The climatic impact of food consumption in a representative sample of Irish adults and implications for food and nutrition policy.

OBJECTIVE: To evaluate the greenhouse gas emissions (GHGE) associated with the diet of Irish adults. DESIGN: GHGE were estimated by applying conversion factors to habitual food consumption data taken from the National Adult Nutrition Survey, which was representative of the population. Descriptive analyses were undertaken for GHGE for the total population, as well as accounting for energy misreporting and across categories of sociodemographic and socio-economic factors and tertiles of emissions. SETTING: Republic of Ireland. SUBJECTS: Adults aged 18-87 years (n 1500). RESULTS: The GHGE derived from daily dietary intakes was estimated as 6·5 kg of CO2 equivalents (CO2eq) per person. Males, younger consumers, those with secondary education and student employment status were associated with significantly higher GHGE. Red meat was the highest contributor to GHGE with 1646 g CO2eq arising from a mean intake of 47 g/d. Dairy and starchy staples were the next largest dietary GHGE sources, with mean daily emissions of 732 g CO2eq and 647 g CO2eq, respectively. The lowest emissions were associated with consumption of vegetables, fruits and legumes/pulses/nuts. CONCLUSIONS: Based on profiling using actual food consumption data, it is evident that one single measure is not sufficient and a range of evidence-based mitigation measures with potential to lower emissions throughout the food chain should be considered. The research contributes towards an improved understanding of the climatic impact of the dietary intakes of Irish adults and can serve to inform a sustainability framework to guide action in food and nutrition policy development.

Victimisation in urban primary schools of high-poverty areas: associations with health-related quality of life, depression and social support.

BACKGROUND: Although a widespread issue, research on victimisation among primary school children in high-poverty regions is limited. The aim of this research was to explore victimisation incidence and associated mental health correlates from first-wave data of the 'Healthy Schools' programme in a high-poverty urban region. METHOD: The study explored victimisation incidences among 458 Irish primary school children and associations with depression, health-related quality of life (HRQoL) and social support. RESULTS: Victimisation (33.8%) was consistent with recent literature. On the stand-alone victimisation question, victims scored lower on all HRQoL subscales compared with nonvictims. Further categorisation revealed that frequent victims scored lower on four of these subscales, compared with nonvictims. Furthermore, over half of children felt that their school was not doing enough to combat school aggression. CONCLUSIONS: Although from a high-poverty area, rates were consistent with data from more affluent areas. Results stress an importance on specific school aggression behaviours when measuring victimisation rates, along with corresponding health consequences. Future research should continue to adopt the behaviour-based assessment of victimisation to provide an overall picture of the problem.

Parenthood, Child Care, and Heroin Use: Outcomes After Three Years.

BACKGROUND: Internationally there is a lack of measurement on the impact of childcare on people who use drugs. OBJECTIVES: The aim of this article was to longitudinally measure drug use, familial and social status and criminal involvement between parents and nonparents who use heroin and have children in their care. METHODS: From 2003 to 2006, 404 participants were recruited to the Research Outcome Study in Ireland Evaluating Drug Treatment Effectiveness (ROSIE) as part of a longitudinal cohort study design. Participants completed the Maudsley Addiction Profile and 88% (n = 356) completed interviews at the 3-year period. One way between groups ANOVA with post hoc tests and backward, stepwise multiple regression were employed for analysis. RESULTS: At follow-up, parents who had children in their care used heroin (p = .004), illicit methadone (p ≤ .001) and cocaine (p = .024) on fewer days than those who had no children, or those who had children but did not have children in their care. These differences were not observed at intake. Living with someone at intake who used drugs was found to be significantly associated with increased heroin (p ≤ .001), benzodiazepine (p = .039), and tobacco (p = .030) use at 3 years. Furthermore, a change in childcare status to caring for a child was associated with increased cannabis use (p = .025). Conclusion/Importance: While caring for children was associated with reduced heroin use at 3 years, living with a person who used at intake removed this effect, thus indicating that while individual based addiction theories reflected observed outcomes, social network connectedness was more influential.

