Data processing The microarray data obtained was analysed by using

the EMMA 2.8.2 software [74]. The mean signal intensity (A i) was calculated for each spot using the formula A i = log2(R i G i)0.5[75]. R i = I ch1(i) − Bg ch1(i) and G i = I ch2(i) − Bg ch2(i), where I ch1(i) or I ch2(i) is the intensity of a spot in channel 1 or channel 2, and Bg ch1(i) or Bg ch2(i) is the background intensity of a spot in channel1 or channel 2, respectively. The log2 value of the ratio of signal intensities (Mi) was calculated for each spot using the formula Mi = log2(Ri/Gi). Spots were flagged as “empty” if R ≤ 0.5 in both channels, where R = (signal mean–background mean)/background standard deviation [76]. The raw data were normalized GS-4997 by the method of LOWESS (locally

weighted scattered plot smoothing). A significance test was performed by the method of false discovery rate (FDR) control and the adjusted p-value https://www.selleckchem.com/products/GSK872-GSK2399872A.html defined by FDR was called q-value [77, 78]. An arbitrary cutoff, fold change (FCH) greater than 1.5, was applied to the genes with a q-value of ≤0.01. Only those genes which meet both filter conditions (q ≤ 0.01 & FCH ≥ 1.5) were regarded to be significantly differentially expressed. Real-time PCR The first-strand cDNA was obtained by reverse transcription with RevertAidTM Premium Reverse Transcriptase (Fermentas, St. Leon-Rot, Germany), using random hexamers as primers. Oligonucleotide Pexidartinib price primers were designed by the software PrimerExpress and listed in supplemental materials (Additional files 1: Table S4). Real-time PCR was performed with SYBR® Green PCR Master click here Mix kit (Carlsbad, California, USA) using 7500 Fast Real-Time PCR System (Carlsbad, California, USA) according to the manufacturers’ instructions. As an internal control, the housekeeping gene gyrA was used as its expression was not significantly altered in all microarray experiments. Three

technical replicates were carried out for each target gene. Quantification was analysed based on the threshold cycle (Ct) values as described by Pfaffl [79]. The raw data of the Micro-array experiments, described here, are available in the ArrayExpress database under the accession numbers: E-MEXP-3421, E-MEXP-3550, E-MEXP-3551, E-MEXP-3553, E-MEXP-3554, respectively (see also Additional file 3: Table S6). Acknowledgements The financial support for FB by the Priority Academic Development Program of Jiangsu Higher Education Institutions and the National Natural Science Foundation of China (No. 31100081) and the German Academic Exchange Service (DAAD) is gratefully acknowledged, as well as, the financial support given to RB in-frame of the competence network Genome Research on Bacteria (GenoMikPlus, GenoMikTransfer) and of the Chinese-German collaboration program by the German Ministry for Education and Research (BMBF).

J Bone Miner Res 27:694–701PubMedCentralPubMedCrossRef”
“Erratum

to: Osteoporos Int DOI 10.1007/s00198-013-2422-6 Incorrect data were given under the heading “Secular trends” in the Results section of this CHIR-99021 mouse article. The corrected text is given here. Secular trends for the period 1989–2008 in the over-70 age group, shown in Fig. 2, AZD8931 order reveal the time trend for incidence of MOS—the first hip, clinical vertebral, distal forearm, and upper arm fractures. The hip fracture rate increased for women in the period 1989–2000. After that, the rate decreased, resulting in 20 % lower rate in the period 2005–2008, compared to 1997–2000 (p = 0.056), and 7 % lower rate than in 1989–1992. In contrast, the rate for men increased (p = 0.076) until 2001 when it leveled off. The rate from 2005 to 2008 was 40 % higher than the rate in 1989–1992, ending in 501 events per 100,000 person years. The women/men ratio changed from 2.6 to 1.7 during the 20-year period. The incidence of other MOS fractures increased until 2001 for both men and women and declined similarly for both sexes during the last

decade, except for upper arm fractures in men. There was 38 % decline (IRR = 0.62, P = 0.11) for men and 31 % decline (IRR = 0.69, P = 0.019) for women in clinical vertebral fracture incidence during the period 1989–2008. For distal forearm fractures, the average incidence among women almost doubled from the first period (1989–1992) until the mid-period (1997–2000) (IRR = 1.62, Dinaciclib price P < 0.001) when a peak in the incidence was seen with a reduction of 17 % (IRR = 0.83, P = 0.11) until the last period (2005–2008). Men followed a similar

