We suggest that such model could be applicable here considering a

We suggest that such model could be applicable here considering a thin native oxide layer on the silicon surface. It is likely that physisorption, chemisorption, or desorption of gas species govern the observed charge dynamics. In a gaseous environment, both the internal and external charge transfer mechanisms occur in PS simultaneously but on different time scales resulting in non-trivial transients. The initial fast process during the light-on transient could be related to the drift of the illumination-induced electrons in Si TH-302 towards the bulk and holes towards

the Si/oxide interface due to the electric field in the space charge region (cf. [8]). On the other hand, the electrons in the trap states at the interface may recombine with the flux of holes from the Si side leading to the initial decrease of the CPD in the light-on transient. The decrease in the band bending reduces the depletion width and the barrier height. More electrons can now cross the barrier, tunnel through the oxide layer and become captured by the physisorbed gas species at the free surface and convert them into chemisorbed ones. This increases the negative charge at the free surface and consequently the band bending yielding a slow increase in the CPD of the light-on

transient. However, when similar measurements were performed in vacuum, slow components were absent in transients (Figure 3). We propose that in vacuum, the physisorbed species Selleck Ilomastat could be removed from the surface during evacuation. Thus, only the PS internal mechanism would contribute to the SPV transients in vacuum during the light-on process, explaining the observed behavior. In addition, our experiments show that there is no

difference between the 17-DMAG (Alvespimycin) HCl SPV transients for the bare and PFT�� datasheet Ni-filled PS. This fact indicates that the metal deposits inside the pores do not influence the optoelectronic transport properties of PS, an important observation considering potential multifunctional (magnetic/chemical/pressure) sensor applications of Ni-filled PS. Conclusions In this work, employing transient SPV, we studied charge dynamics of mesoporous silicon and Ni-filled mesoporous silicon in different gas ambients and vacuum. We found that SPV transients for both types of samples in gaseous environments showed a non-trivial behavior during the light-on and light-off events. Vacuum transients showed a different behavior without the slow component. The time scale of the light-on and light-off events in vacuum and in gaseous ambients differs by three orders of magnitude. We suggest that the observed behavior is related to the charge exchange between the silicon/oxide interface and free-surface adsorbates. Acknowledgements The authors thank the Institute for Electron microscopy at the University of Technology Graz, Austria, for SEM investigations of the samples.

05) These data obviously showed that upresgulation of

05). These data obviously showed that upresgulation of miR-451 might effectively enhance the sensitivity of A549 cells to DDP. Figure 5 Effect of miR-451 upregulation on the in

vitro sensitivity of A549 cells to DDP. A. Effects of various concentrations (0, 5, 10, 15, 20 and 25 μg/ml) of DDP on cells (mock A549, A549/miR-NC or A549/miR-451) for 12 h assessed by MTT assay. B. Effects of 5 μg/ml DDP on cells (mock A549, A549/miR-NC or A549/miR-451) for GSK126 solubility dmso varied time length (0, 12, 24, 36 and 48 h) evaluated by MTT assays. C. Effects of 5 μg/ml DDP on colony formation of cells (mock A549, A549/miR-NC or A549/miR-451). All experiments were performed in triplicate, * P < 0.05. Upregulation of miR-451 enhances DDP-induced apoptosis of A549 cells The precise underlying mechanisms by which upregulation Seliciclib manufacturer of miR-451 enhances chemosensitivity of A549 cells to DDP were further investigated. Then, the apoptosis was detected by flow cytometric assay. As shown in Figure 6A, the apoptotic rare of A549/miR-451 treated with 5 μg/ml DDP was increased by approximately 11.7% in comparison with mock A549 cells treated with 5 μg/ml DDP (P < 0.05). However, the apoptotic rate of A549/miR-NC cells treated with DDP showed no significant difference compared with that of mock A549 cells treated with DDP (P > 0.05). Figure 6B showed the results of AnnexinV-FITC apoptosis

detection assay, which Vadimezan chemical structure confirmed the results of flow cytomeric assay. Finally, the activity of caspase-3 was also determined by colorimetric assay.

