In the WT strain, a transcriptional start site (T) located 140 bp from the start codon (Fig. 4a) was determined by 5′ RACE Daporinad mouse PCR (not shown). Upstream, a potential σA-type promoter was identified with a (TATAAT) −10 box, and a (TTTACA) −35 box, exhibiting high conservation with the Bacillus subtilis consensus sequences. A sequence motif TGAAGAATATA, highly similar to the consensus sequence

of the bacterial cold-box element [TGA (C/A) N (A/T) ACANA, Hunger et al., 2006], was mapped at +25 bp downstream of the transcriptional start (Fig. 4a). Two additional putative boxes, also displaying homology with cold-box consensus sequences, were located upstream of the −10 and −35 promoter regions. The BC0259 gene is followed by an inverted repeat with a ΔG° of −28.3 kcal mol−1. This repeat could be a transcriptional terminator, suggesting PLX4032 nmr a BC0259 transcription as a single unit (Fig. 4a). RT-PCR with RNA from WT and mutant cultures at 10 and 30 °C confirmed that the BC0259 gene was not cotranscribed with the upstream and downstream genes (data not shown). The BC0259 gene

encodes a protein of 533 aa with a calculated molecular weight of 59 400 Da and a pI of 9.58. Alignment of the BC0259 aa sequence with NR-database sequences showed the presence of nine motifs highly conserved in the DEAD-box family of RNA helicases (Fig. 4b). Motif I (Walker A) and motif II (Walker B) are required for NTP/ATP binding and hydrolysis. Motif III has been suggested to couple NTP hydrolysis to helicase activity. Motif VI was shown to function in ATP hydrolysis. Motifs Ia, Ib, IV and V bind to substrate RNA. The Q motif is thought to be specific to DEAD-box RNA helicases and acts as an ATP sensor (Cordin et al., 2006; Bleichert & Baserga, 2007). In addition to this core protein, BC0259 is flanked by a C-terminal domain of approximately 92 aa, rich in glycine and arginine and Thiamet G containing several RNRD (arginine/asparagine/arginine/aspartic acid) repetitions conserved in the BC0259 homologues

of the sequenced genomes of the B. cereus group strains. BC0259 gene expression at 10 and 30 °C in WT and 9H2 cultures at OD600 nm=1.0 was tested by RT-PCR experiments. WT transcripts were detected at 30 and 10 °C and amplicons were also obtained from 9H2 RNA (data not shown), indicating that insertion of the transposon upstream BC0259 gene did not abolish its expression at both 30 and 10 °C. RNAs were then quantified by real-time RT-PCR in cells (1) grown at 30 °C at OD600 nm=1.0 and (2) grown at 10 °C at OD600 nm=0.2 and 1.0. The expression of BC0259 was 1.85-fold higher when WT cells were grown at 30 °C and at OD600 nm=0. 2 than at OD600 nm=1.0. It was 2.1-fold higher when the cells were grown at 10 °C at similar ODs (data not shown). Thus, this gene was more expressed during the lag phase, at both tested temperatures. When compared with WT, BC0259 expression was repressed in 9H2 for cells grown at 10 °C and at OD600 nm=0.

S1. Lesion selleck chemicals llc reconstructions for the animals of the ‘Responders’ group. Fig. S2. Lesion reconstructions for the animals of the ‘Non-Responders’ group. Fig. S3. Coronal section of lesioned pMS cortex. “
“Neuronal activity in the subthalamic nucleus (STN) of patients with Parkinson’s disease (PD) is characterised

by excessive neuronal synchronization, particularly in the beta frequency range. However, less is known about the temporal dynamics of neuronal oscillations in PD. In this respect long-range temporal correlations (LRTC) are of special interest as they quantify the neuronal dynamics on different timescales and have been shown to be relevant for optimal information

processing in the brain. While the presence of LRTC has been demonstrated in cortical data, their existence in deep brain structures Target Selective Inhibitor Library high throughput remains an open question. We investigated (i) whether LRTC are present in local field potentials (LFP) recorded bilaterally from the STN at wakeful rest in ten patients with PD after overnight withdrawal of levodopa (OFF) and (ii) whether LRTC can be modulated by levodopa treatment (ON). Detrended fluctuation analysis was utilised in order to quantify the temporal dynamics in the amplitude fluctuations of LFP oscillations. We demonstrated for the first time the presence of LRTC (extending up to 50 s) in the STN. Importantly, the ON state was characterised by significantly stronger LRTC than the OFF state, both in beta (13–35 Hz) and high-frequency (> 200 Hz) oscillations. The existence