pattern PLEKHB2 albeit with a much lower number of fractures. We did a separate analysis for the time trend of cervical and trochanteric fractures which were very similar.”
“Introduction The use of glucocorticoids, even in low doses, is associated with rapid bone loss and an increased risk of fractures [1–4]. Bisphosphonates have been shown to be the most effective drugs for glucocorticoid-induced osteoporosis prophylaxis (GIOP) [5, 6] and are therefore recommended in (inter)national guidelines for management of GIOP [7–9]. The most important recommendation in the Dutch guideline is to consider starting bisphosphonates in post-menopausal women and men over 70 years who are expected to be treated with >7.5 mg prednisone (equivalents) per day for at least 3 months. In addition, all other patients who are expected to use >15 mg prednisone (equivalents) for more than 3 months should be treated with bisphosphonates. Although the awareness of the importance of osteoporosis prophylaxis seems to have increased [10], the widespread implementation of guidelines remains difficult. Audits have shown that only 10–60 % of patients who are eligible for GIOP receive appropriate treatment [11–14].

In brief, crude ORs and corresponding 95% CIs were preformed to assess the association between ATM D1853N polymorphism and breast cancer risk. The pooled ORs were calculated for

heterozygote comparison (GA buy Quisinostat versus GG), homozygote comparison (AA versus GG), dominant model (GA/AA versus GG) and recessive model (AA versus GA/GG), respectively. The statistical heterogeneity among studies was checked by Q-test and I 2 statistics [23]. If the P value greater than 0.10 for Q-test, indicating absence of heterogeneity, the fixed-effects model (the Mantel-Haenszel method) was used to calculate the pooled OR [24]; otherwise, the random-effects model (the DerSimonian and Laird method) was used [25]. Publication bias of literatures was estimated using Begg’s funnel plot [26]. All statistical analyses were carried out with STATA software, version selleck chemicals 10.0 (STATA Corp., College Station, TX). Results Characteristics of studies Overall, nine studies EPZ015666 cost involving 4,191 cases and 3,780 controls about ATM D1853N polymorphism and breast cancer susceptibility were available for this meta-analysis. The main characteristics of eligible studies are summarized in Table 1. There were six studies of European populations, two studies

of South American populations, and one study of mixed population that included more than one ethnic descent. Several genotyping methods were used, including polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), denatured high performance liquid chromatography (DHPLC), allele-specific oligonucleotide (ASO), PCR-single strand conformation polymorphism (PCR-SSCP), conformation sensitive gel electrophoresis (CSGE), TaqMan, and sequencing. Approximately 67% (6/9) of these studies described quality control for the genotyping assay. The genotype distributions in the Amisulpride controls of all studies were consistent with Hardy-Weinberg equilibrium except for one study

[27]. Main results The main results of this meta-analysis are shown in Table 2. Overall, no significant association between the ATM D1853N polymorphism and breast cancer risk was observed. After subgroup analyses according to ethnicity, significantly increased risk was observed in South American population (GA versus GG: OR = 2.19; 95% CI, 1.38-3.47; and dominant model: OR = 2.15; 95% CI, 1.37-3.38, respectively) but not in European and mixed populations. Table 2 Main results of pooled ORs in the meta-analysis   na Cases/controls GA versus GG AA versus GG GA/AA versus GG (dominant) AA versus GA/GG (recessive)       OR (95%CI) P b OR (95%CI) P b OR (95%CI) P b OR (95%CI) P b Total 9 4,191/3,780 1.18 (0.90-1.53) < 0.001 0.77 (0.58-1.03) 0.50 1.16 (0.89-1.51) < 0.001 0.78 (0.59-1.04) 0.66 Ethnicity                        European 6 3,913/2,852 1.00 (0.77-1.31) < 0.001 0.75 (0.56-1.01) 0.34 0.98 (0.75-1.29) < 0.001 0.77 (0.57-1.02) 0.

