain storm under a Manchineel tree. Manchineel exposure and ingestion can lead to severe and even fatal disease. The Manchineel (Hippomane mancinella) is a member of the Eurphobias

or spurge family and produces one of the most potent tree toxins known.1,2 Exposure to the toxic P-type ATPase sap (which contains Diterpene esters of the tigliane phorbol and daphnane types) can cause severe dermatitis and ophthalmitis consisting of erythema, blistering, swelling, inflammation, pustulation, and conjunctivitis with painful burning sensation typical of Chemical Irritant Contact Dermatitis (CICD).3–5 Exposure most commonly takes place when individuals take refuge from the rain under a Manchineel tree. Ingestion of the Manchineel fruit (Beach Apple or “Manzanilla de la muerte”) can cause severe swelling, ulceration, and hemorrhage of the oral and gastrointestinal mucosa which has been reported to be fatal in extreme cases.5,6 Systemic manifestations can be significant and persistent bradycardia requiring permanent pacemaker insertion has been ascribed to Manchineel toxicity.7 The toxin has been used in bellicosities by aboriginals from Florida to the southern Caribbean by treating arrow tips or poisoning water.8,9 Ponce De Leone is said to have died subsequent to a poisoned arrow wound containing Manchineel toxin in West Florida following an encounter with the hostile Calusa indians.9 The Manchineel is found in Florida, the West Indies, and Central and South America.

Similarly in cases associated with H1N1v (‘Swine flu’) treatment has often been prescribed regardless of symptom duration. Oseltamivir 75 mg bd po for 5 days is currently the preferred neuraminidase inhibitor [134,135]. Inhaled zanamivir 10 mg (two puffs) bid by inhalation device for 5 days is an alternative [136] and has even been suggested as the preferred agent for HIV-seropositive adults with significant immunosuppression in some guidelines on the basis of increased rates of oseltamivir resistance

in this group [137]. Most pandemic IAV strains in 2009–2010 retained susceptibility to neuraminidase inhibitors, but strains with reduced susceptibility to oseltamivir have been reported PARP inhibitor occasionally in individuals living with HIV [138]. In addition, seasonal IAV strains in 2008–2009 were frequently oseltamivir-resistant [139] and the selection of the most appropriate neuraminidase inhibitor must be made in light of the prevailing susceptibility of the strain(s) circulating in a given ‘flu season’ in consultation with local virologists. While many of these strains remain susceptible to zanamivir at present, multi-resistant strains have been reported

in other immunocompromised groups [140]. Some authorities have suggested combination therapy will be required, particularly for immunocompromised patients, in the future and clinical trials are exploring this possibility in patients (not specifically HIV-seropositive individuals) MG-132 cell line with severe infection [141,142]. For critically ill individuals parenteral formulations ROS1 of neuraminidase inhibitors, currently available for compassionate use or through expanded access programmes, include iv peramivir and zanamivir but there are currently no data on their use in HIV-seropositive individuals. Neuraminidase inhibitors have proven efficacy against IAV in individuals considered at high risk of IAV complications [143]. It is recommended that immunocompromised patients also receive doxycycline 200 mg stat then 100 mg od or co-amoxiclav 625 mg tid

po with clarithromycin 500 mg bd po as an alternative, all for 7 days during an episode of IAV but again no specific data are available for HIV-seropositive populations [144]. If pneumonia develops, coverage should be as per the guidelines above for community-acquired pneumonia but if patients fail to respond promptly, there are epidemiological concerns that methicillin-sensitive or -resistant Staphylococcus aureus (MSSA/MRSA) may be causing bacterial super-infection or there is a significant incidence of bacterial super-infection with MSSA/MRSA, then antibacterial therapy should also target these organisms. IAV vaccination should be offered to all HIV-seropositive individuals every ‘flu season (category Ib recommendation) [97,99,145].

