Regarding simple pre-post assessments of QoL in single-arm studies, it is probably unnecessary to state that

they are generally not appropriate for judging influences on QoL, since it is affected by many factors. Concerning survival (Table 3), some of the RCTs show a statistically significant benefit while others find more show a statistical trend or no difference. Most of the non-RCTs (which included larger patient numbers) show a major impact. The validity of the studies is limited because of their small sample size (median only 52 participants per RCT), and because 8 of the 9 RCTs were imbedded in the same (large) epidemiological cohort study. This study was started in the 1970s, before modern standards of data quality control (ICH-GCP, GEP) were established, and it therefore does not fulfil modern standards in this respect. The 9th RCT had enrolled more patients but was conducted even earlier, and suffers from a major attrition rate due to protocol violation [62]; the subsequent analysis followed the “”as treated”" instead of the “”intention-to-treat”" principle [145]. Hence https://www.selleckchem.com/products/lee011.html bias cannot be excluded. None of the survival studies was blinded, but survival is generally not easily affected by observer bias or suggestive effects [138–140]. Seen altogether, although results were consistent, questions regarding survival

remain and validity of evidence is moderate at best. An independent, GCP-conform trial with sufficient power would be desirable to further evaluate potential survival benefit. Regarding tumour behaviour, evidence from RCTs is Niraparib in vivo scanty; most benefits were shown in non-randomized studies. In single-arm studies of patients with no concomitant conventional cancer treatment, high-dose or local application of whole VAE led to substantial remission of tumour or malignant effusion. This was also observed in animal studies: local application resulted in tumour-growth inhibition and increased survival. However, this application and dosage is not standard and cannot be recommended widely

due to potential risks of high dose or local application. With ordinary Ribonucleotide reductase VAE application, schedule and dosage, spectacular tumour remissions tend to be the exception [20, 36]. No tumour remission was observed after application of rMLs. Remission in CIN cannot be distinguished from spontaneous remission rates, which are frequent in this indication. Apart from the discussed issues, the following validity aspects have to be considered: An attrition rate above 10% was present in 10 RCTs. In 5 of these RCTs [49–51, 53], patients were excluded before baseline assessment. Here the patients were provisionally enrolled into the matching and pairwise randomization procedure; subsequently they were asked for informed consent, and were excluded from the study if they declined, together with their matched twin.

The specificity and the efficiency of the primer pairs was verified by melting curves and the construction of standard curves based on a Olaparib ic50 serial two-fold dilution (20 – 2-5) using soil DNA as the template. Template plasmids were used to generate a standard curve that was used as an external

standard. The target DNA sequence was cloned into the pGEM-T vector selleck products (Promega) and the resulting plasmids were purified. All plasmids were quantified by spectrometry using a Nanodrop ND-1000 instrument (Thermo Scientific) and copy numbers were estimated based on the molecular weight of the template. The number of copies of the cloned target DNA in the dilution series ranged from 106 to 101. Real-Time

PCR assays Real-time PCR was performed using the iQ SYBR Green Supermix (Bio-Rad). The reaction mixtures PD-0332991 purchase contained 7.5 μl of iQ SYBR Green Supermix, 1 μl of DNA solution (corresponding to 1 ng of DNA), and 350 nmol of each gene-specific primer. The experiments were conducted in 96-well plates with an iQ 5 Multicolour Real-Time PCR Detection System (Bio-Rad). PCR was always performed with three biological and three technical replicates. The cycling conditions were 10 s at 95°C, 30 s at 55°C or 62°C. Template abundances were determined based on the Ct values (which measure the number of cycles at which the fluorescent signal exceeds the background level and surpasses HA-1077 supplier the threshold established based on the exponential phase of the amplification plot). The significance of differences between the Ct values of different treatments were determined by one way analyses of variance ( p < 0.05) and grouped according to the Tukey HSD test in R (R Core team, 2012). Acknowledgments We thank D. Krüger for advice on fungal PCR primer construction. We thank K. Hommel, I. Krieg and B. Krause for oak micropropagation

and S. Recht for her role in setting up the soil microcosms. Financial support was supplied by the German Science Foundation (DFG) (TA 290/4-1) and by the Helmholtz Gemeinschaft. This work was kindly supported by Helmholtz Impulse and Networking Fund through Helmholtz Interdisciplinary Graduate School for Environmental Research (HIGRADE). The authors thank the Laboratory of Electron Microscopy BC AS CR, v.v.i. – Parasitology Institute České Budějovice for a productive collaboration on scanning electron microscopy. Electronic supplementary material Additional file 1: Experimental setup for quantification of AcH 505 and P. croceum under different culture conditions. (PDF 310 KB) Additional file 2: qRT-PCR melting and standard curves obtained using the AcH107 primer pair. (PDF 362 KB) Additional file 3: qRT-PCR melting and standard curves obtained with the ITS-P primer pair.