Parameters were labeled as apparent (app.) values, since, Y 27632 given the limited spatial resolution, they cannot depict the true trabecular structure. Fuzzy logic Previously, fuzzy logic was applied on magnetic resonance images to characterize trabecular bone structure [19, 21, 26]. The application on our CT images was conducted similarly. For the calculation of the 3D fuzzy logic parameters, no binarization was required. In a first step, which is known as “concentration,” each voxel within a VOI was multiplied by itself to increase contrast. Then each voxel was fuzzily segmented into the bone subset and the marrow subset by using fuzzy c-means clustering. Voxels were allowed

partial memberships in both subsets at the same time. The membership value of the voxel in the bone subset was considered as the amount of bone in the voxel, since the range of values for each voxel was from 0 to 1, where 0 represented a marrow voxel, 1 represented a bone voxel, and any value in between represented the corresponding BF of that voxel. Thus, fuzzy-bone volume fraction (f-BVF) maps could be generated. Based on these f-BVF maps, the fuzzy-bone fraction (f-BF) of the VOI could be calculated. ML323 Furthermore, 3D linear and quadratic indices of fuzziness and 3D logarithmic and exponential fuzzy entropies were computed according

to Carballido-Gamio et al. [19]. SIM-derived parameter The SIM is a tool for the structural characterization of arbitrary-dimensional

point distributions. For trabecular bone structure analysis, tomographic images can be interpreted as four-dimensional point distributions where each point (voxel) is defined by its x-, y-, and z-coordinate and its intensity value. A binarization of the images is not necessary. The 3D-based scaling index α can be calculated for each point of the distribution; α reveals the local dimensionality: rod-like stiripentol structures (α ~ 1), plate-like structures (α ~ 2), and random background (α ~ 3) can be differentiated. Nonlinear texture parameters can be derived from the probability distributions P(α) of the scaling indices α. According to previous studies, we extracted the scaling indices α in our CT images and calculated \( m_P\left( \alpha \right) \) with two sliding windows in the P(α) spectrum [18, 20] (Fig. 1). The position and width of the two windows were chosen to achieve optimal correlations between \( m_P\left( \alpha \right) \) and failure load (FL). Minkowski functionals The MF can be applied to multidimensional objects to characterize the composition of their components. In 3D, the four MFs, namely, volume (V MF), surface area (SurMF), mean integral curvature (CurvMF), and Euler characteristic (EulMF), entirely characterize one HSP inhibitor object.

The adherence assay was done after incubating bacteria with INT-407 cells for 30 min, after which adherence is assumed to be close to maximal, and the invasion assay was begun after 3 h of incubation of bacteria with INT-407 cells [26]. It must be noted that INT-407 cells have been found to contain contaminating HeLa markers. However, they have been used extensively for testing the adherence and invasion of Campylobacter jejuni[8, 10, 12] and have been found GSK1120212 concentration useful in that respect. Use of these cells should provide acceptable information as long as there is no attempt to make inferences regarding in vivo situations. Sentinel site surveillance

C-EnterNet sentinel site surveillance in the Region of Waterloo, Ontario (human population of approximately 500,000) has been described previously [7], http://​www.​phac-aspc.​gc.​ca/​c-enternet/​index-eng.​php. Isolates from both human and non-human (retail meats, on-farm manure, and surface water) sources from the sentinel site were characterized as part of the previous study. For each human case reported to the health unit a public health inspector contacted the patient to complete a comprehensive standardized questionnaire. Answers to the symptomology questions were collated and linked to the patient’s Campylobacter isolate information. Statistical analysis Statistical analysis for cell culture adhesion and invasion assays was

done by using the One-way Analysis of Variance (ANOVA) performed using BVD-523 order the Sigma Stat functions within the SigmaStat 3.5 software (Systat Software Inc.). The significance of each pairwise comparison was evaluated using the Holm-Sidak Test. The number of observations

used for each factor is given in the legend to Figure 2. XAV-939 in vivo Swarming assay (motility) results were also assessed statistically by using the One Way ANOVA within SigmaStat 3.5 software. The association of the presence of the CJIE1 prophage and the prophage + ORF11 with patient symptoms was analyzed using the Chi-Square analysis of contingency or the Fisher Exact Test within Sigma Stat 3.5 software. Acknowledgements We would like to acknowledge the invaluable help and advice provided by Dr. M. Konkel regarding cell culture adherence and invasion assays. The filipin funding source was Government of Canada A-base funds. References 1. Fouts DE, Mongodin EF, Mandrell RE, Miller WG, Rasko DA, Ravel J, Brinkac LM, DeBoy RT, Parker CT, Daugherty SC, Durkin AS, Madupu R, Sullivan SA, Shetty JU, Ayodeji MA, Shvartsbeyn A, Schatz SC, Badger JH, Fraser CM, Nelson KE: Major structural differences and novel potential virulence mechanisms from the genomes of multiple Campylobacter species. PLoS Biol 2005, 3:0072–0085.CrossRef 2. Parker CT, Quiñones B, Miller WG, Horn ST, Mandrell RE: Comparative genomic analysis of Campylobacter jejuni strains reveals diversity due to genomic elements similar to those present in C.

