For the GaAsSb QW sample, an emission peak of 1.242 eV at RT was found, corresponding to an Sb content of approximately 15% according to theoretical and experimental results for such a GaAsSb QW thickness [15]. Regarding the GaAsN QW, a content of N around 2.3% can be estimated when comparing with similar reported QWs [16]. The LT PL from the quaternary QW sample shifted from the GaAs gap

energy a higher value (527 meV) than the addition of shifts in the GaAsSb (216 meV) and GaAsN (255 meV) QW samples. This is selleck in agreement with studies reporting a facilitated incorporation of N by the presence of Sb [17, 18]. Indeed, the difference of 56 mV points to a higher N content corresponding to approximately 2.8%. For these N and Sb contents, the system will still be in the type-I band alignment region [12]. Furthermore, since the Sb/N ratio is larger than 2.6 (the condition for lattice matching to GaAs) it can be assumed that the GaAsSbN layer grows under compressive buy ABT-737 strain on GaAs and will act as a strain-reducing CL. Capping

layer growth temperature First, the study focuses on finding the optimal growth temperature for the GaAsSbN CL. The incorporation of N in GaAs has been found to be temperature independent in a wide range of temperatures from 400°C to 480°C [19] or even higher temperatures [20, 21]. However, for temperatures higher than that, N incorporation is strongly reduced. This is probably induced by the temperature-enhanced desorption of N from the growth surface, as it has been

theoretically predicted [22]. On the other hand, as expected from the fact that Sb has a higher sublimation energy than As [23], increasing the temperature affects substantially the incorporation of Sb [24, 25]. Thus, Sb desorption has been found to increase with temperature, becoming substantial above 490°C [24]. Hence, in order to avoid a significant desorption of both Sb and N as well as a substantial modification of the InAs QDs, we studied the effect of the CL growth temperature in a range between 450°C and 3-oxoacyl-(acyl-carrier-protein) reductase 480°C. A series of four learn more samples was grown with CL growth temperatures set to 450°C, 460°C, 470°C, and 480°C (labeled as A1, A2, A3, and A4, respectively). Figure 1 shows the PL spectra of the four samples. The small peak wavelength shifts observed do not follow any tendency with the growth temperature and are likely within the reproducibility error bar. Nevertheless, an improvement of the luminescence properties can be observed with increasing the growth temperature from 450°C up to 470°C, being more remarkable for the last temperature case. The full width at half maximum (FWHM) is slightly reduced, and the integrated intensity is approximately doubled when raising the temperature within this range. However, above 470°C, the integrated PL intensity is reduced by approximately 65% and the FWHM is slightly increased.

The dominant inheritance PF-02341066 in vivo can be explained by hetero-oligomerization of wild-type/mutant AQP2 proteins and dominant-negative effect of mutant protein on wild-type protein [7]. In a female patient of family 5, a novel

heterozygous 1-nucleotide deletion mutation (750delG) was found. The patient’s sister and father were symptomatic. Her urine osmolality did not respond to vasopressin. This mutation causes a frame shift, with a new amino acids sequence starting from Val251 and ending at codon 334 in the C-terminal of AQP2. In Family 6, a 2-year-old girl was found to have a novel heterozygous 1-nucleotide deletion mutation (775delC) that causes frame shift with a new C-terminus starting at Leu259. The parents did not show NDI symptoms and did not carry the mutation, which indicated that the mutation occurred de novo. find more The girl showed polyuria and polydipsia and NDI was diagnosed by water deprivation and vasopressin administration tests. These identified two deletion mutations cause frame shifts from Val251 and Leu259 and a new C-terminal tail ending at codon 334 (Table 4). We previously reported three small

