Stratification of chemotherapy-treated stage III colorectal cancer patients using multiplexed imaging and single-cell analysis of T-cell populations.

Colorectal cancer (CRC) has one of the highest cancer incidences and mortality rates. In stage III, postoperative chemotherapy benefits <20% of patients, while more than 50% will develop distant metastases. Biomarkers for identification of patients at increased risk of disease recurrence following adjuvant chemotherapy are currently lacking. In this study, we assessed immune signatures in the tumor and tumor microenvironment (TME) using an in situ multiplexed immunofluorescence imaging and single-cell analysis technology (Cell DIVETM) and evaluated their correlations with patient outcomes. Tissue microarrays (TMAs) with up to three 1 mm diameter cores per patient were prepared from 117 stage III CRC patients treated with adjuvant fluoropyrimidine/oxaliplatin (FOLFOX) chemotherapy. Single sections underwent multiplexed immunofluorescence staining for immune cell markers (CD45, CD3, CD4, CD8, FOXP3, PD1) and tumor/cell segmentation markers (DAPI, pan-cytokeratin, AE1, NaKATPase, and S6). We used annotations and a probabilistic classification algorithm to build statistical models of immune cell types. Images were also qualitatively assessed independently by a Pathologist as 'high', 'moderate' or 'low', for stromal and total immune cell content. Excellent agreement was found between manual assessment and total automated scores (p < 0.0001). Moreover, compared to single markers, a multi-marker classification of regulatory T cells (Tregs: CD3+/CD4+FOXP3+/PD1-) was significantly associated with disease-free survival (DFS) and overall survival (OS) (p = 0.049 and 0.032) of FOLFOX-treated patients. Our results also showed that PD1- Tregs rather than PD1+ Tregs were associated with improved survival. These findings were supported by results from an independent FOLFOX-treated cohort of 191 stage III CRC patients, where higher PD1- Tregs were associated with an increase overall survival (p = 0.015) for CD3+/CD4+/FOXP3+/PD1-. Overall, compared to single markers, multi-marker classification provided more accurate quantitation of immune cell types with stronger correlations with outcomes.

Impact of canine-assisted ambulation on hospitalized chronic heart failure patients' ambulation outcomes and satisfaction: a pilot study.

BACKGROUND AND RESEARCH OBJECTIVE: Chronic heart failure (HF) is a prevalent and costly disease process. Early ambulation has been shown to have a positive impact on patient outcomes and length of stay. Animal-assisted therapy is a novel modality that has shown to be a safe and effective adjunct to a number of traditional treatment plans. This study sought to synergistically combine ambulation and animal-assisted therapy by using canine-assisted ambulation (CAA) to improve the ambulation outcomes of HF patients. SUBJECTS AND METHODS: Sixty-nine hospitalized patients with a primary diagnosis of HF were approached to ambulate with a restorative aide. After recording their initial response, they were given the opportunity to participate in CAA (walking with a therapy dog). Initial ambulation refusal rate was compared with a historical population of 537 HF patients. Distance ambulated was recorded using a pedometer and compared with a randomly selected, 64-patient sample from the historical HF patient population, stratified by day of hospital stay. Patient satisfaction was assessed through a 5-item Likert scale survey. RESULTS AND CONCLUSION: The 537-patient historical HF population had an ambulation refusal rate of 28%. When offered the chance to participate in CAA, only 7.2% of the study population refused ambulation (P = .0002). Of the 69-patient study sample, 13 initially refused ambulation then agreed when offered CAA (P = .0009). Distance ambulated increased from 120.2 steps in a randomly selected, stratified historical sample to 235.07 in the CAA study sample (P < .0001). Patients unanimously agreed that they enjoyed CAA and would like to participate in CAA again. Canine-assisted ambulation is a safe and effective adjunct to an early ambulation program for HF patients. Canine-assisted ambulation may decrease hospital length of stay and thereby decrease the costs of HF care. Additional research involving CAA's application to other disease processes in various settings is warranted.