Combining our results with the results from CGM in a previous study, miglitol could reduce glucose fluctuations and hypoglycemic symptoms more effectively than other α-GIs. However, it is still unclear whether glucose fluctuations were lower in type 2 diabetic patients who

were treated longer with miglitol than in those who were treated longer with other α-GIs. Although CGM AG-881 order during the treatment of α-GIs were performed under oral meal loading tests at breakfast, lunch, and dinner in patients hospitalized for 4 days in the previous study [34], the diet during days when SMBG was performed in our selleck chemicals llc trials was dependent on each patient. RCT trials, in which dietary habits are well controlled, should examine whether glucose fluctuations by long-term CGM are lower in find more type 2 diabetic patients treated with miglitol than in those treated with acarbose or voglibose. It should be noted that our trial is a prospective exploratory trial that is not an RCT, which introduces some confounding factors and bias in our trial. It has been reported that blood glucose control is affected by seasonal changes. Indeed, it has been reported that HbA1c has a duration across the year that is highly detected during spring and gradually decreases by autumn in Japan [35]. One of the other possibilities

is that lifestyles such as dietary Pregnenolone and exercise habits in patients were changed during the trial. In this trial, the doctor assigned caloric intake and the suggestion was not changed during the trial. However,

it is possible that the lifestyles of patients were changed by themselves. In addition, miglitol treatment may reduce a patient’s appetite because the change of α-GI to miglitol treatment inhibits symptoms of hypoglycemia and reduction of blood glucose levels during a meal; however, our results indicate that the change of α-GI to miglitol reduced glucose fluctuation but not HbA1c. Thus, the effect is most likely a result of the effects of miglitol because changes in dietary and exercise habits may alter HbA1c levels. Whether miglitol treatment reduces circulating CVD risk factors including MCP-1 and sE-selectin in type 2 diabetic Japanese patients needs to be examined in an RCT. 5 Conclusion The results of this study indicate that switching from acarbose or voglibose to miglitol for 3 months suppressed glucose fluctuations and serum protein concentrations of MCP-1 and sE-selectin more effectively than the prior α-GI. Acknowledgments This study was sponsored by Sanwa Kagaku Kenkyusho Co., LTD, Nagoya, Japan. Conflict of interest Mr. Fuchigami is an employee of Sanwa Kagaku Kenkyusho Co., LTD, Nagoya, Japan.

The associated decrease in intracellular pH is a factor leading to muscle fatigue [23, 24]. Therefore, during maximal exertion blood flow is this website needed not only for oxygen supply to support continued oxidative phosphorylation, but also for H+ removal for muscle pH regulation. It would seem that exogenous ATP would likely have a greater impact on the muscles’ ability to perform fatiguing exercise by increasing substrate availability to the muscle and/or CA4P facilitating waste product removal through increased blood flow through the muscle tissues. Both ATP and adenosine can act through purinergic receptors in endothelial

smooth muscle of the vascular system resulting in vasodilation and increased blood flow [14, 15, 25]. A study by Gonzalez-Alonso showed that intra-arterial infusion of ATP was associated with vasodilation and increased blood flow with a significant reduction in venous ATP levels in the non-exercising limb suggesting utilization of ATP or metabolites [26]. These

observations were confirmed by Calbet et al. who hypothesized learn more that increased delivery of ATP would affect non-exercising vasoconstrictive muscle tissue [20]. These are most likely due to activation of purinergic receptors affecting blood flow [13]. Furthermore, exogenous adenosine administration results in vasodilation [14] and increased glucose and O2 uptake by muscle which provide for an increased substrate pool [12]. The ATP used in the present study was not enterically coated and was fed encapsulated as the disodium salt. The sodium salt would have

provided buffering of the ATP through the stomach and the ATP itself should have been metabolically check details available as soon as it reached the proximal duodenum, which has been shown to be the most biologically active site for ATP metabolism and/or absorption [17]. In France, this chemical form of ATP is approved as a drug for lower back pain [27, 28]. One proposed mechanism of action is through improved oxygenation of the muscle, which could be of similar benefit during exhaustive exercise. Other effects of ATP or its metabolites could also indirectly impact work performance as ATP has immunomodulatory effects [29], and inotropic effects on cardiac muscle [30, 31]. Oral administration of ATP to rabbits for 14 days results in systemic effects through a reduction in peripheral vascular resistance, improvement of cardiac output, reduction of lung resistance, and increased arterial PaO2[32]. A study in humans demonstrated that interstitial infusion of adenosine in muscle induced nitric oxide formation in skeletal muscle and nitric oxide and prostacyclin formation in microvascular endothelial cells [15]. Alternatively, the effects of cbvexogenously administered ATP may also be due to the associated increase in plasma uric acid, which has been proposed to act as an anti-oxidant [33, 34].

