Filamentous phage pI and pIV were shown to interact both in vivo and when co-expressed in isolation from the other phage proteins using crosslinking approaches (Feng et al., 1999). An interaction between BfpB and BfpG was also demonstrated by crosslinking and affinity purification (Daniel et al., 2006). Yeast two-hybrid studies further refined the binding site to the N-terminal third of BfpB (Daniel et al., 2006). While PilP does not consistently affect PilQ stability

or assembly, an interaction between the two proteins has been demonstrated. Far-westerns and cryo-electron microscopy show PilP binds a central region of PilQ (Fig. 3c) (Balasingham et al., 2007). Significant structural rearrangements in the ‘cap’ and ‘arms’ regions were visible in the PilP–PilQ secretin selleck chemical complex compared to the PilQ NVP-BKM120 price secretin complex alone. Nanogold labeling showed that

PilP was localized to the displaced regions of the secretin; the stoichiometry could not be determined as several different surfaces were labeled. Our knowledge of the ways in which secretins and pilotins/accessory proteins interact has grown significantly through the implementation of innovative functional assays and the advances in protein structure determination. Over time, the increasing diversity of mechanisms by which secretins are formed has become evident. While bacteria have a general secretion pathway for the majority of exoproteins, additional systems have evolved to specialize in and accommodate very specific functions: T4P production, the T3S needle-like injectosome, DNA uptake, and secretion of specialized proteins in response to environmental stimuli. Presumably, these systems are costly to maintain in the genome but have been retained to enable survival in niche environments. The fact that filamentous see more phage also use secretins to extrude from their bacterial hosts

certainly prompts speculation about the degree of co-evolution between the host and pathogen. A significant impediment to studying the in vivo interactions within these large membrane-spanning complexes has been the technical barriers to extraction of intact protein complexes from the membrane environment. However, the increasing body of research in membrane proteins and membrane protein complexes shows this is clearly no longer a deterrent. Continued research will undoubtedly lead to the development of novel methods to work with membrane proteins that will allow us to better understand the interactions between secretins and the proteins required for their formation. Despite the accumulation of a significant amount of data on secretin–pilotin and accessory protein interactions to date, many outstanding questions remain. While the Lol system is likely responsible for trafficking a pilotin–secretin subunit complex to the outer membrane, the process by which the secretin is assembled is unknown.

The eyes open/closed paradigm elicited alpha-band modulations in both lighting conditions, manifested in occipital and frontal electrodes. Fig. 2A depicts an example of alpha-band (8–12 Hz) modulations for a single subject, while the averaged results for all subjects can be seen in Fig. 2B. Alpha amplitude

was significantly larger when derived from occipital electrodes compared with frontal electrodes in both dark and light conditions, during eyes open as well as eyes closed (all paired t-tests, P < 0.005). Additionally, occipital alpha amplitude was larger during the eyes-open condition in the dark compared 3-MA clinical trial to light (paired t-test, P < 0.05). In accordance TGF-beta inhibitor with these results, an anova

conducted on alpha amplitude in all conditions (location, lighting and eye state) revealed a significant main effect for both eye state and location (P < 0.0001) but not for light (P < 0.88). Furthermore, a significant interaction was found for eyes × location (P < 0.002) but not for eyes × lighting (P < 0.23). The EEG classifier revealed a significant contribution of the alpha rhythm to eye state classification across subjects in both lighting conditions [P < 0.05, false discovery rate (FDR)-corrected; see Figs. 3A and B]. The weight of the alpha rhythm was 0.27 in the light condition and 0.2 in the complete darkness condition, indicating the amount of variability explained by the alpha rhythm between eyes open/closed in both lighting conditions. The chosen electrode for each subject was mostly located in the occipital regions under the light condition (in 79% of all subjects)

and in frontal regions under complete darkness (in 64% of all subjects). In accordance, highest classification rates were achieved in occipital electrodes under the light condition and in frontal electrodes under the complete darkness condition (for a distribution of classification rates across the scalp see Figs. 3C and D). Furthermore, the classifier revealed two more EEG bands as significantly contributing to eye state inference Etoposide datasheet – a wide beta (24–33 Hz) contribution during the light condition and a contribution of theta (4–7 Hz) during complete darkness. Accordingly, we found a significant correlation (average r = 0.46, P < 0.0002) between the alpha and theta timecourses in the dark condition in 79% of all subjects. This finding suggests a link between alpha and theta modulations mostly evident in the complete darkness condition, which could be related to internal mental context, as discussed later, or reflect changes in subjects’ vigilance state. Nonetheless, theta-related fMRI activation did not reach high statistical significance and therefore is not further discussed in the current paper, which focuses on the alpha band.