of LRTC in subcortical structures such as STN provides further ADP ribosylation factor evidence for their ubiquitous nature in the brain. The weaker LRTC in the OFF state might indicate limited information processing in the dopamine-depleted basal ganglia. The present results implicate LRTC as a potential biomarker of pathological neuronal processes in PD. “
“Striatal-enriched protein tyrosine phosphatase (STEP) is a brain-specific phosphatase that opposes synaptic strengthening by the regulation of key synaptic signaling proteins. Previous studies suggest a possible role for STEP in learning and memory. To demonstrate the functional importance of STEP in learning and memory, we generated STEP knockout (KO) mice and examined the effect of deletion of STEP on behavioral performance, as well as the phosphorylation and expression of its substrates. Here we report that loss of STEP leads to significantly enhanced performance in hippocampal-dependent learning and memory tasks. In addition, STEP KO mice displayed greater dominance behavior, although they were normal in their motivation, motor coordination, visual acuity and social interactions.

Next, the new deletion unit LD3-5-2 was added to Δ17aK to construct Δ18aK. The KmR marker was removed by the addition of the deletion unit OCL37 without the KmR marker using the ‘415S Sm system’

to construct Δ19a (Kato & Hashimoto, 2008). Similarly, Δ20a–Δ28a were constructed using the ‘ApR-415S Sm system. The dps gene was added to Δ28a to construct Δ29a. The DNA fragment, in which the chromosomal regions flanking the regions of the deletion unit 15 were joined to the sides of the ApR-dps fragment, was introduced into Δ28a. The region of the first DNA fragment was replaced with the second DNA fragment, in which the TcR–FRT fragment was flanked by one of the chromosomal regions and Ap. The third DNA fragment, in which the chloramphenicol-resistance PD0325901 purchase (CmR)–FRT

and the dps fragments were joined to the sides of the chromosomal Anti-diabetic Compound Library order region, was cloned into the plasmid pSG76A (ApR) (Posfai et al., 1997; Kato & Hashimoto, 2008). Using this plasmid, the TcR and ApR markers were removed to yield Δ29a. The prophage regions were deleted to construct Δ30a–Δ33a by the ApR-415S Sm system (see Results and discussion). The primers used to construct the deletion units are shown in Supporting Information, Fig. S1, and Tables S1 and S2. The deletion mutants were grown on antibiotic medium 3 plates and then colonies were transferred to 2 mL of antibiotic medium 3 for 24 h at 37 °C with shaking. For aerobic cultures, the precultures were diluted 1/100 into 3 mL of antibiotic medium 3 and incubated for 24 h at 37 °C with shaking. The stationary culture (0.5 mL) was added to a sampling tube, mixed with menadione solution (in ethanol) or ethanol, and incubated for 24 h at 4 °C with rotation. These cultures were diluted, plated on antibiotic medium 3 plates, and the colonies were counted after incubation for 1–4 days at 37 °C. For anaerobic cultures, the precultures were diluted 1/100 into 3 mL of antibiotic medium 3 and, after bubbling with N2, were incubated DOK2 for 24 h at 37 °C with rotation. The

stationary culture (0.5 mL) was added to a sampling tube with an O-ring, mixed with menadione solution (in ethanol) or ethanol and, after flashing with N2, was incubated for 24 h at 4 °C with rotation. These cultures were diluted and plated on antibiotic medium 3 plates, and the colonies were counted after incubation for 1–4 days at 37 °C. The concentrations of menadione were 1.0 mM for Δ1–Δ15a and 0.1 mM for Δ14a–Δ33a (anaerobic culture), and 1.0 mM for Δ1–Δ26a and 0.5 mM for Δ25a–Δ33a (aerobic culture). In order to obtain final concentrations of 1.0, 0.5, and 0.1 mM, 10 μL of 50 mM, 5 μL of 50 mM, and 2 μL of 25 mM menadione in ethanol were added to 0.5 mL cultures, respectively.

