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Elevation of naphthoquinones and estrogen quinones, which are reactive metabolites of naphthalene and estrogen, is thought to be an important indicator of naphthalene- and estrogen-induced carcinogenesis. We compared background levels of naphthalene and estrogen quinone-derived adducts in serum albumin (Alb) from 143 women with breast cancer and 119 healthy controls. Cysteinyl adducts of naphthoquinones, including 1,2-naphthoquinone (1,2-NPQ) and 1,4-naphthoquinone (1,4-NPQ), and estrogen quinones, including estrogen-2,3-quinones (E2-2,3-Q) and estrogen-3,4-quinones (E2-3,4-Q), were characterized after adduct cleavage. Levels of estrogen quinones and naphthoquinones were positively correlated in healthy controls, but not in breast cancer patients (p 2-3,4-Q were elevated by two- to ten-fold in cancer patients (p 2-3,4-Q-2-S-Alb to (E2-2,3-Q-4-S-Alb plus E2-3,4-Q-2-S-Alb) in patients and controls. These two groups were separable using albumin adducts of estrogen quinones and naphthoquinones, with 99.6% overall correct classification rate (overall accuracy). The findings of this study suggest that differences in the disposition of estrogen and naphthalene, and the subsequent elevation of cumulative E2-3,4-Q and 1,2-NPQ may serve as biomarkers of breast cancer risk.

Background Vitiligo is an acquired disease that involves low pigment variation in the skin. The use of cultured melanocytes for treatment of recalcitrant vitiligo has become a well-established treatment modality. In vitro cultured melanocytes present an effective autologous transplantation treatment modality for vitiligo. The present study investigated the in vitro culture of epidermal melanocytes sampled from multiple body parts, as well as the differences in total propagation time. Methods Skin specimens were collected from 30 participants (14 males and 16 females) who supplied normal colored skin sections from various regions of their body, including the face, chest, abdomen, buttock, and extremities. Subsequently, all of these specimens were treated with an identical melanocyte purification and culturing process. Results Melanocytes from the face demonstrated the most rapid growth and longest total propagation time. Melanocytes from the buttock, abdomen, and extremities displayed similar results to one another, and melanocytes from the chest and back had the slowest growth and shortest total propagation time. Conclusion Selecting the most favorable site to obtain epidermal melanocytes will reduce the required quantity of skin and culturing time, and maximize the growth and total propagation time of melanocytes. Therefore, care should be exercised when selecting the region of skin when culturing epidermal melanocytes.

Background The aim of the current study was to report a single-institution experience using breast-conserving surgery after neoadjuvant chemotherapy (NACT), focusing on the association between microscopic resection margin status and locoregional recurrence (LRR). Methods Our institutional prospectively maintained database was reviewed to identify patients who were treated with NACT between January 2008 and April 2018. Results Among the main partial mastectomy specimens available for analysis (n = 161), 28 had margins 2 mm. LRR occurred in 16 patients (9.9%) and distant metastases were detected in 27 (16.8%) patients. There was no significant difference in the LRR between the > 2 mm margin group with a 60-month cumulative survival of 85.2% compared with 76.2% for the ≤2 mm group (P = 0.335) in the Kaplan-Meier analysis. When we stratified patients by margin widths of ≥1 mm or P = 0.161) with a 60-month cumulative LRR-free survival of 84.9% compared with 69.5% for the P = 0.150). Conclusions In the absence of multiple scattered microscopic tumour foci, a negative margin of no ink on tumour maybe sufficient for stage I–III invasive breast cancer treated with NACT and breast-conserving surgery.

Identification of minimal residual disease (MRD) is important in assessing the prognosis of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). The current best clinical practice relies heavily on Flow Cytometry (FC) examination. However, the current FC diagnostic examination requires trained physicians to perform lengthy manual interpretation on high-dimensional FC data measurements of each specimen. The difficulty in handling idiosyncrasy between interpreters along with the time-consuming diagnostic process has become one of the major bottlenecks in advancing the treatment of hematological diseases. In this work, we develop an automatic MRD classifications (AML, MDS, normal) algorithm based on learning a deep phenotype representation from a large cohort of retrospective clinical data with over 2000 real patients’ FC samples. We propose to learn a cytometric deep embedding through cell-level autoencoder combined with specimen-level latent Fisher-scoring vectorization. Our method achieves an average AUC of 0.943 across four different hematological malignancies classification tasks, and our analysis further reveals that with only half of the FC markers would be sufficient in obtaining these high recognition accuracies.