As shown in Figure 6C, the caspase-3 activity in A549/miR-451 Niclosamide cells treated with DDP remarkably increased by approximately 308% compared that mock A549 or A549/miR-NC cells treated with DDP (P < 0.05). Therefore, upregulation of miR-451 might increase DDP chemosensitivity of A549 cells by enhancing DDP-induced apoptosis. Figure 6 Effect of combined miR-451 upregulation with DDP (5 μg/ml) on apoptosis of A549 cells. A. Flow cytometry analysis of apoptosis in mock A549, A549/miR-NC or A549/miR-451 cells. B. Hoechst staining analysis of apoptosis in mock A549, A549/miR-NC or A549/miR-451 cells. C. Analysis of relative caspase-3 activity in mock A549, A549/miR-NC or A549/miR-451 cells. All experiments were performed in triplicate. Upregulation of miR-451 increases in vivo chemosensitivity of A549 cells to DDP To explore whether upregulation of miR-451 on chemosensitivity of A549 cells to DDP in vivo, s.c. tumors were developed in nude mice followed by treatment with DDP or PBS. As shown in Figure 7A, the tumors formed from A549/miR-451cells grew significantly slower than those from A549/miR-NC after the treatment with DDP. At 28 days after inoculation, the average tumor volume of A549/miR-451 cells (212 ± 36 mm3) was significantly lower than that of A549/miR-NC (323 ± 13 mm3) following DDP treatment (P < 0.05; Figure 7B).

2006, 1:5531–5534 PubMed 33 Miller S, Kastner S, Krijnse-Locker

2006, 1:5531–5534.PubMed 33. Miller S, Kastner S, Krijnse-Locker J, Bühler S, Bartenschlager R: The non-structural protein 4A of dengue virus is an integral membrane protein inducing membrane alterations in a 2K-regulated manner. J Biol Chem 2007, 282:8873–8882.PubMedCrossRef 34. Costa RL, Voloch CM, Schrago CG: Comparative evolutionary epidemiology of dengue virus serotypes. Infect Genet Evol 2012, 12:309–314.PubMedCrossRef

35. Zhang C, Mammen MP Jr, Chinnawirotpisan P, Klungthong C, Rodpradit P, Monkongdee P, Nimmannitya S, Kalayanarooj S, Holmes EC: Clade replacements in dengue virus serotypes 1 and 3 are associated with changing serotype prevalence. learn more J Virol 2005, 79:15123–15130.PubMedCrossRef 36. Holmes EC: Patterns of intra- and interhost non-synonymous variation reveal strong purifying selection in dengue virus. J Virol 2003, 77:11296–11308.PubMedCrossRef 37. Ming-Wei S, Chu WC, Yuan HS: Distinguish dengue virus serotypes via codon usage patterns. Proc of the 1st IEEE Intl Conf on Bioinfo and Biomed Eng (iCBBE’07) 2007, 2:1346–1348. 38. Dey S: Benefits of being biased! J Genet 2004, 83:113–115.PubMedCrossRef 39. Lobo FP, Mota BE, Pena SD, Azevedo V, Macedo AM, Franco GR: Virus-host

coevolution: common patterns of nucleotide motif usage in Flaviviridae and their hosts. PLoS One 2009, 4:e6282.PubMedCrossRef 40. Behura SK, Severson DW: Intrinsic features of Aedes aegypti genes affect transcriptional Y 27632 responsiveness of mosquito genes to dengue virus infection. Infect Genet Evol 2012, 12:1413–1418.PubMedCrossRef 41. Behura SK, Gomez-Machorro C, Harker BW, deBruyn B, Lovin DD, Hemme RR, Mori A, Romero-Severson J, Severson DW: ML323 Global cross-talk of genes of the mosquito Aedes aegypti in response to dengue virus infection. PLoS Negl Trop Dis stiripentol 2011, 5:e1385.PubMedCrossRef 42. Doolittle JM, Gomez SM: Mapping protein interactions between Dengue virus and its human and insect hosts. PLoS Negl Trop Dis 2011, 5:e954.PubMedCrossRef 43. Guo X, Xu Y, Bian G, Pike AD, Xie Y, Xi Z: Response of the mosquito protein interaction network

