It facilitates automatic application and scanning in situ. The composition of the mobile phase for the development selleck catalog of the chromatographic method was optimized by testing different solvent mixtures of varying polarity. Acetonitrile-0.2% acetate (6:4, 7:3, v/v) and methanol-0.2% acetate (8:2, 9:1, v/v) were tried. The best results were obtained using acetonitrile-methanol�C0.1% acetic acid (3.5:2.6:3.9, v/v). This mobile phase showed good resolution of losartan potassium peak from other formulation components or excipients tested. Densitometric scanning of all the tracks showed compound with Rf value 0.61 (single violet spot), identified as losartan potassium. The present method is quicker as the time needed for development of plate is reduced considerably to less than half an hour for chamber saturation.

The method was successfully used in the analysis of losartan potassium from the tablet dosage forms, and in case of Cozaar Tablets without interference of the formulation excipients. Figure 1 Typical chromatogram of losartan potassium and internal standard Recovery study Results showed high extraction efficiency of losartan potassium from formulation components. The recovery of losartan potassium ranged from 96.58 to 98.27%, average of 97.33%. This confirms that the proposed method can be used for determination of losartan potassium in tablet formulation. Precision and accuracy Five microliter aliquots of samples containing 5.0, 15.0, and 30.0 ng losartan potassium were analyzed according to the proposed method. In order to control the scanner parameters, one spot was analyzed several times.

By spotting and analyzing the same amount several times the precision of the automatic spotting device and the derivatization technique, was evaluated. The coefficient variation (% C.V.) for the analysis of eight replicates indicated good precision for the proposed TLC method (% C.V. consistently less than 5) and scanning eight spots in one run is the method of choice. Results obtaining were of good accuracy and high precision. The accuracy was found to be in the range of 88.76�C98.88% and % C.V. in range of 1.02-6.81. The results are presented in Tables Tables11�C3. Table 1 Precision and accuracy data of HPTLC method performed on losartan potassium Table 3 Precision data of the HPTLC assay for losartan potassium Table 2 Accuracy and precision of the assay Limit of detection and limit of quantification The limit of detection was 3.

0 ng/ml and the limit of quantification was 16.0 ng/ml and the solutions were stable for the 3 days. CONCLUSIONS The developed HPTLC technique is precise, specific, accurate, and sensitive. It proves that the method is repeatable and selective for the analysis of losartan potassium as bulk drug and in pharmaceutical formulations without any interference from the excipients. ACKNOWLEDGEMENT AV-951 Author wish to thank to J.S.S.Mahavidyapeetha, Mysore for providing this facility to carry out this work.

After the shotgun stage, reads were assembled with parallel phrap (High Performance Software, LLC). Possible mis-assemblies were corrected with gapResolution, Dupfinisher [49], or sequencing cloned bridging PCR fragments with subcloning or transposon bombing (Epicentre Biotechnologies, Madison, WI). Gaps between contigs were closed by editing in Consed, by selleck chemical Ruxolitinib PCR and by Bubble PCR primer walks (J.-F.Chang, unpublished). A total of 60 additional reactions were necessary to close gaps and to raise the quality of the finished sequence. Illumina reads were also used to correct potential base errors and increase consensus quality using a software Polisher developed at JGI [50]. The error rate of the completed genome sequence is less than one in 100,000. Together, the combination of the Illumina and 454 sequencing platforms provided 2,171.

8 �� coverage of the genome. The final assembly contained 384,925 pyrosequence and 63,008,730 Illumina reads. Genome annotation Genes were identified using Prodigal [51] as part of the Oak Ridge National Laboratory genome annotation pipeline, followed by a round of manual curation using the JGI GenePRIMP pipeline [52]. The predicted CDSs were translated and used to search the National Center for Biotechnology Information (NCBI) nonredundant database, UniProt, TIGR-Fam, Pfam, PRIAM, KEGG, COG, and InterPro databases. Additional gene prediction analysis and functional annotation was performed within the Integrated Microbial Genomes – Expert Review (IMG-ER) platform [53]. Genome properties The genome consists of a 2,272,954 bp long chromosome with a GC content of 35.

