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Importance: Immune-related adverse events (irAEs) arising from immune checkpoint inhibitor (ICI) cancer therapy may potentially predict improved outcomes., Objective: To evaluate the association between irAEs and atezolizumab efficacy in patients with advanced non-small cell lung cancer (NSCLC) using pooled data from 3 phase 3 ICI studies., Design, Setting, and Participants: IMpower130, IMpower132, and IMpower150 were phase 3, multicenter, open-label, randomized clinical trials to evaluate the efficacy and safety of chemoimmunotherapy combinations involving atezolizumab. Participants were chemotherapy-naive adults with stage IV nonsquamous NSCLC. These post hoc analyses were conducted during February 2022., Interventions: Eligible patients were randomly assigned 2:1 to receive atezolizumab with carboplatin plus nab-paclitaxel, or chemotherapy alone (IMpower130); 1:1 to receive atezolizumab with carboplatin or cisplatin plus pemetrexed, or chemotherapy alone (IMpower132); and 1:1:1 to receive atezolizumab plus bevacizumab plus carboplatin and paclitaxel, atezolizumab plus carboplatin and paclitaxel, or bevacizumab plus carboplatin and paclitaxel (IMpower150)., Main Outcomes and Measures: Pooled data from IMpower130 (cutoff: March 15, 2018), IMpower132 (cutoff: May 22, 2018), and IMpower150 (cutoff: September 13, 2019) were analyzed by treatment (atezolizumab-containing vs control), irAE status (with vs without), and highest irAE grade (1-2 vs 3-5). To account for immortal bias, a time-dependent Cox model and landmark analyses of irAE occurrence at 1, 3, 6, and 12 months from baseline were used to estimate the hazard ratio (HR) of overall survival (OS)., Results: Of 2503 randomized patients, 1577 were in the atezolizumab-containing arm and 926 were in the control arm. The mean (SD) age of patients was 63.1 (9.4) years and 63.0 (9.3) years, and 950 (60.2%) and 569 (61.4%) were male, respectively, in the atezolizumab arm and the control arm. Baseline characteristics were generally balanced between patients with irAEs (atezolizumab, n = 753; control, n = 289) and without (atezolizumab, n = 824; control, n = 637). In the atezolizumab arm, OS HRs (95% CI) in patients with grade 1 to 2 irAEs and grade 3 to 5 irAEs (each vs those without irAEs) in the 1-, 3-, 6-, and 12-month subgroups were 0.78 (0.65-0.94) and 1.25 (0.90-1.72), 0.74 (0.63-0.87) and 1.23 (0.93-1.64), 0.77 (0.65-0.90) and 1.1 (0.81-1.42), and 0.72 (0.59-0.89) and 0.87 (0.61-1.25), respectively., Conclusions and Relevance: In this pooled analysis of 3 randomized clinical trials, longer OS was observed in patients with vs without mild to moderate irAEs in both arms and across landmarks. These findings further support the use of first-line atezolizumab-containing regimens for advanced nonsquamous NSCLC., Trial Registration: ClinicalTrials.gov Identifiers: NCT02367781, NCT02657434, and NCT02366143.

Background: Esophageal adenocarcinoma is a lethal disease. For locally advanced patients, neoadjuvant chemoradiotherapy followed by surgery is the standard of care. Risk stratification relies heavily on clinicopathologic features, particularly pathologic response, which is inadequate, therefore establishing the need for new and reliable biomarkers for risk stratification., Methods: Thirty four patients with locally advanced esophageal adenocarcinoma were analyzed, of which 21 received a CROSS regimen with carboplatin, paclitaxel, and radiation. Capture-based targeted sequencing was performed on the paired baseline and post-treatment samples. Differentially mutated gene analysis between responders and non-responders of treatment was performed to determine predictors of response. A univariate Cox proportional hazard regression was used to examine associations between gene mutation status and overall survival., Results: A 3-gene signature, based on mutations in EPHA5, BCL6, and ERBB2, was identified that robustly predicts response to the CROSS regimen. For this model, sensitivity was 84.6% and specificity was 100%. Independently, a 9 gene signature was created using APC, MAP3K6, ETS1, CSF3R, PDGFRB, GATA2, ARID1A, PML, and FGF6, which significantly stratifies patients into risk categories, prognosticating for improved relapse-free (p = 4.73E-03) and overall survival (p = 3.325E-06). The sensitivity for this model was 73.33% and the specificity was 94.74%., Conclusion: We have identified a 3-gene signature (EPHA5, BCL6, and ERBB2) that is predictive of response to neoadjuvant chemoradiotherapy and a separate prognostic 9-gene classifier that predicts survival outcomes. These panels provide significant potential for personalized management of locally advanced esophageal cancer., (© 2022. The Author(s).)

Background: Pheochromocytomas are a subset of paragangliomas, which are a rare group of neural crest cell-derived tumors. Malignant cases of both pheochromocytomas and paragangliomas are even rarer, and currently there is no standard of care. This case report details the use of off-label immunotherapy and its efficacy in the management of the aforementioned tumor., Case Presentation: Herein is presented a case of a 60-year-old Caucasian female with a rare malignant pheochromocytoma. The tumor was determined to be unresectable because of involvement of surrounding organs. Radiation therapy was also not a viable option because of concerns over appreciable toxicity in relation to mass size. As there is no standard of care for malignant cases, the patient was started on chemotherapeutic agents but was soon shown to be intolerant to this treatment. As she was ineligible for several clinical trials, the patient was started on the off-label immunotherapeutic agents nivolumab and ipilimumab. Immunotherapy use resulted in decreased tumor size, improved quality of life, and reconsideration for radiation therapy., Conclusions: The use of immunotherapy in pheochromocytoma in this patient clearly demonstrated substantial benefit, as she was able to be reconsidered for radiation therapy. Not only has the patient been tolerant of this treatment, but she has exhibited progression-free survival of over 20 months. As there is no current standard treatment for malignant pheochromocytomas, the success of its use in this patient lends support to the ongoing clinical trials regarding the use of immunotherapy in rare tumors, including pheochromocytomas.