Prognostic significance of tumor budding in rectal cancer biopsies before neoadjuvant therapy.

Tumor budding is an increasingly important prognostic feature for pathologists to recognize. The aim of this study was to correlate intra-tumoral budding in pre-treatment rectal cancer biopsies with pathological response to neoadjuvant chemoradiotherapy and with long-term outcome. Data from a prospectively maintained database were acquired from patients with locally advanced rectal cancer who underwent neoadjuvant chemoradiotherapy. Pre-treatment rectal biopsies were retrospectively reviewed for evidence of intra-tumoral budding. Multivariate logistic regression was used to identify factors contributing to cancer-specific death, expressed as hazard ratios with 95% confidence intervals. Of the 185 patients with locally advanced rectal cancer, 89 patients met the eligibility criteria, of whom 18 (20%) exhibited budding in a pre-treatment tumor biopsy. Intra-tumoral budding predicted a poor pathological response to neoadjuvant chemoradiotherapy (higher ypT stage, P=0.032; lymph node involvement, P=0.018; lymphovascular invasion, P=0.004; and residual poorly differentiated tumors, P=0.005). No patient with intra-tumoral budding exhibited a tumor regression grade 1 or complete pathological response, providing a 100% specificity and positive predictive value for non-response to neoadjuvant chemoradiotherapy. Intra-tumoral budding was associated with a lower disease-free 5-year survival rate (33 vs 78%, P<0.001), cancer-specific 5-year survival rate (61 vs 87%, P=0.021) and predicted cancer-specific death (hazard ratio 3.51, 95% confidence interval 1.03-11.93, P=0.040). Intra-tumoral budding at diagnosis of rectal cancer identifies those who will poorly respond to neoadjuvant chemoradiotherapy and those with a poor prognosis.

Predicting response to vascular endothelial growth factor inhibitor and chemotherapy in metastatic colorectal cancer.

BACKGROUND: Bevacizumab improves progression free survival (PFS) and overall survival (OS) in metastatic colorectal cancer patients however currently there are no biomarkers that predict response to this treatment. The aim of this study was to assess if differential protein expression can differentiate patients who respond to chemotherapy and bevacizumab, and to assess if select proteins correlate with patient survival. METHODS: Pre-treatment serum from patients with metastatic colorectal cancer (mCRC) treated with chemotherapy and bevacizumab were divided into responders and nonresponders based on their progression free survival (PFS). Serum samples underwent immunoaffinity depletion and protein expression was analysed using two-dimensional difference gel electrophoresis (2D-DIGE), followed by LC-MS/MS for protein identification. Validation on selected proteins was performed on serum and tissue samples from a larger cohort of patients using ELISA and immunohistochemistry, respectively (n = 68 and n = 95, respectively). RESULTS: 68 proteins were identified following LC-MS/MS analysis to be differentially expressed between the groups. Three proteins (apolipoprotein E (APOE), angiotensinogen (AGT) and vitamin D binding protein (DBP)) were selected for validation studies. Increasing APOE expression in the stroma was associated with shorter progression free survival (PFS) (p = 0.0001) and overall survival (OS) (p = 0.01), DBP expression (stroma) was associated with shorter OS (p = 0.037). Increasing APOE expression in the epithelium was associated with a longer PFS and OS, and AGT epithelial expression was associated with a longer PFS (all p < .05). Increasing serum AGT concentration was associated with shorter OS (p = 0.009). CONCLUSIONS: APOE, DBP and AGT identified were associated with survival outcomes in mCRC patients treated with chemotherapy and bevacizumab.

The influence of CTGF single-nucleotide polymorphisms on outcomes in Crohn's disease.