An annual offer of a full sexual health screen (regardless of reported history) and the outcome documented in the HIV case notes, including whether declined (IIb). Syphilis serology should be documented at baseline and performed yearly. In individuals or groups at increased risk of syphilis (MSM), syphilis serology E7080 cost should be considered with routine HIV follow-up (2–4 times yearly) (IIb). All women should have cervical smears

performed annually (IV). Screening for anal dysplasia by anal cytology may be beneficial; however, there is insufficient evidence at this time to support its routine introduction (IV). Gender-specific aspects of HIV monitoring will be discussed fully in the BHIVA women’s guidelines currently under

development. Approximately 20% of HIV-infected individuals accessing care in the UK are aged 50 years or more [1]. The prevalence of ageing HIV-infected Obeticholic Acid individuals continues to increase as a result of: (i) greater survival rates among HIV-infected patients; (ii) delayed recognition of the infection in older individuals; and (iii) continued new infections in older individuals. There is a need to adapt the management of HIV-infected individuals to ensure that the clinical needs of these individuals continue to be met as they age. However, very little is known about the likely healthcare needs of these patients. Existing reports on the clinical picture of HIV infection among older individuals are largely anecdotal; HIV may accelerate several

age-related conditions, and HIV-infected individuals may experience accelerated frailty, accelerated bone mineral loss and different levels of drug absorption and metabolism compared with their younger counterparts. Impaired glomerular function, impaired tubular function and proteinuria are all more common in the elderly. While this age-related decline in renal function is unlikely to result in severe kidney failure, it may affect many homeostatic processes, which may have implications for exacerbation of bone mineral loss and/or increased cardiovascular risk. The impact on adherence and potential drug–drug interactions of treatment for age-related comorbidities in patients who may be receiving ART has not been documented. HIV infection GPX6 and ageing are also both associated with changes in immunity and host defence. The potential for full immune restoration among older individuals receiving HAART for prolonged periods of time has not been fully investigated. In older individuals, drug pharmacokinetics (absorption, distribution, metabolism, and elimination) are altered [2] as a result of: (i) changes in gastric pH; (ii) body fat increase and water decrease; (iii) reductions in liver volume, blood flow and metabolic enzyme activity; (iv) decreased renal function.

In all known cases, in normally growing cells, toxins form a stable complex with their cognate antitoxins that blocks the toxin activity. Antitoxin also functions as a repressor for individual TA operons (Gerdes et al., 2005). Under stress conditions, intrinsically unstable antitoxin is lost from the cells, releasing toxin freely and inhibiting various essential cellular functions, such as DNA replication, mRNA stability, protein synthesis, and cell division (Jiang

et al., 2002; Zhang et al., 2003; Tan et al., selleck chemicals 2011; Zhang & Inouye, 2011). This leads to a reversible cell growth arrest, which is implicated in the persister phenotype. The TA system is also shown to be associated with pathogenicity, programmed cell death, and biofilm formation (Pandey & Gerdes, 2005; Nariya & Inouye, 2008; Wang & Wood, 2011). Escherichia coli have two essential bacterial cytoskeletal proteins, FtsZ and MreB. FtsZ is a highly conserved GTPase and is homologous to eukaryotic cytoskeleton protein, tubulin (Mukherjee et al., 1998). It forms a ring structure at the mid-cell and functions as a scaffold for divisome, a multiprotein

complex responsible for cell division. MreB is an actin-like ATPase, essential for maintaining the typical rod shape and cell polarity in E. coli (Osborn & Rothfield, 2007). MreB is also implicated in chromosome segregation, localization of membranous organelles, and coordinating cell division with cell biosynthesis (Kruse et al., 2005; Komeili et al., 2006; Madabhushi & Marians, 2009; Domínguez-Escobar et al., 2011; Selumetinib supplier Rucaparib price Garner et al., 2011). Because both FtsZ and MreB are involved in a number of essential cellular functions, the inhibition of their functions is detrimental to the cells. For example, the inhibition of FtsZ polymerization by SulA or MinCD results in blocking the septum formation, causing the formation of filamentous cells (Mukherjee et al., 1998; Pichoff & Lutkenhaus, 2001). The inhibition of MreB by A22 [S-(3,4-dichlorobenzyl) isothiourea] leads to the loss of its rod shape and eventual cell lysis (Karczmarek et al.,

2007; Bean et al., 2009). Here, we have identified a novel TA system in E. coli genome using RASTA (Sevin & Barloy-Hubler, 2007). The putative toxin, YgfX, inhibits the cell growth and causes significant changes in the cellular morphology of E. coli. Upon induction of YgfX, the cells were first elongated and then subsequently became inflated in the middle. The YgfX toxicity was neutralized by the co-expression of YgfY, indicating that YgfY is an antitoxin of YgfX. YgfX is the first toxin of E. coli TA systems shown to be associated with membrane. We further demonstrated that YgfX physically interacts with FtsZ and MreB and inhibits their polymerization in vitro and that the C-terminal soluble domain of the YgfX is responsible for the inhibition.

