Using results from a large number of studies including a wide ran

Using results from a large number of studies including a wide range of sample characteristics, the minimum number of consumers can be determined as the minimum number that provides stable sample configurations. For each study the average RV coefficient across simulations is determined for different number of assessors and the number required for obtaining an average RV coefficient of 0.95 is determined (Figure 2). This approach has been used for making recommendations on the minimum number of consumers needed for sorting tasks [22••], CATA questions [23] and projective mapping [25]. Despite the potentialities of this approach

for evaluating reliability it is still necessary to evaluate other parameters to evaluate the similarity between sample configurations. In particular, it is important to stress that the RV coefficient depends PD-0332991 manufacturer on the number of samples considered in the study [26] and

therefore it might not be the best parameter for evaluating the similarity of sample configurations. An alternative would be to use the RV2 coefficient, as stressed by Tomic et al. [17]. Another important issue that deserves further research is the threshold considered for determining that sample configuration MAPK inhibitor is stable. As an example, Vidal et al. [25] reported that changing the RV coefficient from 0.95 to 0.90 strongly changed conclusions on the stability of sample configurations but did not decrease sample discrimination. In closing this section it is interesting to highlight that additional MycoClean Mycoplasma Removal Kit statistical tools can be used to evaluate the stability of sample configurations. The adjusted Rand index has been recently proposed to evaluate the agreement of partitions of a set of samples in a sorting task [27]. This statistical tool can be extended to evaluate the stability of sample grouping obtained using cluster analysis on sample coordinates on the configurations gathered with different rapid methodologies. Perhaps one of the most important challenges regarding new methodologies for sensory characterization

is identifying their limitations. It is clear that these methodologies are not a replacement of classic DA with trained assessors. However, it has not been clearly established yet in which situations new methodologies provide equivalent information to DA and when they their application is not recommended if high quality detailed information is sought. Several studies comparing sensory characterizations obtained using DA with trained assessors and new methodologies with non-trained assessors have been performed using samples that show large or medium differences among them 28 and 29. In this sense, studies focusing on the effect of sample complexity and the degree of difference among samples on the discriminative ability of new methodologies are still lacking.

Hence, within one experiment, only one investigator should be ass

Hence, within one experiment, only one investigator should be assigned the tasks of recovering the S-9 fraction. Other sources of variation may be attributed to differences in the analyst, temperatures in the lab and in the spectrofluorometer, timing of thawing and preparation of reaction solutions, and reagent quality. Munkittrick et al. (1993) pointed out large differences among laboratories in reported extinction coefficients of standard resorufin solutions, GSK126 reflecting differences among batches of standard, the instruments used to measure extinction coefficient, and the procedures of each laboratory. To assess the occurrence and extent of variation, we maintain control

sheets showing the variations among assays in activity of standard S-9 fractions, prepared Protein Tyrosine Kinase inhibitor from control rainbow trout or trout exposed to ß-naphthoflavone (BNF), a model CYP1A inducer. These ‘lab standards’ were prepared by mixing the S-9 fractions from numerous control and BNF-exposed fish, dividing the mixed S-9s into small aliquots and storing them frozen at −80 °C. One

of each is analyzed with each experimental set of samples over a 6–18 month period to demonstrate that the analytical method works on each occasion, and to identify occasions when the method generated data that might be higher or lower than normal. After a new batch of S-9s has been prepared and stored, the control chart is prepared from the first five samples of positive and negative control samples analyzed. The chart consists of the 95% confidence limits about the geometric mean EROD activity of the positive and negative controls, and of the induction (positive divided by negative). Subsequent samples are plotted on the same chart, and most of the new values should fall within the 95% confidence limits, and any random or systematic change in Liothyronine Sodium expected activity can be identified quickly. As Fig. 1 demonstrates for one batch of positive and negative control S-9s tested over 16 months, that EROD activities of induced and control

fish, and induction (the ratio of induced to control activities) varied considerably among assays. Because some of this variation could be due to poor mixing of the original S-9 fractions from individual fish, we also analyzed five control and five BNF S-9 standards on one occasion. The coefficient of variations for the positive and negative controls, and for induction based on arithmetic means were 31%, 19%, and 39%, respectively, much lower than the ‘among assay’ variations of 140%, 39%, and 104%, respectively from the first five samples tested in Fig. 1. Therefore, the scatter observed in Fig. 1 was due to ‘among assay’ variance rather than ‘within assay’ variance, and reflected differences in procedures or assay conditions, even though the analyses were done by the same person. The data also suggest an increasing trend in positive and negative control results, but not in induction.