OBJECTIVE: To examine the association between single-nucleotide polymorphisms (SNPs) in CTGF (connective tissue growth factor) and patient outcomes after terminal ileal resection for Crohn's disease. BACKGROUND: The primary indication for intestinal resection in Crohn's disease is fibrostenotic terminal ileal disease. CTGF is a cytokine overexpressed in the intestine of patients with Crohn's disease that influences outcomes in other disease processes. METHODS: DNA was extracted from formalin-fixed, paraffin-embedded tissue from 147 patients with Crohn's disease who had undergone terminal ileal resection between 1981 and 2009. Genotyping was performed for 4 CTGF SNPs (rs9402373, rs12526196, rs6918698, and rs9399005), which modulate nuclear factor binding and CTGF production, and a smad3 SNP (rs17293632) involved in the CTGF pathway. Patients were phenotyped using the Montreal Disease Classification. RESULTS: Sixty-seven of 147 patients (45.6%) were male; the mean age at diagnosis was 30.3 ± 12.6 years and the mean follow-up duration was 8.3 ± 7.1 years. Genotype-phenotype analysis demonstrated that the rs6918698GG genotype was associated with an older age of disease onset [>40 years; 30.6% vs 13.2%; odds ratio (OR): 2.891; 95% confidence interval (CI): 1.170-7.147). The rs9402373CC genotype was positively associated with type B1 disease (50.7% vs 26.3%; OR: 2.876; 95% CI: 1.226-6.743) and negatively associated with B2 disease (37.0% vs 65.0%; OR: 0.317; 95% CI: 0.144-0.699). None of the 5 SNPs assessed influenced clinical or surgical recurrence of Crohn's disease after intestinal resection. On multivariate analysis, male sex odds ratio (OR): 0.235; 95% CI: 0.073-0.755; P = 0.015] and never having smoked tobacco (OR: 0.249; 95% CI: 0.070-0.894; P = 0.033) reduced the risk, whereas having a prior appendectomy increased the risk (OR: 5.048; 95% CI: 1.632-15.617; P = 0.005) of surgical recurrence. CONCLUSIONS: These data implicate the rs6918698GG genotype with an age of disease onset of greater than 40 years in Crohn's disease whereas the rs9402373CC genotype is associated with a nonstricturing, nonpenetrating disease phenotype. CTGF SNPs do not influence the rate of recurrence after terminal ileal resection for Crohn's disease.

A comparative study of short- and medium-term outcomes comparing emergent surgery and stenting as a bridge to surgery in patients with acute malignant colonic obstruction.

BACKGROUND: The use of self-expanding metal stents as a bridge to surgery in the setting of malignant colorectal obstruction has been advocated as an acceptable alternative to emergency surgery. However, concerns about the safety of stenting have been raised following recent randomized studies. OBJECTIVES: The aim of the current study was to compare outcomes. DESIGN: This was an observational, comparative study. SETTINGS: This study was conducted at a tertiary referral center and university teaching hospital. PATIENTS AND INTERVENTIONS: Patients with malignant colonic obstruction (n = 49) treated by either emergency surgery (n = 26) or with stent placement (n = 23) as a bridge to surgery were identified and followed. MAIN OUTCOME MEASURES: Short-term outcomes including stoma rates and postoperative morbidity and medium-term oncological outcomes were compared based on an "intention-to-treat" analysis. RESULTS: Patients in both groups were well matched on clinicopathological parameters. Technical and clinical successful stent deployment was achieved in 91% and 83%. This did not adversely impact cancer-specific and overall survival (log rank = nonsignificant). No difference was observed in stoma rates, primary anastomosis rates, perioperative mortality rates, or reoperation rates between the 2 groups. Significantly fewer patients underwent total colectomy in the stent group in comparison with the emergency surgery group (1/23 vs 6/26: p = 0.027). There was no difference in postoperative morbidity (59% vs 66%: p = 0.09). There was a significant reduction in readmission rates in the stent group (5/26 vs 0/23: p = 0.038). LIMITATIONS: The small sample size of this study could lead to type II error. In addition, the study was nonrandomized and demonstrated a limited length of follow-up. CONCLUSION: Despite a high rate of technical and clinical success in selected patients with colonic obstruction, stenting has no impact on stoma rates. Despite concerns about the rate of stent-associated perforation, stenting does not adversely impact disease progression or survival. Future comparative trials are essential to better define the role of stenting in this setting and to ensure that we are not using costly technology to create an elective operative situation without concomitant patient benefits.

