Effects of combined use of keratinolytic enzymes and sugarcane fibre on the hairball excretion in cats.

Grooming is a natural hygienic behaviour of cats that favours the formation of hairballs. Increased fibre concentration in the diets is a strategy to minimize hairball formation, but it is not fully effective. Because cat hair is formed mostly by keratin, the addition of keratinases in the diets might be an alternative for hairball control. Thus, the objective was to evaluate the effect of the combined use of sugarcane fibre and a protease complex to reduce the hairball excretion in cats. Twenty-four adult cats were divided into four treatment groups (n = 6 per treatment) in a 2 × 2 factorial arrangement. Treatments were as follows: control diet (Control; containing low concentration of insoluble fibre, 5.34% of total dietary fibre), control diet plus enzyme (Co-e), high-fibre diet (HF; containing high amounts of insoluble fibre, 17.8% of total dietary fibre), and HF diet plus enzyme (HF-e). Proteases from Bacillus licheniformis PWD-1 were administered orally (5 mg/day) as gastro-resistant capsules. Total collection of faeces was carried out to determine the number of excreted hairballs and the coefficient of total tract apparent digestibilities (CTTAD) of the diets. Separate addition of insoluble fibre (HF; p = .5947) or enzyme complex (Co-e; p = .3633) had no effect on the hairballs excreted in the faeces. However, the combined use of insoluble fibre and enzymes (HF-e) reduced (p = .0344) the total number of hairballs excreted. The size distribution of hairballs (small, medium, or large) was not affected by treatments (p = .3763). The CTTAD of crude protein was not affected by protease addition (p = .781) but was reduced by HF and HF-e treatments. Sugarcane fibre associated to keratinolytic enzymes reduces the hairball excretion in cats. This strategy can be adopted for this purpose; however, methods for faecal hairball quantification must be improved. If you have not already completed a Copyright Transfer Agreement, please log on to Wiley Author Serivices, https://authorservices.wiley.com/bauthor/, sign-in and complete the License Agreement form".

Relapse in rheumatoid arthritis patients undergoing dose reduction and withdrawal of biologics: are predictable factors more relevant than predictive parameters? An observational prospective real-life study.

OBJECTIVE: To determine predictive/predictable factors of relapse in rheumatoid arthritis (RA) patients undergoing biologic Disease-Modifying Anti-Rheumatic Drugs (bDMARDs) dose reduction/discontinuation. PATIENTS AND METHODS: RA patients receiving the same bDMARD for more than 1 year, in Simplified Disease Activity Index (SDAI) remission, were selected in an observational monocentric real-life study. The 18-month follow-up included spacing (6 months) and withdrawal (12 months) periods of bDMARD. Clinical, biological and ultrasonographic (US) parameters were collected regularly. Relapse was defined by SDAI>11. RESULTS: Fifty-three RA patients (mean age: 58 years; 72% women; median duration: 11 years) were enrolled. Forty-two received anti-cytokinic bDMARD targeting tumour necrosis factor (n=39) or interleukin-6R (n=3) and 11 were treated by abatacept. The number of relapses during the spacing and discontinuation periods were 19 and 20, respectively. After 18 months of follow-up, among the 53 patients, 12 maintained bDMARD-free remission, 39 had relapsed and 2 were lost of follow-up. Median time to relapse was 11.8 months. In multivariate analysis, baseline factors predictive of relapse were corticosteroid intake, female gender, longer disease duration and no methotrexate intake with bDMARD. Concerning the survival analysis, also taking into account the factors of predictability, the main risk factor of relapse after discontinuation was an increase of SDAI >0 during the spacing period (p=0.03). US findings were not contributive. CONCLUSION: In the context of RA in remission under bDMARDs, variation of SDAI during the dose-reduction phase is more relevant than baseline parameters to predict success of drug withdrawal.

Determinants of Undesirable Behaviors in American Quarter Horses Housed in Box Stalls.