The video contained 300 frames and each frame was presented to the model for 40 ms of simulation time. Each image was originally 256 × 256 pixels. Because our cortical model is made up of single columns, however, the input size was reduced to http://www.selleckchem.com/products/MS-275.html 20 × 20 pixels (see Fig. 2B) to approximate the visual space that would drive neurons in a receptive field of a V1 cortical column. This was an assumed approximation given the 100 deg2 receptive field and 36 × 36 (64 × 64 pixel) input from the Goard and Dan experiment.

In the 256 × 256 pixel image, RF1 received input from pixels (121–140) × (121–140) and RF2 received input from pixels (141–160) × (121–140). Figure 3 shows the architecture of RF1 and RF2. It has been shown that retinal neurons remove linear correlations by ‘whitening’ images before they reach the cortex (Simoncelli & Olshausen, 2001). To simulate this, all the images were whitened and normalised before being presented to the network (Fig. 2B). Whitening was achieved by applying a Gaussian filter to the Fourier-transformed image (see http://redwood.berkeley.edu/bruno/npb261b/). This flattens the power spectrum of the image TGF-beta family and is essentially equivalent to convolving the image with an on-center off-surround filter, as is observed in retinal

ganglion cells and the lateral geniculate nucleus (LGN). As we were not interested in modeling orientation selectivity development, we assumed that the simulated V1 columns, RF1 and RF2, were selective to vertical edges. Therefore, the images were convolved with a vertical Gabor filter after whitening.

The Gabor filter was constructed by modulating a Gabor kernel with a sinusoidal wave as shown in Eqn. (1), where σx and σy determine the spatial extent of the Gaussian in x and y and f specifies the preferred spatial wavelength IKBKE (Dayan & Abbott, 2001). Excitatory Poisson spike generators converted the images into spike trains in the input layer. (1) To develop our model, we used a publicly available simulator, which has been shown to simulate large-scale spiking neural networks efficiently and flexibly (Richert et al., 2011). The model contained a TRN, LGN, BF, two prefrontal cortex areas (providing top-down attention) and two, four-layered cortical microcircuits (Fig. 3). The cortical microcircuit architecture was adapted from Wagatsuma et al. (2011), which was able to account for experimental observations of attentional effects on visual neuronal responses and showed that top-down signals enhanced responses in layers 2/3 and 5. All connections that occur between layers in a microcircuit are shown in Fig. 3. Within each layer, there are excitatory–excitatory, excitatory–inhibitory, inhibitory-excitatory and inhibitory–inhibitory connections (data not shown). Connection probabilities in our cortical model were the same as used in Wagatsuma et al. (2011) and are given in Table 1. All subcortical and top-down connection probabilities were set to 0.

No comparative studies have been performed and hence there is no optimal ‘gold-standard therapy’ (level of evidence 1B). We recommend that chemotherapy regimens should be combined with HAART therapy (level of evidence 1B). We recommend Dapagliflozin solubility dmso the addition of rituximab (level of evidence 1C). 4.6.1 Recommendations for IT prophylaxis We recommend

that patients with DLBCL, considered to have a high risk of CNS relapse, should be given CNS prophylaxis (IT and/or IV methotrexate) according to the same criteria as HIV-negative patients (level of evidence 1C). We recommend that prophylactic intrathecal chemotherapy should be offered to all patients with Burkitt lymphoma (level of evidence IB). 4.8.1 Recommendations for patients with relapsed/refractory aggressive ARL We recommend that patients deemed fit

for intensive chemotherapy Selleck Staurosporine should receive a second-line chemotherapy regimen (level of evidence 1C), which may contain platinum (level of evidence 2C). We recommend that those patients responding to second-line chemotherapy (CR or PR) should be considered for HDT with ASCT (level of evidence 1C). 5 Primary central nervous system lymphoma (PCNSL) 5.4 Recommendations We recommend that all patients with PCNSL should be started on HAART if not already on it (level of evidence 1C). We recommend that patients with an adequate performance status should be treated, if possible, with high-dose methotrexate-containing chemotherapy regimen (level of evidence 1D). We recommend that whole brain radiotherapy is a useful palliative treatment modality for control of symptoms or should be considered as an alternative first-line treatment modality in those patients where the risks of toxicity from high-dose intravenous agents are considered unacceptable (level of evidence 1C). 6 Primary effusion lymphoma (PEL) 6.6 Recommendations