9% (Table 3 and Figure 3). Of the 2,181 genes predicted, 2,105 were protein-coding genes, and 76 RNAs; 56 pseudogenes were also identified. The majority of the protein-coding genes (65.5%) were assigned with a putative function while the remaining ones were annotated as hypothetical proteins. The distribution of genes into COGs functional categories is presented in Table 4. Table 3 Genome Statistics Figure 3 Graphical circular map of the chromosome. From outside to the center: Genes on forward strand (color by COG categories), Genes on reverse strand (color by COG categories), Cilengitide RNA genes (tRNAs green, rRNAs red, other RNAs black), GC content, GC skew. Table 4 Number of genes associated with the general COG functional categories Acknowledgements This work was performed under the auspices of the US Department of Energy Office of Science, Biological and Environmental Research Program, and by the University of California, Lawrence Berkeley National Laboratory under contract No. DE-AC02-05CH11231, Lawrence Livermore National Laboratory under Contract No. DE-AC52-07NA27344, and Los Alamos National Laboratory under contract No.

The reference standard, at three different concentrations http://www.selleckchem.com/products/Y-27632.html (50, 100, and 150%), was added to a fixed amount of the preanalyzed sample and the amounts of the drug were analyzed by the proposed method. Results from the recovery studies are given in Tables Tables55 and and66. Table 5 Results of the recovery study of tolperisone hydrochloride Table 6 Results of the recovery study of etodolac Solution stability The stability of TOLP and ETD standard and sample solutions was determined by storing the solutions at an ambient temperature (20 �� 10��C). The solutions were checked in triplicate after three successive days of storage and the data were compared with the freshly prepared samples. In each case, it could be noticed that the solutions were stable for 48 hours, as during this time the results did not decrease below 98%.

This showed that TOLP and ETD were stable in standard and sample solutions for at least two days, at ambient temperature. Robustness The robustness of the method was determined by making slight changes in the chromatographic conditions like flow rate (�� 0.1), temperature (�� 5), and pH (�� 0.2) of the mobile phase. It was observed that there were no marked changes in the chromatograms, which demonstrated that the RP-HPLC method developed was robust. RESULTS AND DISCUSSION The RP-HPLC procedure was optimized with a view to develop an accurate and stable assay method with the pure drugs TOLP and ETD, in a tablet formulation. A Phenomenax C18 (150 mm �� 4.6 mm, 5 ��) column in isocratic mode was used, with a mobile phase phosphate buffer (KH2PO4, pH 5.

5) : Methanol : Acetonitrile : Tri-ethylamine (40 : 40 : 20 : 1.5); pH of the buffer adjusted with orthophosphoric acid. The flow rate was 1 mL / minute and identical components were measured, with detection at 257 nm. Linearity was assessed by plotting concentration versus area, which is shown in Table 1, and it is linear in the range of 3.0 �C 21.0 ��g / ml for TOLP and 8.0 �C 56.0 ��g / ml for ETD, with correlation coefficients of 0.9998 and 0.9995, respectively, with a good linearity response, greater than 0.999. The % recovery was found to be within limits of the acceptance criteria with a recovery range of 99.42 �C 101.15% for TOLP and 99.28 �C 100.94% for ETD. The %RSD for intraday and Interday precision was less than 2% for TOLP and Carfilzomib ETD. The detection limit of the proposed method was 0.16 ��g / ml and 0.58 ��g / ml, and the quantification limit was 0.51 ��g / ml and 1.7 ��g / ml for TOLP and ETD, respectively. A typical overlain chromatogram of the standard solution is shown in Figure 3, chromatogram of the standard solution of TOLP and ETD at the test level is shown in Figure 4, and a chromatogram of the test solution is shown in Figure 5.

Although ECC was the fastest technique, it could not achieve full-thickness repair of the esophageal wall. Moreover, larger gaping defects could not be bridged by the jaws of the clips. In contrast, ECS anchors were deployed across the entire esophageal wall and showed well-healed scares with the smallest remaining gaps. One of the disadvantages inhibitor CHIR99021 of T-bars is that placing them beyond the gastrointestinal wall cannot be performed under direct vision. So, the needle tip may harm or inadvertedly place a T-bar into an unwanted structure as reported in a previous study [30]. The novel over-the-scope clip (OTSC) system showed promising results for gastrostomy closure [31] and has been used in for closure of postoperative leaks following gastrectomy and primary repair after spontaneous acute esophageal perforation [32].

