Quantification of Linrodostat and its metabolites: Overcoming bioanalytical challenges in support of a discovery Indoleamine 2,3 dioxygenase program.

BMS-986205 (Linrodostat) is a small molecule inhibitor of Indoleamine 2, 3 dioxygenase (IDO) that is currently being evaluated in clinical trials for the oral treatment of advanced cancer. Initially, there were concerns regarding possible toxicity following administration, since BMS-986205 undergoes metabolism to form 4-chloroaniline. However, it was later determined that the downstream metabolites of 4-chloroaniline might be a greater concern. To evaluate the potential toxicity of these metabolites, a sensitive LC-MS/MS analytical method was needed to quantify both the parent compound and multiple metabolites. This presented a challenge since the method required the analysis of multiple analytes while still retaining the analytical sensitivity required to support studies. By utilizing a multi-function analytical method, we were able to quantify the necessary analytes using a complex LC-MS/MS-based method including the application of both negative and positive electrospray ionization.

Targeted Metabolomics With Ultraperformance Liquid Chromatography-Mass Spectrometry (UPLC-MS) Highlights Metabolic Differences in Healthy and Atopic Staffordshire Bull Terriers Fed Two Different Diets, A Pilot Study.

Background: While anecdotal evidence has long claimed that a raw meat-based diet (RMBD) improves the metabolic health of canines, no rigorous scientific study has clarified this issue. Canine atopic dermatitis (CAD) has also been linked to metabolic health, but its relation to diet remains poorly understood. This study investigates whether dietary choice is linked to metabolic health in healthy and CAD-diagnosed canines via targeted serum and urine metabolomic analysis of polar, non-ionic metabolites, as well as whether the underlying CAD condition modulates the response to nutritional intake. Materials and Methods: Serum metabolites of client-owned Staffordshire bull terriers, divided into CAD-diagnosed (n = 14) and healthy (n = 6) cohorts, were studied. Urine metabolites of a subset of the CAD-diagnosed canines (n = 8) were also studied. The canines were split into two cohorts based on diet. The first cohort were fed a commercially available high-fat, moderate-protein, low-carbohydrate RMBD (n = 11, CAD diagnosed n = 8, healthy n = 3). Those in the second cohort were fed a commercially available moderate-fat, moderate-protein, high-carbohydrate kibble diet (KD) (n = 9: CAD diagnosed n = 6, healthy n = 3). The diet intervention period lasted approximately 4.5 months (median 135 days). Statistical analyses of the serum profiles across all dogs (n = 20) and the urine profiles of the CAD-diagnosed subset (n = 8) were performed. Results and Discussion: The KD cohort was found to have higher concentrations of methionine than the RMBD cohort, both in serum (all dogs, p < 0.0001) and in urine (CAD-only cohort, p < 0.0002), as well as cystathionine and 4-pyridoxic acid. Methionine plays important roles in homocysteine metabolism, and elevated levels have been implicated in various pathologies. The CAD (n = 14) cohort dogs showed starker metabolic changes in response to diet regarding these pathways compared to the healthy (n = 6) cohort. However, there was no significant change in CAD severity as a result of either diet. Likely due to the higher meat content of the RMBD, higher concentrations of several carnitines and creatine were found in the RMBD cohort. Citrulline was found in higher concentrations in the KD cohort. Our findings provide insight into the relationship between diet and the serum and urine metabolite profiles of canines. They also suggest that neither diet significantly affected CAD severity.

The effect of a raw vs dry diet on serum biochemical, hematologic, blood iron, B12 , and folate levels in Staffordshire Bull Terriers.

BACKGROUND: To date, very few studies have compared the effects of different types of feeding practices on canine physiology, such as feeding exclusively dry, raw, or homemade foods. OBJECTIVES: We aimed to report the changes in hematologic, serum biochemical, plasma folate, B12 , and whole blood iron levels in dogs fed two different diets. METHODS: A pilot study was developed to compare the effects of a heat-processed high carbohydrate (HPHC) and nonprocessed high-fat (NPHF) diet. A total of 33 client-owned Staffordshire Bull Terriers were used; 18 had canine atopic dermatitis, seven were healthy, and eight were grouped as "borderline" dogs since they did not fulfill at least six of Favrot's criteria. The comparisons were made between the diet groups at the end visit of the diet intervention, as well as within the diet groups during the study. RESULTS: Significant differences between and within the diet groups were observed, although the majority of outcomes remained within the RIs. The median time of diet intervention was 140 days. Red blood cell counts, mean cell hemoglobin concentrations, and platelet counts were significantly higher, and mean cell hemoglobin, mean cell volume, alkaline phosphatase, inorganic phosphorus, and cholesterol were significantly lower in the dogs fed the NPHF diet compared with those fed the HPHC diet after the diet trial was completed. In addition, folate, B12 , and iron decreased significantly in the NPHF diet group. CONCLUSIONS: This pilot study indicated that diet had an impact on blood values, although most remained within RIs, pointing out the need for further studies.

Selective Cleavage of Lignin ß-O-4 Aryl Ether Bond by ß-Etherase of the White-Rot Fungus Dichomitus squalens.