We suggest that first-line treatment of PEL in HIV-infected individuals includes CHOP-like regimens. No comparative studies have been performed and there is no optimal gold-standard therapy (level of evidence 2C). Patients, where possible, should be entered into clinical trials that are testing novel targeted approaches (GPP). Lck We recommend that chemotherapy regimens should be combined with HAART (level of evidence 1C). 7 Plasmablastic lymphoma 7.6 Recommendation We recommend that patients should receive HAART with systemic anthracycline-containing chemotherapy as first-line therapy (level of evidence 1C). 8 Cervical intraepithelial neoplasia (CIN) and cervical cancer 8.6 Key recommendations We recommend that all women newly diagnosed with HIV should have cervical surveillance performed by, or in conjunction with, the medical team managing their HIV infection (level of evidence 1B). An initial colposcopy and annual cytology should be performed if resources permit (level of evidence 2C).

The activity of the soluble protein kinases, 2′3′ cAMP phosphodiesterase (cyclic phosphodiesterase), total phosphodiesterases, AC and the phosphatases was measured in cells recovering from γ radiation effects (Fig. 3). The AC activity increased rapidly following γ irradiation and reached a maximum in 0.5 h PIR (Fig. 3a), during which the activity of phosphodiesterases and phosphatases was low. Whereas the AP did not change significantly during PIR, the acid phosphatase increased nearly 1.5-fold from 1 h PIR (5.146 μmol min−1 mg−1 protein) to 4 h PIR (8.243 μmol min−1 mg−1

protein) (Fig. 3b). The levels of cyclic phosphodiesterase decreased rapidly in 1 h PIR followed by an increase of nearly threefold in 4 h PIR (Fig. 3c). These click here results might support the argument that the net increase in the cAMP levels was due to differential regulation of AC and cyclic phosphodiesterase activities in response to DNA damage. Although, D. radiodurans R1 genome does not annotate the selleck chemicals llc classical bacterial AC and 2′3′ cAMP phosphodiesterase, it encodes for protein with a phosphodiesterase-type functional domain with nearly 30% genome without annotated functions, leaving the strong possibility

that unknown proteins are responsible for these activities. The amino acid sequence of AC from Escherichia coli was subjected to multiple sequence alignment, which showed different levels of amino acid similarities with some of the deinococcal ORFs. Among them, DR_1433 showed close to 75% match with E. coli protein in psiblast analysis. The presence of AC and cyclic phosphodiesterase activities in cell-free extracts of this bacterium suggested the strong possibility of AC and cyclic phosphodiesterase activities containing uncharacterized proteins in bacterial genome and it will be interesting to investigate these activities separately. Aliquots of γ-irradiated cells were collected during PIR and nucleotide-binding proteins were purified by heparin-sepharose affinity chromatography. Fractions

were tested for nucleolytic activity on dsDNA substrate. Results showed the presence of nucleolytic activity in unirradiated and zero PIR-irradiated samples. This Casein kinase 1 activity was completely absent in 1- and 2-h PIR samples (Fig. 4a) but reappeared in 3- and 4-h PIR samples. This indicated that the bacterium has an as yet unidentified mechanism to regulate the nuclease activity during different stages of PIR. It may be speculated that during early PIR, i.e. before 2 h PIR, the bacterium needs to protect its shattered genome and very low nuclease activity might be required for DSB end-joining, whereas at a later stage, i.e. after 2 h PIR, high recombinase functions are needed, which requires the high nuclease activity observed at 3 and 4 h PIR. Except for the unirradiated control, all the samples, including 1 and 2 h PIR, showed inhibition of nucleolytic function with 2 mM ATP (Fig.