Both HIV-1 and HIV-2 are associated with similar opportunistic infections and AIDS. Natural history studies indicate HSP inhibitor that HIV-2 is less pathogenic than HIV-1 [16–18]. Although the mortality rate in individuals infected with HIV-2 is two-to-three times that seen in HIV-negative populations, this compares with a 10-fold higher mortality rate in those

infected with HIV-1 than in those who are HIV negative. HIV-2 infection has a longer asymptomatic phase than HIV-1 infection and some patients with HIV-2 may never develop AIDS [19]. A cohort study of seroconverter women in Senegal found that the incidence of AIDS-defining illness was 0.95 [95% confidence interval (CI) 0.2–3.8] per

selleckchem 100 person-years among HIV-2-infected women as compared with 5.6 (95% CI 3.3–9.8) in HIV-1-infected women [16]. In practice, it is not unusual to see patients who remain asymptomatic for 10–20 years without treatment [20]. There are, however, patients in whom disease progresses as rapidly as in those who have HIV-1. AIDS-defining illnesses have been noted to occur at higher CD4 cell counts in individuals infected with HIV-2 than in those infected with HIV-1, although this is unusual [21]. Plasma viral loads are lower in HIV-2-infected individuals, suggesting that HIV-2 replication is restricted in comparison to that of HIV-1. An in vivo study has clearly demonstrated that, like HIV-1, HIV-2 can establish a stable, integrated proviral infection but that HIV-2 produces less mRNA, which may attenuate HIV-2 replication and pathogenesis [22]. HIV-2 is less infectious than HIV-1 early in the course of infection and, although infectivity increases as the disease advances, in general HIV-2 has significantly lower infectivity than HIV-1 [23]. HIV-2 infection does not protect against HIV-1 infection and dual

infection is well documented [24–26] although it is still uncommon in the United Kingdom. Studies from West Africa demonstrate that dual infection is more common in older women [25]. Dually infected patients tend to present at a more advanced stage of disease than those with HIV-2 only. Parvulin Infection with both HIV-1 and HIV-2 generally carries the same prognosis as HIV-1 monoinfection [19]. It is important to note that HIV-2 has a different capsid antigen from the HIV-1 p24 antigen and that this capsid antigen may result in a prolonged seroconversion window period for HIV-2, but there is no current evidence from human studies that it is longer than the 3-month period described for HIV-1. Detection of HIV-2 infection is based on the demonstration of virus-specific antibodies using enzyme-linked immunosorbent assay-based techniques.

, 2011). For information on the commercial value and application of cold-active enzymes, BMS-354825 concentration we suggest reading Marx et al. (2007). One of the major adaptations of cold-proteins includes modifications of structural features that increase flexibility, and specific amino acids have emerged as key elements (Marx et al., 2007). Glycine has been reported as an important residue to improve the flexibility of protein structure, providing more amplitude to the relative movements between elements of the secondary structure. In pioneering work, Saunders et al. (2003) compared the global proteomes of two cold-adapted Archaea (Methanogenium frigidum

and Methanococcoides burtonii) with mesophilic proteomes. They found that these cold-adapted prokaryotes displayed higher frequencies of charged polar residues (mainly Gln and Thr) and a lower frequency of hydrophobic amino acids, mainly Leu. Using a different approach, check details Gianese et al. (2001) showed that, among psychrophilic enzymes, Ala and Asn were increased and Arg decreased at exposed sites, and some other differences

were found within α-helices and β-strands. More recently, Grzymski et al. (2006) showed that the most significant changes found in Antarctic bacterial protein sequences were a reduction of Pro, stabilizing hydrophobic clusters, and in salt-bridge-forming residues (Arg, Glu, and Asp). The availability of more genome sequences from psychrophilic microorganisms will be crucial

for understanding the adaptation of proteins to a cold environment, which in turn will have an obvious biotechnological application. Relevant biotechnological cold-active bacterial enzymes have been identified using culture-dependent studies (Margesin & Schinner, 1994; Vazquez et al., 2004; Martínez-Rosales & Castro-Sowinski, 2011; among many others). Currently, however, the most promising approach is based upon metagenomics, a culture-independent genomic Nintedanib (BIBF 1120) analysis. Functional metagenomics relies on the extraction of environmental DNA and subsequent cloning to eventually identify the entire genetic set of a habitat. This allows the analysis of a wide diversity of genes and their products as well as the study of their potential for biotechnological use (Schmeisser et al., 2007). Through metagenomics, several cold-active enzymes with many potential biotechnological applications have been identified, cloned in heterologous hosts and characterized. Examples include lipases and esterases (Cieslinski et al., 2009; Heath et al., 2009; Yuhong et al., 2009; Berlemont et al., 2011; Yu et al., 2011; Hu et al., 2012), proteases (Berlemont et al., 2011; Zhang et al., 2011), cellulases (Berlemont et al., 2011), and glycosyl hydrolases (Berlemont et al., 2009, 2011).