deletion mutations in the C-terminus that cause similar frame shifts and show dominant inheritance [12] (Table 4). These frame-shift mutations share the loss of the last tail of the AQP2 protein, the site where PDZ proteins and ubiquitines interact, and the GSK1210151A mouse presence of extended C-terminal tails that contain missorting signals. As a result of these effects, these mutant AQP2 proteins making tetramers with wild-type proteins are incorrectly translocated to the basolateral membrane instead of the apical membrane [20, 30, 31]. This missorting is confirmed in knockin mice harboring a human C-terminal deletion mutation (c.763–772del) [32]. It is interesting that these deletion mutations are observed more often that missense mutations in Japanese patients, which is different from the frequencies in a total global

summary [3, 20]. We could not detect mutations in the two genes in seven families (9 %, Table 1). It is said that causative gene mutations cannot be found in Epothilone B (EPO906, Patupilone) approximately 5 % of all congenital NDI patients [4]. Possibilities such as the presence of mutations in the promoter regions of the AVPR2 or AQP2 genes are a likely explanation [4]. Our mutational analysis does not usually cover the promoter regions; thus, this possibility remains to be examined. To date, no genes other than AVPR2 and AQP2 have been attributed to NDI. However, it is possible that mutations in the genes encoding signaling cascade molecules connecting these two key membrane proteins cause NDI. Progress in gene mutational analysis methods such as whole-exome sequencing will address this possibility. Acknowledgments We thank Mieko Goto for technical assistance and Dr. Daniel Bichet for help in mutation analysis. We thank Drs. M. Asai, A Ashida, T. Aso, T. Hamajima, T. Hasegawa, M. Hayashi, D. Hirano, K. Ichida, E. Ihara, M. Iketani, T. Imanishi, H.

This growth phase of Aspergillus fumigatus is inhibited by lactoferrin-mediated iron depletion [28]. In contrast, inhibition of the hyphal form of Aspergillus fumigatus requires NADPH oxidase [28, 30]. Aspergillus PND-1186 cell line nidulans lacking the catalase genes are capable of causing disease in gp47phox KO mice, which suggested that reactive oxygen intermediates

might not be inhibiting the organism directly [30]. It has been suggested that activation of intracellular proteases by reactive oxygen intermediates is important for killing Candida and several types of bacteria [31]. There is one report that MK-8931 nmr administration of pentraxin 3 protected gp47phox mice from experimental Aspergillus fumigatus infection, suggesting that this molecule in important for resistance to Aspergillus fumigatus and may be lacking in CGD mice [32]. The only evidence that primary pathogenic fungi are more virulent in CGD mice is a study with Sporothrix schenckii [33]. These investigators found that gp91phox KO mice infected with Sporothrix schenckii intradermally died within three months, whereas control mice survived this infection. They also found that PMN from gp91phox KO mice were not able to control the growth of Sporothrix schenckii as well as the controls. We have not been able to find any published data on Blastomyces dermatitidis, C. immitis or

Histoplasma capsulatum MLN2238 in vivo experimental infections in CGD mice. People with chronic granulomatous disease have increased susceptibility to Aspergillus infections and, to a lesser extent, infections due to other opportunistic fungi [34]. There have been no reports of increased susceptibility to the primary pathogenic fungi Coccidioides, Histoplasma

capsulatum, Blastomyces dermatitidis or Sporothrix schenckii. One expert states that these infections are not a problem in chronic granulomatous disease [34]. One CGD patient has been observed to recover uneventfully from pulmonary coccidioidomycosis without anti-fungal therapy (J. Galgiani, very personal communication). The observation that NADPH oxidase is not required for a protective immune response to experimental coccidioidomycosis raises the question of what immune mechanisms used to kill spherules and endospores in vivo. One potential protective immune effector mechanism is oxidative stress due to nitric oxide. We have previously reported that IL-10 exacerbates the course of experimental coccidioidomycois and inhibits nitric oxide synthase [35]. On the other hand, a very recent study suggests that Coccidioides is resistant to killing by NO and that mice with a deletion mutation in inducible nitric oxide synthase are able to kill Coccidioides [36]. Coccidioides spherules can be very large (more than 60 μM in diameter) and therefore difficult to phagocytose. Perhaps inhibiting the growth of the endospore controls the growth of the organism. Understanding the mechanisms of protective immunity is important for optimally preventing and treating infections with this pathogenic fungus.