The objective of this study was to investigate the management practices that affect undesirable behaviors in American Quarter Horses (AQH) confined to box stalls. A total of 105 adult AQH, comprising 29 females, 22 intact males, and 54 castrated males, housed in box stalls for at least 30 days, were included in the study. Behavioral activities were recorded through visual inspection at 5-min intervals over a 24-h period by two observers who took turns in shifts of 4 hours. Parameters related to animal characteristics, stall conditions, handling, training, nutrients supplied, and feeding behaviors were considered independent variables, and the time spent in undesirable behaviors was the dependent variable. Data were subjected to exploratory factor analysis and hierarchical cluster analysis. Four principal components were extracted. Two clusters were identified based on the time spent in undesirable behaviors: low-incidence group (25 ± 13 minutes) and high-incidence group (97 ± 52 minutes). Nutritional status (P = .1447), physiological status (P = .1372), and training routine (P = .1160) had no significant effects on the dependent variable; however, the effect of feeding behavior (P = .0444) was significant. A low dry matter intake/requirement ratio (P = .0470) and a high heart rate at rest (P = .0444) indicated that inadequate nutrient supply is a determinant of the time spent in undesirable behaviors in AQH. Aspects related to management practices such as horse nutrition, feeding behaviors, and training routine should be considered to reduce undesirable behaviors in AQH.

Rheumatoid arthritis, insulin resistance, and diabetes.

Recent progress in the management of rheumatoid arthritis (RA) is turning attention toward comorbidities, such as diabetes. The objectives of this review are to clarify the links between RA and diabetes and to assess potential effects of disease-modifying antirheumatic drugs (DMARDs) on diabetes. The increased insulin resistance seen in RA is closely linked to the systemic inflammation induced by certain proinflammatory cytokines such as tumor necrosis factor α (TNFα) and interleukin-6. The prevalence of type 2 diabetes is increased in patients with RA. Furthermore, certain DMARDs including hydroxychloroquine, methotrexate, TNFα antagonist, and interleukin-1ß antagonists seem to improve the markers of glucose metabolism. In contrast, glucocorticoids tend to adversely affect glycemic control, particularly when taken chronically. Consequently, a crucial yet insufficiently applied rule is that cardiovascular risk factors must be sought and treated routinely, particularly as the choice of the DMARD may affect glucose metabolism.

Validation of the 2010-ACR/EULAR -classification criteria using newly EULAR-defined erosion for rheumatoid arthritis on the very early arthritis community-based (VErA) cohort.

OBJECTIVE: To validate the 2010-ACR/EULAR criteria for rheumatoid arthritis (RA), taking into account the recent EULAR definition of "erosive disease", on the 310 patients comprising the very early arthritis cohort (VErA). METHODS: 2010-criteria performances were tested by first strictly applying its three items successively: ≥ 1 clinical synovitis/another disease(s)/score ≥ 6/10), then the typical erosion grid without obtaining a score of ≥ 6 to diagnose RA. We tested successively: no erosion (S1), ≥ 1 erosion(s) (S2), EULAR-defined erosive disease (S3). Two gold standards were used: expert diagnosis at six years and EULAR erosive disease at two years. RESULTS: At inclusion, median age was 52 years; median RA duration 4.2 months. 2010-ACR/EULAR criteria, including EULAR-defined erosive disease applied at baseline, classified comparable numbers of patients as the 1987 criteria (P=0.27). Using expert diagnosis at six years, more patients were classified as RA with S2 than 1987-ACR criteria (P<0.04). In contrast, sensitivity and specificity indicated that 2010-ACR/EULAR-S3 criteria performed slightly but not significantly better than 1987-ACR criteria. On ROC curves, a score ≥ 6 correctly classified RA. When EULAR-defined erosion at two years was the gold standard, the 1987-ACR, the 2010-S1, -S2 and -S3 criteria performed comparably. CONCLUSIONS: Using the very early community-based, conservatively treated VErA cohort, the strict application of 2010-ACR/EULAR criteria using the new EULAR definition of erosive disease or not performed slightly but not significantly better than the 1987-ACR criteria.