Background: Esophageal adenocarcinoma (EAC) is a deadly disease with limited treatment options. STING is a transmembrane protein that activates transcription of interferon genes, resulting in stimulation of APCs and enhanced CD8+ T-cell infiltration. The present study evaluates STING agonists, alone and in combination with radiation to determine durable anticancer activity in solid tumors., Materials and Methods: Esophagojejunostomy was performed on rats to induce reflux leading to the development of EAC. At 32 weeks post operatively, rats received intratumorally either 50 μg STING (ADU-S100) or placebo (PBS), +/- 16Gy radiation. Drug activity was evaluated by pre- and post- treatment MRI, histology, immunofluorescence and RT-PCR., Results: Mean MRI tumor volume decreased by 30.1% and 50.8% in ADU-S100 and ADU-S100 + radiation animals and increased by 76.7% and 152.4% in placebo and placebo + radiation animals, respectively ( P < 0.0001). Downstream gene expression, pre- to on- and post- treatment, demonstrated significant upregulation of IFNβ, TNFα, IL-6, and CCL-2 in the treatment groups vs. placebo. On- or post- treatment, radiation alone, ADU-S100 alone, and ADU-S100 + radiation groups demonstrated enhanced PD-LI expression, induced by upregulation of CD8+ T-cells ( p < 0.01)., Conclusions: ADU-S100 +/- radiation exhibits potent antitumor activity and a promising immunomodulatory profile in a de novo EAC., Competing Interests: CONFLICTS OF INTEREST Authors have no conflicts of interest to declare., (Copyright: © 2021 Zaidi et al.)

Background: The standard of care for non-metastatic squamous cell carcinoma of the anal canal (SCCA) is concurrent chemoradiotherapy. It is postulated that chemotherapy could be omitted for the earliest stages without worsening outcomes., Methods: We queried the NCDB from 2004-2016 for patients with cT1N0M0 SCCA treated non-operatively with radiation, with and without chemotherapy, and at least two months of follow-up. Of the 2,959 patients meeting eligibility, 92% received chemotherapy (n = 2722) and 8% (n = 237) did not. Most patients were white (n = 2676), female (n = 2019), had private insurance (n = 1507) and were treated in a comprehensive cancer center (n = 1389). Average age was 58.5 years., Results: Predictors of chemotherapy omission were age > 58 years (OR 0.66, 95% CI [0.49-0.90], P = 0.0087), higher comorbidity score (OR 0.62, 95% CI [0.38-0.99], P = 0.0442), African American race (OR 0.57, 95% CI [0.36-0.90], P = 0.0156) and treatment at the start of the study period (OR 1 for years 2004-2006). HR for single-agent chemotherapy was 0.70 (95% CI [0.50-0.96], P = 0.0288) and 0.48 for multi-agent (95% CI [0.38-0.62], P <0.0001). Overall survival was 86% in those that received chemotherapy vs 65% in those who did not (P <0.0001)., Conclusions: In conclusion, patients with early-stage squamous cell cancer of the anus who are treated with combination chemoradiation continue to demonstrate better overall survival than those who undergo radiotherapy alone., (Copyright © 2021. Published by Elsevier Ltd.)

Purpose: Large cell neuroendocrine carcinoma (LCNEC) is a rare pulmonary malignancy with clinicopathologic features of both non-small cell lung cancer (NSCLC) and small-cell lung cancer (SCLC). Given the paucity of available data regarding LCNEC management, we queried the National Cancer Database (NCDB) to describe trends in management, identify predictors of treatment receipt, and compare outcomes in patients receiving chemotherapy (ChT) and chemoradiotherapy (CRT)., Methods: We identified patients with locally advanced (Stage III) LCNEC of the lung treated with definitive ChT or CRT between the years of 2004-2015. Odds ratios were calculated to determine predictors of CRT receipt. Multivariable cox regression was used to determine predictors of overall survival., Results: Using the above criteria, 5797 patients were identified, 54 % of whom received CRT (n = 3153) while 46 % (n = 2644) received ChT alone. Most patients had T4 (35 %) and N2 (59 %) disease. Median overall survival was 11.9 months (11.3-12.6) in patients receiving ChT compared to 16.1 months (15.4-16.9) in patients receiving CRT (p < 0.0001). Overall survival at 1, 3, and 5 years was 50 %, 20 %, and 13 % versus 60 %, 27 %, and 18 %, in patients receiving ChT and CRT, respectively. Older patients and those with higher comorbidity scores were less likely to receive CRT; whereas patients with higher education level, treatment receipt at an academic/research program facility, N2 disease, and later treatment year were more likely to receive CRT. On multivariable analysis, older age, greater comorbidity score, presence of N2 disease, and presence of T4 disease were all associated with decreased OS. CRT receipt was an independent predictor of increased overall survival., Conclusions: Definitive CRT was an independent predictor of increased overall survival in patients with locally advanced LCNEC of the lung. Findings from our study may help guide potential areas of future investigation to help define an ideal treatment approach for LCNEC., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Many genomic alterations have been identified that are critical to the malignant phenotype. Some of these, termed "driver mutations," are critical for tumor proliferation and progression. The landscape of targeted therapy has expanded as well. Next-generation sequencing (NGS) of tumors reveals cancer-related genomic alterations and provides therapeutic recommendations for specific targeted therapy. We analyzed our experience with FoundationOne, a validated NGS genomic profiling test, in a community oncology network. Methods: NGS results from May 2014 to September 2016 from a community oncology network in Western Pennsylvania were analyzed. Medical records were reviewed for primary site, stage, biopsy site, time of testing, prior treatment, FDA-approved therapy in patient's and other tumor types and potential clinical trials based upon mutations detected. Two co-primary endpoints for this study were to determine the percentage of patients having mutations with a FDA-approved targeted agent and the percentage of patients in whom a treatment decision was made based on these NGS results. Results: One Fifty-Seven NGS results were available for analysis. 82% patients had a mutation with a FDA-approved targeted agent available while 18% patients had no FDA-approved targeted agent for the mutation detected. Clinical trials were available for 93% cases. The NGS results were utilized in treatment decisions in 18% patients ( n = 28) with, 7% ( n = 11) initiating a targeted agent, 6% ( n = 9) were on an appropriate targeted agent prior to testing and 5% ( n = 8) being unable to start a targeted agent because of insurance denial, clinical deterioration or patient preference. 38% cases were tested early in the disease course (at diagnosis, during or shortly after first-line treatment) and 62% at progression. Conclusions: NGS is a valuable tool to identify molecular targets for personalizing cancer care. From our experience, the actual number of patients starting a targeted agent based on NGS results is low but it provides substantial information in terms of providing additional treatment options, identifying resistance conferring mutations and facilitating clinical trial enrollment. Optimal time of testing, early or late in disease course, financial implications of testing and using targeted therapy and survival benefit of targeted therapy need further studies., (Copyright © 2020 Tayshetye, Miller, Monga, Brem, Silverman and Finley.)