Antonie van Leeuwenhoek 1994, 65:227–243.CrossRefPubMed 33. García-Estrada C, Ullán RV, Velasco-Conde T, Godio RP, Teijeira F, Vaca I, Feltrer R, Kosalková K, Mauriz E, Martín JF: Post-translational enzyme modification by the phosphopantetheinyl

transferase is required for lysine and penicillin biosynthesis but not for roquefortine or fatty acid formation in Penicillium chrysogenum. Biochem J 2008, 415:317–324.CrossRefPubMed 34. Keller NP, Hohn TM: selleck screening library Metabolic Pathway Gene Clusters in Filamentous Fungi. Fung Genet Biol 1997, 21:17–29.CrossRef 35. Spröte eFT508 order P, Hynes MJ, Hortschansky P, Shelesty E, Scharf DH, Wolke SM, Brakhage AA: Identification of the novel penicillin biosynthesis gene aatB of Aspergillus nidulans and its putative evolutionary relationship to this fungal secondary metabolism gene cluster. Mol Microbiol 2008, 70:445–461.CrossRefPubMed 36. Klein AT, van den Berg M, Bottger G, Tabak HF, Distel B:Saccharomyces cerevisiae acyl-CoA oxidase follows a novel, non-PTS1, import pathway into peroxisomes that is

Selleckchem CH5424802 dependent on Pex5p. J Biol Chem 2002, 277:25011–25019.CrossRefPubMed 37. Seemüller E, Lupas A, Baumeister W: Autocatalytic processing of the 20S proteasome. Nature 1996, 382:468–470.CrossRefPubMed 38. Tobin MB, Cole SC, Kovacevic S, Miller JR, Baldwin JE, Sutherland JD: Acyl-coenzyme A: isopenicillin N acyltransferase from Penicillium chrysogenum

: effect of amino Cytidine deaminase acid substitutions at Ser227, Ser230 and Ser309 on proenzyme cleavage and activity. FEMS Microbiol Lett 1994, 121:39–46.CrossRefPubMed 39. Aplin RT, Baldwin JE, Roach PL, Robinson CV, Schofield CJ: Investigations into the post-translational modification and mechanism of isopenicillin N:acyl-CoA acyltransferase using electrospray mass spectrometry. Biochem J 1993, 294:357–363.PubMed 40. Laich F, Fierro F, Cardoza RE, Martín JF: Organization of the gene cluster for biosynthesis of penicillin in Penicillium nalgiovense and antibiotic production in cured dry sausages. Appl Environ Microbiol 1999, 65:1236–1240.PubMed 41. Laich F, Fierro F, Martín JF: Production of penicillin by fungi growing on food products: Identification of a complete penicillin gene cluster in Penicillium griseofulvum and a truncated cluster in Penicillium verrucosum. Appl Environ Microbiol 2002, 68:1211–1219.CrossRefPubMed 42.

However, this approach detects the viral nucleic acids of both infectious and ubiquitin-Proteasome degradation non-infectious viruses. Therefore, it is important to develop and evaluate simple and efficient tools which make it possible to overcome the limitations of the traditional cell culture and PCR assays [9]. An approach based on an enzymatic treatment with RNAse combined with a proteinase K treatment was found to be successful in some cases in distinguishing between infectious and non-infectious viruses [10–12]. For bacteria, a relatively recent approach is the treatment of samples with the DNA-intercalating dyes ethidium

monoazide (EMA) or propidium monoazide (PMA) [13–17]. EMA and PMA are closely related ITF2357 nmr DNA intercalating dyes with a photo-inducible azide group that covalently cross-link to DNA through visible-light photoactivation. PMA has the advantage of being more selective than EMA for dead cells as it is more membrane-impermeant [18]. Recently, promising PMA / EMA treatments have also been tested for distinguishing between infectious and non-infectious RNA viruses [19, 20]. A study concluded that PMA-RT-PCR assays that include pretreatment of enteroviruses