to dengue infection. BMC Genomics 2010, 11:380.PubMedCrossRef 44. Coffey L, Vasilakis N, Brault AC, Powers AM, Tripet F, Weaver S: Arbovirus evolution in vivo is constrained by host alternation. PNAS 2008, 105:6970–6975.PubMedCrossRef 45. Vasilakis N, Deardorff ER, Kenney JL, Rossi S, Hanley K, Weaver S: Mosquitoes Put the Brake on Arbovirus Evolution:Experimental Evolution Reveals Slower MutationAccumulation in Mosquito Than Vertebrate Cells. PLoS Pathog 2009, 5:1–18.CrossRef 46. Mendez JA, Usme-Ciro JA, Domingo C, Rey GJ, Sanchez JA, Tenorio A, Gallego-Gomez JC: Phylogenetic history demonstrates two different lineages of dengue type 1 virus in Colombia. Virol J 2010, 7:226.PubMedCrossRef 47. Rico-Hesse R: Dengue virus evolution and virulence models. Clin Infect Dis 2007, 44:1462–1466.PubMedCrossRef 48.

Overall the observed induction of exo genes is in agreement with

Overall the observed induction of exo genes is in agreement with the mucoid phenotype observed for S. meliloti after growing on low pH plates (data not shown). In low pH soils this response could be a strategy of the cell to establish a more favourable microenvironment by secreting succinoglycan. It was shown that an EPS I overproduction results in a reduced nodulation efficiency [54], therefore Selleck Temsirolimus the induction of EPS I biosynthesis genes could also be one of the reasons for the observed limited nodulation efficiency of rhizobia in low pH soils [2]. Figure 4 Map of genes in the EPS I biosynthesis region on pSymB and their expression in response

to acidic pH. The EPS I biosynthesis gene region on pSymB is schematically displayed with its genes given by open arrows coloured according to the K-means cluster distribution. Gene names are given below. Black arrows indicate known operon structures in this region. The graph above shows on the Y-axis the time after pH-shift and on the Z-axis for each time point the expression of the corresponding genes by the M value. Whereas the exo gene expression was increased, several mTOR inhibitor genes of chemotaxis and flagellar biosynthesis (flgB, flgG, flgL, flgF, flgC, flgE, fliE, flbT, motA, mcpU) were decreased in their expression levels. After 63 minutes of low pH treatment

the genes have reached the highest level of repression. VisR is the main activator of the flagellar genes and forms together with VisN the top layer of a hierarchy of three expression classes. Since the visN gene expression was decreased early in the time course experiment (therefore visN was grouped into cluster E) the other flagellar genes follow the repression of their activator [55]. The gene coding for the subordinated regulator Rem [56] was also decreasingly expressed with time, but did not reach the threshold for clustering. A detailed

consideration of the expression levels of the flagellar biosynthesis genes on the chromosome (Fig. 5) reveals a repression of the complete region, with some parts responding stronger than others. The decreased expression level of motA, flgF and flgE is likely to be a result of their first position in an operon. It is noticeable that among the 10 down-regulated and strongly responding Exoribonuclease flagellar genes in cluster F five are coding for parts of the rod (flgF, flgB, flgC, fliE and flgG) and two for parts of the hook (flgE and flgL) of the Epacadostat supplier flagellum. The genes motA, fliM, fliN and fliG are proposed to form an operon [55]. While the expression of motA, which is coding for a transmembrane proton channel protein, was decreased in the time course experiment, the other three genes which encode flagellar switch proteins did not respond to the shift to acidic pH. If this behaviour is caused by a specific regulation or is due to mRNA degradation processes cannot be answered.