Cardiac septal occluders might be a valuable alternative. Repici et al. have recently reported the first human case of esophagus-tracheal fistula closure by using a cardiac septal occluder with good results [33]. Other prototype suturing/apposition devices might be of future use in esophagotomy closure, namely, Padlock-G clips (Aponos Medical, Kingston, NH, USA) [34], NDO Plicator (NDO Surgical Inc., Mansfield, MA, USA) [35], g-Cath/g-Prox (Usgi Medical Inc, San Clemente, CA, USA) [36], flexible Endostich (Covidien, North Haven, Conneticut, USA) [37], OverStich (Apollo Endosurgery, Austin, TX, USA) [38], Direct Drive Endoscopic System (DDES Boston Scientific, Natick, MA, USA) [39], Anubis-scope (Karl Storz, Tuttlingen, Germany) [40],and Endo-Samurai (Olympus, Tokyo, Japan) [41].

Von Reitein et al. presented a prototype self-expanding metal stent (SX-ELLA stent, ELLA-CS, Hradec Kralove, Czech Republic) for direct incision esophagotomy closure without any suture [22]. Fifteen-millimeter direct incision esophagotomies were created in 12 domestic pigs using a prototype endoscopic Maryland dissector (Ethicon Endosurgery, Cincinnati, OH, USA). Six animals were randomly assigned to open surgical repair and six animals to endoscopic closure using the self-expanding, covered, nitinol stent in a nonsurvival setting. Pressurized leak test results were not different for stent compared to surgical closures. Six animals underwent transesophageal endoscopic mediastinal interventions and survived for 17 days. Stents were extracted at day 10.

All survival animals were found to have complete closure and adequate healing of the esophagotomies, without leakage or infectious complications. Finally, the hybrid approach presented Dacomitinib by Rolanda et al. might be useful for safe esophagotomy closure. Using a thoracoscope with a 5mm working channel, the authors inserted a needle-holder and performed an end-to-end esophageal anastomosis with gastroscopic intruments assistance [18]. 4.

6.2. Exclusion Criteria Universal exclusion criteria varied with pregnancy, ref 3 previous bariatric or gastric surgery, hiatal hernia, uncontrolled diabetes cardiovascular risks, a history of eating disorders, such as bulimia, medical therapy for weight loss within the previous 2 months, or any other condition that constitutes a significant risk of undergoing the procedure. A BMI > 50 was defined as an exclusion criterion for the Brethauer et al. and Skrekas et al. series. 6.3. Preoperative Preparation In most studies, patients underwent upper GI endoscopy, blood tests, and abdominal ultrasound preoperatively. Anticoagulants were given 12h preoperatively, and chemoprophylaxis with antibiotics was given with the induction of anesthesia [9]. Esophageal pH-metry was also performed in the Khazzaka and Sarkis study of the Obese-GERD group.

6.4. Surgical Technique Patient positioning on the operating table is standard in all cases, in an anti-Trendelenburg position at 30-degree French position (operator between legs) and two assistants on each side of the patient. Trocar placement is also standard in all cases. Closed pneumoperitoneum is achieved using a five-trocar port technique similar to that employed in laparoscopic Nissen fundoplication. Trocar placement is as follows: one 10mm trocar above and slightly to the right of the umbilicus for the 30�� laparoscope; one 10mm trocar in the upper left quadrant (ULQ) for passing the needle, for suturing, and for the surgeon’s right hand; one 5mm trocar also in the upper right quadrant (URQ) below the 10mm trocar at the axillary line for the surgeon’s assistant; one 5mm trocar below the xiphoid process for liver retraction; and one 5mm trocar in the upper left quadrant (ULQ) for the surgeon’s left hand [10].

Ramos et al. preferred dissection of the angle of His as the first step of the operation, whereas in the larger studies of Skrekas et al. and Andraos et al. it was the final step of the dissection of the greater curvature of the stomach. Mobilization of the greater curvature is performed using either a LigaSure Vessel Ligation System (Covidien) or a Harmonic scalpel (Ethicon Endo-Surgery, Inc., Cincinnati, Ohio) initially by opening the greater omentum at the transition between the gastric antrum and gastric body. Once access to the posterior wall is achieved, the greater curvature vessels are dissected distally up to the pylorus and proximally up to the angle of His.

Occasionally, posterior gastric adhesions are also dissected to allow optimal freedom for creating and sizing the invagination properly. The next step is the introduction of a bougie which was of a Brefeldin_A diameter of 36Fr in the Skrekas et al. study with 135 patients, and of 32Fr in the studies of Andraos et al. and Ramos et al. with a total of 166 patients.