Background: Surgery remains the standard of care in rectal cancer. Select patients will not undergo surgery for reasons such as medical inoperability or a watch-and-wait approach and instead are managed with definitive chemoradiation., Objective: We used the National Cancer Database to identify overall survival and predictors thereof in the nonoperative management of patients with rectal cancer., Design: This was a retrospective review., Settings: This study used deidentified data from the National Cancer Database., Patients: We queried the national cancer database from 2004 to 2014 for stage 1 to 3 rectal adenocarcinoma treated with only chemotherapy and radiation to definitive doses. Dose escalated therapy was defined as >54 Gy., Main Outcome Measures: Univariable and multivariable analyses were performed to identify sociodemographic, treatment, and tumor characteristics predictive of dose escalation and overall survival. Propensity-adjusted Cox proportional hazard ratios for survival were used to account for indication bias., Results: Among the 6311 patients eligible for the study, 11% were treated with doses >54 Gy. Earlier stage and increased age/comorbidity patients were more likely to receive dose escalation, and patients with more recent treatment and treatment at an academic facility were less likely. The median follow-up time was 31 months (range, 2-154 mo). Three- and 5-year overall survival rates for all patients were 60% and 46%. Patients treated with dose escalation had a median survival of 33 months compared with 56 months for those treated with ≤54 Gy (p < 0.0001)., Limitations: The main limitation is the inherent selection bias present in National Cancer Database studies. Important treatment details and outcomes as they relate to a definitive chemoradiation approach in rectal cancer are lacking. Salvage therapy was also not recorded, which in this population could be surgery., Conclusions: In this analysis, dose escalation in the nonoperative management of rectal cancer was associated with a lower overall survival compared with more conventional doses. Careful patient selection and enrollment on appropriate clinical trials may be warranted in the nonoperative setting. See Video Abstract at http://links.lww.com/DCR/B15. LA QUIMIORRADIACIÓN DEFINITIVA PARA EL CÁNCER RECTAL: ¿HAY LUGAR PARA EL AUMENTO DE LA DOSIS? UN ESTUDIO DE BASE DE DATOS NACIONAL DEL CÁNCER:: La cirugía sigue siendo el estándar en el tratamiento del cáncer rectal. Algunos pacientes no son quirúrgicos por razones como, no ser operables o con el enfoque de ver y esperar, y en su lugar son tratados con la quimiorradiación definitiva.Utilizamos la base de datos nacional del cáncer para identificar la supervivencia general y los factores predictivos de la misma, en el tratamiento no quirúrgico de pacientes con cáncer rectal.Esta fue una revisión retrospectiva.Utilizamos los datos identificados en la base de datos nacional del cáncer.Se consultó la base de datos nacional del cáncer del 2004-2014, para adenocarcinoma rectal en estadio 1-3, tratada únicamente con quimioterapia y radiación hasta la dosis definitiva. La terapia de aumento de la dosis se definió como >54 Gy.Se realizaron análisis univariables y multivariables para identificar características sociodemográficas, de tratamiento y predictivas del aumento de la dosis y supervivencia en general. Los índices de riesgo proporcionales de Cox ajustados a la propensión para la supervivencia, se utilizaron para tener en cuenta el sesgo de indicación.Entre los 6311 pacientes elegibles para el estudio, el 11% fue tratado con dosis >54 Gy. Los pacientes en estadios tempranos y con mayor edad/comorbilidad, tenían más probabilidades de recibir aumento de la dosis, y menos propensos los pacientes con tratamientos recientes y de centros académicos. El tiempo medio de seguimiento fue de 31 meses (2-154 meses). Las tasas de supervivencia global de tres y cinco años para todos los pacientes, fueron respectivamente del 60% y 46%. Los pacientes tratados con aumento de la dosis, tuvieron una supervivencia media de 33 meses, en comparación con los 56 meses para los pacientes tratados con ≤54 Gy (p < 0,0001).La principal limitación es el inherente sesgo en la selección, presente en los estudios de la base de datos nacional del cáncer. Faltan los detalles importantes del tratamiento y los resultados en relación con el enfoque definitivo de quimiorradiación en cáncer rectal. Tampoco se registró la terapia de rescate, que en esta población podría ser la cirugía.En este análisis, el aumento de la dosis en el manejo no quirúrgico del cáncer rectal, se asoció con una menor supervivencia global, en comparación con la dosis más convencional. La cuidadosa selección del paciente y la inscripción en los apropiados ensayos clínicos, pueden estar justificados en el entorno no quirúrgico. Vea el Resumen del Video en http://links.lww.com/DCR/B15.

