Twin characterisation using 2D and 3D EBSD

IntroductionTwin characterisation plays an important role inunderstanding the deformation and annealing behaviourof many metals,particularly those with hcp crystalstructures[1~8].Often,the aim is to determine the vol-ume fraction of twins within a material,perhaps as afunction of strain.However,many materials twin onseveral differentsystems and formsecondary or tertiary,as well as primary,twins.Furthermore,the variants ofa particular twin type may be activated to differing ex-tents.Analysis o…     

α-Emitting cancer therapy using 211At-AAMT targeting LAT1

α-Methyl-l -tyrosine (AMT) has a high affinity for the cancer-specific l -type amino acid transporter 1 (LAT1). Therefore, we established an anti-cancer therapy, with ²¹¹At-labeled α-methyl-l -tyrosine (²¹¹At-AAMT) as a carrier of ²¹¹At into tumors. ²¹¹At-AAMT had high affinity for LAT1, inhibited tumor cell growth, and induced DNA double-stranded breaks in vitro. We evaluated the accumulation of ²¹¹At-AAMT in vivo and the role of LAT1. Treatment with 0.4 MBq/mouse ²¹¹At-AAMT inhibited tumor growth in the PANC-1 tumor model and 1 MBq/mouse ²¹¹At-AAMT inhibited metastasis in the lung of the B16F10 metastasis model. Our results suggested that ²¹¹At would be useful for anti-cancer therapy and that LAT1 is suitable as a target for radionuclide therapy.     

Direct to implant breast reconstruction by using SERI?, preliminary report

Background

There has been a “rising tide” in mastectomy utilization that can be attributed to more skin-sparing mastectomies (SSMs) performed concurrently with immediate breast reconstruction. We report our experience of the first use of SERI® Surgical Scaffold (SERI®; Allergan, Inc.) in 21 cases of direct to implant (DTI) breast reconstruction after SSM.

Methods

Our retrospective experience, from April 2013 to May 2014, is based on 21 cases of direct to implant (DTI) breast reconstruction after SSM (9 monolateral 6 bilateral). All the patients were oncological with a preoperative cancer stage was into 0–2 stage. In order to assess the level of satisfaction with the aesthetical result, on 4–13 months post-operative patients were asked to complete a questionnaire that evaluated various parameters by means of a Visual Analogue Scale (V.A.S.).

Results

Over a 13-months period, a total of 15 patients underwent 21 immediate breast reconstructive procedures with Allergan Natrelle 410 style implants plus SERI® after SSMs. Definitive histological examination give evidence of 5 patients intraductal carcinoma, 6 patients multifocal carcinoma and 4 patients carcinoma in situ. 6 bilateral cases of direct to implant (DTI) breast reconstruction after SSM had a monolateral oncological treatment and on the other side a prophylactic treatment. At the end of the short follow up (minimum 6 months) all the patient were cancer free with an excellent outcome. Complication rate presents just one implant exposure followed by a revised surgery. At V.A.S. the mean patient satisfaction was 5,77 (good), 4,09 (fair) for sensitivity of the nipple areola complex, 6,33 (good) assessment of implant position, 6,28 (good) self esteem, 5,2 (good) attraction ability, 4,99 (fair) intimate life, 6,81 (good) overall feelings about breast reconstruction, 6,71 (good) simmetry.

Conclusions

The really encouraging results of our early experience will help surgeons introducing SERI® into their practice to select appropriate patients for direct-to-implant single-stage immediate breast reconstruction. A larger study cohort and longer follow-up times are required to identify additional predictors and indications.     

First French experiences of total body irradiations using helical TomoTherapy®

Purpose

Dynamic conformal radiotherapy with helical TomoTherapy^® (HT) offers a more quantitative paradigm for total body irradiation. Treatment planning, delivery, dose verification of the first French experiences of total body irradiation using helical TomoTherapy^® are presented.

3D computer vision using Point Grey Research stereo vision cameras

This paper provides an introduction to stereo vision systems designed by point grey research and describes the possible application of these types of systems. The paper presents an overview of stereo vision techniques and outlines the critical aspects of putting together a system that can perform in the real world. It also provides an overview of how the cameras can be used to facilitate stereo research.     

What can we expect from dose escalation using proton beams?