.. Table 1 Classification and general features of L. arenae DSM 19593T according to the MIGS recommendations [16] published by the Genome Standards Consortium FTY720 order [17]. Morphology and physiology Cells of strain GA2-M15T are Gram-negative short rods (0.7-1.2 ��m in width and 1.2-2.4 ��m in length) and contain a polar flagellum for motility [1], [Figure 2]. Polyhydroxybutyrate is accumulated in the cells. Colonies are deep-brown, circular and contain clear margins. Cells are catalase and oxidase positive [1]. Growth occurs at 5-35 ��C with an optimum at 30 ��C. Cells were successfully grown on marine agar (MA), nutrient agar (NA, weak growth), salt tolerance agar (STA, containing 1% (w/v) tryptone, 0.3% (w/v) yeast extract and 1.5% (w/v) agar supplemented with salts) as well as on basal medium agar (BMA, recipe after [1]).

No growth was observed on Reasoner��s 2A agar (R2A), trypticase soy agar (TSA) or MacConkey agar. The salinity range for growth is 0.85�C8% NaCl (w/v), but the strain does not grow below 0.34% or at above 10% NaCl (w/v). The pH range for growth is pH 6�C9 with an optimum at pH 7 [1]. Figure 2 Micrograph of L. arenae DSM 19593T. Cells hydrolyze aesculin and tyrosine weakly, but do not show any hydrolysis of alginic acid, casein, chitin, CM-cellulose, DNA, gelatin, pectin, starch or urea [1]. They assimilate citrate, D-fructose, D-galactose, D-glucose, L-glutamate, glycerol, ��-hydroxybutyrate, D-mannitol, D-mannose, melibiose, propionate, pyruvate, L-serine, L-tyrosine and D-xylose, but not L-alanine, L-arabinose, L-aspartate, cellobiose, glycine, L-histidine, lactose, L-leucine, maltose, L-rhamnose, D-ribose, sucrose, L-threonine or trehalose.

Cells are positive for ��-galactosidase, esterase (C4), esterase lipase (C8), leucine arylamidase, naphthol-AS-BI-phosphohydrolase, ��-glucosidase and ��-glucosidase, but negative for indole production, arginine dihydrolase, alkaline phosphatase, lipase (C14), valine arylamidase, cystine arylamidase, trypsin, ��-chymotrypsin, acid phosphatase, ��-galactosidase, ��-glucuronidase, N-acetyl-��-glucosaminidase, ��-mannosidase and ��-fucosidase [1]. The substrate utilization and resistance patterns of L. arenae DSM 19593T were also determined for this study using Generation-III microplates in an OmniLog phenotyping device (BIOLOG Inc., Hayward, CA, USA).

The microplates were inoculated at 28��C with a cell suspension at a cell density of 95-96% turbidity and dye IF-A. Further additives were vitamin, micronutrient and sea salt solutions. Brefeldin_A The exported measurement data were further analyzed with the opm package for R [26,27], using its functionality for statistically estimating parameters from the respiration curves such as the maximum height, and automatically translating these values into negative, ambiguous, and positive reactions.

Additionally, higher curing light intensities may lead to superior physical and mechanical properties.8 Light-emitting diode (LED) curing lights are used to polymerize necessary resin-based restorative materials, and their effect on the physical properties of resin-based restorative materials has been investigated.9 Because contemporary LED lights yield superior irradiance (approximately 1,000 mW/cm2) to QTH light (approximately 800 mW/cm2), LED lights are expected to polymerize resin materials as or even more effective than QTH.9 Post-cure heating of resin composite materials is a extremely popular restorative technique. This method subjects a light-cured composite inlay to an immediate heat treatment for the purpose of increasing material cure and thus enhancing clinical properties.

10 In Rueggeberg��s study, the degree of color change was influenced more by the amount of resin content in the composite systems than by the particular resin composition. Microfill composite resins showed greater potential for color change than did the other types of materials. If clinicians are considering using conventional light-cured composite materials for inlays, the choice of material as well as the post-cure temperature will influence the ability to match the inlay with the initial shade of composite chosen.11 An important factor is the possibility of using post-polymerization mechanisms associated with heat, pressure, or high light intensity in indirect composite systems.12 With respect to materials, indirect and direct composites have similar compositions.