Upon her return to Oxford from her USA sabbatical she also studie

Upon her return to Oxford from her USA sabbatical she also studied the patterns of RNA metabolism in the different functional states of bone cells. These investigations of the dynamics of cell differentiation and the hormonal effects on bone were investigated in great detail, by the tedious method of cell and autoradiographic grain Selumetinib cost counting, in ground-breaking publications. In particular the effects of parathyroid extract were shown in vivo on both mature osteoblasts and osteoclasts and also on osteogenic and osteoclastic bone progenitor

cellular activity. An important observation was that the there were different and rapid effects of the hormone on uptake of RNA precursors in the osteoblasts and osteoclasts. In Linsitinib cell line addition to her studies with radioactively labelled amino acids, labelled glucosamine as a precursor of glycoproteins was used to show the autoradiographic distribution with time of labelled material found associated with osteoclasts and the resorbing bone surfaces. The osteoclast was suggested to synthesise and subsequently to extrude such labelled material on the resorbing bone surface to aid resorption. This uptake was enhanced by parathyroid hormone. Another important outcome from her work was the observation that the high

labelling by thymidine of the ‘preosteoblastic layer’ on the periosteal surfaces of the young rabbits studied, most probably was in a layer immediately adjacent to the local stem cells of this tissue from which other cells are derived. In addition the cell kinetics of the fibroblast–pre-osteoblast–osteoblast–osteocyte system was investigated and this generated many important conclusions concerning cell transitions through these

compartments and functional matrix synthesis. At that time, however, in the early 1960s, the osteogenic and osteoclastic cell lineages were considered to be different functional states of the same cell rather than having distinctly separate origins postnatally, as was proven later. Following Dame Janet’s retirement Maureen, now a permanent member of the MRC External Scientific Staff, succeeded her as head of the newly-named MRC Bone Research Laboratory firstly at the Churchill Hospital Oxford, and subsequently, DAPT solubility dmso in 1974, at the Nuffield Orthopaedic Centre. At this point in her career her focus on osteoblastic cell differentiation became paramount. With remarkable insight, she recognised that the progenitor cells of musculoskeletal stromal tissues would be central to future investigations of bone diseases and their treatments and to normal musculoskeletal physiology. In an editorial in Calcified Tissue Research in 1978, Maureen drew attention to the importance of the two separate cell systems present in bone marrow, the stromal and haemopoietic systems.

In addition, FLI-1 is also involved in various malignancy formati

In addition, FLI-1 is also involved in various malignancy formation and progression in vitro and/or in vivo, including Ewing’s sarcoma [7], melanoma [8], breast cancer [9], lymphoma [10] and [11] and head and neck squamous cell cancer (HNSCC) [12], and tumor micro-angiogenesis [13]. Studies on the role of FLI-1 expression in NPC are rare. FLI-1 was found to be over-expressed in the metastatic Venetoclax NPC cell line, the 5-8F cell line, in the research by Yang et al [14]. However, little is known about the FLI-1 expression and prognostication of NPC patients. Therefore, this study aims to detect FLI-1 expression in NPC tissue

samples by immunohistochemistry (IHC), analyze the associations between FLI-1 expression and clinicopathological characteristics, and evaluate the prognostic value of FLI-1 for NPC patients.

This study was approved by the Clinical Ethics Review Board of Sun Yat-sen University Cancer Center. All the patients signed informed consent documents before participating in the study. Patients were recruited according to the following criteria: histologically diagnosed NPC with available biopsy sample; newly proven and non-metastatic NPC; no other malignancy or prior anti-cancer treatment; selleck inhibitor continuously finished at least radiotherapy at the Cancer Centre of Sun Yat-sen University with complete and detailed medical records and regular follow-ups. A total of consecutive 198 patients were eligible, who were diagnosed between May 2005 and December 2006. Medical files were reviewed retrospectively and patients were restaged based on the American Joint Committee on Cancer (AJCC) staging