Background: Neoadjuvant chemoradiotherapy (nCRT) followed by resection and postoperative multi-agent chemotherapy (maChT) is the standard of care for locally advanced rectal cancer. Using this approach, maChT administration can be delayed for several months, leading to concern for distant metastases. To counteract this, a novel treatment approach known as total neoadjuvant therapy (TNT) has gained popularity, in which patients receive both maChT and nCRT prior to resection. We utilized the National Cancer Database to examine temporal trends in TNT usage, and any potential effect on survival., Aim: To study the temporal trends in the usage of TNT and evaluate its efficacy compared to neoadjuvant chemoradiation., Methods: We queried the National Cancer Database for patients with locally advanced rectal cancer, Stage II-III, from 2004-2015 treated with nCRT or TNT. TNT was defined as maChT initiated ≥ 90 d prior to nCRT initiation. Overall survival was calculated from the date of diagnosis to the date of last contact or death using Kaplan-Meier curves to present the cumulative probability of survival, with log-rank statistics to assess significance. Multivariable cox regression was used to identify predictors of survival and propensity score analysis accounted for bias., Results: We identified 9066 eligible patients, with 8812 and 254 patients receiving neoadjuvant chemoradiation followed by maChT and TNT, respectively. Nodal involvement, stage III disease, and treatment in recent years were predictive of TNT use. There was greater use of TNT with more advanced stage, specifically > 1 node involved (odds ratio [OR] = 2.88, 95% confidence interval [CI]: 2.11-3.93, P < 0.01) and stage III disease (OR = 2.88, 95%CI: 2.11-3.93, P < 0.01). From 2010 to 2012 the use of TNT increased (OR = 2.41, 95%CI: 1.27-4.56, P < 0.01) with a greater increase from 2013 to 2015 (OR = 6.62, 95%CI: 3.57-12.25, P < 0.01). Both the TNT and neoadjuvant chemoradiation arms had a similar 5-year survival at 76% and 78% respectively. Multivariable analysis with propensity score demonstrated that increased age, high comorbidity score, higher grade, African American race, and female gender had worse overall survival., Conclusion: Our data demonstrates a rising trend in TNT use, particularly in patients with worse disease. Patients treated with TNT and nCRT had similar survival. Randomized trials evaluating TNT are underway., Competing Interests: Conflict-of-interest statement: The authors have no conflicts of interest to declare., (©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Management options for unresected malignant pleural mesothelioma (MPM) are largely limited to palliative chemotherapy and best supportive care. This study sought to delineate subgroups most likely to benefit from chemotherapy., Patients and Methods: The National Cancer Database was queried for newly-diagnosed unresected sarcomatoid, biphasic, and/or metastatic (M1) MPM. Statistics included Kaplan-Meier overall survival (OS) analysis with and without propensity matching, landmark Kaplan-Meier analysis to address immortal time bias, and multivariable Cox proportional hazards modeling in all patients as well as within histologic/M-classification-based subcohorts., Results: Of 4655 patients (48% chemotherapy, 52% best supportive care), 15%, 27%, and 40% had epithelioid, biphasic, and sarcomatoid disease, respectively; 41% had M1 disease. The median OS in the chemotherapy and BSC cohorts was 10.4 versus 4.8 months (P < .001). OS differences persisted following landmark analysis (P = .038) and propensity matching (P < .001). Chemotherapy was associated with higher OS in M1 cases with unknown histology and M1 epithelioid patients (P < .001 for both). For non-epithelioid cases, chemotherapy was associated with higher OS for M0 (P < .001 for sarcomatoid and biphasic) but not M1 (P > .05 for both) disease., Conclusions: Chemotherapy may benefit metastatic epithelioid and non-metastatic non-epithelioid MPM to a greater degree than metastatic non-epithelioid disease. Causation, however, is not implied, and careful patient selection in this population cannot be understated., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: The purpose of the current study is to determine the efficacy of a PI3K/mTOR dual inhibitor, LY3023414, on established EAC in an in vivo model., Background: Esophageal adenocarcinoma (EAC) is a highly lethal cancer with limited treatment options. The PI3K/mTOR pathway is upregulated in EAC and may be a target for novel therapies., Methods: Esophagojejunostomy was performed on Sprague-Dawley rats to induce carcinogenesis, and LY3023414 was cyclically administered intraperitoneally between 32 and 40 weeks postsurgery to treatment animals. Magnetic resonance imaging (MRI) and histology were used to determine clinical response. Immunohistochemistry, immunofluorescence, and Western blot were used to validate apoptosis by cleaved caspase-3, proliferation by Ki67, and pathway inhibition, respectively., Results: Mean MRI tumor volume increased by 109.2% in controls (n = 32) and decreased by 56.8% in treatment animals (n=17) (P < 0.01). Treatment with LY3023414 demonstrated tumor volume increase in 0% (control = 46.4%) (P < 0.01), decrease in 58.8% (control = 7.1%) (P < 0.01), and stable volume in 41.2% (control = 46.4%) (P = 0.77). EAC prevalence in controls increased by 25%; whereas, prevalence in treatment animals decreased by 29.4% (P < 0.01). Approximately, 75% of treatment animals presenting with residual masses on MRI had a histological response >50%. Increased apoptosis by cleaved caspase-3 (P = 0.03) and decreased proliferation by Ki67 (P < 0.01) were demonstrated in the treatment arm, when compared with the control arm. On Western blot analysis of pathway checkpoints, p-mTOR (p=0.03) and PI3K-α (P = 0.04) were downregulated in treatment responsive residual tumors, when compared with controls., Conclusions: LY3023414 demonstrates efficacy against EAC in a preclinical model, establishing the rationale for clinical testing.