Stage II colonic adenocarcinoma: a detailed study of pT4N0 with emphasis on peritoneal involvement and the role of tumour budding.

AIMS: Evaluation of peritoneal involvement in colonic cancer (CC) can be difficult. We studied pT4N0 cancers and their association with pathological prognostic markers, including tumour budding. METHOD AND RESULTS: Tumours were classified as (i) at the peritoneal surface or free in the peritoneal cavity (pT4a subgroup n = 44); (ii) directly invading adjacent organ (pT4b subgroup n = 8); or (iii) showing inflammatory involvement of the peritoneum (pT4I subgroup n = 25). A published pT3N0 cohort was used to compare Stage II subgroups. Standard pathological markers including tumour budding were assessed. Elastin staining was performed in the pT4I subgroup. Seventy-seven Stage II CCs met inclusion criteria. There was no significant difference in survival across subgroups. pT4b tumours were larger than pT4a tumours (P < 0.001). Over-represented features in pT4a versus pT4b tumours were tumour budding (P = 0.02) and infiltrative margin (P = 0.02). Tumour budding did not predict survival. Using multivariate analysis, neural invasion was the only parameter predictive of survival (hazard ratio = 2.8; 95% CI 1.2-6.4; P = 0.02). CONCLUSION: Stage II pT4I CCs have a similar outcome to T4a/b tumours. Elastin staining is useful in defining this group. Tumour budding may facilitate peritoneal invasion in pT4a tumours, but does not predict outcome in pT4N0 disease. Only neural invasion independently predicted poor outcome.

A prospective analysis of patient outcome following treatment of T3 rectal cancer with neo-adjuvant chemoradiotherapy and transanal excision.

BACKGROUND: Local excision is an alternative to anterior or abdomino-perineal resection in patients with early rectal cancer. In more advanced disease, neo-adjuvant therapy (CRXT) can result in significant disease regression such that local excision may be considered. The primary aim was to assess oncological outcome in patients with T3 rectal cancer treated with CRXT and local excision due to unsuitability for or aversion to anterior resection and stoma. The secondary aim was to examine oncological outcomes in patients treated in a similar way in the published literature. METHODS: Between July 2006 and July 2009, patients with rectal cancer staged T3, N0/N1, M0 who were deemed unfit for or who refused anterior resection were offered long-course CRXT. Patients were restaged 8 weeks following completion. If there was a good response (regression grade 2 or 3 clinically and radiologically), full thickness transanal excision was performed. All patients were followed regularly (monthly CT abdomen/pelvis and annual endoscopy) to assess for recurrence of disease. A literature search of PubMed was performed to identify all prospective data available of T3 rectal cancers managed with CRXT and local excision. RESULTS: Ten patients were treated over 3 years. Six patients had complete pathological response, while four patients had a partial response. The resection margins following local excision were clear in all. There was no local recurrence (median follow-up 24 months, range 9-42 months). CONCLUSION: Neo-adjuvant chemoradiotherapy and local excision is an option in patients unfit for or averse to major surgical resection if there is a good response to CRXT.

Consumer intention towards the phosphate-reduced processed meat products using the extended theory of planned behaviour.