At the same time, one should be aware of the fact that multivariate approaches selleckchem may also be sensitive to confounds that systematically

covary with the conditions of interest. The fact that the GLM identified regions that overlapped with those found by the multivariate approach provides support that the multivariate approach is also driven by neural correlates of shifts in object-based attention. Furthermore, we analysed if the decoding was driven by highly localized activity patterns or by distributed cortical activations by training and testing decoders on individual clusters detected in the GLM. Because decoding on these small individual clusters yielded poor decoding performance compared with the whole-brain or GLM-restricted decoders, it suggests that faces and places are encoded in the brain using distributed patterns cortical activations, and as such detection of these patterns requires a multivariate decoder with input features spread across the brain. Finally, because the MVA-W classifier – trained only on pictures of separately presented faces and places – could not recruit any regions related to attention, we conducted a reverse MVPA to find regions associated with attention.

We trained two classifiers: one on the feedback condition; and the other on the non-feedback condition. Subsequently, these classifiers were tested on the localizer. We not only found activations in the same brain regions learn more responsible for processing faces and places as we found in MVA-W, but also detected

additional brain regions associated with attention and cognitive control. We found activation in superior frontal, middle frontal and superior medial frontal 4-Aminobutyrate aminotransferase gyri. These are part of the frontal-parietal network that have been known to become active in top-down attentional control paradigms (Li et al., 2010) and during bistable perception in which the observer’s perception can fluctuate between competing stimuli (Knapen et al., 2011). We also found activation in crus I of the left cerebellum. The cerebellum not only plays an important role in motor coordination, but has also been shown to be involved in higher cognitive functions such as selective visual attention (Allen, 1997). Moreover, activations in middle and anterior cingulate were also detected. Previous studies have shown that these regions play a crucial role in attention-demanding tasks by competition monitoring and goal-directed selective attention (Danckert et al., 2000; Davis et al., 2000). Activation in bilateral precuneus was also found, but only in the classifier trained on the non-feedback condition. Activation in this region has been shown in a previous study (Hahn et al., 2006) during engagement of top-down spatial selective attention. This may imply that subjects were engaged in both object-based and space-based visual attention during the non-feedback condition.

4 mM each of dNTP (Bangalore Genei), 0.2 U of Taq DNA polymerase (Bangalore Genei), 10 pM each of forward and reverse primers, and 10 ng of genomic DNA was used as template in PCR tubes. PCR program was as initial denaturation at 95 °C for 3 min, subsequently, five touch-down PCR cycles comprising of 94 °C for 20 s, 60/55 °C (depending on the marker as given in Table 3) for 20 s, and 72 °C for 30 s were performed. These cycles were followed by 40 cycles of denaturation at 94 °C for 20 s with constant annealing

temperature of 56/51 °C (depending on marker) for 20 s, and extension at 72 °C for 20 s, and a final extension at 72 °C for 20 min. All PCR amplicons were www.selleckchem.com/PARP.html resolved by electrophoresis on 3% agarose gel to identify the informative SSR loci across all the isolates. GeneRuler 100-bp DNA ladder (MBI Fermentas) was used to estimate the allele size. The amplification data generated

by SSR markers were analyzed using SIMQUAL route to generate Jaccard’s similarity coefficient (Jaccard, 1908) using ntsys-pc, software version 2.1 (Rohlf, 1998). These similarity coefficients were used to construct a dendrogram depicting genetic relationships among the isolates by employing the Unweighted Paired Group Method of Arithmetic Averages (UPGMA) GSK2126458 clinical trial algorithm and SAHN clustering. The robustness of the dendrogram was evaluated with a bootstrap analysis performed on the binary dataset using winboot software (version. 2.0). The allelic diversity or polymorphism information content (PIC) was measured as described by Botstein et al. (1980). PIC is defined as the probability that two randomly chosen copies of gene will be different alleles within a population. The PIC value was calculated with the formula as follows: where Pij represents the frequency of the jth pattern for marker i, and summation extends over n patterns. The frequency of repeat motifs in the consensus EST sequences and annotated transcripts was assessed, and both perfect and compound SSRs were selected with a minimum acceptable length of 12 bp for di, tri, and tetra-nucleotide motifs Orotidine 5′-phosphate decarboxylase (Garnica et al., 2006). Only SSRs with a minimum