and noroviruses with PMA prior to RT-PCR enable rapid differentiation between infectious and non-infectious enteric viruses when the virus particles are inactivated by heating at 72°C or 37°C or by using hypochlorite. However, unlike poliovirus, PMA treatment did not affect detection of heat-inactivated Norwalk virus by quantitative RT-PCR [21]. Another study found that EMA did not distinguish between infectious and non-infectious GDC-0449 nmr avian influenza virus particles [22]. Sánchez et al. [23] showed that PMA treatment previous to RT-qPCR detection is a promising alternative for assessing Celecoxib HAV infectivity. The usefulness of EMA or PMA for distinguishing between infectious and non-infectious RV and HAV was investigated. Both viruses were chosen for their cultivability and their differences in genomic organization. RV, the leading cause

of severe dehydrating diarrhea in infants and young children worldwide, are non-enveloped viruses that possess a genome with 11 segments of double-stranded RNA contained in a triple-layered protein capsid and belong to the Reoviridae. Hepatitis A virus (HAV) infection is the leading worldwide cause of acute viral hepatitis. HAV is a positive single-stranded non-enveloped RNA virus classified in the Hepatovirus genus of the Picornaviridae family. The purpose of this study was to develop a method based on pre-treatment-RT-qPCR assays in order to discriminate between infectious and non-infectious viruses (HAV, RV) following thermal inactivation. To this end, the binding of EMA and PMA to RV and HAV RNA was investigated. Then, a pre-treatment based on “PMA or EMA +/− surfactant RT-qPCR” was optimized for each virus. Finally, this method was applied to establish viral thermal inactivation kinetics through three RT-qPCR assays.

Inserts from each DNA clone were PCR-amplified directly from bacteria. Amplification reactions were performed in 96-well plates,

with each well carrying a 50-μl volume containing 0.2 μM of each primer (T7 and SP6), 200 μM of each dNTP, 1× PCR buffer, and 1.25 units of Taq polymerase (AmpliTaq® DNA polymerase, Promega Corporation). An MJ Research thermal cycler was used for 35 PCR cycles, as follows: 95°C for 45 s, 56°C for 45 s, and 72°C for 1 min. We also amplified a selected set of conserved effector and hrp genes (e.g. XopX, avrXa7, XopD, avrRxv, avrXv3, hpaF, and hrpx), housekeeping AZD5153 research buy genes, and other conserved bacterial genes from genomic DNA of Xoo MAI1. Random PCR samples were visualized on agarose gels. All PCR Rabusertib clinical trial products were transferred to a 384-well plate and a volume of 2× betaine solution was added. The PCR products were arrayed once on poly-L-lysine slides (TeleChem International, Inc., Sunnyvale, CA, USA), using an SPBIO™ Microarray Spotting Station (MiraiBio, Inc., Alameda, CA, USA). The microarray contained 4708 elements. Bacterial inoculation and quantification The Xoo strain MAI1 was grown on PSA medium (10 g l-1 peptone,

10 g l-1 sucrose, 1 g l-1 glutamic acid, 16 g l-1 agar, and pH 7.0) for 2 days at 30°C. The bacterial cells were re-suspended in sterilized water at an optical density of 600 nm (OD 600) (about 10-9 cfu ml-1). Bacterial blight inoculation was carried out on the two youngest, fully expanded leaves on each tiller of 6-week-old rice plants (var. Nipponbare), using Orotidine 5′-phosphate decarboxylase the leaf-clipping method [67]. Experiments were conducted under greenhouse conditions at 26°C and 80% relative humidity. We determined Xoo MAI1 multiplication in planta at seven time points after infection by leaf clipping (0 and 12 h, and 1, 3, 6, 10, and 15 days after inoculation) in 8-week-old plants of the susceptible rice cultivar Nipponbare. The number of cells