The prognosis management is crucial for highrisk disease like Acute Myeloid Leukemia (AML) in order to support decisions of clinical treatment. However, the challenges of accurate and consistent forecasting lie in the high variability of the disease outcomes and the complexity of the multiple clinical measurements available over the course of the treatment. In order to capture the multi-dimensional and longitudinal aspect of these comprehensive clinical parameters, we utilize an attention-based bi-directional long shortterm memory (Att-BLSTM) network to predict AML patient's survival and relapse. Specifically, we gather a 10-year worth of real patient's clinical data including blood test, medication, HSCT status, and gene mutation information. Our proposed Att-BLSTM framework achieves 77.1% and 67.3% AUC in tasks of predicting the next 2-year mortality and disease relapse with these comprehensive clinical parameters, and our further analysis demonstrates that a next 0 to 3 months prediction performs equally well, i.e., 74.8% and 67% AUC for mortality and relapse respectively.

Background and purposeEngorged medullary vein (EMV) in patients with intracranial dural arteriovenous fistula (DAVF) suggests venous congestion. The aim of this study is to investigate its prevalence, pattern, and correlation with clinical findings.Materials and methodsCT angiography (CTA) raw data of DAVF were used for multiplanar reconstruction and then analyzed for the presence and pattern of EMV, which is defined as a dilated vein in the cerebral white matter. Patients with EMV were divided into two groups: regional and extensive. Regional type is defined as EMV limited to one cerebral hemisphere or cerebellum without evidence of subcortical calcification. Extensive type is defined as EMV involvement of more than one cerebral hemisphere or both the cerebrum and cerebellum. Descriptive analysis of clinical information, DAVF characteristics, and other imaging findings was conducted. Clinical information, including demographic data, clinical presentation, and hemorrhage, were correlated with both types of EMV.ResultsAmong 192 eligible patients with DAVF, 71 (37%) had EMV. Patients with EMV were older (63 years vs 56 years, P=0.02), with DAVF more often at the transverse and sigmoid sinus (PConclusionsEMV in patients with DAVF is associated with an aggressive manifestation. Regional type EMV is associated with a higher risk of a hemorrhagic presentation.

Background Proper patient selection criteria for treatment of carotid chronic total occlusion (CTO) are unclear. This study was designed to predict procedure successful rate and 1-year patency after carotid artery stenting (CAS) for carotid CTO using pre-procedural CTA. Methods Patients with CTO detected on CTA who underwent recanalization within 3months were divided into those with occlusions at (or distal to) the clinoid segment of the internal carotid artery (group A) and those with occlusions proximal to the clinoid segment (group B) and outcomes were compared between groups. Results Technical success rates, major complications, and re-occlusions within 1-year were 52%, 22%, 91% in group A (N=23), and 89%, 0%, 0% in group B (N=19), respectively. Diabetes was more frequent in group A (43%) compared with group B (11%). Conclusion CTA may play a role in predicting successful rate and 1-year patency for endovascular recanalization in carotid CTO.

Introduction Current risk stratification of Acute Lymphoblastic Leukemia (ALL) approach has significantly improve the outcome for pediatric and adolescent ALL. However, the outcome for adult ALL remains poor. With the emerging technologies that could drastically increase the parameters and scales of clinical data for treatment strategy formulation and more new therapeutic agents approved for ALL treatment, a better integrated and more systematic risk stratification method is needed for ALL. In our study, we propose to use artificial intelligence to develop an integral outcome prediction models that include all clinical exam and treatment history, aiming to further improve current risk stratification strategies. Method Retrospective clinical data of patients with ALL, including demographic (age & gender), laboratory results including complete blood count (CBC), white blood count (WBC), molecular genetics, cytogenetics, flow cytometry (FC) and treatment history at National Taiwan University Hospital were collected. A total of 513 newly diagnosed ALL from 2005 to 2017 were enrolled this study. The median age at diagnosis was 20 years and the median follow up duration was 49.1 months, 78.9% of them achieved complete remission (CR), 27.5% had relapsed, 31.6% had received HSCT (Table 1). In total, 243,464 CBC & WBC records, 32 types of anti-neoplastic medications in L01, L03 and L04 ATC code category with a total of 20,043 medication records from these 513 patients were utilized in developing the ALL outcome prediction model. A vectorized representation that captures the static-dynamic clinical aspects of an ALL patient can be learned directly from the collected data. The representation includes both static personal attributes (demographic, and genetic) and time-varying progression of patient's clinical assessment across time (laboratory results, HSCT, and medication). The time-dependent representation was derived from training a deep network architecture of bi-directional long short-term memory network (BLSTM). By taking 7 days as a time step, the BLSTM took the input of Fisher-vector encoded time series of a patient's CBC and WBC, medication, and HSCT records separately. The last output layer, which summarized the relapse/mortality risk exhibited in the measurements of patient's clinical conditions over time, of each separately-learned BLSTM was used as the encoded dynamic clinical representation. The concatenation of the ALL patient's static features and the time-dependent representations were fed into a deep neural network followed by a support vector machine to carry out the final prediction. The prediction models were conducted in 5-fold cross-validation experiments and further evaluated using metrics of accuracy (ACC). Results The median leukemia free survival and median overall survival of these 513 ALL patients are 29.3 months and 61.9 months respectively. The outcome prediction models developed achieved accuracy of next 3-month relapse, relapse or mortality, and mortality prediction reached 86.2%, 74.9% and 64.3% respectively (Table 2). Conclusions Our team has demonstrated the BLSTM-DNN model's potential that is capable in taking multiple and longitudinal clinical measurements into integrated relapse or mortality prediction in AML in 2018 (ASH 2018 #2811) and we have observed the same potential in ALL relapse and mortality prediction. More external and multi-national center studies are needed to further improve the performance of such integrated outcome prediction models. Disclosures Ko: Roche: Honoraria.