13 Currently, easy and inexpensive techniques have been enhanced for direct composites in indirect restorations. Coupled with the use of devices that are always found in dentists�� offices to apply these techniques, better performance can now be obtained and harnessed from material properties, resulting in improved dental health.14 Presently, electronic shade-matching instruments have been used for dental practice.15 In order to determine objectively the color changes on composite restorations, some methods have been experienced, among them the spectrophotometry, which makes the study of several parameters related to color stability of composite resins possible. In this method reflected wavelength by a body is changed in values expressed in ��E* units.

The ��E* values can be used so that present the color changes provided by the composite resin after treatment or period of time.16 Instrumental color analysis offers a potential advantage over visual Brefeldin_A color determination, as instrumental readings are objective, quantifiable, and more rapidly obtainable.17 The Comission Internationale de l��Eclairage Lab (CIE L*a*b) color system has been used for determination of color difference.18�C21 Chroma is calculated as C*ab= (a*2+ b*2) ?.

After exclusion of 23 cases from Group 2, 222 cases underwent molecular investigations for MSI, KRAS and BRAF. The kinase inhibitor MEK162 aim of this study group was to determine the prognostic value of TOPK in CRCs, with subgroup analysis by KRAS and BRAF mutation. Multivariable cancer-specific survival time models were evaluated by including candidate variables such as age, sex, pT and pN classification, vascular invasion and MSI status. A total of 23 cases were excluded from Group 3 (Figure 1B). The remaining 71 Lynch syndrome-associated CRCs underwent molecular analysis for KRAS and BRAF. The association of TOPK expression with mutational status of KRAS and BRAF, clinicopathological features and cancer-specific survival time, was assessed.

One case of metastatic CRC was excluded from Group 4 because of insufficient material for adequate assessment of TOPK expression (Figure 1C). Immunohistochemistry for PTEN and molecular investigations of MSI, KRAS and BRAF were previously performed (Frattini et al, 2007). The prognostic and predictive value of TOPK in this group of patients was analysed, with specific end points of interest being cancer-specific survival time and objective tumour response to anti-EGFR agents. The use of all patient material was approved by local Ethics Committees. Statistical analysis methods Associations of TOPK with categorical features were investigated by Chi-Square and Fisher’s Exact tests where appropriate, and by Student’s t-test for age. Survival analysis was performed using the Kaplan�CMeier method, log-rank test and by multiple Cox regression analysis after verification of the proportional hazards assumption.

The appropriate number of variables to be included in regression models was dependent on the frequency of patient deaths in each analysis. We included 1 variable per 10 deaths, to prevent overfitting. Differences in TOPK expression between normal colonic mucosa and tumour were determined using Wilcoxon’s rank-sum test for medians. The most clinically relevant cutoff score for TOPK was determined on subgroup A by receiver operating characteristic (ROC) curve analysis for end point survival/death. To prevent overfitting, re-sampling of data was performed by bootstrapping 200 times. The inter-observer variability of TOPK staining was assessed using the intra-class correlation coefficient (ICC), with values of 0.8 indicating excellent agreement.

Missing clinicopathological data were assumed to be at random. No imputation was performed; rather, only patients with complete Dacomitinib data for all features were included in multivariable analyses. P-values <0.05 were considered to be statistically significant. Results TOPK expression in normal colon versus sporadic CRC T-cell-originated protein kinase expression in 57 normal colonic mucosa samples was compared with sporadic CRCs from Group 1 (n=1044).

Enforced expression of miR-205 was shown to inhibit cell invasion and suppress lung metastasis of breast cancer cells in nude mice, possibly through targeting ErbB3 [35]. MiR-205 also exerts inhibitory effects on cellular invasiveness and migration http://www.selleckchem.com/products/Pazopanib-Hydrochloride.html in prostate cancer and glioblastoma cells, through down-regulation of the protein kinase C�� and low-density lipoprotein receptor-related protein 1, respectively [36,37]. Using miR target prediction algorithms, ErB3, E2F1, E2F5, ZEB1, ZEB2, and protein kinase C�� have been indentified as putative miR-205 targets [36]. In the present study, knockdown of miR-205 expression substantially enhanced cellular expression of ZEB2 in ESCC cells. In fact, previous and present studies employing a reporter gene assay confirmed miR-205 binding to the ZEB2 3′-UTR [15,17].