It has been demonstrated without doubt in the literature, including elsewhere in this issue, that much better conformal dose distributions in radiation therapy can be obtained with proton beams than with photons (X-rays) or electrons. It is also clear that this remains entirely true--for the fundamental reason of particle range--even after the latest and projected developments in computer-generated IMRT (intensity-modulated radiation therapy) photon dose escalation are fully considered. We consider several examples of tumour dose-response curves that illustrate the quite large gains to be obtained when dose escalation can be achieved, if normal tissue complications can also be avoided. Two contrasting types of tumour are considered in detail, prostate tumours and non-small-cell lung carcinomas. There is a considerable way to go yet to achieve really high non-recurrence rates, especially in the lung tumours. Proton beams would make this progress much safer and more effective than any variants with photons.     

Clinical analysis of intracranial germinoma＇s craniospinal irradiation using helical tomotherapy

Objective： To evaluate the short-term clinical outcomes of intracranial germinoma patients treated with craniospinal irradiation （CSI） using helical tomotherapy （HT） system in our center. Methods： Twenty-three patients who were treated with CSI in our center from January 2008 to July 2012 were collected, with an average age of 20. M1 of the patients＇ CSI used the HT system. The total doses were 27-36 Gy/15-20 F （1.5-2 Gy per fraction）, and total local doses were 46-60 Gy/30-50 F （5 fractions per week）. M1 female patients for CSI were treated with left-right parallel-opposed field irradiation to protect their ovarian functions. Median follow-up time was 30.9 months （range, 5-67 months）. The SPSS19.0 software was used, and the overall survival （OS） was calculated using the Kaplan-Meier method. Results： Among 17 patients with assessable tumors, 9 cases （52.9%） were CR, 7 cases （41.2%） were PR, and 1 case （5.9%） was SD. Hematological toxicity was the severest side-effect occurred in the procedure of CSI. The level 1-4 acute leukopenia were 8.7%, 30.4%, 34.8% and 21.7% and the level 1-4 acute thrombopenia were 8.7%, 30.4%, 21.7% and 8.7%, respectively. Conclusions： For primary intracranial germinomas, HT can be used to implement CSI for simplifying radiotherapy procedures, improving radiotherapy accuracy, enhancing protection of peripheral organs at risk （ORA） and guaranteeing therapeutic effects. With the acceptable acute and long-term toxicity, CSI using HT in intracranial germinoma patients can be a safe and alternative mode.     

In vitro culture of human dendritic cells by using a HydroCell™

Cancer Immunotherapy using dendritic cells would be a feasible and useful tool for cancer treatment. However, no immunotherapy has been approved in Japan because of a lack of any randomized clinical studies. We are now trying to develop an automatic dendritic cell culture system in order to perform a large-scale randomized clinical trial. In this study, we investigated the utility of a HydroCell? for in vitro culture of human dendritic cells induced from peripheral blood monocytes. The dendritic cells grew one and a half times when they were cultured in a HydroCell?. All the cells were floating and harvested easily without any enzymes. The cells expressed the CD80 and CD83 molecules on their surface and still had strong phagocytosis. This results demonstrated that a HydroCell? was a useful tool for in vitro culture of dendritic cells.     

Characterization of Lin−ALDHbright population using Ehrlich ascites tumor cells in mice

Cancer stem cells (CSCs)/tumor initiating cells have been shown to exist in recent studies; however, it is challenging to isolate these cells. The latest evidence suggests that elevated aldehyde dehydrogenase (ALDH) activity is a hallmark of CSCs. In this study, mice implanted with Ehrlich ascites tumor (EAT) cells were used to isolate cancer stem cells. Femoral bone marrow aspirations were performed 15 days after the injection of EAT cells and Lin^?ALDH^bright and Lin^?ALDH^low cell populations were isolated. Lin^?ALDH^bright cells isolated from EAT-bearing mice accounted for 11.08?±?10.52 % of all the Lin^? cell population. Analysis of hematopoietic stem cell markers showed that Sca-1, c-kit, and CD38 were expressed higher in the Lin^?ALDH^bright population compared with Lin^?ALDH^low. The Lin^?ALDH^bright population expressed P-glycoprotein, a product of the multidrug resistance (MDR) gene. P-gp activity measured by rhodamine 123 (Rh123) and blocked by verapamil. Among the cells treated with doxorubicin for 48 h, the Lin^?ALDH^bright cell groups were more resistant and had higher overexpression of Bcl-2 protein than Lin^?ALDH^low.     