in the leaves was determined at the top 10 cm of each leaf which was cut into five 2-cm sections, and labelled A, B, C, D, and E, with A being the inoculation point. The leaf pieces were then ground in 1 ml of sterilized water. Serial dilutions were made and spread onto PSA agar plates. The plates were incubated at 28°C until single colonies could be counted. The number of colony-forming units (cfu) per leaf (equivalent to about 2 cm2) was counted and standard deviations calculated. The experiment was repeated independently three times. RNA extraction To obtain RNA from cells growing in planta, 30 rice leaves were inoculated by the leaf-clipping method. At each time point, leaves extending 2 cm from the tip were collected and, to facilitate exudation of bacterial cells, vortexed for 30 s with RNAprotect EPZ015938 purchase bacteria Reagent (QIAGEN, Inc., Courtaboeuf, France). The leaves were removed and bacterial cells were collected in a 15-ml tube by centrifuging at 4000 rpm for 30 min at 4°C.

Hafner et al [9] suggested that E6 expression was linked to lymph node status but, as in previous studies [27, 28], there was a high overlapping of values between positive and negative lymph nodes. Coutant et al reported that HPV DNA screening in SLN by means of PCR might help to identify patients at risk of lymph node metastases and recurrence although HPV DNA was noted in only 46.7% of positive SLN and in 13.6% of negative SLN [29]. While molecular techniques (such as RT-PCR) may be more sensitive than IHC, they carry a high false positive rate [30]. Indeed, Van Trappen et al underlined that specific tumour DNA found in

histologically normal lymph nodes may originate from dead cell material or macrophages and that viral DNA can be found in various Pevonedistat research buy cell types thus limiting its usefulness as a molecular marker for micrometastases

[27]. Marchiolé et al noted that even RT-PCR had a better sensitivity than IHC though this is counterbalanced by a lack of specificity [12]. Moreover, it is not possible to differentiate macrometastasis from benign glandular inclusion using only RT-PCR. In addition, even if a correlation has been established between the number of copy cells and the size of metastases, RT-PCR lacks accuracy in differentiating true macrometastases with proved prognostic value from multiple micrometastases or submicrometastases with questionable clinical relevance. In endometrial cancer few data are selleck chemicals available on the contribution of molecular techniques to detect lymph node metastases. Fishman et al were the first to report a high CK-20 expression by RT-PCR in primary tumours www.selleckchem.com/products/tariquidar.html and in pelvic lymph nodes. Among the 18 patients with negative pelvic lymph nodes by routine H&E histology, six (33%) were CK-20 positive suggesting

a potential contribution of molecular biology in assessing lymph node status. So far, no data are available on CK-20 expression by RT-PCR in SLN in patients with endometrial cancer [31]. Incidence of micrometastases and potential clinical implications in patients with uterine Isotretinoin cancers The definition of micrometastases is rarely clearly mentioned in published reports representing a potential bias in the interpretation of their prognostic relevance. Moreover, as previously noted, the incidence of micrometastases can differ significantly according to the histological and biological technique used. In cervical cancer, whatever the histological technique used for detecting lymph node involvement, the rate of macrometastases varied from 7.1% to 42% (table 1, 2). Table 1 Ultrastaging of sentinel lymph node using H&E and IHC in patients with cervical cancer Study Year Method of analysis Nb of patients FIGO stage Macrometastatic SLN (%) Micrometastatic SLN (%) Lambaudie 2003 H&E +IHC 12 IA2-IB1 2 (18.2) 0 Niikura 2004 H&E +IHC 20 IB1-IIA 2 (10) 0 Martinez Palones 2004 H&E +IHC 23 IA2-IIA 3 (13) 0 Kraft 2006 H&E +IHC 54 IB1-III 21 (42) na Total     109   28 (25.

