Metabolic signature of Mycobacterium avium subsp. paratuberculosis infected and infectious dairy cattle by integrating nuclear magnetic resonance analysis and blood indices

The early diagnosis of Mycobacterium avium subsp. paratuberculosis (MAP) is one of the current challenges of farmers and veterinarians. This work aimed to investigate the changes in metabolic levels associated with natural MAP infection in infected and infectious dairy cattle. The study included sera from 23 infectious/seropositive, 10 infected but non-infectious/seronegative, and 26 negative Holstein Fresian cattle. The samples were selected from a collection of samples gathered during a prospective study. The samples were analyzed by quantitative nuclear magnetic resonance (NMR) spectroscopy and routine blood chemistry. The blood indices and the 1H NMR data were concatenated by low-level data fusion, resulting in a unique global fingerprint. Afterwards, the merged dataset was statistically analyzed by the least absolute shrinkage and selection operator (LASSO), which is a shrinkage and selection method for supervised learning. Finally, pathways analysis was performed to get more insights on the possible dysregulated metabolic pathways. The LASSO model achieved, in a 10 time repeated 5-fold cross-validation, an overall accuracy of 91.5% with high values of sensitivity and specificity in classifying correctly the negative, infected, and infectious animals. The pathway analysis revealed MAP-infected cattle have increased tyrosine metabolism and enhanced phenylalanine, tyrosine and tryptophan biosynthesis. The enhanced synthesis and degradation of ketone bodies was observed both in infected and infectious cattle. In conclusion, fusing data from multiple sources has proved to be useful in exploring the altered metabolic pathways in MAP infection and potentially diagnosing negative animals within paratuberculosis-infected herds

Tryptophan catabolism and response to therapy in locally advanced rectal cancer (LARC) patients

In locally advanced rectal cancer patients (LARC), preoperative chemoradiation improves local control and sphincter preservation. The response rate to treatment varies substantially between 20% and 30% and it is an important prognostic factor. Indeed, non-responsive patients are subjected to higher rates of local and distant metastases, and worse survival compared to patients with complete response. In the search of predictive biomarkers for response prediction to therapy in LARC patients, we found increased plasma tryptophan levels in non-responsive patients. On the basis of plasma levels of 5-hydroxy-tryptophan and kynurenine, the activities of tryptophan 5-hydroxylase 1 (TPH1) and indoleamine-2,3-dioxygenases 1 (IDO1)/tryptophan-2,3-dioxygenase (TDO2) have been obtained and data have been correlated to gene expression profiles. We demonstrated that TDO2 overexpression in non-responsive patients correlates with kynurenine plasma levels. Finally, through the gene expression and targeted metabolomic analysis in paired healthy mucosa-rectal cancer tumour samples, we evaluated the impact of tryptophan catabolism at tissue level in responsive and non-responsive patients

Investigating the physiological response and antibody concentration of gilthead sea bream (Sparus aurata) following Vibrio anguillarum vaccination depending on the stress coping style

Stress coping styles (SCSs) are defined as coherent sets of individual physiological and behavioral differences in stress response consistent across time and context and are described in a wide range of taxa, including fishes. These differences in behavior and physiology are of great interest because they may have direct implications on animal health, welfare, and performance in farming systems, including aquaculture. In this study, the physiological responses of sea bream (Sparus aurata) from different SCSs following Vibrio anguillarum vaccination were monitored. Fish were first screened either bold or shy (proxy of proactive and reactive SCSs, respectively) using group risk-taking tests and were then injected with a vaccine against V. anguillarum. Following vaccination, the fish were implanted with an accelerometer tag to monitor their swimming activity (proxy of energy expenditure), and blood sampling was carried out to measure health and welfare parameters (e.g., cortisol, glucose, hemoglobin) and aspecific immunity (e.g., protease, total proteins). In addition, blood was also collected at three different sampling times to screen antibody levels and, thus, to evaluate the efficiency of the vaccine. Following vaccination, bold fish displayed lower swimming activity values, indicative of lower energy expenditure, and also displayed higher levels of hematocrit, total proteins, and lysozyme in the plasma than the shy ones, which could be indicative of better health/welfare status and greater aspecific immunity. Finally, the V. anguillarum vaccination appeared to be more efficient in bold fish since the number of total antibodies was found higher than in shy fish 1 month after vaccination. Such results could help improve both health/welfare and productivity of farmed sea breams by selecting more robust fish, better adapted to farming conditions