Background Magnetic resonance (MR) enterography has emerged as a new imaging modality in evaluating patients with Crohn's disease. However, whether interpretations of MR enterography findings are consistent among radiologists has not yet been fully investigated. The purpose of this study is to evaluate the intraobserver and interobserver agreement for identifying extraluminal manifestations of Crohn's disease with MR enterography. Methods MR enterography was performed with a 3-tesla system. We retrospectively enrolled patients with Crohn's disease that had MR enterography between November 2011 and March 2013. Three radiologists reviewed the images independently. Extraluminal findings of these patients were recorded and described. Intraobserver and interobserver agreement were calculated using the kappa statistic. Results A total of 22 sessions of MR enterography were performed on 15 patients (12 men and 3 women) with Crohn's disease. The mean age was 38.7 ± 11.4 years. We identified extraluminal manifestations such as engorged vasa recta, enlarged mesenteric lymph nodes, fibrofatty proliferation, fistulas, and abscesses. In addition, avascular necrosis of the femoral head as a complication of long-term steroid treatment was also depicted. The intraobserver agreement was almost perfect (κ = 0.83; p

Introduction The development of complex renal cysts after crizotinib treatment for non–small-cell lung cancer (NSCLC) is a reported side effect. However, its occurrence and characteristics have not been reported. Methods Medical records and computed tomography images of crizotinib-treated patients in three prospective clinical trials were reviewed. The size and Bosniak category of the renal cysts before and after crizotinib treatment were determined. Patients' clinical characteristics, tumor stage, treatment response, renal function, and outcomes were analyzed. Results During December 2010 to March 2013, we enrolled 32 patients who received crizotinib. There were 23 patients who had renal cysts before crizotinib. The median follow-up time was 493 days. Seven patients (22%, six with baseline renal cyst and one without baseline renal cyst) had significant renal cyst change. Four (13% of all) had new complex renal cysts. The median time from crizotinib treatment to first recognization of significant renal cyst change was 77 days. After stopping crizotinib, complex renal cysts regressed significantly. Patients with significant renal cyst change received more previous anticancer therapy (median, 5 lines versus 3 lines, p = 0.04) and received crizotinib for longer duration (median, 956 days versus 248 days, p = 0.007) compared with those without significant renal cyst change. Conclusions Change of renal cysts after crizotinib treatment is not uncommon. Development of complex renal cysts reverses after stopping crizotinib.

Background Tumor characteristics was sought to be related to axillary lymph node metastasis (ALNM), the paramount prognostic factor in patients with invasive breast cancer. This study was aimed to identify the ALNM-associated tumor characteristics and to determine the predictive clinical pathway. Material/Methods Data from 1325 patients diagnosed with invasive breast cancer between January 2004 and January 2010 were retrospectively reviewed. The structure equation model (SEM) was used to build the predictive clinical pathway. Results Among the factors found in the final model, the status of human epidermal growth factor receptor 2 is the primary influence on ALNM through histology grade (β=0.18), followed by tumor size (β=0.16). Tumor size was highly relevant to lymphovascular invasion (LVI) and influenced ALNM through LVI (β=0.26), the strongest predictor of ALNM in the final model (β=0.46) and the highest risk of ALNM (odds ratio=9.282; 95% confidence interval: 7.218–11.936). Conclusions The structure equation model presented the relation of these important predictors, and might help physicians to assess axillary nodal condition and appropriate surgical procedures.

Objective To observe the clinical efficacy and safety of Shuanghu Qinggan Granule (双虎清肝颗 粒, SQG) plus Yigan Yiqi Jieyu Granule (乙肝益气解郁颗粒, YYJG) combined with lamivudine (LAM) on chronic hepatitis B (CHB) patients.

Tumors located in the upper/upper inner quadrant of the breast warrant more attention. A small lesion relative to the size of breast in this location may be resolved by performing a level I oncoplastic technique. However, a wide excision may significantly reduce the overall quality of the breast shape by distorting the visible breast line. From June 2012 to April 2015, 36 patients with breast cancer located in the upper/upper inner quadrant underwent breast-conservation surgery with matrix rotation mammoplasty. According to the size and location of the tumor relative to the nipple-areola complex, 11 patients underwent matrix rotation with periareolar de-epithelialization (donut group) and the other 25 underwent matrix rotation only (non-donut group). The cosmetic results were self-assessed by questionnaires. The average weights of the excised breast lumps in the donut and non-donut groups were 104.1 and 84.5 g, respectively. During the 3-year follow-up period, local recurrence was observed in one case and was managed with nipple-sparing mastectomy followed by breast reconstruction with prosthetic implants. In total, 31 patients (88.6%) ranked their postoperative result as either acceptable or satisfactory. The treated breasts were also self-evaluated by 27 patients (77.1%) to be nearly identical to or just slightly different from the untreated side. Matrix rotation is an easy breast-preserving technique for treating breast cancer located in the upper/upper inner quadrant of the breast that requires a relatively wide excision. With this technique, a larger breast tumor could be removed without compromising the breast appearance.