of three repeats were included in the analyses of penta- and hexa-nucleotide repeats. Maximum number of SSR (1679) was identified in Fol followed by Foc (313) and Fom (204). The higher number of SSRs in Fol was expected because the total size covered by transcripts sequences of Fol (21.7 Mb) was much higher than that of ESTs of Fom (1.3 Mb) and Foc (2.4 Mb). To compare the SSR count between all three formae speciales, the complete length of each set of sequences was analyzed, and thus, total relative abundance and total relative density were calculated and depicted in Table 1. It was found that relative abundance of SSRs in Fom (157) was higher than Foc (130) and Fol (77). Similarly, the relative density of SSR was also higher in Fom (2117) in comparison with Foc (1680) and Fol (1071) (Table 1; Fig. 1a and b).

9 Mosquito bite protection is an essential component of malaria prevention, and N,N-diethyl-3-methybenzamide (DEET) repellents can be used for infants aged >2 months.10 Generally, pediatric malaria case numbers are increasing as more children travel and the profile of migration http://www.selleckchem.com/products/VX-765.html is changing.11–13 In the study from Stäger et al.,14 returning to the country of origin to visit friends and relatives was a significant risk factor for the acquisition of malaria. A recent analysis suggests that it is cost-effective to subsidize malaria chemoprophylaxis for low-income travelers visiting high-risk malaria endemic areas, and this may encourage use of malaria prophylaxis in VFR travelers.15

School-, sport-, and community-based strategies to reach VFR children need to be evaluated.16 A relation between the place of exposure and the spectrum of disease can help in diagnostic approaches and empiric therapies.17,18 Nontravel medicine practitioners should be reminded to ask the question “did you travel recently?” when taking a history. Depending on the travel destination, travelers may be exposed to a number of infectious diseases; exposure depends on the presence of infectious agents in the respective area. The risk of becoming infected will vary according to the purpose of AZD8055 cell line the trip, the itinerary within the area, the standards

of accommodation, hygiene, and sanitation, as well as the behavior of the traveler and the reason for travel—whether it is for ID-8 tourism, VFR travel, or for immigration.19 VFR travelers are exposed to an increased risk of travel-related health problems.20–22 General practitioners should be aware of possibly serious travel-related disease in VFR risk groups in their community. VFR travel to Africa is associated with malaria, while VFR travel to Asia including Turkey is associated with typhoid fever. Two cases of tuberculosis in VFR

children were acquired in Turkey and Kosovo. Physicians attending to returned ill children need to be aware of and to diagnose a complete range of diseases from commonplace to serious. Parents can be provided with a simple range of pediatric medications and instructions on how to treat self-limiting conditions. The pre-travel consultation is an opportunity to provide concise preventive advice for pediatric travelers. The country of origin of settled migrants has an important role to play in the diagnosis profile. VFR children will present with potentially more serious illnesses such as typhoid fever, hepatitis A, and malaria. We thank the members of the secretariat especially Mrs Lopez from the University of Zürich Children’s Hospital, Division of Infectious Diseases. P. S. has received research grants and consultancy fees from F. Hoffmann La Roche, speaker’s honorary from GSK, and is an advisory board member of sigma tau. The other authors state that they have no conflicts of interest to declare. All authors have seen and approved the final version of the paper. T. H.

The ability to selectively target specific subpopulations of GIRK channels may prove effective in the treatment of disorders of excitability. “
“Abnormally large tremor during movement is a symptom of many movement disorders and significantly impairs activities of daily living. The aim of this study was to investigate whether repetitive magnetic brain stimulation (rTMS) can reduce tremor size during human movement. We hypothesised that inhibitory rTMS over motor cortex would reduce tremor size during subsequent movement. The study involved 26 healthy young adults