Table_1_Metabolic signature of Mycobacterium avium subsp. paratuberculosis infected and infectious dairy cattle by integrating nuclear magnetic resonance analysis and blood indices.DOCX

The early diagnosis of Mycobacterium avium subsp. paratuberculosis (MAP) is one of the current challenges of farmers and veterinarians. This work aimed to investigate the changes in metabolic levels associated with natural MAP infection in infected and infectious dairy cattle. The study included sera from 23 infectious/seropositive, 10 infected but non-infectious/seronegative, and 26 negative Holstein Fresian cattle. The samples were selected from a collection of samples gathered during a prospective study. The samples were analyzed by quantitative nuclear magnetic resonance (NMR) spectroscopy and routine blood chemistry. The blood indices and the 1H NMR data were concatenated by low-level data fusion, resulting in a unique global fingerprint. Afterwards, the merged dataset was statistically analyzed by the least absolute shrinkage and selection operator (LASSO), which is a shrinkage and selection method for supervised learning. Finally, pathways analysis was performed to get more insights on the possible dysregulated metabolic pathways. The LASSO model achieved, in a 10 time repeated 5-fold cross-validation, an overall accuracy of 91.5% with high values of sensitivity and specificity in classifying correctly the negative, infected, and infectious animals. The pathway analysis revealed MAP-infected cattle have increased tyrosine metabolism and enhanced phenylalanine, tyrosine and tryptophan biosynthesis. The enhanced synthesis and degradation of ketone bodies was observed both in infected and infectious cattle. In conclusion, fusing data from multiple sources has proved to be useful in exploring the altered metabolic pathways in MAP infection and potentially diagnosing negative animals within paratuberculosis-infected herds.</p

Interobserver reproducibility of cytologic p16INK4a/Ki-67 dual immunostaining in human papillomavirus-positive women

BACKGROUND: The accumulation of cyclin-dependent kinase inhibitor 2A (p16(ink4a)) protein in a cell is associated with neoplastic progression in precancerous cervical lesions. Dual staining for p16(ink4a) and Ki-67 has been proposed as a triage test in cervical cancer screening for women who test positive for human papillomavirus DNA. In this study, interobserver reproducibility of the interpretation of this test was assessed. METHODS: Forty-two immunostained, liquid-based cytology slides were divided into 2 sets and were interpreted by 17 to 21 readers from 9 different laboratories, yielding a total of 816 reports. Immunostaining results were classified as positive, negative, inconclusive, or inadequate. After evaluation of the first set of slides and before circulation of the second set, the results were discussed in a plenary meeting. The 10 slides with the most discordant results were evaluated again by selected expert cytopathologists. RESULTS: The overall kappa value was 0.612 (95% confidence interval [CI], 0.523-0.701), it was higher for the positive and negative categories (kappa=0.692 and kappa=0.641, respectively), and it was almost null for the inconclusive category (kappa=0.058). Considering only readers from laboratories with documented experience, the kappa value was higher (kappa=0.747; 95% CI, 0.643-0.839) compared with nonexperienced centers (kappa=0.498; 95% CI, 0.388-0.616). The results were similar in both sets of slides (kappa=0.505 [95% CI, 0.358-0.642] and kappa=0.521 [95% CI, 0.240-0.698] for the first and second sets, respectively). Reinterpretation of the slides with the most discordant results did not provide any improvement (first evaluation, kappa=0.616 [95% CI, 0.384-0.866]; second evaluation, kappa=0.403 [95% CI, 0.182-0.643]). CONCLUSIONS: Dual staining for p16(ink4a) and Ki-67 demonstrated good reproducibility, confirming its robustness, which is a necessary prerequisite for its adoption as a triage test in cervical cancer screening programs that use human papillomavirus DNA as a primary test. Cancer Cytopathol 2017; 125:212-20. (C) 2016 American Cancer Society