Phosphates are essential for maintaining various quality attributes of processed meat products such as water-binding properties, texture and sensory properties and their removal would drastically change the products' technical and sensory qualities. Currently, meat industries are faced with the challenge of removing phosphates to address the consumers' demand to remove the negatively perceived synthetic additives from processed meat products. This study measured these consumers' purchase intention of phosphate-reduced processed meat products with different quality, using the extended theory of planned behaviour (TPB). An online survey was conducted among the consumers (n = 548) of the Republic of Ireland (ROI) to predict their knowledge and attitude towards phosphate additives. Analysis of the survey responses showed that about two-third of the participants consumed processed meat products 5-6 times per week. The results of multiple linear regression showed that the theory constructs attitude, subjective norms, perceived health risks significantly (P < 0.05) influenced the consumer behavioural intention whereas the perceived behavioural control (PBC) produced insignificant impacts. The results also revealed that the extended TPB model predicted the consumers' intention with better explanatory power (adjusted R2 = 0.46) than the original TPB model. In conclusion, various recommendations and implications were developed based on the results to improve the consumers' purchase intention of these products.

Surgery, Crohn's disease, and the biological era: has there been an impact?

INTRODUCTION: The management of Crohn's disease (CD) has changed considerably over the last 20 years. Immunomodulators and biological therapies now play a role in treating patients with CD, but little is known of their influence on surgical rates. AIM: To review the surgery rates for CD in an Irish university hospital over a 20-year period and to determine whether newer therapies had an impact on surgical rates. METHOD: Seven hundred twenty-two patients attending St Vincent's University Hospital, Dublin, with CD over a 20-year period (January 1986 to December 2005) were identified. The patients were divided into quartiles. Resection rates were determined in all the quartiles, at both 1 and 3 years from diagnosis. RESULTS: A decline in surgery, 3 years from diagnosis, was noted between the first quartile (72 patients, 40%) and the second quartile (58 patients, 32%; P=0.03). No significant change in surgical rates at 3 years occurred between the other 3 quartiles (32%, 30%, and 35%, respectively; P=NS). The patients who required a resection within 3 years were diagnosed at a younger age in later years. There was a similar predominance of 60% of female patients requiring surgery in all groups. The patients requiring surgery were twice as likely to be ex-smokers or current smokers in all groups. Use of infliximab, within 3 years from diagnosis, increased from 0, 0, and 16 patients (8.8%) to 40 patients (22.1%) in the last quartile. The majority of patients were treated with infliximab on an "on demand" basis. Use of infliximab earlier within the course of the disease was seen in later quartiles (ie, within 1 y of diagnosis): 0, 0, 6, and 21 patients. CONCLUSION: Despite the introduction of infliximab over the past 10 years, no demonstrable difference has been seen in the rates of patients requiring resection surgery within 3 years of diagnosis. The reasons for this are unclear, but may relate to episodic treatment, rather than regular maintenance treatment. Female patients and smokers seem to be particularly at risk of resection surgery.

Treatment and outcomes of anorectal melanoma.

INTRODUCTION: anorectal melanoma is an uncommon disease constituting less than 3% of all melanomas. Due to its rarity, there are a lack of randomized control trials regarding appropriate management and current evidence is based mainly on retrospective studies. METHODS: in view of the controversial surgical treatment of anorectal melanoma, we review the most published literature in an attempt to elucidate its typical clinical features along with current thinking with respect to management approaches to this aggressive disease. Using the keywords "anorectal" and "malignant melanoma", a medline search of all articles in English was performed and the relevant articles procured. Additional references were retrieved by cross reference from key articles. RESULTS: anorectal melanoma affects the elderly with a slight preponderance for females. It commonly presents disguised as benign disease with local bleeding or suspicion for haemorrhoidal disease. There is no convincing evidence to indicate that radical resection of primary anorectal melanoma is associated with improvement in local control or survival, and local excision is an acceptable treatment option. CONCLUSION: optimum management depends on several factors and the therapeutic goals should be to lengthen survival and preserve quality-of-life. Given that wide local excision is a more limited intervention with comparable survival it should be considered as the initial treatment choice. Unfortunately prognosis for patients with this disease remains poor despite choice of treatment strategy with overall five year disease-free survival less than twenty percent in most studies.