In a 1997 study of 595 patients with melanoma, Joseph et al evalu

In a 1997 study of 595 patients with melanoma, Joseph et al evaluated the contribution of serial sectioning, immunohistochemistry (IHC) and a molecular technique with reverse transcriptase polymerase chain reaction (RT-PCR) to routine SB525334 mouse hematoxylin and eosin (H&E) histology to detect lymph node metastases. The study showed that routine H&E histology identified 73.8% of all metastases [3]. The remainder was detected by serial sectioning (7.8%) and IHC staining (18.4%) [3]. Moreover, RT-PCR upstaged 47% of the negative sentinel lymph nodes (SLN) [3]. In breast cancer, Selleck NVP-HSP990 Cote et al reported that serial sectioning and IHC were

able to detect respectively 7% and 20% of metastases in negative lymph nodes on H&E histology

[1]. In 2001, a multicenter study of stage I-III colorectal cancer by Saha et al. reported Thiazovivin in vitro that serial sectioning and IHC detected lymph node micrometastases in 14% of patients [4]. The concept of ultrastaging implies that lymph nodes be systematically analysed using serial sectioning and IHC. However, histological and/or molecular techniques used to assess ultrastaging on all nodes are time consuming and expensive thus limiting its routine use. Hence, the concept of ultrastaging is inseparable from that of SLN biopsy [5]. In melanoma, breast cancer, vulvar and colon cancers, the relevance of SLN biopsy has been validated and is considered an alternative to comprehensive lymphadenectomy to assess lymph node status. Although accumulating data on SLN in uterine cancers

are available, its validation remains a matter of debate especially for endometrial cancers due to the absence of consensus on the SLN technique. Moreover, few data are available on ultrastaging in uterine cancers. Therefore, the objective of the present review is to evaluate the contribution of ultrastaging in uterine cancers and its potential therapeutic implications. Concept of ultrastaging in uterine cancers Despite favourable prognostic features, pelvic recurrence occurs in up to 15% of patients with early stage cervical cancer and histologically negative pelvic lymph nodes by routine examination using H&E staining 6-phosphogluconolactonase [6, 7]. Holmgren et al. suggested that some of these recurrences could be due to metastases not detected by routine H&E histology of lymph nodes, so-called “”dormant”" or “”occult”" metastases [8]. Hafner et al. reported that using routine H&E histology, the chances of identifying a tumour cell cluster of less than 3 cell diameters was only 1% [9]. In 2003, Dargent and Enria evoked the concept of micrometastases without clear histological definition in cervical cancer. They reported that the use of serial sectioning and IHC gave a possible tenfold increase in detecting micrometastases [10].

In the Netherlands, a study by Tilburg et al [28] sampled ST20 f

In the Netherlands, a study by Tilburg et al. [28] sampled ST20 from cattle and ST33 from humans, sheep and goats. Huijsmans et al. [21] also genotyped recent samples from the Netherlands, albeit not with MST. However, overlapping reference samples, the results from Tilburg et al. [28] and a comparison to the phylogenetic relationships of MST genotypes, suggests that the Huijsmans [21] genotypes 1, 2, 4, 6 and 8 are likely to be (or be closely related to) MST genotypes

ST33, ST20, ST20, ST8 and ST18 respectively. While likely ST8 samples MM-102 clinical trial have been associated with recent livestock and human clinical samples, such associations with likely ST20 samples are rare (for example see [29]) and it is not clear if any of the Spanish ST20 samples were from animals with clinical manifestations [21, 27, 28, 30]. From the recent outbreak in a UK dairy goat herd [29] and historical

collections, it is clear that ST20 can cause disease in Epacadostat research buy humans and livestock [19, 20]. The scarcity of ST20 among clinical samples, despite being the dominant genotype among cow milk samples, suggests that U.S. ST20 strains have a reduced ability to cause disease in humans or cause a very mild form. Prevalence of C. burnetii on goat and cow farms has been previously assessed, but comparisons across studies are difficult due to different serological or DNA-based detection methods. Sampling individual animals, herds, or products pooled across herds also confounds comparisons although as expected, selleck screening library prevalence generally increases as bulk samples become inclusive of more individuals [6, 8, 13, 34–37]. Similarly, we