(21 ± 2 years) this website and began with application of single TMS stimuli to measure baseline corticospinal excitability. The response to stimulation was recorded in hand muscles with electromyography. Subjects then performed a 3-min task to measure baseline tremor during movement. This involved matching index finger position with a moving target on a computer screen. Tremor was recorded with an accelerometer on the fingernail. Finger acceleration was analysed with fast-Fourier transform to quantify tremor in the physiological range (7.8–12.2 Hz). Subjects then received 10 min of real (n = 13) or sham (n = 13) inhibitory rTMS. Tremor and corticospinal

excitability were then remeasured. CYC202 supplier Real rTMS significantly decreased corticospinal excitability by ~30% (P = 0.022) and increased tremor size during movement by ~120% (P = 0.047) relative to sham rTMS. However, the direction of tremor change was opposite to that hypothesised for inhibitory rTMS. The results suggest that rTMS over (-)-p-Bromotetramisole Oxalate human motor cortex can modulate action tremor and the level of corticospinal

excitability may be important for setting the amplitude of action tremor in healthy young adults. “
“In adult mice, classical conditioning in which whisker stimulation is paired with an electric shock to the tail results in a decrease in the frequency of head movements, induces expansion of the cortical representation of stimulated vibrissae and enhances inhibitory synaptic interactions within the ‘trained’ barrels. We investigated whether such a simple associative learning paradigm also induced changes in neuronal excitability. Using whole-cell recordings from ex vivo slices of the barrel cortex we found that layer IV excitatory cells located in the cortical representation of the ‘trained’ row of vibrissae had a higher frequency of spikes recorded at threshold potential than neurons from the ‘untrained’ row and than cells from control animals. Additionally, excitatory cells within the ‘trained’ barrels were characterized by increased gain of the input–output function, lower amplitudes of fast after-hyperpolarization and decreased effect of blocking of BK channels by iberiotoxin.

7% agar and containing 100 μL of overnight bacterial culture was spotted after solidification with 5 μL suspensions of the 16 isolated bacteriophages. The plates were incubated at 25 °C, and the occurring lysis was investigated after 18–24 h. Propagation of phages for genome isolation was initiated according to the double agar layer method (Adams, 1959). After incubation at 25 °C for 18 h, the top layers were collected and placed into 10 mL SM buffer. Belnacasan cost After gentle agitation for 4 h at 25 °C, 2 mL chloroform was added, mixed, and incubated at 4 °C for 18 h. The resulting suspension was decanted over the chloroform and centrifuged at 5000 g for 40 min at 10 °C to eliminate the bacterial cells;

the supernatant was centrifuged again at 16 000 g for 60 min at 10 °C to collect the phage particles. The pellet was resuspended in 500 μL EDTA (pH 8.0) and 500 μL TES (10 mM Tris–HCl, 10 mM EDTA, 2% SDS, pH 8.0). After vigorous vortexing

for 30 s, the solution was incubated for 30 min at 65 °C. The proteins were precipitated with protein precipitation solution (Sigma-Aldrich), incubated on ice for 5 min, and centrifuged at 15 000 g for 4 min at 10 °C. The nucleic acid was precipitated with 0.1 volume 3 M Na-acetate and 1 volume ethanol. The pellet was washed with 70% ethanol; dried, and resuspended in 20 μL TE buffer (10 mM Tris–HCl, 1 mM EDTA, pH 8.0). To determine the type of the genome nucleic acid, RNase and DNase treatments were carried out according to the manufacturer’s (Sigma) instructions. Restriction fragment pattern differences of the investigated phage DNAs were examined SB203580 research buy with 21 different restriction endonucleases: AatI, AluI, ApaI, BamHI, BcuI, BglI, BglII, Bsh1236I, ClaI, DraI, EcoRI, HaeIII, Hin6I, HindIII, KpnI, MspI, NotI, PstI, RsaI, SacI, TaqI (Fermentas, Thermo Scientific). The growth Methane monooxygenase characteristics of the Bf7 were investigated using the double layer method

(Adams, 1959) on its host, incubated for 18–48 h at different temperatures (5, 10, 20, 25, 30, and 35 °C), and the resulting plaque numbers and morphologies were compared. The single-step growth curve experiments were carried out according to the protocol of Keel et al. (2002), with minor modifications. The LB liquid medium was supplemented with glucose (0.3%), CaCl2 (0.075 mM), MgSO4 (2 mM), and FeCl2 (0.004 mM) according to the suggestions of Sambrook et al. (1989), for higher phage titer. Exponential-phase culture of P. tolaasii 2342T was treated with bacteriophage solution to have a multiplicity of infection (MOI) of 0.06. To visualize phage morphology with transmission electron microscopy, phage plaques were picked and placed in SM buffer. Aliquots were mounted on a carbon-coated formvar film supported by a 300 mesh copper grid. Samples were negatively stained with 1% uranyl acetate and examined by a Zeiss CEM 902 electron filtering electron microscope.