After a mean follow-up of 41.4 months, there have been 2 cases of ASBO recurrence in the icodextrin group and 10 cases in the control group (p < 0.05). Only one patient in the first group was submitted to surgery showing an Adhesion Severity Score = 2,

whereas three patients in the latter Ferrostatin-1 group were operated, and the ASS was respectively 3,2 and 3. In accordance with this data, the use of icodextrin 4% solution seems to be safe and effective to prevent intra-abdominal adhesion formation and the risk of re-obstruction [100]. Intergel solution (Lifecore Biomedical, Inc, Chaska, MN), which contains .5% ferric hyaluronate, is another product used for adhesion prevention. In preliminary studies it has been shown to reduce the number, severity, and extent of adhesions

in peritoneal surgery [101]. However, the use of Intergel in abdominal surgery in which the Blasticidin S in vivo gastrointestinal tract was opened still led to an unacceptably high rate of postoperative complications [102]. An interesting experimental finding is the reduction of both number and type of adhesions after postoperative stimulation of gastrointestinal motility by a prokinetic agent [103]. Finally merits mention that peritoneal infusion Aurora Kinase inhibitor with cold saline has shown to decrease the degree of postoperative intra-abdominal adhesion formation in an animal model [104]. Adhesions quantification Among the different adhesions scoring

systems which have been proposed mainly by gynecologists, the more complete and easy to use one is the PAI score proposed by Coccolini et al. [105]. In fact, specific attention should be paid to uniformity of measurement. We therefore triclocarban suggest a regimented classification system for adhesions in an effort to standardize their definition and subsequent analysis. In this way, different surgeons in different treatment centers can more effectively evaluate patients and compare their conditions to past evaluations using a universal classification system (Figure 3). This classification is based on the macroscopic appearance of adhesions and their extent to the different regions of the abdomen. Using specific scoring criteria, clinicians can assign a peritoneal adhesion index (PAI) ranging from 0 to 30, thereby giving a precise description of the intra-abdominal condition [105]. Figure 3 Peritoneal adhesion index: by ascribing to each abdomen area an adhesion related score as indicated, the sum of the scores will result in the PAI. Conclusions ASBO is a common disease. Non operative management should be attempted in absence of signs of peritonitis or strangulation.

Conclusions Evaluating scattering and near field properties of metallic and dielectric nanoparticles, we firstly found that the scattering cross sections can, in both cases, reach a value of several times the geometrical cross sections. For the dielectric nanoparticles, no parasitic absorption exists, whereas for the metallic ones, non-zero absorption cross sections are present, which however can be reduced by increasing the particle radius. The nanoparticle radius can be

BAY 80-6946 concentration used to tune the resonance position to the desired wavelengths. Scattering cross section maps, calculated here with Mie theory, give a fast overview of the parameter field and quickly show that dielectric nanoparticles with a refractive index around 2 require significantly larger radii (approximately 1.5 times) than metallic ones from, e.g., Ag in order to obtain similar resonance wavelengths. The electromagnetic near fields around the two different

nanoparticle types also significantly differ; whereas for the metallic nanoparticles, the field vanishes inside and builds up a strong localized field around the surface, the dielectric nanoparticles have strong fields inside, which however are not absorbed but preferentially scattered to the forward direction. These observations of both typical dielectric and metallic near-fields are found for semiconducting materials. On the one hand, they have a selleck inhibitor region of constant refractive index and zero absorption and thus a dielectric-like scattering behavior, but on the other Sodium butyrate hand, they can also show significant charge

carriers and thus metallic plasmon resonances. However, since the semiconductor also has a band gap and according high absorption for wavelengths below, it may only be of interest when the band to band absorption is outside the wavelength range in focus. Although semiconductors show the scattering properties of both dielectrics and metals, it was not possible to combine the two effects constructively. Depending on the application, one or the other type of material by itself may be preferred to a combination of both. Aside from the scattering ability and the near field distribution, also the angular distribution of the scattered light plays a crucial role for applications. Considering in particular the application to ultra-thin solar cells, both an enhanced near field and a particular scattering of the nanoparticle may contribute to enhance the absorption. In a homogeneous medium, the near field is stronger around the metallic nanoparticle, the scattering efficiency (scattering over scattering plus absorption) is stronger for A-1155463 in vitro non-absorbing dielectric nanoparticles, so that up to that point, no decision about the ideal choice of material can be made.