Introduction While continuous advancement has been made in the treatment of AML, the overall 5-year survival rate achieved in current days is still less than 40%. The ability to accurately assess AML patient's risk of mortality and relapse is crucial to act on the most appropriate comprehensive treatment strategy. However, the surge of clinical parameters and the scale of data collection are becoming too complex for us to stratify risk and deriving predictive model systematically with conventional statistical methods. In our study, we propose to utilize artificial intelligence techniques in creating risk stratification and outcome prediction model through retrospective data analysis. Method Retrospective clinical data of patients with AML, including basic demographics (age & gender), laboratory results of complete blood count (CBC), white blood composition (WBC) and cytogenetics, and treatment history such as Hematopoietic Stem Cell Transplant (HSCT) and other medication history, was collected at the National Taiwan University Hospital. A total of 479 patients with newly diagnosed de novo AML were enrolled in this study. The median age at diagnosis was 50 years, and the median follow-up duration was 26.6 months, 77.7% of them achieved CR and, 53.4% had relapsed, and 47.2% had received HSCT. In total, 479 cytogenetics tests at diagnosis, 43,518 CBC & WBC records, 28 types of anti-neoplastic medications in L01 & L03 ATC code category with a total of 69,546 medication records from these 479 AML patients were used in deriving the outcome prediction model. A vectorized representation that captures the static-dynamic clinical aspects, i.e., demographics, laboratory results, and treatments, of an AML patient can be learned directly from the collected data. The representation includes both static personal attributes (demographic, and cytogenetic) and time-varying progression of patient's clinical assessment across time (laboratory results, HSCT, and medication). The time-dependent representation was derived from training a deep network architecture of bi-directional long short-term memory network (BLSTM). By taking 10 days as a time step, the BLSTM took the input of Fisher-vector encoded time series of a patient's CBC and WBC, medication, and HSCT records separately. The last output layer, which summarized the relapse/mortality risk exhibited in the measurements of patient's clinical conditions over time, of each separately-learned BLSTM was used as the encoded dynamic clinical representation. The concatenation of the AML patient's static features and the time-dependent representations were fed into a deep neural network followed by a support vector machine to carry out the final prediction. The prediction models were conducted in 5-fold cross-validation experiments and further evaluated using metrics of accuracy (ACC). Results The median leukemia-free survival and median overall survival of these 479 patients are 7.2 months and 49.0 months respectively. By using the CBC & WBC data, the accuracy of next 3-months relapse and mortality prediction at any time point reached 77.9 and 77.0% respectively, while models built with CBC, WBC combined with medication records reached accuracies of 78.4% and 84.6% respectively. Furthermore, incorporating static features, i.e., demographics and cytogenetics, together with time series representation of CBC & WBC, and medication records in our prediction model, we achieved further improved performances of 82.2% and 85.0% accuracies in next 3-month relapse and mortality prediction respectively (Figure 2). Conclusions The BLSTM-DNN model is a novel approach that is capable of jointly taking into account multiple heterogeneous clinical measurements throughout the clinical courses of AML patients in order to derive relapse and mortality prediction. It could be applied at any given time point using past 3-month data to predict the next 3-month relapse or mortality. We also observed the model accuracy increased as we increased the number of clinical exams included in our model. Our study results demonstrated a potential model in facilitating precise and personalized risk assessment that could support physicians for better risk-averse intervention in the future. We plan to integrate other exams including genomic and pathology reports with a larger patient's cohort into our model in the next phase of model development. Disclosures Ko: GNT Biotech & Medicals Crop.: Research Funding; Roche: Research Funding; Abbevie: Research Funding; Mumdipharma Taiwan: Consultancy.