7% agar and containing 100 μL of overnight bacterial culture was spotted after solidification with 5 μL suspensions of the 16 isolated bacteriophages. The plates were incubated at 25 °C, and the occurring lysis was investigated after 18–24 h. Propagation of phages for genome isolation was initiated according to the double agar layer method (Adams, 1959). After incubation at 25 °C for 18 h, the top layers were collected and placed into 10 mL SM buffer. selleck kinase inhibitor After gentle agitation for 4 h at 25 °C, 2 mL chloroform was added, mixed, and incubated at 4 °C for 18 h. The resulting suspension was decanted over the chloroform and centrifuged at 5000 g for 40 min at 10 °C to eliminate the bacterial cells;

the supernatant was centrifuged again at 16 000 g for 60 min at 10 °C to collect the phage particles. The pellet was resuspended in 500 μL EDTA (pH 8.0) and 500 μL TES (10 mM Tris–HCl, 10 mM EDTA, 2% SDS, pH 8.0). After vigorous vortexing

for 30 s, the solution was incubated for 30 min at 65 °C. The proteins were precipitated with protein precipitation solution (Sigma-Aldrich), incubated on ice for 5 min, and centrifuged at 15 000 g for 4 min at 10 °C. The nucleic acid was precipitated with 0.1 volume 3 M Na-acetate and 1 volume ethanol. The pellet was washed with 70% ethanol; dried, and resuspended in 20 μL TE buffer (10 mM Tris–HCl, 1 mM EDTA, pH 8.0). To determine the type of the genome nucleic acid, RNase and DNase treatments were carried out according to the manufacturer’s (Sigma) instructions. Restriction fragment pattern differences of the investigated phage DNAs were examined Ipilimumab price with 21 different restriction endonucleases: AatI, AluI, ApaI, BamHI, BcuI, BglI, BglII, Bsh1236I, ClaI, DraI, EcoRI, HaeIII, Hin6I, HindIII, KpnI, MspI, NotI, PstI, RsaI, SacI, TaqI (Fermentas, Thermo Scientific). The growth Meloxicam characteristics of the Bf7 were investigated using the double layer method

(Adams, 1959) on its host, incubated for 18–48 h at different temperatures (5, 10, 20, 25, 30, and 35 °C), and the resulting plaque numbers and morphologies were compared. The single-step growth curve experiments were carried out according to the protocol of Keel et al. (2002), with minor modifications. The LB liquid medium was supplemented with glucose (0.3%), CaCl2 (0.075 mM), MgSO4 (2 mM), and FeCl2 (0.004 mM) according to the suggestions of Sambrook et al. (1989), for higher phage titer. Exponential-phase culture of P. tolaasii 2342T was treated with bacteriophage solution to have a multiplicity of infection (MOI) of 0.06. To visualize phage morphology with transmission electron microscopy, phage plaques were picked and placed in SM buffer. Aliquots were mounted on a carbon-coated formvar film supported by a 300 mesh copper grid. Samples were negatively stained with 1% uranyl acetate and examined by a Zeiss CEM 902 electron filtering electron microscope.

Introns were detected in the cox1, cox2, nad5, rns and rnl genes. All of these are type I introns, except for the single type II intron in rns. All type I introns contained endonuclease-like gene sequences with the conserved LAGLIDADG motif, except for the first intron in the cox1 gene, which had the GIY-YIG motif. The endonuclease in cox1 intron-12 appears to be truncated and does not have the full LAGLIDADG domain. Of the genes found in the mitochondrial genome of T. cingulata, the structure of cox1 is the most complex. Of the 16 exons that

make up the cox1 gene, five are smaller than 20 nt long, with the smallest two being only 11 nt. All 15 introns have at least one ORF larger than 100 codons. ORFs encoding endonuclease-like sequences Birinapant nmr were also seen in all other introns, except for intron-1 of cox2. The reading frames of the exons 1 and Selleckchem Bcl2 inhibitor 2 of nad5 continue well beyond the predicted splice sites into the respective introns. These extended reading frames also encode endonuclease-like sequences within an ORF (Fig. 1). While the coding regions have been well conserved among the Agaricomycotina