In this study, GSK1210151A we were able to assess the usefulness of VNTRs for the study of Xam populations. Remarkably, only 5 VNTR loci offered a very similar panorama of the pathogen populations to that obtained by 57 AFLP loci.

This finding is relevant for further studies on the population dynamics of Xam, because VNTR markers provide a faster and less expensive characterization of bacterial isolates, as has been reported for several pathogenic microorganisms [22, 24, 25, 49]. The fact that amplification of VNTRs requires neither a complex DNA extraction procedure, nor compounds different from those used in a regular PCR, makes VNTRs ideal when a large PND-1186 solubility dmso number of isolates are considered and when funding is limiting. Moreover, sharing information between laboratories would be considerably more straightforward with VNTRs than with AFLPs, because results from VNTRs can be more easily coded [17]. For future Xam survey studies we recommend the use of VNTRs. The rising number of sequenced

genomes available nowadays, provides an additional advantage to identify new VNTR loci, hence improving the characterization of several pathogens [19, 21, 50, 51]. Recently, 65 partial genomes of Xam strains have been released [52], providing a valuable opportunity to detect VNTRs with high discriminatory power. Currently, we are focusing on the prediction and evaluation of new VNTR loci into a core of the representative Xam strains using the information obtained from the 65 draft genome sequences. Our goal is to obtain a small sets of VNTRs with a high discriminatory Ribonucleotide reductase Tanespimycin concentration power, aiming to implement them in studies that involve a large number of isolates to provide a more accurate description of evolving processes taking place in Xam populations. Conclusions This study represents the first attempt to type populations of Xam using VNTRs as molecular markers. Here we demonstrated that a small number

of VNTR loci could offer a similar panorama of the status of the pathogen to that offered by AFLPs markers. Because VNTRs represent a fast and simple tool to type Xam populations, their implementation will allow a constant and adequate surveillance of the pathogen, which could provide information to improve the efficiency of strategies for disease control, such as the deployment of resistant varieties. Availability of supporting data The data sets supporting the results of this article are available in the Dryad Digital Repository: http://​doi.​org/​10.​5061/​dryad.​t173v. DNA sequences are available in Genbank database: (Accession numbers XaG1_02: KJ736838 – KJ736944; XaG1_29: KJ736945 – KJ737053; XaG2_52: KJ737163 – KJ737268; XaG1_67: KJ737269 – KJ737369; XaG1_73: KJ737054 – KJ737162). Ethics statement This study did not involve any human material, or human data.

However, during nanocutting process of materials, this assumption is not reasonable since the cutting tool edge radius is on the same scale as the undeformed chip thickness. Thus, the simulation has been done with the cutting edge radius of 2

nm. The spherical indenter contained 36,259 atoms with AZD2014 a radius of 50.0 Å. The motions of the atoms in the Newton and thermostat atoms are assumed to follow Newton’s law of motion which can be computed from the interatomic forces as follows: (1) where a ix represents the i atom’s acceleration in the X direction, m i is the mass of the i atom, F ix is the interaction force between the i atom by the j atom in the X direction, x i indicates the i atom’s X-coordinate, and V is the potential energy. The temperature of atoms during the machining simulation can be calculated using the conversion between the kinetic energy and temperature as ARRY-438162 mw follows: (2) where N is the number of atoms in

groups, v i represents the velocity of the i atom, k b is the Boltzmann constant which is equal to 1.3806503 × 10−23 J/K, and T represents the temperature on atoms. In order to keep the temperature constant during the nanocutting process and nanoindentation process, in other words, ensuring reasonable heat conduction outwards from the Newtonian atom zone [10], the thermostat atom zone is set to absorb the heat from the specimen. When the temperature of the thermostat atom zone is higher than the preset one of 296 K, the velocity rescaling method as shown in Angiogenesis inhibitor Equation 3 [11] is used to control the temperature of the thermostat atom zone and