Purpose: This study aimed to identify molecular determinants of sensitivity of non–small cell lung cancer (NSCLC) to anti–insulin-like growth factor receptor (IGF-IR) therapy. Experimental Design: A total of 216 tumor samples were investigated, of which 165 consisted of retrospective analyses of banked tissue and an additional 51 were from patients enrolled in a phase II study of figitumumab, a monoclonal antibody against IGF-IR, in stage IIIb/IV NSCLC. Biomarkers assessed included IGF-IR, epidermal growth factor receptor, IGF-II, IGF-IIR, insulin receptor substrate 1 (IRS-1), IRS-2, vimentin, and E-cadherin. Subcellular localization of IRS-1 and phosphorylation levels of mitogen-activated protein kinase and Akt1 were also analyzed. Results: IGF-IR was differentially expressed across histologic subtypes (P = 0.04), with highest levels observed in squamous cell tumors. Elevated IGF-IR expression was also observed in a small number of squamous cell tumors responding to chemotherapy combined with figitumumab (P = 0.008). Because no other biomarker/response interaction was observed using classical histologic subtyping, a molecular approach was undertaken to segment NSCLC into mechanism-based subpopulations. Principal component analysis and unsupervised Bayesian clustering identified three NSCLC subsets that resembled the steps of the epithelial to mesenchymal transition: E-cadherin high/IRS-1 low (epithelial-like), E-cadherin intermediate/IRS-1 high (transitional), and E-cadherin low/IRS-1 low (mesenchymal-like). Several markers of the IGF-IR pathway were overexpressed in the transitional subset. Furthermore, a higher response rate to the combination of chemotherapy and figitumumab was observed in transitional tumors (71%) compared with those in the mesenchymal-like subset (32%; P = 0.03). Only one epithelial-like tumor was identified in the phase II study, suggesting that advanced NSCLC has undergone significant dedifferentiation at diagnosis. Conclusion: NSCLC comprises molecular subsets with differential sensitivity to IGF-IR inhibition. Clin Cancer Res; 16(18); 4654–65. ©2010 AACR.

Bevacizumab, an antiangiogenic monoclonal anti- body against vascular endothelial growth factor, was designed to normalize tumor vasculature and reduce intratumoral pres- sure. It can create a 'normalization window' during which the cancer can be attacked the most effectively, and the effects of chemotherapeutic drugs are enhanced. Representative trials (E2100, AVADO, RIBBON-1, RIBBON-2 and TURANDOT) have shown that the addition of bevacizumab to chemotherapy has significant benefits on progression‑free survival for metastatic breast cancer, but not on overall survival. The present study describes two patients with meta- static triple-negative breast cancer who received 6 courses of bevacizumab-containing chemotherapy. Each course comprised 5-7.5 mg /kg bevacizumab administered on days 1 and 15, and 20-24 h after bevacizumab delivery, 80 mg/m 2 paclitaxel was administered for 3 weeks on days 2, 9 and 16, followed by 1 week of rest. Following sequential treatment with bevacizumab and paclitaxel, the results of computed tomog- raphy showed that the tumors were rapidly reduced in size. Based on the imaging findings from three‑dimension power Doppler ultrasonography in one of the breast cancer patients who received neoadjuvant chemotherapy with bevacizumab, the possible timing of the normalization window was 20-24 h after the administration of bevacizumab. The normalization window may provide an opportunity to enhance the effect of chemotherapy with the aid of bevacizumab.

Pylephlebitis (septic thrombophlebitis of the portal venous system) is a rare complication of intra-abdominal infection. We aimed to investigate the recent trend of its etiology, clinical manifestation, and prognosis.We retrospectively studied the etiology, clinical manifestation, and outcome by reviewing the medical records of all imaging-confirmed pylephlebitis cases diagnosed during the period 2002-2011 in a university hospital in Taiwan. To identify the risk factors for pylephlebitis, we randomly selected 160 patients with intra-abdominal infections but without pylephlebitis as the comparison group.We identified 35 cases of pylephlebitis. Most patients were men [29/35 (83 %)]. The median age of the patients was 57 years (range 35-90 years). Unspecified abdominal pain (18/35) and fever (10/35) were the most common clinical manifestations. Klebsiella pneumoniae liver abscess (7/35) and cholangitis (7/35) were the most common etiologies. Liver abscess was a risk factor for pylephlebitis (13/35 vs. 27/160, P = 0.01). With antibiotic therapy, there was no in-hospital mortality, but pylephlebitis was still associated with an excess hospital stay (22.2 ± 17.6 vs. 9.8 ± 7.1 days, P 0.001).Our study results suggested a different pattern of pylephlebitis from previous Western literature. K. pneumoniae liver abscess (7/35) is an emerging etiology of pylephlebitis in Taiwan.

We have previously demonstrated that PRL acts as an autocrine growth factor in the pituitary tumor GH3 cell line. In this study we present evidence that the 235-1 cell line also uses PRL as an autocrine growth factor, and that in this case interaction between PRL and its receptor occurs intracellularly. First, the PRL produced by 235-1 cells was shown to be capable of initiating a proliferative response by placement in the Nb2 bioassay. Second, PRL receptors were demonstrated to be present, but, unusually, primarily in the Golgi complex by light and electron microscope immunocytochemistry. Incubation of the cells in anti-PRL had no effect on 235-1 cell proliferation until interleukin-1 treatment caused the PRL receptors to move to the cell surface, whereupon cell proliferation was inhibited. Specific interference with PRL biosynthesis in noninterleukin-1-treated cells also inhibited cell proliferation. Thus, the essential elements of an autocrine loop are present and can be demonstrated to be functional by antibody inhibition of growth after movement of the receptors to the cell surface and by antisense inhibition of growth with the receptors in their normal intracellular location.