Esophageal adenocarcinoma (EAC) is an aggressive cancer necessitating the development of improved risk stratification tools for personalized care. Previously, microRNAs have been shown to correlate with the progression and prognosis of various cancer types; however, the value in EAC remains largely unexplored. We performed global microRNA profiling on 32 formalin-fixed, paraffin-embedded EAC specimens to identify microRNAs associated with progression. Literature search and pathway analysis further refined output to five significantly deregulated candidate biomarkers. Four of the five microRNAs (miR-652-5p, miR-7-2-3p, miR-3925-3p, and miR-219-3p) were validated by qRT-PCR. Survival outcomes were evaluated in testing set of 26 stage II/III EAC patients to determine the prognostic relevance of the selected microRNAs. In the testing set, miR-652-5p and miR-7-2-3p expressions were significantly associated with progression-free survival (p-value = .00771 and p-value = .00293). The highest area under receiver operating characteristic (ROC) curve was 0.8212 for the combination of miR-652-5p and miR-7-2-3p. Collectively, our findings demonstrated that the miR-652-5p/miR-7-2-3p signature may serve as a promising prognostic marker in patients with locally advanced EAC.

Despite many years of testing mink for serum antibodies against the Aleutian mink disease virus (AMDV) by counterimmunoelectrophoresis (CIEP) and elimination of reactors, this virus has remained the number one disease threat for the mink industry in Nova Scotia (NS). The objective of this study was to analyze CIEP test results to determine the success of the AMDV-control strategy in NS. A total of 2,964,920 CIEP test results from 82 ranches, spanning an eight-year period between 1998 and 2005, were analyzed. This survey included approximately 60% of the active ranchers in the province. The number of ranchers that tested their animals was 42 in 1998, gradually increased to 58 in 2003 and then showed some decline. The overall proportion of CIEP-positive mink was 3.34%, and varied between 5.22% in 1999 and 1.35% in 2005. The proportion of infected ranches ranged between 23.8% in 1998 and 70.7% in 2003. The overall trend was for a smaller proportion of infected animals but a larger proportion of infected ranches during this time period. Of the 82 ranches, 24 (29.3%) had negative CIEP in all tests, 15 (18.3%) had CIEP positive animals in every year tested, and 43 (52.4%) had positive and negative results in different years, indicating that AMDV infection was widespread in NS. There were 23 infected ranches with 8 years of uninterrupted testing. These ranchers performed 75.8% of the total samples tested (2,246,711), implying that they have diligently been trying to eradicate the virus. Infection persisted on three of these ranches for the entire 8 year period, and only two of the ranches remained CIEP negative for longer than four years. The average percentage of CIEP-positive mink on these ranches was 2.2, which was lower than 6.35% for the 33 infected ranches with occasional testing, and 73.6% and 82.4% for two ranches that had never used the CIEP test, showing that persistent test-and-removal strategy has been effective in reducing the prevalence of infected animals but has failed to eradicate the virus from most of the infected ranches., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Wnt family members are critical to many developmental processes, and components of the Wnt signaling pathway have been linked to tumorigenesis in familial and sporadic colon carcinomas. Here we report the identification of two genes, WISP-1 and WISP-2, that are up-regulated in the mouse mammary epithelial cell line C57MG transformed by Wnt-1, but not by Wnt-4. Together with a third related gene, WISP-3, these proteins define a subfamily of the connective tissue growth factor family. Two distinct systems demonstrated WISP induction to be associated with the expression of Wnt-1. These included (i) C57MG cells infected with a Wnt-1 retroviral vector or expressing Wnt-1 under the control of a tetracyline repressible promoter, and (ii) Wnt-1 transgenic mice. The WISP-1 gene was localized to human chromosome 8q24.1-8q24.3. WISP-1 genomic DNA was amplified in colon cancer cell lines and in human colon tumors and its RNA overexpressed (2- to >30-fold) in 84% of the tumors examined compared with patient-matched normal mucosa. WISP-3 mapped to chromosome 6q22-6q23 and also was overexpressed (4- to >40-fold) in 63% of the colon tumors analyzed. In contrast, WISP-2 mapped to human chromosome 20q12-20q13 and its DNA was amplified, but RNA expression was reduced (2- to >30-fold) in 79% of the tumors. These results suggest that the WISP genes may be downstream of Wnt-1 signaling and that aberrant levels of WISP expression in colon cancer may play a role in colon tumorigenesis.