system 2010 clinical classification (the seventh edition). All 198 patients were histologically diagnosed with differentiated non-keratinized carcinoma or undifferentiated non-keratinized carcinoma. Forskolin in vitro The tumor specimens were obtained by biting biopsy from primary NPC, prior to treatment, and processed through formalin fixation for at least 8 hours and paraffin embedment. Patients underwent a routine pretreatment evaluation including history, physical examination of the head and neck, optic fiber nasopharyngoscopy, nasopharynx and neck magnetic resonance imaging (MRI), chest X-ray, the abdominal ultrasonography, bone scanning, a complete blood count and biochemical profile. The serological titer of Epstein-Barr virus immunoglobulin A antibodies against viral capsid antigen (EBV VCA-IgA) was measured using an immunoenzymic assay. The serum titer of Epstein-Barr virus immunoglobulin A antibodies against early antigen (EBV EA-IgA) was further measured using an immunoenzymic assay by Raji cell line.

After 24 weeks, mean change in bodyweight from baseline


After 24 weeks, mean change in bodyweight from baseline

was +1.4 kg in the BIAsp BID + Sit group (difference vs. BIAsp QD + Sit: 1.51 [95% CI 0.82; 2.21], P < 0.001), +2.1 kg for BIAsp BID (difference vs. BIAsp QD + Sit: 2.19 [95% CI 1.49; 2.89], P < 0.001) and Rapamycin datasheet −0.1 kg for BIAsp QD + Sit. No significant difference was reported between the two BID groups. Final total daily dose was 0.66 U/kg, 0.72 U/kg and 0.39 U/kg, respectively, from a baseline of 0.16 U/kg. In the BID groups, the morning dose increased from 0.08 U/kg to 0.35 U/kg (BIAsp BID + Sit) and 0.39 U/kg (BIAsp BID), while the evening dose increased from 0.08 U/kg to 0.31 U/kg (BIAsp BID + Sit) and 0.34 U/kg (BIAsp BID). There were no significant differences in TRIM-D scores among the treatment groups. The overall TRIM-D score after 24 weeks was 76.64, 77.79 and 76.46 in the BIAsp BID + Sit, BIAsp QD + Sit and BIAsp BID groups, respectively, with baseline values

of 70.28, 72.40 and 69.30. Average total medicine costs in each arm (in subjects exposed ≥20 weeks) were GBP 345.7 for BIAsp BID + Sit, GBP 287.9 for BIAsp QD + Sit and GBP 160.0 for BIAsp BID. No further cost analyses were conducted. Clinicians need to balance risks, costs and benefits of different treatment approaches when choosing a suitable treatment plan for patients with diabetes. Moreover, individual circumstances should be considered, i.e. age, comorbidities, baseline HbA1c, ability to find more adhere to complex regimens, to optimize outcomes when choosing an antihyperglycaemic strategy [2]. Sit2Mix included a relatively homogenous population at baseline and investigated three distinct intensification regimens in patients with T2D failing to be controlled on sitagliptin and metformin in combination with other OADs. All regimens were efficacious and well tolerated after 24 weeks of treatment, Interleukin-3 receptor but each presented a different profile in terms of treatment benefits

and risks. The BIAsp BID + Sit regimen showed greater improvement in glycaemic control versus BIAsp QD + Sit and BIAsp BID. Nevertheless, HbA1c change in both the BIAsp QD + Sit and BIAsp BID groups was ≥1.0% and a change of this magnitude is associated with reduced risk for microvascular and macrovascular complications [14]. The improvement observed in mean SMPG after breakfast and lunch, and before lunch and dinner, in the BID groups is likely a reflection of the different dose-administration timings with BID and QD regimens (dosing before breakfast and dinner vs. dosing before dinner only, respectively). Although a greater proportion of patients achieved the recommended HbA1c target <7.0% in the BIAsp BID + Sit group (approximately 60%) versus the other groups, this trend was not maintained upon examination of those patients who achieved target without hypoglycaemia. For this endpoint, the proportions of responders in the BIAsp BID + Sit and BIAsp QD + Sit groups were comparable (39–41.