Although the ESCC cells examined in this study did not express ZEB1 sufficiently, direct interaction of miR-205 with ZEB1 3′-UTR was shown in other cell types but not in ESCC cells examined in this study [15,17]. ZEB1 and ZEB2 are related homeodomain-containing transcriptional factors that repress E-cadherin transcription [17,38,39]. E-cadherin is a central component of the adherens junction complex responsible for cell-cell adhesion and maintenance of cytoskeleton organization [15]. It is known that loss of E-cadherin expression is a key event in the EMT, which can be recapitulated during tumor progression, constituting an early step in tumor metastasis including ESCC [15,40-43].

In line with this, cellular E-cadherin expression was substantially reduced, whereas N-cadherin expression emerged in ESCC cells transfected with anti-miR-205 inhibitor to suppress ZEB2, and they were endowed with properties allowing augmented invasion through EMT. Gregory et al described that the miRNA-200 family (miR-200a, miR-200b, miR-200c, miR-141, and miR-429), as well as miR-205, was markedly downregulated in breast and colon cancer cells that had undergone EMT [15]. Collectively, miR-205, along with members of the miR-200 family, can be a key regulator of EMT to widely enforce the indolent epithelial-like phenotype, not limited to ESCC. In clinical settings, lower levels of miR-205 were significantly associated with loco-regional recurrence and poor survival of patients with head and neck squamous cell carcinoma [41].

Further studies are warranted to assess whether miR-205 expression levels could be a predictive biomarker for clinical outcomes in ESCC. Conclusions MiR-205 expression was specifically increased in ESCC cells. Brefeldin_A MiR-205 is likely to control cell invasion and migration in ESCC cells through its repression of ZEB2, a repressor of E-cadherin. These findings establish the tumor-suppressive role of miR-205, which may serve as a unique therapeutic target for ESCC. Competing interests The authors declare that they have no competing interests.

The levels were assayed by Network Pathology (Melbourne, Australia), using the Chromogranin A ELISA kit (DakoCytomation, Sydney, Australia). Statistical methods The sample size of 63 was Pacritinib designed to give >80% power to detect an 8-month difference in median survival comparing the 40% patients expected to have a positive octreoscan vs the remainder expected to have a negative octreoscan (estimated survivals 12 and 4 months, respectively, as per the treatment and control arms in the Kouroumalis study). Survival times were calculated from the day of registration. Survival curves were calculated using the method of Kaplan�CMeier. The log-rank test was use to compare the relationship between receptor expression by scintigraphy and survival duration. Proportions were compared using ��2 tests.

Quality-of-life results were described and analysed with simple descriptive and comparative methods. For each item in the Patient Benefit Form, we recorded the proportions of patients reporting that each aspect was much better, a little better, the same, a little worse, or much worse than before starting treatment, and used ��2 tests to compare the proportions of patients who reported feeling better vs those who reported feeling worse. For the Patient DATA Form and FACT-Hep, we used Wilcoxon’s rank-sum tests for paired data to compare baseline and 1-month scores for each domain. Responses from the Patient Benefit Form were compared with change scores from the Patient DATA Form using Spearman’s rank-correlation coefficient. RESULTS The study accrued 63 patients with otherwise untreatable HCC between April 2001 and January 2002.

Study sites are listed in Appendix A. The patients’ baseline characteristics are shown in Table 1. The median age was 67 years (range 28�C81 years) and most of them were male. Most had good performance status. Eleven had Child�CPugh class B cirrhosis and 52 had well-compensated disease (Child�CPugh class A). Viral hepatitis was the main cause of cirrhosis, with 28 patients (44%) having evidence of hepatitis B or C infection. A small number of patients had cirrhosis from other causes. Approximately half had received other treatment for HCC. Table 1 Patient characteristics Administration of treatment Four patients (6%) did not receive any octreotide because their disease progressed so rapidly they were unable to start treatment.

These patients were not eligible for analysis Dacomitinib of response or toxicity, but are included in the survival analysis. Three patients (5%) received one dose, 12 (19%) two doses, 10 (16%) three doses, 10 (16%) four doses, seven (11%) five doses, four (6%) six doses, five (8%) seven doses, and seven (18%) eight doses or more. The main reason for stopping treatment was disease progression in 19 patients (30%). A further five patients (8%) withdrew and nine (14%) died during treatment, all from progressive disease.