Background Epidemiological evidence suggests that moderately elevated levels of circulating insulin-like growth factor-I (IGF-I) are associated with increased risk of breast cancer in women. How circulating IGF-I may promote breast cancer incidence is unknown, however, increased IGF-I signaling is linked to trastuzumab resistance in ErbB2 positive breast cancer. Few models have directly examined the effect of moderately high levels of circulating IGF-I on breast cancer initiation and progression. The purpose of this study was to assess the ability of circulating IGF-I to independently initiate mammary tumorigenesis and/or accelerate the progression of ErbB2 mediated mammary tumor growth. Methods We crossed heterozygous TTR-IGF-I mice with heterozygous MMTV-ErbB2 mice to generate 4 different genotypes: TTR-IGF-I/MMTV-ErbB2 (bigenic), TTR-IGF-I only, MMTV-ErbB2 only, and wild type (wt). Virgin females were palpated twice a week and harvested when tumors reached 1000 mm3. For study of normal development, blood and tissue were harvested at 4, 6 and 9 weeks of age in TTR-IGF-I and wt mice. Results TTR-IGF-I and TTR-IGF-I/ErbB2 bigenic mice showed a moderate 35% increase in circulating total IGF-I compared to ErbB2 and wt control mice. Elevation of circulating IGF-I had no effect upon pubertal mammary gland development. The transgenic increase in IGF-I alone wasn't sufficient to initiate mammary tumorigenesis. Elevated circulating IGF-I had no effect upon ErbB2-induced mammary tumorigenesis or metastasis, with median time to tumor formation being 30 wks and 33 wks in TTR-IGF-I/ErbB2 bigenic and ErbB2 mice respectively (p = 0.65). Levels of IGF-I in lysates from ErbB2/TTR-IGF-I tumors compared to ErbB2 was elevated in a similar manner to the circulating IGF-I, however, there was no effect on the rate of tumor growth (p = 0.23). There were no morphological differences in tumor type (solid adenocarcinomas) between bigenic and ErbB2 mammary glands. Conclusion Using the first transgenic animal model to elevate circulating levels of IGF-I to those comparable to women at increased risk of breast cancer, we showed that moderately high levels of systemic IGF-I have no effect on pubertal mammary gland development, initiating mammary tumorigenesis or promoting ErbB2 driven mammary carcinogenesis. Our work suggests that ErbB2-induced mammary tumorigenesis is independent of the normal variation in circulating levels of IGF-I.

Charge isomers of monomer PRL have been described for all species thus far examined. For the rat, cow, and chicken, at least a major proportion of the more acidic isomers have been shown to be phosphorylated. The physiological relevance of this posttranslational phosphorylation, however, remains unclear. In this study we have compared the growth-promoting activities of dephosphorylated and standard rat PRL (partially phosphorylated) in the widely used Nb2 bioassay. Dephosphorylated PRL was produced by treatment of standard rat PRL with acid phosphatase, resulting in the conversion of a 91.9% nonphosphorylated/8.1% phosphorylated preparation to a 98.6% nonphosphorylated preparation. These two preparations were added separately or in combination to stationary Nb2 cells. Cell number was assessed 3 days later using a colorimetric assay. Dephosphorylated PRL showed significantly higher growth-promoting activity than standard PRL at concentrations up to 10 ng/ml. In the 1-5 ng/ml concentration range, dephosphorylated PRL was twice as active. Given that the conversion of 6.7% phosphorylated PRL to nonphosphorylated PRL resulted in a doubling of activity, one can deduce not only that phospho-PRL acted as an antagonist to nonphosphorylated PRL in this assay, but also that it did not do so on an equimolar basis. Titration of the two PRL preparations produced data confirming this latter deduction. These data are of importance in our understanding of PRL-promoted cell proliferation.

Insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) have been shown to have a role in breast cancer, although the specific importance of hybrid IR/IGF1R (Hybrid-R)is unknown. I proposed to use an inducible dimerization system to study Hybrid-R, and then optimize an assay to detect Hybrid-R in breast tumor samples. I constructed chimeric IR and IGF1R plasmids with different dimerization domains and developed stable MCF10A clones expressing only chimeric IR or chimeric IGF1R allowing homodimerization. Homodimerizer AP20187 treatment induced activation of chimeric IGF1R and IR in a dose dependent manner and activated AKT and ERK. However, I encountered technical difficulties in establishing cells expressing both chimeric IR and chimeric IGF1R and was unable to induce specific Hybrid-R. To measure the IR/IGF1R hybrid receptor on paraffin embedded specimens, a proximity ligation assay (PKLA) was optimized using both anti-IR and anti- IGF1R antibodies. The presence of PLA signals was increased in cells had IR or IGF1R overexpression and decreased in cells with IGF1R knock down. The variability of signals was also observed in a tissue microarray with normal breast tissue and breast tumor. This study shows the presence of Hybrid-R in breast cancer cell lines and human tumors and suggests that more studies are needed to determine its function and importance in breast cancer.