found that milk from four of 20 sampled cows were positive while all 3 samples from the bulk milk holding tank (containing milk from 120 cows) were positive. Our milk samples from retail brands bottled in commercial processing plants likely include milk pooled from different (and much larger) dairy farms, making it impossible to know the extent and distribution of infections among cows and herds. However, our detection of C. burnetii DNA in every goat and cow milk sample from the same brands (i.e. processing plants) over time and >95% of milk samples from processing plants across the USA shows high BCKDHB prevalence at either or both the individual and herd levels. Indeed, the prevalence rate reported here is comparable to the high rates reported in other studies [8, 12, 13]. Notwithstanding existing immunity, infectious diseases are density dependent, leading us to suspect that the ratio of infected to uninfected cows on some farms may be greater than our single farm results. Nonetheless, while a small number of infected animals may contaminate a large quantity of milk, it is probable that a significant portion of the 9.2 million dairy cows in the USA [38] are infected with C. burnetii at any given time [13]. Across the ~2.5 year period of sample collection, there was no variation in prevalence of C.

Cell survival assay Cells were seeded in 96-well plates and treat

Cell survival assay Cells were seeded in 96-well plates and treated on the second day with the given concentration of PTL for another 48 hours and then subjected to SRB or MTT assay. For SRB assay, live cell number was estimated as described earlier [33]. After treatment, the medium was discarded firstly. In order to fix the adherent cells,

100 μ1 of cold trichloroacetic acid (10% (w/v)) were adding to each well and incubating at 4°C for at least 1 hour. The plates were then washed five times with deionized water and dried in the air. Each well were then added with 50 μ1 of SRB solution (0.4% w/v selleck kinase inhibitor in 1% acetic acid) and incubated for 5 min at room temperature. The plates were washed five times with 1% acetic acid to remove unbound SRB and then air dried. The residual bound SRB was solubilized with 100 μ1 of 10 mM Tris base buffer (pH 10.5), and then read using a microtiter plate ICG-001 in vitro reader at 495 nm. The MTT assay was executed following the manufacturer’s protocol of Cell Proliferation Kit I (Roche Applied Science, Brandford, CT, USA). 20 μl MTT (5 mg/ml) were added to each sample and incubate at 37° for 4 h, then 100 μl solubilization

solution were added. Cell viability was determined at 595 nm. Cell cycle analysis Cell cycle was evaluated by DNA flow cytometry analysis. Cells were treated with different concentrations of PTL (0, 5, 10, 20 μM) for 24hours. After Tipifarnib concentration treatment, the cells were harvested and washed twice with ice PBS, then fixed in 70% ethanol at -20°C overnight. Before analysis, cells were washed again with ice PBS, incubated with PI (100 μg/ml) and RNase (50 μg/ml) in the dark for 30 min. Then samples were analyzed by FACScan flow cytometer (Becton Dickinson, San Jose, CA) [34]. Western blot analysis Whole cell protein lysates were prepared and analyzed by Western blot according to the protocol described previously [35]. Cells were harvested and rinsed with pro-cold PBS. Then cell extracts were lysed and centrifuged at 4°C for 15 minutes. Whole cell protein lysates (40 μg) were electrophoresed through 12%

denaturing polyacrylamide slab gels and then transferred to a Hybond enhanced chemiluminescence (ECL) membrane by electroblotting. The proteins were probed with the appropriate primary antibodies and subsequently with secondary antibodies. The antibody below binding was detected by the ECL system (Millipore, Billerica, MA, USA), according to the manufacturer’s protocol. siRNA transfection siRNAs targeting sequences of TNFRSF10B, ATF4 and DDIT3 have been described previously and synthesized by GenePharma (Shanghai, China) [36]. The target sequence of PMAIP1 is 5′-GGAAGUCGAGUGUGCUACU-3′. The transfection of siRNA was following the manufacturer’s protocol of X-tremeGENE Transfection Reagent (Roche Molecular Biochemicals, Mannheim, Germany). Cells were seeded in 6-well plates and transfected with control or target siRNA on the second day.