Background: Prostate cancer is the most common cancer in American men and the second leading cause of cancer death. All clinical observations correlate poorly differentiated high-grade prostate cancer with disease-specific mortality. The HER2 cell membrane tyrosine kinase, a member of the epidermal growth factor receptor family that is the transcription product of the erbB2neu oncogene, and HER3, a receptor protein of the same family, are overexpressed in prostate cancer and prostatic intraepithelial neoplasia. The ligand for these receptors and another related family member, HER4, has recently been identified by independent investigator groups and called neu differentiation factor (NDF) or heregulin. In vitro treatment of HER2- and HER3- or HER2- and HER4-expressing breast cancer cells stimulates tyrosine phosphorylation of HER2 and produces changes in the rate of proliferation, degree of cellular differentiation, and synthesis of physiologic secretion products. There are no published reports on the expression of NDF and HER4 in prostate cancer or the in vitro effects of NDF in prostate cancer cells., Methods: Expression of NDF, HER2, HER3, and HER4 was studied in 24 frozen prostatectomy specimens by immunohistochemistry. The biologic effect of human recombinant NDF was studied in vitro, using the LNCaP, PC3, and DU145 human prostate cancer cell lines. HER and NDF protein expression was studied by immunohistochemistry. NDF mRNA was analyzed using reverse transcriptase polymerase chain reaction from whole RNA. The biologic effects of NDF on prostate cancer cells in vitro included cell proliferation, thymidine synthesis, induction of prostate-specific antigen mRNA, anchorage-dependent and anchorage-independent cell growth, and ploidy analysis. Data analysis was performed using Student's t test., Results: Observations in clinical prostatectomy specimens: Immunohistochemistry studies in clinical prostatectomy specimens demonstrate absence of significant NDF expression in prostate cancer, whereas it is expressed in 100% of the stroma, 100% of basal epithelial cells, and 58% of luminal cells in normal and benign hyperplastic prostatic tissue. The HER4 receptor protein is strongly expressed by normal prostate luminal cells, but not prostate cancer. Benign prostate tissue exhibits strong expression of HER2, HER3, and HER4 by basal cells, but only luminal cells significantly express HER4. Only 23% of prostate cancer specimens express HER4, while 95% express HER3 and 82% HER2. Prostatic intraepithelial neoplasia stained similarly to cancer for all proteins studied. Observations in prostate cancer cell lines: In vitro treatment with NDF significantly reduces aneuploidy and proliferation and growth of androgen-sensitive prostate cancer cells. Incubation with NDF also induces prostate-specific antigen mRNA in prostate cancer cells. In spite of displaying NDF mRNA, prostate cancer cells do not produce detectable NDF protein, but express HER2 and HER3 proteins., Discussion: These data suggest that NDF may be a paracrine differentiation factor involved in normal adult prostate physiology and that functional loss of the NDF/HER ligand/ receptor loop may be an early event associated with prostate tumorigenesis.

In 1990-1991, a national survey was conducted to estimate the prevalence of Salmonella species among Canadian commercial turkey flocks. Two hundred and seventy flocks were randomly selected across Canada. The proportion sampled from each province was selected according to each province's share of the national turkey market. Samples, consisting of 12 pooled litter and four pooled dust samples, were used to determine the Salmonella status of the environment of each flock. Additionally, a one kilogram sample of feed was taken from each flock premise. Salmonella was recovered from environmental samples in 234/270 (86.7%) of flocks and from feed samples in 26/266 (9.8%) of flocks. Forty-eight different Salmonella serovars were isolated from flock environmental samples. The most prevalent serovars were S. anatum, S. hadar, S. agona, S. heidelberg and S. saintpaul which were isolated from 53/270 (19.6%), 49/270 (18.1%), 49/270 (18.1%), 42/270 (15.6%) and 34/270 (12.6%) flocks, respectively.

Inflammatory malignant fibrous histiocytomas (IMFH) are rare tumors and are frequently associated with leukocytosis. In rare cases, leukemoid reactions were attributed to tumor production of unidentified hematopoietic factors. In this study, we used immunohistochemical techniques to show cytokine immunoreactivity in the malignant cells of two cases of IMFH presenting with leukemoid reactions and compared them with two malignant fibrous histocytomas, noninflammatory type. All four tumors stained positively for stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF), interleukin-2 (IL-2), IL-4, IL-5, interferon-alpha (IFN-alpha), and insulin-like growth factor-I. Other cytokines detected only in the two IMFH included IL-6, IL-7, IL-8, IFN-gamma, and keratinocyte growth factor. Granulocyte-macrophage-CSF, IL-3, and transforming growth factor-beta staining was present in one of the two IMFH tumors and was not present in the noninflammatory tumors. The immunohistochemical staining was localized to the malignant cells, suggesting deregulated cytokine expression consistent with their monocytic/histocytic origin. Expression of certain cytokines in the IMFH may account for the local inflammatory infiltrate, tumor fibrosis, and the aggressive nature of the malignant cells. We also detected elevated serum levels of SCF, G-CSF, IL-6, and tumor necrosis factor in one or both of the IMFH patients. These latter observations may explain the bone marrow hypercellularity and other paraneoplastic symptoms, including fever, malaise, and weight loss, observed in both patients. Different cytokines present in the two IMFH tumors appear to be responsible for the eosinophilic leukemoid reaction observed in one case and for the granulocytic leukemoid reaction observed in the other patient. They may also be responsible for expansion of the tumor-cell population, fibroblast proliferation, and enhanced secretion of extracellular collagen.

Gastrin, produced in the G-cells of the gastric antrum and regulating acid secretion in the stomach, also acts as a trophic factor in the gastrointestinal tract. Because of its possible role in colon cell proliferation and differentiation, evidence for its presence in normal colorectal mucosa and adenocarcinoma was sought. Utilizing tumors and matched normal mucosa from 26 patients, mature gastrin and progastrin were studied by immunohistochemistry. In normal colonic mucosal crypts, occasional cells stained concordantly for gastrin, progastrin, and chromogranin A, suggesting that they are of neuroendocrine origin. Adenomatous polyps stained neither for gastrin nor chromogranin A. In 22 of 23 adenocarcinomas, more than 50% of tumor cells stained for gastrin and progastrin. The expected gastrin transcript was demonstrable by polymerase chain reaction and RNase protection in tumors and by polymerase chain reaction in normal mucosa. Its identity was confirmed by sequencing the polymerase chain reaction product. A larger transcript containing Intron II was present in both cancers and normal mucosa but was barely discernible in the gastric antrum. Aberrant expression of gastrin may contribute to deregulated proliferation of many colorectal carcinomas.