absorb the heat towards the Newtonian atom zone. The direct velocity scaling method ID-8 was employed to maintain the total kinetic energy at a constant value. The velocity of every atom in the thermostat atom zone needed to be scaled at every integrating step, and the velocity scaling factor is as follows: (3) Selection of potential energy function In this paper, there are two kinds of atoms in the MD simulation model, which are C and Cu atoms. Therefore, there are three different atomic interactions between them, which are the interaction between single-crystal copper atoms (Cu-Cu), the interaction between diamond atoms (C-C), and the interaction between copper atoms and diamond atoms (Cu-C) or (C-Cu). The potential energy function affects the accuracy of the simulation which governs the reliability of results. Between copper atoms in the specimen, the embedded atom method (EAM) potential [12] was applied to describe the Cu-Cu interaction. The EAM potential, which evolved from the density function theory, is based on the recognition that the cohesive energy of a metal is governed not only by the pair-wise potential of the nearest neighbor atoms, but also by embedding energy related to the ‘electron sea’ in which the atoms are embedded.

Synthesis of cDNA were performed from 150 ng of total RNA confirmed free of DNA after an additional DNase treatment, 6 μg hexamers, 10 mM of dNTP with Superscript III and supplied reagents as described above. The primers used in real-time quantitative PCR are listed in Table 1. Real-time PCR was performed with a cDNA dilution in triplicates, representing 0.75 ng RNA, 0.1 μM of each primer with FastStart SYBR Green master included ROX (Roche Applied Science) on an ABI Prism 7700 Sequence Detection System (Applied Biosystems).

After denaturation at 95°C the program was 40 KU55933 mouse cycles, including 95°C for 15 seconds, 30 seconds at 62°C and 72°C for 30 seconds. Standard curves were made for each primer pair to calculate amplification efficiency of the target genes and the endogenous control gene (EF0013). Differential expression was determined by calculating the change in threshold cycles for each gene with the ΔΔCt-method, with RNA isolated from resistant mutants and wild type bacteria. DNA manipulations and sequencing Isolation of DNA from E. faecalis V583 buy EPZ-6438 and mutants was done using Advamax-beads (Advanced Genetic Technologies Corp.). PCR products were generated with Phusion DNA polymerase (Finnzymes). Other enzymes for DNA manipulation were from New England Biolabs. DNA fragments were purified by use of agarose gel electrophoresis and Qiaquick PCR purification columns (Qiagen).

Plasmids were isolated using Qiagen miniprep columns. Standard procedures [32] were used for restriction cutting of DNA, ligation and cloning in E. coli. DNA was sequenced using the ABI Prism BigDye terminator sequencing ready reaction kit version 3.1 and analyzed with the ABI Prism 3100 genetic analyzer according to the supplier’s procedures (Applied Biosystems). Results Isolation and characterization of bacteriocin resistant mutants Four class IIa bacteriocin resistant mutants of E. faecalis V583 were obtained. Mutants MOP1 and MOP5 were isolated after exposure to two different

concentrations of pediocin PA-1. A third spontaneous mutant (MOP2) was obtained by selecting colonies resistant to 2-DG. The MOP2 mutant was also resistant to pediocin (Table 2). Pediocin PA-1 resistant mutants were Histamine H2 receptor isolated at a frequency of 3 10-4, consistent with reported resistance frequency in Enterococcus and Listeria [6, 7]. Previous studies have shown that pediocin resistance can be obtained by mutations in the mannose PTS operon, mpt [33, 34], therefore we GSI-IX supplier constructed a resistant E. faecalis V583 (MOM1) disrupted in mptD. Mutants MOM1 and MOP5 were highly resistant to pediocin PA-1, while MOP1 and MOP2 were less resistant (Table 2). The pediocin resistance phenotype was stably maintained in all mutants in the absence of bacteriocin. All mutants were resistant to 2-DG (results not shown). In exponential phase up to an optical density of 0.