Herein, the hepatotoxicity in rats exposed

Herein, the hepatotoxicity in rats exposed Pevonedistat nmr to SWCNTs by intratracheal instillation was explored using a 1H NMR-based metabonomic approach to examine blood

https://www.selleckchem.com/TGF-beta.html plasma and liver tissue extracts obtained from rats treated with different SWCNTs concentrations. Concurrently, the toxic threshold and identification of potentially useful toxicity biomarkers of SWCNTs-induced hepatotoxicity were also studied by conventional clinical chemistry and histopathological examinations. Methods Single-walled carbon nanotubes and suspension preparation Non-functionalized SWCNTs, produced by CoMoCAT® (Sigma-Aldrich, St. Louis, MO, USA) catalytic CVD process, were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Their diameter of 0.8 to 1.2 nm and a length of several microns were determined by transmission electron microscopy (TEM, JEM-2010FEF, JEOL, Ltd., Tokyo, Japan) selleck compound (Figure 1A). Raman spectroscopy had been used to assess purity (Raman spectrometer, RM200, Renishaw, Gloucestershire, UK) (Figure 1B). The carbon content and the proportion of carbon as SWNT were above 90% and 70%, respectively. Figure 1 The non-functionalized SWCNTs. (A) TEM of SWCNTs. (B) Raman spectra of SWCNTs. SWCNTs samples at 150, 300, and 450 mg were dispersed in 20-mL

volumes of 0.9% sodium chloride solution, followed by ultrasonication at <50°C for 5 h. The resulting SWCNTs concentrations were 7.5, 15, and 22.5 mg/mL, respectively. Ethics statement All experiments involving the care and use of animals were performed in accordance with the guidelines and regulations concerning the ethics of science research in the Institute of Health and Environmental Medicine and approved by the Ethics Review Board of the Institute of Health and Environmental Medicine Sodium butyrate (approval number JKYSS-2009-018). Animals

and treatment Thirty healthy male Wistar rats (8 weeks of age, weight 180 to 220 g) were obtained from the Academy of Military Medical Sciences (Beijing, China). All procedures concerning animal usage were reviewed and approved by the Institutional Animal Care and Use Committee of the Academia. All rats were housed individually in metabolic cages and, throughout the study period, allowed food and tap water ad libitum, with light/dark cycles altering every 12 h, environment at 18°C to 22°C, and humidity from 40% to 60%. After 1 week of acclimatization, weight-matched rats were divided randomly into four groups (n = 6 per group) comprising a sodium chloride group (control) and low-, medium-, and high-concentration groups (7.5, 15, and 22.5 mg/kg body weight and named SWCNTs-L, M, and H, respectively). The rats were exposed to SWCNTs by intratracheal instillation of the corresponding SWCNTs suspensions once a day for 15 consecutive days, with the control group treated concurrently with 0.9% sodium chloride solution.

The images were obtained using portable X-ray equipment with a fi

The images were obtained using portable X-ray equipment with a film–focus distance of 0.45 m, and the right hand was used. The study included municipal school children from five communities in Northern Sjaelland for whom the parents gave consent, resulting in images from 97% of all children, which makes this data set a true representation of the population. The images used are a random subset of the 3,600 images that make up the original study. The Erasmus

study: 531 healthy Caucasian subjects, including 255 boys (median age 12.4 years, range 3.8–20.1 years) Tubastatin A concentration and 276 girls (median age 12.6 years, range 3.8–20.0 years) from the Erasmus Gymnasium in Rotterdam were studied in 1997 by researchers at the Erasmus Medical Centre (EMC) [13]. The younger children were children of employees at the EMC institutions. Institutional Review Board approval was given to obtain radiographs of www.selleckchem.com/products/CX-6258.html the left hand and use these data for subsequent analysis. Informed consent was obtained from the parents or custodians and, for children above 12, also from the