He did not attend hospital for subsequent follow-up imaging, but on telephone review remains well one year post-procedure with no recurrence of any PARP activity of his symptoms. In this case, follow up imaging would have been useful to

examine for involution of the pseudoaneurysm and continued exclusion, as well as resolution of splaying of the vessels. Discussion This unique case comprises both the first description of a variant of SMA syndrome caused by a traumatic SMA pseudoaneurysm, and the first account of successful treatment of both the aneurysm and duodenal obstruction by endovascular stent placement. Two similar cases were described in 1990 [11], however, in these cases, obstruction was caused by rupture of an SMA pseudoaneurysm, treated with open surgery. Barium meal examination is useful for the diagnosis of SMA syndrome [9]. It demonstrates both narrowing of the fourth part of the duodenum with increased transit time, proximal dilatation and uncoordinated peristaltic activity. Such functional information is not readily obtainable from CT. CT proved to be the

key modality for diagnosis in this patient. It enabled detection of the pseudoaneurysm and its relationship to the SMA. CT with 3D reconstruction has been used in SMA syndrome to demonstrate reduction of the angle between the SMA and the aorta [12]. Despite the paucity of cases of SMA pseudoaneurysm, several reports describe successful endovascular treatment of this condition. not Open surgery is often rendered difficult by the underlying cause of the psuedoaneurysm DMXAA (such as pancreatitis) or by Trichostatin A in vivo adhesions, which increase the risk of failure

of open vascular reconstruction and of anaesthesia in the unstable patient [1]. Other options for treatment of this condition include placement of coils, injection of thrombin or N-butyl-2-cyanoacrylate (glue) [1]. This case presented an unusual challenge, as two problems needed addressing; stenting of the aneurysm to prevent subsequent rupture, and exclusion of the aneurysm sac to encourage involution and thus relieve the SMA syndrome. The immediate resolution of this patient’s symptoms was most likely due to loss of pressure within the aneurysm sac by exclusion of arterial inflow. Data on possible shrinkage of aneurysm sacs post-stenting are conflicting, with one large series of 90 endovascular repairs of a range of visceral artery aneurysms demonstrating no shrinkage at follow-up imaging [1]. However, one study reported shrinkage of abdominal aortic aneurysms post-stent placement [13]. This phenomenon, in addition to decreased pressure within the sac, may be helpful in the treatment of aortoduodenal syndrome, which has hitherto only been treated by open repair. Conclusions A unique case of a variant of SMA syndrome secondary to a pseudoaneurysm is presented. Exclusion of the aneurysm and relief of the obstruction were simultaneously achieved by placement of a stent.

Microbiol Immunol 2002, 46:195–205.PubMedCrossRef 12. Rimoldi M, Chieppa M, Larghi P, Vulcano M, Allavena P, Rescigno M: Monocyte-derived dendritic cells activated by bacteria or by bacteria-stimulated epithelial cells are functionally different. Blood 2005, 106:2818–2826.PubMedCrossRef 13. Banchereau J, Steinman RM: Dendritic cells and the control of immunity. Nature

1998, 392:245–252.PubMedCrossRef 14. Huang Q, Liu D, Majewski P, Schulte LC, Korn JM, Young RA, Lander ES, Hacohen N: The plasticity of dendritic cell responses to Autophagy Compound Library purchase pathogens and their components. Science 2001, 294:870–875.PubMedCrossRef 15. Christensen HR, Frokiaer H, Pestka JJ: Lactobacilli differentially modulate expression of cytokines and maturation surface markers in murine dendritic cells. J Immunol 2002, 168:171–178.PubMed 16. Hart AL, Lammers K, Brigidi P, Vitali B, Rizzello F, Gionchetti PCI-34051 manufacturer P, Campieri M, Kamm MA, Knight SC, Stagg AJ: Modulation of human dendritic cell phenotype and function by probiotic bacteria. Gut 2004, 53:1602–1609.PubMedCrossRef 17. Medina M, Izquierdo E, Ennahar S, Sanz Y: Differential immunomodulatory properties of Bifidobacterium longum strains: relevance