In the first 48 weeks, new AIDS events were observed in 2.7% of late presenters, 0.8% of late starters and 0.9% Afatinib clinical trial of ideal starters (P=0.0001; χ2 test). In contrast, among those who remained alive beyond week 48 and were still under follow-up, the rate of new AIDS events between weeks 48 and 96 was similar in the three groups at 1.3, 1.0 and 0.5%, respectively (P=0.11; χ2 test). Deaths were more frequent in late presenters in the first 48 weeks (2.0%vs. 1.0 and 0.5% in late and ideal starters, respectively; P=0.0003; χ2 test) but among those surviving the first 48 weeks, death rates between weeks 48 and 96 were similar in the three groups (1.0%vs. 1.1 and 1.0% in late and ideal starters, respectively; P=0.96; χ2 test). Overall,

clinical progression (new AIDS events or death) occurred in 4.6% of late presenters, 1.8% of late starters and

1.4% of ideal starters in the first 48 weeks (P=0.0001). The rates of new clinical progression after a year of HAART (i.e. among those who remained alive AZD6738 and under follow-up beyond week 48) were 2.2, 1.9 and 1.3% in late presenters, late starters and ideal starters, respectively (P=0.21). Multivariable analyses (Table 2) suggested that late presenters were at increased risk of a new AIDS event or death in the first 48 weeks (P=0.01) compared with late starters, but that this excess risk was lost if patients survived beyond week 48 (P=0.83). In contrast, clinical progression rates in late starters and ideal starters did not differ significantly, either at 48 (P=0.64) or 96 (P=0.40) Anidulafungin (LY303366) weeks. The differences in virological and immunological endpoints between late presenters and late starters were unchanged after additionally controlling for the pre-HAART CD4 cell count and viral load. However, the difference in clinical progression rate in late presenters compared with late starters at 48 weeks was reduced and nonsignificant [adjusted odds ratio (OR) 1.69; 95% confidence interval (CI) 0.93, 3.06; P=0.09]. When we assumed that all individuals who were lost to follow-up or who had missing viral load values at week 48 had not achieved

virological suppression (a missing equals failure approach), virological suppression rates were 55.3, 58.6 and 56.1% in late presenters, late starters and ideal starters, respectively. Differences between late presenters (adjusted OR 1.08; 95% CI 0.91, 1.28; P=0.37) and late starters, and between ideal starters (adjusted OR 0.94; 95% CI 0.79, 1.12; P=0.47) and late starters were not significant in multivariable analyses. Similar results were obtained at 96 weeks. When similar analyses were performed for the clinical outcome, in which patients who were lost to follow-up over the first 48 weeks (and between week 48 and week 96 for the 96-week outcome) were assumed to have experienced clinical failure, failure rates were 16.2, 12.9 and 20.5% in late presenters, late starters and ideal starters, respectively, at week 48 (P=0.0001), and 18.8, 17.7 and 18.8%, respectively (P=0.

cinerea, as well as its effects on PCR. To achieve these goals, 2-mL samples were spiked with 8 × 106 cells of Y. lypolitica, a microorganism that is absent from grapes before nucleic acid extraction. The LIP4 gene from Y. lipolytica was used as an internal control. From the calibration curve of Y. lypolitica obtained previously, DNA extracted from 8 × 106 CFU per 2 mL of the yeast Y. lypolitica yielded a Ct of 29.4 ± 0.631. We used this Ct value as a www.selleckchem.com/products/pirfenidone.html normalizer for the quantification of B. cinerea DNA concentration on grapes. Ct values obtained from B. cinerea were normalized according to the following equation: The resultant Ct values were

converted into DNA concentrations by extrapolation to a standard curve generated from qPCR analysis using 10-fold dilutions of between 102 and 106 pg B. cinerea DNA (Fig. 1). A total GSK J4 nmr of 14 strategies, which included various fungicide treatments for controlling B. cinerea, were applied to grapes at different growing stages: flowering, bunch closure, 10 days after bunch closure and veraison (colour change) (Table 1). In each experimental plot, microbial communities on grape berries were assessed at harvest. Our qPCR method was used to assess the level of B. cinerea contamination in each treatment (spore and