Small local rivers originating in the Siwalik Hills, including th

Small local rivers originating in the Siwalik Hills, including the Turia, Jharia and Bhaluhi, dissect the floodplain in a North-South orientation (Pathak and Rao, 1998). The plain is situated on the Rapti–Gandaki interfan region and is mainly comprised of Holocene alluvium (NASC, 2004). Unlike other regions of Terai, where finer of sediments typically increase toward the south, fines predominate in the north and sand and gravels are found near the Nepal–India border (Shrestha et

al., 2004). In the areas with fine grained sediments, elevated concentrations of As are typically recorded (Brikowski et al., 2004 and Brikowski et al., 2013). All glass and plasticware used during sampling and laboratory analysis were soaked in 5% HCl for 24 h and then rinsed with deionized Selleckchem HSP inhibitor water (Milli-Q) for at least 24 h. All reagent solutions were prepared with Milli-Q water having a resistivity of 18.2 MΩ/cm. In October 2012, tubewell water samples were collected along the floodplain LBH589 of the Bhaluhi River, Nawalparasi district

(Fig. 1). The sampling area was divided into three topo-gradient regions along the flow path of the Bhaluhi River referred herein as (i) the Upper region, (ii) the Middle region and (iii) the Lower region. The upper region lies on the edge of the Bhabar zone, while the middle and lower region are situated on the Terai plain. This division was based on the recognition that geomorphic and landform features can exert a vital control on aquifer stratigraphy and corresponding geochemistry and As concentrations (McArthur et al., 2011, Nath, 2012, Shamsudduha et al., 2008, Weinman et al., 2008 and Winkel et al., 2008) and the distribution of As concentrations in the aquifer derived from prior testing of the region. Seventy-three water samples from tube wells

and eight samples from the Bhaluhi River were collected for detailed aqueous phase characterization (Fig. 1). Most of the investigated tube wells were currently used by local people for domestic drinking or irrigation purposes. Information such as depth, age, screen interval, method of drilling and construction of tube well were collected via interview with the owner or nearest CYTH4 household user of the well. Each tube well was subjected to 5–10 min of continuous pumping, during which time the redox potential, pH, temperature, dissolved oxygen (DO) (luminescence probe) and electrical conductivity of the water was measured with HACH multimeters (HQd) and freshly calibrated probes. After 5–10 min of pumping and stabilization of physico-chemical parameters, water samples were collected into a clean high-density polyethylene (HDPE) bottle flushed with sample water three times and filled without any headspace.

(2012) The core of the Pelops anticyclonic eddy (Figures 2f–j) d

(2012). The core of the Pelops anticyclonic eddy (Figures 2f–j) displays insignificant warming relative to the surrounding area, indicating an insignificant change in the intensity of the Pelops eddy. Moreover, the grouping of eddies in the western Levantine basin (Millot, 2005 and Poulain et al., 2012) is less obvious, as there is only one anticyclonic eddy south of Crete in autumn (warm core, 21.8 °C). The core of this anticyclonic eddy displays more significant (insignificant) warming than does the surrounding area in summer, autumn and winter (spring), indicating the dominance of this eddy

and suggesting that it may become more intense in the future. In addition, about three obvious anticyclonic eddies are attributable selleck products to the seasonal warming gradient over the western Levantine, especially in summer and autumn, indicating that the western Levantine eddies may become more significant in the

near future. The eastern Levantine eddies (Poulain et al. 2012) are not obvious from the seasonal average SST gradient, as a 1/4° projection grid was used. Poulain et al. (2012) described the eastern Levantine eddies using altimetry data with 1/8° gridded resolutions. There is an obvious grouping of eddies in the eastern Levantine attributable to the seasonal warming gradient, especially in summer and autumn, indicating more intense eastern Levantine eddies in the future. The

Ionian sub-basin SST increases zonally from north (Gulf of Taranto) to south (west of Gulf of Sidra, Libya) in winter (13.9–17.4 °C) and autumn (18.1–22.2 °C), and from north-east (24.2 °C) to south-west (28 °C; Gulf of Gabes, Tunisia) in summer. In spring, however, the Ionian SST displays a mixed zonal and meridional gradient, ranging from 18.2 °C off the north-western Ionian coast to 20.8 °C in the Gulf of Carbohydrate Gabes, Tunisia. Ionian mesoscale structures do not generally become more obvious with the seasonal SST increase, although the Ionian mesoscale eddies do become more obvious with the seasonal warming. The latter may indicate a significant increase in the intensity of Ionian mesoscale eddies in the near future. The Mid-Ionian Jet (Poulain et al. 2012) is generally obvious in the annual SST distribution (SST, 20.2 °C), most markedly in summer (SST, 25.5 °C). There is a significant difference in the SST gradient between the northern (meridional distribution) and southern (zonal distribution) Tyrrhenian sub-basin, partly due to the surface water circulation. The northern Tyrrhenian SST increases from north-east to south-west as follows: 13.6–14.6 °C (winter), 17.6–19 °C (spring), 23.2–26 °C (summer), and 16.4–19.8 °C (autumn). However, the southern Tyrrhenian SST increases zonally from south to north. The northern Tyrrhenian gyre (Poulain et al.