To combat interferences seen using absorbance as endpoint readout, a cytotoxicity assay using resazurin and its fluorescent product this website was applied. AuNP-only controls suspended in EMEM medium were included and interference was detected. We observed a concentration-dependent decrease in the levels of fluorescence as a result of AuNP interference (Figure 9c). At the highest

concentration of AuNP, levels decreased by 11% to 24% depending on the AuNP in question. Au[(Gly-Tyr-TrCys)2B] exhibited the highest level of interference. The results were interpreted with care in order to avoid drawing erroneous conclusions. Cytotoxicity was assumed only when the decrease in fluorescence was lower than possible interference levels. We also examined whether the AuNPs used in this study interacted with the glutathione assay. AuNPs selleck chemicals absorbed at the wavelength used in this assay (405 nm). A dose-dependent increase appeared for some of them at concentrations of 1.56 μg/ml (data not shown) or higher. Additionally, when glutathione was incubated with a range of AuNP concentrations for 2 h the

level of free glutathione decreased as the concentration of AuNPs increased (Figure 9d). Therefore, this assay was not considered suitable for studying the oxidative stress potential of the AuNPs. However, no interference was observed with the ROS production assay (data 3Methyladenine not shown). Figure 9 PBH-capped AuNP interference with the toxicity assays. (a) MTT, (b) neutral red uptake (NRU), (c) resazurin-based cytotoxicity assay and (d) glutathione detection. Cytotoxicity Methyl thiazol tetrazolium and neutral red uptake assays The MTT and NRU assays could not be performed as there was AuNP interference at the wavelengths used in these tests (570 and 550 nm, respectively) (Figure 9a,b). Resazurin assay Cytotoxicity assays were performed Cell press with cells incubated

in EMEM/S+ and EMEM/S- after 24- and 48-h exposure periods. Only results with cells incubated in EMEM/S- are shown in Table 3, as clear evidence of cytotoxicity in cells exposed to AuNPs in EMEM/S+ could not be determined because of high interference levels in this assay under these conditions (Figure 9c). Cytotoxicity is expressed as percentage of live cells (viability) compared to the untreated control (100%). At the highest concentration (100 μg/ml), all AuNP preparations caused approximately 10% decrease in viability. This was the highest decrease in viability recorded after 24 h of incubation for the AuNP preparations tested. This decrease in viability was not higher than that recorded for the cell-free AuNP-only controls in the interference studies (11% to 24% decrease). Therefore, the reduction in viability is perceived to be a result of NP interference and cannot be reported as cytotoxicity. After 48 h of incubation, the level of cytotoxicity for Au[(Gly-Tyr-Met)2B], Au[(Met)2B] and Au[(TrCys)2B] increased significantly for the two highest doses of 50 and 100 μg/ml (p < 0.01).

37 [20] FFIVC131 CDC2412-93 O139   1 1 0 1995 Human USA 1 1 1 1 1 1 2.43 [20] FFIVC133   O139   1 1 0 2003 unknown unknown 1 1 1 1 1 1 2.49 [20] 080025/FR Vib31 O141   1 1 1 1993 Human Spain singleton 8 7 3 2 9 2.24 [18] FFIVC050   non O1/O139   0 0 0   Mussels Norway singleton 8 9 9 11 5 2.28 [20] FFIVC084   non O1/O139   0 0 0 2003 Mussels Norway singleton 4 2 4 3 3 2.45 [20] FFIVC114   non O1/O139   0 0 0 2004 Water Norway 4 6 1 6 6 6 2.29 [20] FFIVC115   non O1/O139  

0 0 0 2004 Water Norway 4 6 1 6 6 6 2.39 [20] FFIVC137   non O1/O139   0 0 0   Human Norway singleton 7 5 8 10 4 2.41 [20] 2/110/2006   non O1/O139   0 0 0 1998 Water selleck Poland 5 10 4 2 12 4 2.25 [18] 3/110/2006   non O1/O139   0 0 0 1998 Water Poland 5 10 4 2 12 4 2.42 [18] 4/110/2006   non O1/O139   0 0 0 2004 Water Poland singleton

11 0 13 0 11 2.38 [18] 14/110/2006   non O1/O139   0 0 0 1998 Water Poland singleton 5 3 10 4 7 2.37 [18] 17/110/2006   non O1/O139   0 0 0 1998 Water Poland 6 3 6 7 5 10 2.47 [18] 22/110/2006   non O1/O139   0 0 0 2004 Water Poland 6 3 6 7 5 10 2.26 [18] 070256/J V. mimicus ATCC 33655 –   1 0 0     10 14 10 12 1 14 1.71 [18] a“0” means no PCR product was obtained. bMSP value: highest logarithmic value of the four generated MS-spectra score value compared to Biotyper reference library. cReference(s), in which the isolate EPZ5676 manufacturer is described previously. Confirmation of strain identification Identification of the isolates at species level was confirmed by MALDI-TOF MS using Biotyper 2.0 (Bruker Daltonics GmbH, Bremen, Germany) [11]. Serogroup and serotype were confirmed using the Vibrio cholerae E Agglutinating Sera kit containing specific antisera O1 polyvalent agglutination serum, Inaba agglutination serum, and Ogawa agglutination serum (Remel Europe Ltd. Darford, Kent, United Kingdom) according to the manufacturer’s guidelines. Genotyping of isolates with multilocus sequence typing (MLST) analysis MLST analysis was performed