The inhibition of c-FLIP expression can down-regulate HCC cell viability and up-regulate drug-induced cell apoptosis. Our data suggest that targeting c-FLIP in conjunction with anticancer therapies may have therapeutic potential by enhancing

HCC cell death. Acknowledgements This study was supported in part by a grant from National Natural Scince Foundation of China (No. 30700810). The authors would like to thank Dr Yi Wan(Department of medical statistics, FMMU, China) for his help with statistical work and Dr Haichao Wang(Chief, Basic Science Research Program, Department of Emergency Medicine, NSUH-NYU School of Medicine, Manhasset, NY) check details for linguistic revision of the manuscript. References 1. Igney FH, Krammer PH: Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer 2002, 2: 277–88.CB-839 price CrossRefPubMed 2. Bouchet D, Tesson L, Ménoret S, Charreau B, Mathieu P, Yagita H, Duisit G, Anegon I: Differential sensitivity of endothelial cells of various species to apoptosis induced by gene transfer of Fas ligand: role of FLIP levels. Mol Med 2002, 8: 612–23.PubMed 3. Ishioka T, Katayama R, Kikuchi R, Nishimoto M, Takada S, Takada R, Matsuzawa S, Reed JC, Tsuruo T, Naito M: Impairment of the ubiquitin-proteasome system by cellular FLIP. Genes Cells 2007, 12: 735–44.PubMed 4. Rogers KM, Thomas M, Galligan L, Wilson TR, Allen WL, Sakai

H, Johnston PG, Longley DB: Cellular FLICE-inhibitory protein regulates chemotherapy-induced apoptosis in breast cancer cells. Mol Cancer Ther

2007, 6: 1544–51.CrossRefPubMed 5. Mezzanzanica D, Balladore E, Turatti F, Luison E, Alberti P, Bagnoli M, Figini M, Mazzoni A, Raspagliesi AR-13324 order F, Oggionni M, Pilotti S, Canevari S: CD95-mediated apoptosis is impaired at receptor level by cellular FLICE-inhibitory protein (long form) in wild-type p53 human ovarian carcinoma. Clin Cancer Res 2004, 10: 5202–14.CrossRefPubMed 6. Hyer ML, Sudarshan S, Kim Y, Reed JC, Dong JY, Schwartz DA, Norris JS: Downregulation ifenprodil of c-FLIP sensitizes DU145 prostate cancer cells to Fas-mediated apoptosis. Cancer Biol Ther 2002, 1: 401–6.PubMed 7. Krueger A, Baumann S, Krammer PH, Kirchhoff S: FLICE-inhibitory proteins: regulators of death receptor-mediated apoptosis. Mol Cell Biol 2001, 21: 8247–54.CrossRefPubMed 8. Kataoka T, Ito M, Budd RC, Tschopp J, Nagai K: Expression level of c-FLIP versus Fas determines susceptibility to Fas ligand-induced cell death in murine thymoma EL-4 cells. Exp Cell Res 2002, 273: 256–64.CrossRefPubMed 9. Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, Steiner V, Bodmer JL, Schröter M, Burns K, Mattmann C, Rimoldi D, French LE, Tschopp J: Inhibition of death receptor signals by cellular FLIP. Nature 1997, 388: 190–5.CrossRefPubMed 10. Wilson TR, McLaughlin KM, McEwan M, Sakai H, Rogers KM, Redmond KM, Johnston PG, Longley DB: c-FLIP: A Key Regulator of Colorectal Cancer Cell Death. Cancer Res 2007, 67: 5754–62.CrossRefPubMed 11. Wajant H: Targeting the FLICE Inhibitory Protein (FLIP) in cancer therapy.

cerevisiae with a much higher number. This yeast seems therefore to differ clearly from filamentous fungi in the sense that it possesses quite a lower number of O-glycosylated proteins (Table 1), only partially explained by the smaller genome size, but they are more extensively O-glycosylated (Figure 2). Figure 2 Frequency distribution of the number of O -glycosylation sites per protein predicted by NetOGlyc. Inset displays the average number of O-glycosylated

residues per protein, corrected by multiplying by 0.68 to compensate the overestimation of O-glycosylated sites produced by the server on fungal proteins. See details in the text. If we look at individual proteins we can find some with an AG-881 cell line extremely high number of O-glycosylation sites (Additional file 2). The protein with the highest LY3039478 in vivo proportion of predicted O-glycosylated residues is the M. grisea protein MG06773.4, of unknown function, with about half of its 819 amino acids being predicted to be O-glycosylated. Next is the S. cerevisiae protein YIR019C (Muc1), a mucin-like protein necessary for the yeast to grow with a filamentous pseudohyphal form [15]. Muc1 is a 1367-amino acids protein, of which 42% are predicted to be O-glycosylated.