child. A detailed description of this cohort was published by Lequin et al. [13]. Radiographs of the left hand were recorded on mammography film (Philips Diagnost H, Imation GTU film, Alfa-II Trimax intensifying screens, small 0.6 mm focus, film–focus distance 1.5 m, 45 kV, 16 mAs) to obtain excellent quality. The Seiiku study followed ten boys and ten girls with growth hormone deficiency treated with growth hormone Decitabine datasheet and gonadotropin-releasing hormone analogue for a period of 1.75–6.75 years. The data consist of 284 images recorded in the period ca. 1984–2001. The children were followed from an age of 4–11 years to an age of 15–21 years. The images were obtained approximately once every 6 months. The films were digitised in 300 dpi with 12 bits per pixel using a Vidar Diagnostic Pro Advantage scanner (Vidar, Hemdon, VA, USA) using software version TWAIN 5.2. However, the Seiiku study and one third of the Sjælland images were digitised with a UMAX Powerlook

1100 scanner (Umax Data Systems Inc, Taipei, Taiwan) in 300 dpi with 8 bits per pixel, using MagicScan 4.5 software. Method The method is based on the BoneXpert system for automatic determination of bone age [4–7] (Visiana, Holte, Denmark, www.​BoneXpert.​com). The images are first reduced to 150 dpi and 8 bits, and then the boundaries of the metacarpals (and other bones) are determined. For more mature bones, the boundary includes both the diaphysis and the fused epiphysis, while for the less mature bones there are separate boundaries for the diaphysis and the epiphysis. The boundary of the diaphysis is computed as 64 points, which correspond to the same P505-15 concentration anatomical locations across subjects [4, 14]. Two of the points correspond to the proximal and distal ends of the diaphysis, and they are used to define the bone axis (see Fig. 1). The length, L, of the bone is measured along this axis, and it includes the epiphysis.

2010 [36] • ≥65 years • Pharmacists trained by investigators • Ad

2010 [36] • ≥65 years • Pharmacists trained by investigators • Ads in local newspaper Control 133 • Usual care

and print material from OP Canada RCTc, Canada (Alberta) • 50–64 years with ≥1 major risk factord   • Notices in participating pharmacies Intervention 129 • 30-min appointment on clinic day: 15 community pharmacies • No BMD test in prior 2 years   • Participants called to book appointment      • Print material from OP Canada   • No current OP treatment   • Pharmacist identification in pharmacy      • Pamphlet designed by study investigators   • English speaking          • Pharmacist counseling (tailored OP education)              • Heel QUS measurement and interpretation       LY3039478        • Patients encouraged to follow-up with their primary care physician        

     • Physicians provided with study details, QUS results, Salubrinal and information regarding patient eligibility for central BMD testing              • Follow-up              • Telephone: 2 and 8 weeks              • Patients asked to return to pharmacy at 16 weeks RCT randomized controlled trial (in cluster RCT, PRN1371 cost groups randomized by pharmacy), BMD bone mineral density, DXA dual-energy X-ray absorptiometry, OP osteoporosis, QUS quantitative ultrasound, n number of participants aOf all pharmacists agreeing and eligible to participate, one was randomly selected from each of six suburban and six rural areas. These 12 pharmacists were then randomized into one of two groups with three suburban and three rural pharmacies in each of the two groups bPharmacies from a specialized provider network consisting of pharmacists with previous training and/or certification in drug

therapy monitory and research Neratinib chemical structure participation cRandomized by secure internet randomization services (sequence stratified by site with block size of 4) dMajor risk factor included: family history of osteoporosis, previous fracture, systemic glucocorticoids >3 months, or early menopause eSample size after exclusion of missing data or participants who did not complete the study Table 2 Summary of potential risk of bias based on threats to internal validity Study Selection Bias Information Bias Allocationa Attritionb Performancec Detectiond Crockett et al. [34] High High High High • Better recruitment success in BMD group in rural regions (n = 60 vs. n = 43) • 3-month follow-up, 87% • Definition of risk differed between groups • Self-report assessment based on patient recall of pharmacist recommendations and whether or not they complied with the pharmacist’s recommendations • Non-BMD group had larger proportion with history of low-trauma fracture (21% vs. 11%) • 6-month follow-up, 10%; only “high-risk” followed • Group 1: questionnaire only • Group 2: questionnaire and forearm BMD results McDonough et al.