according to Teh et al. [21]. Internal gene fragments of dnaE, lap, recA, gyrB, and cat were PCR amplified and sequenced. The gmd gene was not included in the analysis due to low discriminatory power [21]. Each sequence variant of a locus was assigned a distinct allele number. In the case that no PCR product could be obtained for a specific allele, the Selleck BI-2536 number zero was assigned. The allele profiles next were entered into BioNumerics version 6.6 software (Applied-Maths, Belgium) as character values, and the genetic relationship between isolates was constructed using the categorical coefficient and the Minimum Spanning Tree algorithm. Isolates that differed at two or fewer loci were considered genetically closely related, while single locus variants (SLV) were defined as having at least three alleles that were different from all other tested isolates.

Furthermore, patients treated with upfront ZOL had a significantly higher risk of bone pain than patients with delayed ZOL. More attentions should be paid to patients with musculoskeletal disorders. For patients with low risk of osteoporosis, immediate ZOL may be not needed due to additional adverse effects in some conditions. Or it can be stopped after the occurrence of these adverse events. Further randomized clinical trials with large sample size should

be taken to evaluate the side effects of ZOL, especially for musculoskeletal disorders. Conflict of interest The authors declare that they have no competing interests. Acknowledgements We are grateful to Dr. Jifu Wei (Clinical Experiment Center, the First Lonafarnib ic50 Affiliated Hospital with Nanjing JSH-23 molecular weight Medical University) for critical discussion in our study. This work was supported in part by Wu Jie-Ping Foundation (320.670010009), the National Natural Science Foundation of China (81071753), the Six Kinds of Outstanding

ARS-1620 clinical trial Talent Foundation of Jiangsu Province (To Wei He), the Science and Education for Health Foundation of Jiangsu Province (RC2007054), the Natural Science Foundation of Jiangsu Province (BK2008476, BK2009438 and BK2010581), the Program for Development of Innovative Research Team in the First Affiliated Hospital of NJMU (IRT-008), and A project Funded by the Priority Academic Program Development of Jiangsu higher Education Institutions (PAPD). References 1. Elmore JG, Armstrong K, Lehman CD, Fletcher SW: Screening for breast cancer. JAMA 2005, 293:1245–1256.PubMedCrossRef 2. Early Breast Cancer Trialists’ Collaborative Group (EBCTCG): Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet 2005, 365:1687–1717.CrossRef 3. Forbes JF, Cuzick J, Buzdar A, Howell A, Tobias JS, Baum M: Effect of anastrozole and tamoxifen as adjuvant treatment for early-stage breast cancer: 100-month analysis of the ATAC trial. Lancet Oncol 2008, 9:45–53.PubMedCrossRef 4. Coates AS, Keshaviah A, Thurlimann B, Mouridsen H, Mauriac L, Forbes JF, Paridaens R, Castiglione-Gertsch

M, Gelber RD, Colleoni M, Lang I, Del Mastro L, Smith I, Chirgwin J, Nogaret JM, Pienkowski T, Wardley A, Jakobsen EH, Price KN, Goldhirsch A: Five years of letrozole Etofibrate compared with tamoxifen as initial adjuvant therapy for postmenopausal women with endocrine-responsive early breast cancer: update of study BIG 1–98. J Clin Oncol 2007, 25:486–492.PubMedCrossRef 5. Di Cosimo S, Alimonti A, Ferretti G, Sperduti I, Carlini P, Papaldo P, Fabi A, Gelibter A, Ciccarese M, Giannarelli D, Mandalà M, Milella M, Ruggeri EM, Cognetti F: Incidence of chemotherapy-induced amenorrhea depending on the timing of treatment by menstrual cycle phase in women with early breast cancer. Ann Oncol 2004, 15:1065–1071.PubMedCrossRef 6.