The Impact of the Covid-19 Pandemic on Food Consumers' Awareness of Antimicrobial Resistance, OneHealth, and Animal Welfare Information on Food Labels.

Covid-19 is a OneHealth crisis with far-reaching and unexpected impacts on many aspects of society. Previous OneHealth issues, such as antimicrobial resistance (AMR), have not received a similar level of attention or action from the public despite representing significant public health and economic threats to society. The current study aimed to explore whether the Covid-19 pandemic may act as a catalyst to increase public awareness related to OneHealth issues, in particular, AMR. This short paper presents overview findings from a survey carried out in September 2020 with a representative sample of food consumers on the island of Ireland (n = 972). The survey revealed Covid-19 had increased awareness of AMR amongst 47% of respondents; increased awareness of connected animal and human health amongst 43% of respondents; and increased awareness of animal welfare information on food labels amongst 34% of respondents. A cluster analysis revealed five distinct consumer segments impacted differently by Covid-19. These segments differed in their levels of objective and subjective knowledge of antibiotic use practises in farming, AMR risk perception, and attributions of responsibility for action on AMR. Findings are discussed with respect to future efforts by the agri-food sector to communicate with the public about AMR and responsible antibiotic use in farming, with particular emphasis on the implications for strategies that incorporate front-of-pack labelling.

Variables associated with successful performance on the National Board of Chiropractic Examiners Part IV examination.

OBJECTIVE: The objectives of this study were to (1) identify factors predictive of performance on the National Board of Chiropractic Examiners Part IV exam and (2) investigate correlations between the scores obtained in the Part I, Part II, Physiotherapy, and Part III exams and the Part IV examination. METHODS: A random sample of 1341 records was drawn from National Board of Chiropractic Examiners data to investigate the relationships between the scores obtained on the National Board of Chiropractic Examiners exams. A hierarchical multiple regression analysis related the performance on Part IV to examinee's gender, Part IV repeater status, and scores obtained on the Part I, Part II, Physiotherapy, and Part III exams. RESULTS: The analyses revealed statistical relations among all National Board of Chiropractic Examiners exams. The correlations between Part IV and Part I ranged from r = .31 to r = .4; between Part IV and Part II from r = .34 to r = .45. The correlation between Part IV and Physiotherapy was r = .44; between Part IV and Part III was r = .46. The strongest predictors of the Part IV score were found to be examinees' scores in Diagnostic Imaging, ß̂ = .19, p < .001; Chiropractic Practice, ß̂ = .17, p < .001; Physiotherapy, ß̂ = .15, p < .001; and the Part III exam ß̂ = .19, p < .001. CONCLUSIONS: Performance on the National Board of Chiropractic Examiners Part IV examination is related to the performance in all other National Board of Chiropractic Examiners exams.

The transition to digital presentation of the diagnostic imaging domain of the Part IV examination of the National Board of Chiropractic Examiners.

OBJECTIVE: This article introduces changes made to the diagnostic imaging (DIM) domain of the Part IV of the National Board of Chiropractic Examiners examination and evaluates the effects of these changes in terms of item functioning and examinee performance. METHODS: To evaluate item function, classical test theory and item response theory (IRT) methods were employed. Classical statistics were used for the assessment of item difficulty and the relation to the total test score. Item difficulties along with item discrimination were calculated using IRT. We also studied the decision accuracy of the redesigned DIM domain. RESULTS: The diagnostic item analysis revealed similarity in item function across test forms and across administrations. The IRT models found a reasonable fit to the data. The averages of the IRT parameters were similar across test forms and across administrations. The classification of test takers into ability (theta) categories was consistent across groups (both norming and all examinees), across all test forms, and across administrations. CONCLUSION: This research signifies a first step in the evaluation of the transition to digital DIM high-stakes assessments. We hope that this study will spur further research into evaluations of the ability to interpret radiographic images. In addition, we hope that the results prove to be useful for chiropractic faculty, chiropractic students, and the users of Part IV scores.