mycelium). The DNA concentration of B. cinerea present in each sample (200 berries) for each strategy is given Fig. 3. The type of treatment had a clear

impact on B. cinerea contamination. In our case, the best strategy appeared to be AB6, which led to a significant decrease in B. cinerea contamination. This treatment used at least two chemical products during grape development with thinning out of leaves. This prophylactic method increases the efficiency of the treatment strategy as compared with AB5, in which the same chemical product was used Urocanase (fenhexamid and pyrimethanil) but without thinning out of leaves. Nevertheless, the AB10 treatment, in which only one chemical product was used, also appeared to be efficient, i.e. a low level of B. cinerea DNA was detected. The low significant level of B. cinerea DNA concentration observed for strategy AB8 demonstrated that the association of a chemical product together with Bacillus subtilis improves anti-Botrytis treatment. Our trial underlined that bentonite clay (AB14) did not protect grapes from B. cinerea contamination. We developed a highly specific and sensitive qPCR protocol for the detection and quantification of B. cinerea contamination in grapes. This method was developed to serve as an alternative to the various conventional methods: (1) counting spores with a microscope, which is time-consuming and has a low detection limit; (2) spread plate culture method, which underestimates the number of spores (Martinez et al.

, 2010). Considering that the Drosophila P0 protein has DNase and endonuclease activities (Yacoub et al., 1996), it is reasonable to suspect that phosphorylated C. cucullus p33 may be involved in such macronuclear events. Furthermore, the P0 protein may play a role in regulating metabolism during pupal diapause of the flesh fly (Craig & Denlinger, 2000). The C. cucullus p33 may be involved in

the regulation of metabolic activity directly see more or through gene expression, because mitochondrial membrane potential disappeared in the early stage of encystment (Funatani et al., 2010). It is also likely that C. cucullus p33 plays a role in the regulation of encystment-specific gene expression, as was reported in Drosophila. In fact, the expression of encystment-specific proteins in C. cucullus was recently found to be regulated at the NVP-BKM120 nmr transcriptional level (in preparation). In many organisms, the ribosomal S5 protein consists of 190–230 amino acid residues (blast Search) and its free form is phosphoprotein (Matragkou et al., 2009). Taking into account that in mammalian cells, the ribosomal S5 protein has been reported to

be involved in the arrest of cell cycle and the initiation of differentiation (Matragkou et al., 2008),and the p24 must also be involved in the cell cycle arrest and differentiation into resting cyst form in C. cucullus. “
“A large number of novel bioactive compounds were discovered from microbial secondary metabolites based on the traditional bioactivity screenings. Recent fermentation studies indicated that the crude extract of marine Streptomyces sp. W007 possessed great potential in agricultural fungal disease control against Phomopsis asparagi, Polystigma deformans, Cladosporium cucumerinum, Monilinia fructicola, and Colletotrichum lagenarium. To further evaluate the biosynthetic potential of secondary metabolites, we sequenced the genome of Streptomyces sp. W007 and analyzed the identifiable secondary metabolite gene clusters. Moreover, one gene

cluster with type II PKS implied the possibility of Streptomyces sp. W007 to produce aromatic polyketide of angucyclinone antibiotics. Therefore, two novel compounds, 3-hydroxy-1-keto-3-methyl-8-methoxy-1,2,3,4-tetrahydro-benz[α]anthracene and kiamycin with Etoposide potent cytotoxicities against human cancer cell lines, were isolated from the culture broth of Streptomyces sp. W007. In addition, other four known angucyclinone antibiotics were obtained. The gene cluster for these angucyclinone antibiotics could be assigned to 20 genes. This work provides powerful evidence for the interplay between genomic analysis and traditional natural product isolation research. Microbial natural products are an important source of new drugs (Solanki et al., 2008). Among the producers of commercially important metabolites, actinomycetes have proven to be a prolific source with a surprisingly small group of taxa accounting for the vast majority of compounds.