The effects of different concentrations of various K-current blockers on prolactin-induced proliferation and membrane K-currents in malignant lymphocytes (Nb2 cells) were investigated. Membrane currents were measured with the whole cell patch-clamp technique, and lymphocyte density was quantified by both spectrophotometric and conventional methods. K-current blockers tested (quinidine, 4-aminopyridine, barium, and tetraethylammonium) exhibited similar rank order potency for K-current block and inhibition of prolactin-induced proliferation of malignant lymphocytes. Because Nb2 cells proliferate independently of a transmembrane Ca-influx, these results suggest that K-currents per se rather than K-current modulation of Ca-influx is an essential event for lymphocyte proliferation.

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), a significant pungent ingredient in a variety of red peppers of the genus Capsicum, is a type of vanilloid. It has been shown to exert biological activities (anticarcinogenic, antimutagenic and chemopreventive) in many cancer cell lines. It was found that capsaicin induces dose-dependent growth inhibition of MCF-7 cells, which does not express caspase-3. In this study, we investigated the molecular mechanism of capsaicin-induced apoptosis in MCF-7 cells. Treatment with capsaicin for 24 h resulted in dose-dependent apoptosis in these cells. After the addition of capsaicin, the levels of reactive oxygen species were reduced slightly in the earlier stage of treatment. Interestingly, an elevation of intracellular calcium ion concentration was detected in the MCF-7 cells. In time course and dosage studies, the mitochondrial membrane potential of MCF-7 cells decreased. However, the change was not significant. It is worth noting that the apoptosis-inducing factor translocated into the cytosol and nucleus from the mitochondria. Our results suggest that capsaicin induces cellular apoptosis through a caspase-independent pathway in MCF-7 cells, and that reactive oxygen species and intracellular calcium ion fluctuation has a minimal role in the process.

w d L t t t A 71-year-old man had a history of gastric adenocarcinoma status after subtotal gastrectomy with Billroth I gastrodudenostomy for 6 years without clinical recurrence during an nnual follow-up evaluation. During the past month, he expeienced gradual onset of icteric sclera and skin, and dark yelowish urine. Laboratory data revealed increased liver function tudies, including an aspartate aminotransferase level of 185 U/L (normal, 36 IU/L), alanine aminotransferase level of 270 U/L (normal, 41 IU/L), an alkaline phosphatase level of 526 /L (normal, 104 IU/L), a -glutamyl transpeptidase level of 490 U/L (normal, 52 IU/L), and a total/direct bilirubin level of 2.94/1.58 mg/dL (normal, 1.2/0.4 mg/dL). Abdominal ultrasonography was negative for focal lesions. With a presumptive diagnosis of obstructive jaundice, magnetic resonance cholangiopancreatography (MRCP) was performed. Thick-slice MRCP shows slight external compression of the distal common bile duct (CBD) by the juxtapapillary diverticulum (Figure A, arrow), leading to CBD dilatation. On thin-slice MRCP, a duodenal diverticulum (Figure B, arrow) with internal debris was noted in the juxtapapillary region. The distal CBD was displaced slightly and compressed by the diverticulum; no internal filling defect could be identified in the distal CBD. The main pancreatic duct (MPD) was intact. Given the patient’s clinical manifestations and results of the MRCP, Lemmel’s syndrome was diagnosed. The patient’s bilirubin level decreased rapidly (total/direct bilirubin level, 0.8/0.24 mg/dL) after removal of the diverticular debris by endoscopy. Postprocedural MRCP revealed a smaller-caliber CBD (Figure C). The duodenum is the second-most common site of gastrointestinal diverticula after the colon. Most duodenal diverticula occur within 2.0 cm of the ampulla of Vater and usually remain asymptomatic, therefore, they rarely require treatment.1 Lemel’s syndrome usually is seen in cases of juxtapapillary divericula with pancreatobiliary diseases excluding cholelithiasis.1 External compression of the distal CBD in conjunction with sphincter of Oddi dysfunction may lead to pancreatobiliary disease as a result of duodenobiliary and duodenopancreatic reflux.2 Among patients with juxtapapillary diverticula, those ho are older and have larger diverticula are more likely to evelop Lemmel’s syndrome.3 The therapeutic goal in cases of emmel’s syndrome is to relieve the pancreaticobiliary obstrucion, either by excision of the diverticulum, endoscopic sphincerotomy, placement of a biliary stent, or by simple removal of he diverticulum’s content.3