Does clinical validation and the implementation of new models of outpatient service delivery have the potential to reduce waiting lists? A pilot study in Letterkenny University Hospital.

INTRODUCTION: The traditional outpatient paradigm of seeing patients prior to diagnostic tests and treatment is unsustainable without additional funding. New models of service delivery such as "one-stop clinics", direct access to diagnostics and advanced nurse practitioner (ANP)-led clinics have the potential to improve the efficiency of existing services. METHODS: To determine the most effective changes to improve service provision, the reasons for encounter (RFE) to a urology clinic were assessed using the International Classification Primary Care. To test these changes, a clinical validation process was performed on existing waiting patients waiting ≥ 15 months. Direct access to diagnostics and an ANP-led clinic were introduced. The impact of this validation process was measured prospectively using independently-collated National Treatment Purchase Fund waiting list data. RESULTS: From January to December 2017, 1114 new patients were referred. The 3 most frequent RFEs were haematuria, urinary frequency/urgency and cystitis and accounted for 48% of referrals overall. A new outpatient pathway, combining direct access to diagnostics and an ANP-led clinic, was implemented on 508 existing patients waiting ≥ 15 months. The validation process resulted in referral directly to a consultant-led clinic in 36%, to an ANP-led clinic in 12%, direct access to diagnostics in 38% and removal in 13%. This change was implemented in July 2017 and there was a 76% reduction in the number of patients waiting ≥ 12 months by December 2017. CONCLUSION: New models of outpatient service delivery have the potential to reduce existing waiting lists and could be implemented in other Irish hospital groups.

Localization of nuclear cathepsin L and its association with disease progression and poor outcome in colorectal cancer.

Previous in vitro studies have identified a nuclear isoform of Cathepsin L. The aim of this study was to examine if nuclear Cathepsin L exists in vivo and examine its association with clinical, pathological and patient outcome data. Cellular localization (nuclear and cytoplasmic) and expression levels v of Cathespin L in 186 colorectal cancer cases using immunohistochemistry. The molecular weight and activity of nuclear and cytoplasmic Cathepsin L in vivo and in vitro were assessed by Western blotting and ELISA, respectively. Epithelial nuclear staining percentage (p = 0.04) and intensity (p = 0.006) increased with advancing tumor stage, whereas stromal cytoplasmic staining decreased (p = 0.02). Using multivariate statistical analysis, survival was inversely associated with staining intensity in the epithelial cytoplasm (p = 0.01) and stromal nuclei (p = 0.007). In different colorectal cell lines and in vivo tumors, pro- and active Cathepsin L isoforms were present in both the cytoplasm and nuclear samples, with pro-Cathepsin L at 50 kDa and active Cathepsin L at 25 kDa. Purified nuclear and cytoplasmic fractions from cell lines and tumors showed active Cathepsin L activity. The identification of nuclear Cathepsin L may play an important prognostic role in colorectal disease progression and patient outcome. Moreover, these findings suggest that altering active nuclear Cathepsin L may significantly influence disease progression.

High clusterin expression correlates with a poor outcome in stage II colorectal cancers.

The role of clusterin in tumor growth and progression remains unclear. Overexpression of cytoplasmic clusterin has been studied in aggressive colon tumors; however, no correlation between clusterin expression and survival in colorectal cancer has been identified to date. We assessed levels of clusterin expression in a group of stage II colorectal cancer patients to assess its utility as a prognostic marker. The study included 251 patients with stage II colorectal cancer. Tissue microarrays were constructed and immunohistochemistry done and correlated with clinical features and long term outcome. Dual immunofluorescence and confocal microscopy were used with terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling probes and clusterin antibody to assess the degree of co localization. Percentage epithelial cytoplasmic staining was higher in tumor compared with nonadjacent normal mucosa (P < 0.001). Within the stromal compartment, percentage cytoplamic staining and intensity was lower in tumor tissue compared with normal nonadjacent mucosa (P < or = 0.001). Survival was significantly associated with percentage epithelial cytoplasmic staining (P < 0.001), epithelial cytoplasmic staining intensity (P < 0.001), percentage stromal cytoplasmic staining (P = 0.002), and stromal cytoplasmic staining intensity (P < 0.001). Clusterin levels are associated with poor survival in stage II colorectal cancer.