and to a lesser extent with U. maydis, the introns show less similarities (Fig. 1). Trametes cingulata intronic ORFs show greater sequence similarity to P. ostreatus and M. perniciosa than to the more distantly related U. maydis. Schizophyllum commune and C. neoformans do not have introns in the same genes as T. cingulata. The DNA and RNA polymerases dpo and rpo, which have been reported in P. ostreatus and M. perniciosa, are not present in the T. cingulata mitochondrial genome nor were they annotated or obvious in the S. commune, C. neoformans or U. maydis mitochondrial genomes. The 25 identified tRNAs genes

represent all 20 amino acids and include three copies encoding tRNAMet and two each of tRNAArg, tRNASer and tRNALeu. Single genes encode the other 16 tRNAs. We analyzed codon usage for the 15 protein-encoding annotated genes (Supporting Information, Table S1) and found that all of these genes use TAA Phospholipase D1 as the stop codon, except for nad5, which uses TAG. nad5 is also the only gene that uses GAG as a codon for glutamic acid and AGG for arginine, which is otherwise encoded exclusively by AGA. Other codons for arginine, CGG, CGT and CGC, are not used. The glycine codon GGC is also not used. At least one of these four otherwise unused codons is used one or more times in all five of the unidentified ORFs, lending additional support to the hypothesis that these ORFs are not expressed genes. The alternate codon for tryptophan TGA that differs from the standard codon table is not used either as a codon for tryptophan or to represent a stop codon in the 15 protein-encoding annotated genes. However, it is present twice in ORF111 and once in ORF158. Nad4L is the only gene that uses AAG to code for lysine and does it only at one position. Only 13% of the codons for tyrosine use TAC, with the rest using TAT.

Pharmacists perceive NMS to be of value to patients and believe that providing this service should promote their professional reputation. However, the requirement to consent patients and, the language and behaviour adopted by pharmacists when recruiting and providing these services

may result in the profession being unable to fully realise this opportunity. These findings represent the views of a small convenience sample of pharmacists and are not generalisable. 1. Pharmaceutical Services Negotiating Committee. NMS. Available from www.psnc.org.uk/pages/nms.html. Accessed 22nd April 2013. Amelia Taylor, Murray D Smith, Li-Chia Chen University of Nottingham, Nottinghamshire, UK Development of an adherence measure suitable for use with UK primary care general practice prescribing data. Applied Apoptosis inhibitor to measure the use of inhaled corticosteroids (ICS) by asthma patients. The adherence measure, a Prescription Possession Ratio (PPR), was calculated using five alternative strategies. On comparison, the results consistently demonstrate excessive proportions of patient-years were either over- or under-prescribed. PPR may be a useful tool to signal adherence issues and measure changes in adherence over time. Medication adherence1 is a key factor in the efficacy of pharmacotherapy, especially for long-term conditions. For example, poor adherence to ICS is known

as the main cause for therapeutic failure in asthma treatment and is associated with increased morbidity. Despite several techniques being available (e.g. pill counts, electronic Dorsomorphin in vitro measuring devices, questionnaires), there is no gold standard offering cheap and practical adherence measures in clinical practice. In this study, the aim is to use retrospective prescribing data from UK primary care to develop a PPR measure for evaluating asthma patients’ adherence to ICS. This is a retrospective cohort study over a 1997–2010 sample frame involving asthma patients Phosphatidylinositol diacylglycerol-lyase aged between 12 and 65 years who are without a diagnosis of chronic obstructive pulmonary disease. Data are sourced from the Clinical Practice Research Datalink database.

Approval for use of the data was granted by the Independent Scientific Advisory Committee. Patients’ ICS prescriptions are used to calculate individual PPR2 in each annual interval by dividing ‘number of days prescribed during calendar year’ by ‘number of days in the interval’ and converting into a percentage. To develop the PPR, several alternative definitions are considered when calculating the numerator ([a] including or [b] excluding overlap in prescribed days, [c] carryover or [d] proportionally sharing number of prescription days to the next interval) and the denominator ([e] interval started from entry date and calculate by sum of prescription intervals, or [f] set as 365 days). Five scenarios are selected to test the consistency of the PPR measures.