Cancer patients’ intentions towards receiving unsolicited genetic information obtained using next-generation sequencing

Next-generation sequencing (NGS) can be used to generate information about a patient’s tumour and personal genome. This powerful diagnostic tool provides solicited and unsolicited hereditary genetic (risk) information that could have consequences for cancer patients and their quality of life. A well-defined approach for returning appropriate genetic risk information is needed in personalized cancer care. A qualitative design with semi-structured interviews was used. We conducted interviews with 24 Dutch patients with different types of cancer, both NGS-experienced and NGS-inexperienced, to learn their intentions, needs and preferences towards receiving unsolicited genetic information obtained using NGS. Almost all participants had a positive attitude towards receiving unsolicited findings. After receiving comprehensive background information on NGS, including a binning model of four categories of unsolicited findings, most participants preferred to receive only subsets of genetic information. Their main concern was their own and others’ (including family members) ability to cope with (the increased risk of having) a genetic disorder. Providing background information gave cancer patients the opportunity to select subsets of findings and increased their ability to make an informed choice. Special attention is needed for social and emotional factors to support the patients themselves and when communicating test results with their family members.     

Development of a high risk pancreatic screening clinic using 3.0 T MRI

Selective screening for pancreatic cancer (PC) has been proposed. We describe the establishment of a comprehensive multidisciplinary screening program using 3.0 T MRI. Criteria for screening included the presence of PC in: ≥ 2 first degree relatives (FDR), 1 FDR and 1 s degree relative (SDR), ≥ 3 any degree relatives (ADR), or any known hereditary cancer syndrome with increased PC risk. Imaging with 3.0 T MRI was performed routinely and endoscopic ultrasound was used selectively. Screening was completed in 75 patients (pts). Hereditary cancer syndromes were present in 42 (56%) of the 75 pts: BRCA2 (18), ATM (8), BRCA1 (6), CDKN2A (4), PALB2 (3), Lynch (2), and Peutz-Jeghers (1). A family history of PC was present in ≥?2 FDR in 12 (16%) pts, 1 FDR and 1 SDR in 5 (7) pts, and ≥?3 ADR in 16 (21%) pts. Of the 65 pts who received screening MRI, 28 (43%) pts had pancreatic cystic lesions identified, including 1 (1%) patient in whom a cholangiocarcinoma was diagnosed as well. No patient underwent surgical resection. Using a 3.0 T MRI to screen patients at high risk for developing PC identified radiographic abnormalities in 43% of patients, which were stable on subsequent surveillance. Specific guidelines for the frequency of surveillance and indications for surgery remain areas of active investigation as the global experience with high risk screening continues to mature.     

Is there a future for cell kinetic measurements using IdUrd or BdUrd?

PURPOSE: The analysis of causes of radiation failure both in retrospective series of patients with head and neck cancer and in several randomized clinical trials suggests a loss of local control as the overall treatment time increases for the same total dose. This is attributed to tumor cell proliferation during fractionated radiotherapy. As longer treatment times lead to loss of local control, it has been suggested that shorter treatment times could lead to an increase in local control. For this reason accelerated treatment regimens have been and are being designed. However, these treatments may cause severe acute reactions. Due to this, lower total doses are sometimes given. Slowly proliferating tumors, therefore, may do worse when treated with accelerated schedules compared with conventional schedules. In addition, it is not desirable to subject all patients to the more intense acute reactions of accelerated schedules. It would thus be useful to predict which tumors will rapidly proliferate during treatment and are likely to benefit from accelerated radiotherapy. The potential doubling time (Tpot) is defined as the time within which the cell population of a tumor would double if there were no cell loss. The hypothesis is that the median Tpot measured before treatment might correlate with the effective doubling time (Tp) during treatment. CONCLUSION: Tpot can be calculated knowing the labeling index (LI; proportion of cells incorporating the DNA precursor IdUrd or BdUrd) and Ts (the DNA synthesis time) measured by flow cytometry. A recent multicenter analysis has shown that the only pretreatment kinetic parameter for which some evidence is found for an association with local control is LI, not Tpot. Pitfalls associated with cell kinetic measurements such as assay variability, intratumor and intertumor variability, interlaboratory variability and the problem of an admixture of normal and malignant cells make Tpot not accurate and reproducible enough for a robust predictive assay. It therefore appears that pretreatment Tpot measurements using flow cytometry, provide only a relatively weak predictor of outcome after radiotherapy in head and neck cancer. Immunohistochemistry allows a simple measure of LI and may give additional independent information from labeling patterns, suggesting that this method is the (short term) future for clinical cell kinetic measurements using BdUrd or IdUrd.     