Similar examples can be found in the rest of the Blasticidin S cell line genomes, with at least a few proteins predicted to have more than 25% of their residues O-glycosylated. Fungal proteins are rich in pHGRs The glycosylation positions

obtained from NetOGlyc were analyzed with the MS Excel macro XRR in search of O-glycosylation-rich regions. The Glutamate dehydrogenase raw results can be found in Additional file 3 and a summary is presented in Table 2. All the genomes analyzed code for plenty of secretory proteins with pHGRs. Between 18% (S. cerevisiae) and 31% (N. crassa) of all proteins with predicted signal peptide contain at least one pHGR. The average length of pHGRs was similar for the eight genomes, varying between 32.3 residues (U. maydis) and 66.9 residues (S. cerevisiae), although pHGRs could be found of any length between the minimum, 5 residues, to several hundred. All genomes coded for proteins predicted to have quite large pHGRs, the record being the 821-aa pHGR found in the S. cerevisiae protein Muc1 discussed above. Globally, we could summarize these data by saying that among the set of secretory fungal proteins predicted by NetOGlyc to be O-glycosylated, about one fourth shows at least one pHGR having a mean length of 23.6 amino acids and displaying, on average, an O-glycosylated Ser or Thr residue every four amino acids.

After that, if we lift up the tip, the curves in Figure 3 indicate that the manipulated atom will stay in the well near the tip. That is, the atom will follow the tip and be extracted from the surface, as the simulation above shows. From Figure 3, we can also estimate the reliability of the extraction process; the energy curve of 6.1 Å shows that the energy barrier for the manipulated atom escaping from the tip is about 0.25 eV, which indicates that the picking up process is robust against the disturbances

such as thermal diffusion of atoms. Figure 3 Variation of potential energy relative to height of https://www.selleckchem.com/products/pf-03084014-pf-3084014.html manipulated atom. At different tip heights, the relative potential energy varies with the height of the manipulated atom from the Al (111) step surface. The next step of substitutional doping is to position a dopant atom to the HDAC inhibitor vacancy site where the Al atom is extracted. Here, we consider HSP990 cell line two kinds of dopants: Ag and Au atoms. For this purpose,

sharp Ag and Au tips with single apex atom are considered; such sharp tip can be fabricated by electroplating and then annealing, or touching a certain metal surface [17, 18]. In our simulation, the sharp Ag tip is modeled by a heterogeneous one which contains both Ag and Al atoms, as shown in Figure 4. Blue balls indicate the Ag atoms. The apex of heterogeneous tip is mimicked by three layers of Ag atoms, and our test calculations show that three layers of Ag atoms are equivalent to four layers or more. In other words, three layers of Ag atoms

are sufficient for simulation of the sharp Ag tip which is also suitable for the Au tip. Figure 4 The process of positioning Ag dopant to the Galeterone step site by Ag single-apex tip. (a) The tip is located upon the site. (b) As the tip approaches the surface, the dopant atom relaxes toward the up terrace. (c) Move the tip laterally in the X direction. (d) In the end, the dopant atom is released successfully from the tip and adsorbed at the step site. As shown in Figure 4a, the tip is initially placed above the vacancy site with the tip height of 8 Å at which the tip-surface interaction is almost negligible. As the tip approaches the surface step by step, the tip apex atom, i.e., the dopant atom, relaxes toward the up terrace due to the strong attraction. When the tip reaches the height of 7.1 Å, as demonstrated in Figure 4b, the dopant atom shows an obvious movement toward the up terrace since the attraction is strong enough. At this moment, two up-terrace atoms are pulled up slightly and in contact with the dopant atom (see Figure 4b). After that, we move the tip laterally in the X direction in a step of 0.2 Å at a constant height. As the tip moves forward, as shown in Figure 4c, the dopant atom drops gradually because of the decreasing vertical attraction from the tip. In the end, the dopant atom is released successfully from the tip and adsorbed at the step site (see Figure 4d).