Evaluation of pre-induction temperature, cell growth at induction and IPTG concentration on the expression of a leptospiral protein in E. coli using shaking flasks and microbioreactor.

BACKGROUND: Leptospirosis is a zoonose that is increasingly endemic in built-up areas, especially where there are communities living in precarious housing with poor or non-existent sanitation infrastructure. Leptospirosis can kill, for its symptoms are easily confused with those of other diseases. As such, a rapid diagnosis is required so it can be treated effectively. A test for leptospirosis diagnosis using Leptospira Immunoglobulin-like (Lig) proteins is currently at final validation at Fiocruz. RESULTS: In this work, the process for expression of LigB (131-645aa) in E. coli BL21 (DE3)Star™/pAE was evaluated. No significant difference was found for the experiments at two different pre-induction temperatures (28 °C and 37 °C). Then, the strain was cultivated at 37 °C until IPTG addition, followed by induction at 28°C, thereby reducing the overall process time. Under this condition, expression was assessed using central composite design for two variables: cell growth at which LigB (131-645aa) was induced (absorbance at 600 nm between 0.75 and 2.0) and inducer concentration (0.1 mM to 1 mM IPTG). Both variables influenced cell growth and protein expression. Induction at the final exponential growth phase in shaking flasks with Abs(ind) = 2.0 yielded higher cell concentrations and LigB (131-645aa) productivities. IPTG concentration had a negative effect and could be ten-fold lower than the concentration commonly used in molecular biology (1 mM), while keeping expression at similar levels and inducing less damage to cell growth. The expression of LigB (131-645aa) was associated with cell growth. The induction at the end of the exponential phase using 0.1 mM IPTG at 28 °C for 4 h was also performed in microbioreactors, reaching higher cell densities and 970 mg/L protein. LigB (131-645aa) was purified by nickel affinity chromatography with 91% homogeneity. CONCLUSIONS: It was possible to assess the effects and interactions of the induction variables on the expression of soluble LigB (131-645aa) using experimental design, with a view to improving process productivity and reducing the production costs of a rapid test for leptospirosis diagnosis.

Optimization of medium formulation and seed conditions for expression of mature PsaA (pneumococcal surface adhesin A) in Escherichia coli using a sequential experimental design strategy and response surface methodology.

PsaA, a candidate antigen for a vaccine against pneumonia, is well-conserved in all Streptococcus pneumoniae serotypes. A sequence of two-level experimental designs was used to evaluate medium composition and seed conditions to optimize the expression of soluble mature PsaA in E. coli. A face-centered central composite design was first used to evaluate the effects of yeast extract (5 and 23.6 g/L), tryptone (0 and 10 g/L), and glucose (1 and 10 g/L), with replicate experiments at the central point (14.3 g/L yeast extract, 5 g/L tryptone, 5.5 g/L glucose). Next, a central composite design was used to analyze the influence of NaCl concentration (0, 5, and 10 g/L) compared with potassium salts (9.4 g/L K(2)HPO(4)/2.2 g/L KH(2)PO(4)), and seed growth (7 and 16 h). Tryptone had no significant effect and was removed from the medium. Yeast extract and glucose were optimized at their intermediate concentrations, resulting in an animal-derived material-free culture medium containing 15 g/L yeast extract, 8 g/L glucose, 50 µg/mL kanamycin, and 0.4% glycerol, yielding 1 g/L rPsaA after 16 h induction at 25°C in shake flasks at 200 rpm. All the seed age and salt conditions produced similar yields, indicating that no variation had a statistically significant effect on expression. Instead of growing the seed culture for 16 h (until saturation), the process can be conducted with 7 h seed growth until the exponential phase. These results enhanced the process productivity and reduced costs, with 5 g/L NaCl being used rather than potassium salts.