Analysis of T-cell receptor-γ gene rearrangements using heteroduplex analysis by high-resolution microcapillary electrophoresis

Determination of T-cell clonality has an important additional value for diagnosis of T-cell lymphomas. Various molecular methods have been developed, including polymerase chain reaction (PCR) of T-cell receptor γ (TCRγ). The detection of PCR products usually relies commonly on either GeneScan (GS) analysis or heteroduplex (HD) analysis by polyacrylamide gel electrophoresis (PAGE). These techniques have their disadvantages, being relatively time-consuming and laborious or requiring expensive equipment. Here, we propose an alternative method that combines multiplex PCR and HD analysis by microcapillary electrophoresis (ME) on the Agilent 2100 Bioanalyzer. The sensitivity of the method was determined with clonal PEER T-cell line DNA dilution in polyclonal DNA and was evaluated as 1-5%. Fifty-three samples from patients with T-cell lymphoproliferative disorders were analyzed by HD analysis using ME and GS analyses. Comparison of the two techniques showed them to be highly concordant (93% similarity). The rate of clonality detection by GS analysis was higher than HD analysis by ME, but none of the discordant patients (n=5) has yet developed lymphoma. HD analysis by ME to reveal TCRγ gene rearrangements in clinical specimens was consistent with clinical data and the outcome of patients. Detection of T-cell clonality by HD analysis with ME is sensitive, practical, safe and represents a potential alternative to PAGE and GS analysis.     

A highly sensitive enzyme-linked immunosorbent assay for etoposide using β-d-galactosidase as a label

Summary A highly sensitive enzyme-linked immunosorbent assay (ELISA) for etoposide (EP) was developed, which is capable of accurately measuring as little as 40 pg EP/ml. Anti-EP sera were obtained by immunizing rabbits with EP conjugated with mercaptosuccinyl bovine serum albumin (MS.BSA) usingN-[-(4-diazophenyl)ethyl]maleimide (DPEM) as a heterobifuntional coupling agent. An enzyme marker was similarly prepared by coupling EP with -d-galactosidase (-Gal; EC 3.2.1.23) via DPEM. This ELISA was specific for EP and showed a very slight cross-reactivity with its major metabolite,cis-hydroxy acid of EP (0.91%), but none with 4-demethylepipodophyllotoxin and drugs commonly used with EP in combination chemotherapy for cancer treatment. The values for EP concentration detected by this assay were comparable with those detected by the highperformance liquid chromatography (HPLC) method. However, the ELISA was about 1,250 times more sensitive in detecting EP at lower concentrations. Using this assay, drug levels were easily determined in the blood and urine of rats for 7 h after i.v. administration of EP at a single dose of 3 mg/kg. Due to its sensitivity and specificity for EP, the ELISA should prove to be a valuable new tool for use in clinical pharmacological studies.     

Noninvasive diagnosis of urothelial cancer in urine using DNA hypermethylation signatures—Gender matters

Urothelial cancer (UCa) is the most predominant cancer of the urinary tract and noninvasive diagnosis using hypermethylation signatures in urinary cells is promising. Here, we assess gender differences in a newly identified set of methylation biomarkers. UCa-associated hypermethylated sites were identified in urine of a male screening cohort (n = 24) applying Infinium-450K-methylation arrays and verified in two separate mixed-gender study groups (n = 617 in total) using mass spectrometry as an independent technique. Additionally, tissue samples (n = 56) of mixed-gender UCa and urological controls (UCt) were analyzed. The hypermethylation signature of UCa in urine was specific and sensitive across all stages and grades of UCa and independent on hematuria. Individual CpG sensitivities reached up to 81.3% at 95% specificity. Albeit similar methylation differences in tissue of both genders, differences were less pronounced in urine from women, most likely due to the frequent presence of squamous epithelial cells and leukocytes. Increased repression of methylation levels was observed at leukocyte counts ≥500/μl urine which was apparent in 30% of female and 7% of male UCa cases, further confirming the significance of the relative amounts of cancerous and noncancerous cells in urine. Our study shows that gender difference is a most relevant issue when evaluating the performance of urinary biomarkers in cancer diagnostics. In case of UCa, the clinical benefits of methylation signatures to male patients may outweigh those in females due to the general composition of women's urine. Accordingly, these markers offer a diagnostic option specifically in males to decrease the number of invasive cystoscopies.     

Hepatocellular carcinoma: is there a potential for chemoprevention using cyclooxygenase-2 inhibitors?

Inhibitors of cyclooxygenase-2 (COX-2) have proapoptotic and antiangiogenic effects on malignant tumors and inhibit their invasion to surrounding tissues. These properties are derived from COX-dependent and/or COX-independent signaling via peroxisome proliferator-activated receptor gamma. Although the role of COX-2 involvement in human hepatocarcinogenesis has not been determined yet, selective COX-2 inhibitors with COX-independent properties may potentially suppress hepatocarcinogenesis. This hypothesis should be confirmed in in vivo studies using animal models. These studies may provide insights into any application of the COX-2 inhibitor for primary and/or secondary chemoprevention.