to probiotic selection and clinical applications. Clin Exp Immunol 2007, 150:531–538.PubMedCentralPubMedCrossRef 18. Menard O, Batel MJ, Gaboriau-Routhiau V, Waligora-Dupriet AJ: Gnotobiotic mouse immune response induced by Bifidobacterium sp. strains straind from Crenolanib manufacturer infants. Appl Environ Microbiol 2008, 74:660–666.PubMedCentralPubMedCrossRef 19. D’Arienzo R, Maurano F, Lavermicocca P, Ricca E, Rossi M: Modulation of the immune response by probiotic strains in a mouse model of gluten sensitivity. Cytokine 2009, 48:254–259.PubMedCrossRef 20. D’Arienzo R, Bozzella G, Rossi M, De Bellis P, Lavermicocca P, Sisto A: Distinct immunomodulatory properties of Lactobacillus paracasei strains. J Appl Microbiol 2011, 111:1482–1491.PubMedCrossRef 21. Selle K, Klaenhammer TR: Genomic and phenotypic

evidence for probiotic influences Branched chain aminotransferase of Lactobacillus gasseri on human health. FEMS Microbiol Rev 2013, 37:915–935.PubMed 22. Sashihara T, Sueki N, Ikegami S: An analysis of the effectiveness of heat-killed lactic acid bacteria in alleviating allergic diseases. J Dairy Sci 2006, 89:2846–2855.PubMedCrossRef 23. Baruzzi F, Poltronieri P, Quero GM, Morea M, Morelli L: An in vitro protocol for direct isolation of potential probiotic lactobacilli from raw bovine milk and traditional fermented milks. Appl Microbiol Biotechnol 2011, 90:331–342.PubMedCrossRef 24. Vidal K, Grosjean I, Revillard JP, Gespach C, Kaiserlian DJ: Immortalization of mouse intestinal epithelial cells by the SV40-large T gene. Phenotypic and immune characterization of the MODE-K cell line. Immunol Methods 1993, 166:63–73.CrossRef 25.

control (n = 70 vs. n = 26) • Follow-up: • Little evidence that the “usual care” group differed outside the intervention • Although outcomes are based on self-report, evidence suggests that self-report of DXA testing and bisphosphonate use is very good [49, 50] • Intervention group at higher risk, e.g.: • 87% intervention • All participating pharmacists had training or certification in research participation   a. Female (74% vs. 58%)

• 73% control   b. Fracture history (30% vs. 12%) Yuksel et al. [36] Low Low Low Low • Intervention group had significantly more participants with family history of OP (47% vs. 34%) • Attrition: n = 26 (20%) in intervention and n = 23 (17%) in control • All participating pharmacists received training • Self-report confirmed by DXA report from physician (test performed) and pharmacy records

(prescription dispensed) • However, analyses adjusted for age, sex, and family history of OP • However, all were accounted Savolitinib mw for in the analyses (intention to treat analysis) • Control (“usual care”) group also given educational material, and thus, the effect may be larger than what was see more observed in the trial when compared to true “usual care” Low risk of bias means that there is little evidence that this type of bias impacted study results. High risk of bias means that some evidence indicates that this type of bias may have impacted study results BMD bone mineral density group (peripheral DXA), DXA dual-energy X-ray absorptiometry, OP osteoporosis aAllocation bias occurs when randomization fails such that comparison groups differ on important AMN-107 clinical trial prognostic variables bAttrition bias may occur if patients who continue to be followed are systematically different from those who are lost mafosfamide to follow-up in ways that effect outcomes

cPerformance bias results from differences in the provision of care between comparison groups other than differences that relate to the main intervention dDetection bias results from differential outcome assessment between comparison groups 1. Cluster RCT in Australia Crockett et al. completed a cluster RCT in New South Wales, Australia whereby all 86 community pharmacists in six suburb and six rural communities were invited to participate [34]. Of the pharmacists that were willing and had suitable space and staffing to participate, one pharmacist within each of the six suburban and six rural areas was randomly selected for participation. Each of the 12 randomly selected pharmacists was then randomized into one of two groups: (1) non-BMD group, pharmacists offered education, counseling, and risk assessment based on patient questionnaire responses only and (2) BMD group, pharmacists offered education, counseling, and risk assessment based on questionnaire responses and forearm BMD test results. The forearm BMD tests were performed by a radiographer using peripheral dual-energy X-ray absorptiometry (DXA).