IGF-I signaling is involved in tumor progression and drug resistance and many anti-IGF-IR therapeutic agents are currently in clinical trials. Moreover, growing evidence also suggests that insulin signaling may play an important role in cancer development and progression. However, little is known regarding similarities and difference in IGF-I and insulin signaling in cancer, and there are currently no biomarkers to predict patient response to IGF targeted therapy. To gain a better understanding of these signaling pathways in human breast cancer, we measured the levels of insulin receptor (IR) and IGF-IR, and the activity of insulin and IGF-I, in a large panel of human breast cancer cell lines. Comparative gene expression analysis was performed using publicly available gene expression data and IR and IGF-IR mRNA levels in 17 breast cancer cell lines were measured by Q-RT-PCR. IGF-IR and IR levels were found to be highly variable in breast cancer cell lines. MCF7 and MDA-MB-134 have high levels of IGF-IR expression and MDA-MB-468 and ZR-75-1 have high levels of IR expression, relative to other breast cancer cell lines. Reverse phase protein array (RPPA) analysis using 134 different antibodies was performed on MCF7 cells stimulated with increasing doses of insulin or IGF-I. One Way ANOVA was performed to identify proteins affected by insulin and/or IGF stimulation. Maximal response to insulin and IGF-I stimulation was observed at a 10nM concentration of both ligands. Therefore, we further performed RPPA on 21 breast cancer cell lines treated with 10nM of insulin and IGF-I for 6 time points (5min, 10min, 30 min, 6hrs, 24hrs, and 48hrs). Spearman rank correlation was performed on IR and IGF-IR protein levels to mRNA levels from Q-RT-PCR. There was a significant correlation between IGF-IR protein and mRNA levels (r=0.559, p=0.024), but not between IR protein and mRNA levels. Principle component analysis showed that MCF7, MDA-MB-134, T47D and ZR-75-1 are insulin and IGF-I responsive. Results from ANOVA with contrast in MCF7 cells showed that insulin and IGF-I affected similar set of proteins at 10 minute and 24 hours time points, however, there were several proteins affected only by IGF-I or insulin stimulation. For example, phospho-estrogen receptor (p-ER) was found to be increased by 10 minutes IGF-I stimulation, but not by insulin stimulation. Beta-catenin and c-Myc were significantly suppressed by IGF stimulation while PARP was significantly suppressed by insulin stimulation at the 24 hour time point. Moreover, IGF affected twice the number of proteins compared to insulin at the 48 hour time point. For instance, Bcl2 and SMAD3 were suppressed and Raf and FOXO3a were increased by IGF stimulation after 48 hours of stimulation. Further statistical and system biology analysis of the temporal activation of signaling by IGF-I and insulin in the 21 cell lines is ongoing and will be presented. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4133.

Because PRL has growth factor activities in several tissues, we have asked whether it also has autocrine growth factor activity in pituitary GH3 cells. GH3 cells were grown at increasing densities in the presence or absence of antirat PRL (polyclonal and monoclonal) or nonspecific antibodies. Cell proliferation increased with increasing cell density, as did the concentration of PRL in the medium. Antirat PRL, but not control antibody, markedly inhibited but did not eliminate cell proliferation, and this effect was diminished with increasing PRL concentration in the medium. PRL receptors were demonstrated on 40-50% of the cells by indirect immunofluorescence using a specific antirat PRL receptor monoclonal antibody. Cell surface PRL was colocalized to the same 40-50% of the cells and copatched or cocapped along with the receptors. Absence or presence of PRL receptors did not correlate with stage of the cell cycle, as judged by ethidium bromide dual labeling. Cell surface PRL was found to be on PRL-containing cells. These data have fulfilled four criteria necessary for establishment of a substance as a secreted autocrine growth factor: 1) the factor must be secreted; 2) in log growth phase, increased cell proliferation should occur at increased cell densities; 3) the cells must display a receptor for the factor; and 4) there must be a growth response to the factor. Thus we have established that PRL is an autocrine growth factor for at least 40-50% of the GH3 cell population. This, to our knowledge, is the first example of autocrine growth factor activity of a major hormone normotopically expressed.

The expression of the cytochrome P450 CYP3A5 enzymes shows a wide variation across the general population and ethnic groups. This wide disparity implies interracial differences in drug clearance and susceptibility to diseases such as cancer. CYP3A5 polymorphisms were rapidly determined using polymerase chain reaction-restriction fragment length polymorphism analysis in 113 Taiwanese patients with hepatoma, 70 with cervical cancer, 92 with breast cancer, 82 with oral cancer, 90 with thyroid cancer, 133 with lung cancer, and 270 healthy controls. The allelic frequencies of CYP3A5*1 were 25% in hepatoma patients, 33% in cervical cancer patients, 31% in breast cancer patients, 22% in oral cancer patients, 23% in thyroid cancer patients, 20% in lung cancer patients, and 27% in healthy subjects. Lung cancer patients had a significantly lower frequency (20%) of CYP3A5*1 expression than healthy controls (p = 0.028, odds ratio = 1.49, 95% confidence interval = 1.04-2.13), but there was no statistically significant difference between healthy controls and other cancers. We suggest that CYP3A5*1 may play an important role in individual predisposition to lung cancer in Taiwan.