, 2011 and references therein)

Regulation of gene expres

, 2011 and references therein).

Regulation of gene expression by chromatin is in part regulated by a class of enzymes which methylate or demethylate histone proteins. The first lysine specific histone demethylase discovered is LSD1. This enzyme a member of the monoamine oxygenase family (EC: and catalyzes the demethylation of mono- and di-methylated lysine through reduction of FAD. The reaction proceeds through the formation of a positively charged imine intermediate which degrades to produce formaldehyde and the amine. In this process FAD is reduced BMS-354825 in vitro to FADH2 which is subsequently reoxidized by molecular oxygen with the production of hydrogen peroxide. Therefore a number of enzymatic products are available and assays have been developed using LC/MS to detect peptide product (Metzger et al., 2010) and coupled enzymatic reactions have been used to detect either hydrogen peroxide or formaldehyde (Forneris et al., 2005). High-throughput mass spectrometry methods, such as the RapidFire mass spectrometry system

from Biotrove (Hutchinson et al., 2012) can enable HTS on libraries as large as ~200 K in size (Ozbal et al., 2004 and Roddy Roxadustat ic50 et al., 2007). A TR-FRET assay operating in a signal decrease mode, using an antibody that recognizes H3K4me1 but not the unmethylated product, has been recently described (Yu et al., 2012). Additionally, an AlphaScreen-based assay has also been developed using an antibody

to an H3K4me1 peptide (Gauthier et al., 2012). A sensitive assay using TR-FRET-based detection of an unmethylated histone-3 peptide by a fluorescent europium-chelate labeled monoclonal antibody which binds specifically to the H3K4me0 site has been used in HTS (Wang et al., 2011). As the antibody in this assay recognizes the unmethylated product, an increase in signal upon LSD1 inhibition is obtained which is more desirable than a signal decrease mode where compounds which interfere with the signal would tuclazepam be detected. Generic assays for HMTs have also been developed. Some HMTs that catalyze the transfer of a methyl group to either lysine or arginine require the co-factor adenosyl-l-methionine (SAM). A generic assay for this class methyl transferases has been described (Ibanez et al., 2012). In this assay biotinylated peptides are methylated with a [3H-Me]-SAM cofactor and streptavidin-coated SPA beads are used for detection. When histone H3 is employed as a common substrate, this SPA format provides a generic read-out for HMTs.

Ar), and cortical thickness

Ar), and cortical thickness (Ct.Wi) (Table 2B). However, in Haversian canals, haversian labeled surfaced (H.L.Pm/Ec.Pm), mineral apposition rate (H.MAR) and bone formation rate (H.BFR/BS) were dose-dependently decreased, and a significant change was observed in H.L.Pm/Ec.Pm and H.BFR/BS with 0.3 μg/kg eldecalcitol treatment. Activation frequency in Haversian canals (H.Ac.f) of cortical bone was suppressed as was observed in trabecular bone (Ac.f). The reduced Haversian remodeling was consistent with the non-significant reduction in cortical porosity noted with eldecalcitol treatment.

At the periosteal and endocortical bone surfaces, treatment with 0.1 μg/kg eldecalcitol tended to suppress periosteal and endocortical label surfaces

Akt inhibitor (Ps.L.Pm/Ec.Pm; Ec.L.Pm/Ec.Pm) mineral apposition rates (Ps.MAR, Ec.MAR) and bone formation rates (Ps.BFR/BS, Ec.BFR/BS). On the other hand, all of those parameters (Ps.MAR, Ec.MAR, Ps.BFR/BS, Ec.BFR/BS) slightly increased with 0.3 μg/kg eldecalcitol treatment. These results suggest treatment with 0.3 μg/kg eldecalcitol stimulates periosteal and endocortical bone formation, while 0.1 μg/kg eldecalcitol suppresses periosteal and endocortical bone formation. Although, no significant changes from OVX-vehicle control in these parameters were found in either treatment group, at least daily treatment with either 0.1 or 0.3 μg/kg of eldecalcitol for 6 months did not overly suppress periosteal and endocortical bone formation in ovariectomized monkeys. In whole lumbar vertebrae, eldecalcitol treatment improved all bone strength parameters compared to OVX-vehicle controls. Statistical significance was attained for peak load, apparent strength, yield load, yield stress,