Radiation and chemotherapy bystander effects induce early genomic instability events: telomere shortening and bridge formation coupled with mitochondrial dysfunction.

The bridge breakage fusion cycle is a chromosomal instability mechanism responsible for genomic changes. Radiation bystander effects induce genomic instability; however, the mechanism driving this instability is unknown. We examined if radiation and chemotherapy bystander effects induce early genomic instability events such as telomere shortening and bridge formation using a human colon cancer explant model. We assessed telomere lengths, bridge formations, mitochondrial membrane potential and levels of reactive oxygen species in bystander cells exposed to medium from irradiated and chemotherapy-treated explant tissues. Bystander cells exposed to media from 2Gy, 5Gy, FOLFOX treated tumor and matching normal tissue showed a significant reduction in telomere lengths (all p values <0.018) and an increase in bridge formations (all p values <0.017) compared to bystander cells treated with media from unirradiated tissue (0Gy) at 24h. There was no significant difference between 2Gy and 5Gy treatments, or between effects elicited by tumor versus matched normal tissue. Bystander cells exposed to media from 2Gy irradiated tumor tissue showed significant depolarisation of the mitochondrial membrane potential (p=0.012) and an increase in reactive oxygen species levels. We also used bystander cells overexpressing a mitochondrial antioxidant manganese superoxide dismutase (MnSOD) to examine if this antioxidant could rescue the mitochondrial changes and subsequently influence nuclear instability events. In MnSOD cells, ROS levels were reduced (p=0.02) and mitochondrial membrane potential increased (p=0.04). These events were coupled with a decrease in percentage of cells with anaphase bridges and a decrease in the number of cells undergoing telomere length shortening (p values 0.01 and 0.028 respectively). We demonstrate that radiation and chemotherapy bystander responses induce early genomic instability coupled with defects in mitochondrial function. Restoring mitochondrial function through overexpression of MnSOD significantly rescues nuclear instability events; anaphase bridges and telomere length shortening.

Tumour vasculature immaturity, oxidative damage and systemic inflammation stratify survival of colorectal cancer patients on bevacizumab treatment.

Despite treatment of patients with metastatic colorectal cancer (mCRC) with bevacizumab plus chemotherapy, response rates are modest and there are no biomarkers available that will predict response. The aim of this study was to assess if markers associated with three interconnected cancer-associated biological processes, specifically angiogenesis, inflammation and oxidative damage, could stratify the survival outcome of this cohort. Levels of angiogenesis, inflammation and oxidative damage markers were assessed in pre-bevacizumab resected tumour and serum samples of mCRC patients by dual immunofluorescence, immunohistochemistry and ELISA. This study identified that specific markers of angiogenesis, inflammation and oxidative damage stratify survival of patients on this anti-angiogenic treatment. Biomarkers of immature tumour vasculature (% IMM, p=0.026, n=80), high levels of oxidative damage in the tumour epithelium (intensity of 8-oxo-dG in nuclear and cytoplasmic compartments, p=0.042 and 0.038 respectively, n=75) and lower systemic pro-inflammatory cytokines (IL6 and IL8, p=0.053 and 0.049 respectively, n=61) significantly stratify with median overall survival (OS). In summary, screening for a panel of biomarkers for high levels of immature tumour vasculature, high levels of oxidative DNA damage and low levels of systemic pro-inflammatory cytokines may be beneficial in predicting enhanced survival outcome following bevacizumab treatment for mCRC.