Colorectal cancer provides a unique model for the study of molecular changes that are associated with tumorigenesis. The cancer evolves as an apparent ordered sequence from a benign to a malignant lesion in histopathological recognizable stages. Since it is relatively easy to isolate tissue representing each of these stages, studies of molecular events associated with tumor progression are feasible. Such studies have shown that multiple changes in gene structure, expression and activity occur during tumorigenesis.

The myc gene family encodes nuclear phosphoproteins that are thought to play a role in the control of cellular proliferation and differentiation. We have undertaken an immunohistochemical study assessing the expression of myc gene family proteins in individual cells of normal colonic mucosa, colorectal polyps, and colorectal adenocarcinomas. We screened a panel of mouse monoclonal antibodies that we raised against recombinant human c-myc and N-myc proteins for recognition of myc proteins in paraffin tissue sections. Two of these antibodies, H120C69 and H8C150, were selected for indirect immunoperoxidase staining of tissue sections from 16 normal mucosas, 24 polyps, and 30 adenocarcinomas. In normal colon, about 25% of the cells in the lower one-third of the crypts of Lieberkühn stain for myc-related protein. This distribution resembles that of proliferating cells in the crypt. Benign hyperplastic polyps resemble normal mucosa in their myc staining pattern, with about 25% of the cells positive. In adenomatous polyps, the putative precursors of adenocarcinomas, from 50 to 100% of the cells stain positively for myc protein. In these cases, stained cells extend to the luminal surface, consistent with the previously reported expansion of the proliferation zone in these lesions. All adenocarcinomas examined had increased levels of myc protein relative to normal mucosa. The tumor cells exhibited markedly heterogeneous myc staining patterns, both among different tumors and, in some cases, within a single tumor. Comparison with Ki-67 monoclonal antibody staining indicates that myc protein expression in many tumors is uncoupled from cellular proliferation. Surprisingly, we observed increased numbers of myc-expressing cells and increased levels of myc protein in histologically normal colon directly adjacent to tumor, suggesting that many colorectal carcinomas secrete growth factors that activate gene expression in neighboring normal mucosa.

All animals that are to be killed, whether for food, for humane reasons, or because they are homeless, must receive a quick and painless death. In some smaller communities, veterinary or humane society expertise may not be readily available to humanely kill stray and unwanted animals. An alternative that provides for a humane death for the animal is by shooting. The following guidelines are intended to assist persons who must perform this usually distasteful task; they contain recommended techniques that will help to ensure that any animals killed by shooting will die in a humane way.

A nation-wide survey was conducted to estimate the prevalence of Salmonella enteritidis and other salmonellas among Canadian commercial broiler flocks. Environmental (litter and/or water) samples from 226 of 294 (76.9%) randomly selected flocks were contaminated with salmonellas. Litter samples were more often contaminated with salmonellas than water samples (47.4 v. 12.3%). Fifty different salmonella serovars were isolated. The most prevalent serovars were S. hadar, S. infantis, and S. schwarzengrund; they were isolated from samples of 98/294 (33.3%), 26/294 (8.8%), and 21/294 (7.1%) flocks, respectively. Feed samples of 39/290 (13.4%) flocks were contaminated with salmonellas. Salmonella enteritidis was isolated from the environmental samples of 9/294 (3.1%) flocks. Salmonella enteritidis phage type (PT) 8 was isolated from seven flocks, and PT 13a from two flocks.

Musculature from 198 Canadian cattle with suspected lesions of eosinophilic myositis were examined histologically and by pepsin digestion. Sera from 51 of the 198 animals were also examined by enzyme-linked immunosorbent assay (ELISA) for anti-Trichinella antibodies. Viable larvae of Trichinella were not recovered from any of the cattle but one animal from Ontario tested positive for anti-Trichinella antibodies. Histologically, focal and/or diffuse eosinophilic myositis lesions were observed in 149 (75.2%) of the animals studied. Other conditions identified were sarcocystiosis, abscesses, cysticercosis, steatosis, fibrosis, granuloma, lymphosarcoma and necrosis. Sarcocystiosis was identified in 105 of the 198 animals in both normal and affected musculature. The study indicates that trichinosis is not a primary cause of eosinophilic myositis in cattle.

Trichinella spiralis spiralis infections were established in cattle by gavage and by feeding infected musculature in the ration. Trichinae were present in greatest numbers in masseter, tongue and diaphragm. Trichinella spiralis nativa had a low infectivity to cattle although a light infection was established in one cow by a heavy challenge. Cattle had an aversion to eating musculature unless it was camouflaged with molasses. Clinical signs of reluctance to eat and masticate were observed between 10 and 30 days postinfection. Eosinophil counts started to increase at seven days and peaked at about 30 days postinfection. By day 60 eosinophil counts returned to near preinfection levels but in animals examined greater than 90 days postinfection, the counts were variable. Focal lesions of eosinophilic myositis were observed up to about 90 days postinfection. Little cellular reaction was observed surrounding trichinae after muscle invasion and cyst development was completed except for cysts undergoing disintegration. Seroconversion occurred in all cattle examined between 7 and 14 days postinfection. Seroconversion was associated with IgG1 and IgG2 immunoglobulins. Peak levels of antibody occurred between 30 and 60 days. Cattle examined at 182 and 369 days postinfection showed a gradual decrease in antibody levels over time.

Enzyme linked immunosorbent assay (ELISA) using a fraction of larval Taenia hydatigena cyst fluid antigen was carried out on 469 bovine sera collected at slaughter from feedlot cattle for the presence of anticysticercosis antibodies. Cysticerci, in low numbers, were found in the heart, tongue and/or masseter muscles of 84 of the 469 cattle at postmortem inspection. Only nine sera gave positive ELISA reactions and in only one of these nine animals were cysticerci found. Within the limitations of this study, the high rate of false negative and false positive reactions suggests that the ELISA with the antigen used is not a satisfactory procedure to diagnose cysticercosis in cattle, at least in animals with light infections.