More frequent monitoring of renal function (every 4 weeks during the first year, and every 3 months thereafter)

is recommended in the SPC for tenofovir. Referral to a renal physician should be considered for patients suspected to have a glomerulonephritis (haematuria and/or uPCR >100 mg/mmol) and those with a severe or progressive decline in renal function, advanced renal failure (eGFR <30 mL/min) or severe hypertension associated with renal injury (uPCR >100 mg/mmol or eGFR <60 mL/min) (IV). HIV infection is associated with increased levels of triglycerides and decreased levels of high-density lipoprotein (HDL) cholesterol. ART may affect lipid levels and independently increase cardiovascular risk [22-26]. CVD is an increasingly important cause of mortality and morbidity in patients with HIV infection in the UK [27], emphasizing the importance learn more of assessing lipid profiles and managing dyslipidaemia selleck chemicals (as part of the overall cardiovascular risk) in those with HIV infection. Lipid levels should be assessed in the context of overall CVD risk. CVD risk assessments generally incorporate

age, gender, smoking, blood pressure, diabetes, the ratio of total:HDL cholesterol, and the presence or absence of left ventricular hypertrophy on electrocardiogram [28]. The Framingham CVD risk calculator works reasonably well in HIV-positive populations, although it is worth noting that it was not developed for use in non-White groups. Other algorithms may be better suited to these populations. A CVD risk calculator has been developed for use in HIV-positive populations (www.chip.dk/TOOLS) [29], although it should be noted that this provides 5-year risk estimates rather than the usual 10-year estimates. isothipendyl This calculator includes abacavir exposure as a CVD risk factor; the data regarding abacavir as a CVD risk factor, however, remain inconsistent. Alternatively,

the QRISK calculator (www.qrisk.org) or the QIntervention tool (http://qintervention.org), which also provide an estimate of the risk of developing type II diabetes, can be used. CVD risk can be reduced by smoking cessation, blood-pressure management (including nonpharmacological measures) and lipid-lowering interventions. Smoking cessation should be repeatedly encouraged. Weight reduction, diet and exercise may improve blood pressure and HDL-cholesterol levels. Decisions on lipid-lowering therapy should be based on overall cardiovascular risk rather than lipid levels in isolation. D-dimer levels, highly sensitive CRP, and IL-6 have recently been correlated with cardiovascular events and death [30]. While these biomarkers may become useful in identifying high-risk patients and contribute to the debate regarding when to start ART, they remain research tools and are not recommended for routine evaluation at present (IV).

Guidelines recommend that all patients with ED as part GDC-0068 supplier of a minimum assessment should have testosterone measured. By adhering to NICE guidance recommending an annual enquiry in regard to sexual health, diabetologists are already screening for hypogonadism in the diabetic clinic. There is currently no recommendation that testosterone be checked in all diabetic men. The recently updated clinical practice guideline of the American Endocrine Society does say that they suggest measurement

of testosterone in men with type 2 diabetes.22 The benefits of TRT on sexual function and on body composition in hypogonadal men have been recognised for several years and this therapy is a recognised and established treatment for the condition. There is accumulating evidence that TRT may have specific benefits on metabolic and cardiovascular parameters http://www.selleckchem.com/products/forskolin.html in men with type 2 diabetes. When replacing testosterone the aim should be to try and achieve as near normal

physiological replacement as possible. The importance of this is underlined by a recent publication of a study designed to determine the effects of the hormone on frailty where testosterone doses used in frail elderly men with established co-morbidities exceeded those used in normal clinical practice.23

It is important to recognise that this study was not powered to detect a significant increase in cardiovascular events but did report more cardiovascular-related symptoms/events in the testosterone treatment group. The cardiovascular-related events were heterogeneous and included oedema, which would be expected in high testosterone dose therapy, and self-reported symptoms such as syncope. A similar study using normal testosterone gel dosing did not show an increase in cardiovascular events.24 These findings, however, demonstrate that larger and longer-term P-type ATPase studies are needed to verify the cardiovascular and metabolic action of testosterone replacement in men with diabetes. It also underlines the importance of making a correct diagnosis of hypogonadism and, if indicated, treating with testosterone replacement to attain serum testosterone levels usually in the mid-normal to upper normal range.25 THJ is a consultant for ProStrakan as a chief investigator of the TIMES2 study. He has also been a member of advisory boards and has received honoraria for educational lectures from Bayer-Schering Pharma, ProStrakan and Ferring. He has received no funding for the preparation of this article. References are available online at www.practicaldiabetesinternational.com.