stiffness, elastic modulus, and work to failure with 0.3 μg/kg eldecalcitol treatment and for stiffness with 0.1 μg/kg eldecalcitol treatment (Table 3A). click here Vertebral core compression revealed significant increases in yield load, yield stress, stiffness and elastic modulus with 0.3 μg/kg eldecalcitol treatment (Table 3B). In the femoral neck, a statistically significant increase in peak load was observed for the animals treated with 0.3 μg/kg eldecalcitol compared to OVX-vehicle controls (Table 3C), with non-significant increases in stiffness and work to failure (Table 3C). There were no statistically significant differences between the eldecalcitol-treated groups and OVX-vehicle controls for any bone strength parameters in 3-point bending at the femur diaphysis (Table 3D) or cortical beams (Table 3E). In this study, as in previous studies [15] and [16], bone turnover markers increased following ovariectomy (Fig. 1). Eldecalcitol treatment at 0.1 and 0.3 μg/kg for 6 months suppressed bone turnover markers and maintained them within baseline levels (Fig. 1). Bone histomorphometric analysis revealed that bone resorption parameters (ES/BS, Oc.S/BS) and bone formation parameters (OS/BS, MS/BS, Ob.

Three separate endpoints were analysed; detection of K65R, detect

Three separate endpoints were analysed; detection of K65R, detection of M184V and ZD1839 mw detection of either K65R or M184V. Person-time was calculated from the start date of the regimen to detection of the mutation(s) being analysed. Follow-up was censored at the earliest of the stop-date of the regimen and the last visit date. The number of events (detection of mutation(s)) was divided by the person-time to calculate the rate of an event according to whether the regimen contained 3TC or FTC. Rates were also stratified by demographic variables (age, sex,

exposure and ethnicity), current CD4 count, most recent viral load (VL), VL at entry and year of starting regimen. Associations between these variables and the event of interest were determined using Poisson regression, allowing for multiple observations from each patient. Univariable and multivariable analyses were performed; multivariable analyses were adjusted for variables mentioned above. In sensitivity analyses, only first treatment episodes of either the 3TC or the FTC regimen were analysed. In a further

sensitivity analysis we restricted to those who had experienced virological failure (1 viral load >400 copies/ml) whilst receiving the regimen and had a resistance test available. Logistic find more regression was used to determine whether there were any significant associations between 3TC and FTC containing regimens and detection of resistance mutations. In total, 5455 patients received either (or both) 3TC, TDF and EFV or FTC, TDF and EFV through the course of follow up, contributing a total of 6465 episodes over 9962 person-years. Forty-seven of these episodes were preceded by a resistance test showing evidence of the K65R (n = 4) or M184V (n = 43) mutations and were hence excluded from the analyses. Table 1 shows the baseline (at start of regimen) characteristics of the remaining 6418 episodes, contributed by 5414 patients. The majority of episodes consisted of FTC- (n = 5190) rather than 3TC- (n = 1228) MRIP based regimens. Age, sex, ethnicity and exposure were similarly distributed between the two groups. FTC-based episodes

were associated with higher median CD4 counts at the start of regimen (297 vs. 276 cells) compared to 3TC-based episodes (p = 0.01), though the median viral load at start of the regimen was lower in the 3TC-based episodes (53 vs. 312 copies/ml) (p = 0.27). Two hundred and thirty nine of 5140 patients receiving FTC (4.6%)had resistance tests performed compared to 65/1228 patients receiving 3TC (5.3%) (p = 0.31). A higher number of patients failing on 3TC containing regimens had resistance tests performed at the time of failure with 53/277 (19.1%) patients failing on 3TC having resistance tests performed, compared to 148/1060 (14%) of patients receiving FTC (p = 0.03). Over the course of follow up 21 cases of K65R were detected, giving a K65R event rate of 0.21 (95% CI: 0.12, 0.31)/100 person-years follow up (PYFU).