An outbreak of cysticercosis (infestation with the larvae of Taenia saginata) occurred in feedlot cattle in Ontario in 1986. Two hundred and thirty-three of 271 steers were confirmed histologically to be positive for cysticerci. Nineteen (8.2%) animals had viable cysticerci, 87 (37.3%) had degenerated cysticerci, 77 (33.0%) had mineralized cysticerci, and 50 (21.5%) steers had lymphoid granulomas consistent with cysticercosis. Three viable cysticerci were partly evaginated and one degenerate cysticercus was fully evaginated.

Six cesarean-derived lambs were inoculated either with 4.5 X 10(4), 4.5 X 10(6) or 4.5 X 10(8) Mycoplasma mycoides subsp. mycoides intratracheally. One animal receiving the intermediate dose died four days post-inoculation, the two receiving the high dose died six days postinoculation, while one receiving the low dose died eight days postinoculation. The two surviving lambs were challenged on day 20 postinoculation with 1 X 10(8) organisms subcutaneously and 2 X 10(9) organisms intravenously. One animal died eight days following this challenge while the other survived and was killed. Six conventionally reared lambs challenged with 90 to 8500 organisms by intranasal and intraocular instillation failed to become infected. Three conventionally reared calves were each inoculated with 1 X 10(8) organisms by each of intratracheal, subcutaneous and intravenous routes. They were killed 20 days post-inoculation without having shown any clinical signs.

Colorectal carcinoma serves as a model system for the study of changes in gene expression and structure relating to tumorigenesis. In this study, the levels of c-myc, N-myc and L-myc mRNAs were assessed in normal human colonic mucosa in 33 cases representing different stages of adenocarcinoma and in 36 adenomatous polyps, the presumed premalignant stage. Consistent with the findings of Erisman et al. (1985), we found that the c-myc gene was overexpressed (3-24-fold) in approximately two-thirds of the tumors examined. Amplification of the gene (3-4-fold) was observed in 2 of 12 tumors examined and did not correlate with the level of expression. Greater amounts of c-myc-specific mRNA than occur in normal tissue was also found in about two-thirds of the polyps examined demonstrating that premalignant lesions also overexpress the gene. N-myc and L-myc specific transcripts can be detected at low abundance in normal colonic mucosa. These genes were found to be frequently overexpressed in tumors and polyps, but in most cases the level of overexpression was modest. A single case of adenocarcinoma showed an approximately 30-fold increase in the level of N-myc mRNA without gene amplification. Adenomatous polyps more frequently overexpressed the L-myc gene than tumors.

The clinical, macroscopic, and microscopic features of 10 isolated cases of myodegeneration in foals were compared. Low values for selenium and vitamin E content were found in the hay and oats from one breeding stable. Serum selenium concentrations in mares at this stable were also low. Creatinine phosphokinase and serum glutamic oxaloacetic transaminase activities were increased in 2 young foals at this stable; in 1 of these foals, both enzymatic activities were markedly reduced after treatment with vitamin E and selenium. Nutritional myodegeneration was suggested as a diagnosis in this stable, on the basis of the histologic findings, feed analyses, serum selenium values, response to treatment, and enzymatic determinations. Nine other isolated cases of nutritional myodegeneration were tentatively diagnosed on the basis of macroscopic and microscopic findings and the young age of the animal. The gross lesions included pale areas in the myocardium and skeletal muscle masses. Histologically, lesions were characterized by fragmentation and hyaline and granular changes in swollen muscle fibers in widely distributed skeletal muscle masses.

Sheep pulmonary adenomatosis has recently been reported in Canada. The literature is briefly reviewed and an account of the present status of the disease in Canada is described.Sheep pulmonary adenomatosis was first diagnosed in Canada in December 1979 in a first generation descendent of sheep imported from Great Britain. In March 1980 two further cases of sheep pulmonary adenomatosis were diagnosed in a second flock. A total of 43 sheep involving eight flocks from five provinces have been observed from December, 1979 to May, 1981. The clinical signs and laboratory findings were similar to those described in sheep from other countries. It is estimated that 30% of Canadian flocks may contain sheep imported from Great Britain during the 1970's.

A mail survey of disease occurrence in Canadian sheep flocks was conducted. The survey, which covered the period from September 1982 to August 1983, utilized flocks on the Record of Performance (ROP) sheep program and relatively complete data were available from 116 flocks. Data about lambing rates, incidence of a variety of lamb and ewe diseases and reasons for culling were obtained. At the same time a retrospective evaluation of records of diagnoses of sheep diseases recorded at diagnostic laboratories across the country was performed. Data from the years 1978 to 1982 were obtained and summarized. A lambing percentage of 153% (1.53 lambs live born per ewe lambing) was observed and an additional 0.05 lambs were stillborn. The major identified causes of mortality amongst lambs were starvation, pneumonia, scours and accidents. Pasteurella spp. were the etiological agents most commonly associated with pneumonia in lambs and Escherichia coli had the same predominant position with regards to nonparasitic scours. A large discrepancy existed between the proportional mortality rates for internal parasites and coccidiosis as determined from the farm survey data compared to diagnostic laboratory data. This suggests that clinical parasitism may not be adequately recognized at the farm level. Abortions in ewes occurred in approximately half the flocks, but generally at a low level and no severe abortion storms occurred. Pneumonia was the most commonly identified cause of mortality in ewes and although Pasteurella spp. appear to be the most important etiological agents, regional differences were apparent.(ABSTRACT TRUNCATED AT 250 WORDS)