Production of value-added compounds from a renewable aromatic polymer, lignin, has proven to be challenging. Chemical procedures, involving harsh reaction conditions, are costly and often result in nonselective degradation of lignin linkages. Therefore, enzymatic catalysis with selective cleavage of lignin bonds provides a sustainable option for lignin valorization. In this study, we describe the first functionally characterized fungal intracellular ß-etherase from the wood-degrading white-rot basidiomycete Dichomitus squalens. This enzyme, Ds-GST1, from the glutathione-S-transferase superfamily selectively cleaved the ß-O-4 aryl ether bond of a dimeric lignin model compound in a glutathione-dependent reaction. Ds-GST1 also demonstrated activity on polymeric synthetic lignin fractions, shown by a decrease in molecular weight distribution of the laccase-oxidized guaiacyl dehydrogenation polymer. In addition to a possible role of Ds-GST1 in intracellular catabolism of lignin-derived aromatic compounds, the cleavage of the most abundant linkages in lignin under mild reaction conditions makes this biocatalyst an attractive green alternative in biotechnological applications.

Quantification of human mAbs in mouse tissues using generic affinity enrichment procedures and LC-MS detection.

BACKGROUND: The disease state can modulate the penetration of large antibody-sized therapeutic molecules into affected tissues. Suitable bioanalytical methods are required for the quantitative analysis of drug tissue levels to enable a better understanding of the parameters influencing drug penetration and target engagement. RESULTS: Described is a sensitive and selective LC-MS/MS assay for the quantification of human mAb molecules in mouse tissues. By homogenizing tissues directly into serum, a common serum calibration curve can be used for multiple tissues. A generic procedure was used for affinity enrichment. An analytical range of 20 - 20,000 ng/ml was achieved in serum. CONCLUSION: The method described here can be applied for the quantitative analysis of mAb and Fc-fusion therapeutic molecules in a variety of animal tissue matrices.

Childcare outdoor renovation as a built environment health promotion strategy: evaluating the preventing obesity by design intervention.

PURPOSE: To evaluate the effectiveness of Preventing Obesity by Design (POD), a childcare center outdoor renovation intervention. DESIGN: Pre-post intervention evaluation. SETTING: North Carolina licensed childcare centers (N = 27). SUBJECTS: Preschool children. INTERVENTION: Outdoor renovation, teacher training. MEASURES: Behavior mapping, Preschool Outdoor Environment Measurement Scale (POEMS), center director interview. ANALYSIS: Descriptive statistics, ordinary least squares and logistic regressions calculated to assess levels of association between environmental change, children's physical activity (PA), social behaviors, and environmental quality. Qualitative interview data analyzed to help understand intervention impact. RESULTS: Behavior mapping showed that site layout attributes, such as the form (i.e., "single loop" and "double loop") of pathways (functioning as circulation routes and wheeled toy settings), are associated with higher levels of PA. Teacher interaction was associated with decreased children's PA. Absence of teacher or lack of child/child interaction was associated with increased PA. POEMS assessment of environmental quality was higher after renovation. POEMS domains (Physical Space and Teacher/Caregiver Roles) were positively associated with PA. After renovation, 68% of center directors reported positive changes in children's behavior and 40% mentioned edible plant installations as greatest success. CONCLUSION: Built environment renovation of childcare center outdoors, including looped pathways installation, coupled with teacher training, may support increased PA. Renovation, including food gardens, may be a key to success for preschool health promotion and support change in childcare policy.

Use of lean and six sigma methodology to improve operating room efficiency in a high-volume tertiary-care academic medical center.

BACKGROUND: Operating rooms (ORs) are resource-intense and costly hospital units. Maximizing OR efficiency is essential to maintaining an economically viable institution. OR efficiency projects often focus on a limited number of ORs or cases. Efforts across an entire OR suite have not been reported. Lean and Six Sigma methodologies were developed in the manufacturing industry to increase efficiency by eliminating non-value-added steps. We applied Lean and Six Sigma methodologies across an entire surgical suite to improve efficiency. STUDY DESIGN: A multidisciplinary surgical process improvement team constructed a value stream map of the entire surgical process from the decision for surgery to discharge. Each process step was analyzed in 3 domains, ie, personnel, information processed, and time. Multidisciplinary teams addressed 5 work streams to increase value at each step: minimizing volume variation; streamlining the preoperative process; reducing nonoperative time; eliminating redundant information; and promoting employee engagement. Process improvements were implemented sequentially in surgical specialties. Key performance metrics were collected before and after implementation. RESULTS: Across 3 surgical specialties, process redesign resulted in substantial improvements in on-time starts and reduction in number of cases past 5 pm. Substantial gains were achieved in nonoperative time, staff overtime, and ORs saved. These changes resulted in substantial increases in margin/OR/day. CONCLUSIONS: Use of Lean and Six Sigma methodologies increased OR efficiency and financial performance across an entire operating suite. Process mapping, leadership support, staff engagement, and sharing performance metrics are keys to enhancing OR efficiency. The performance gains were substantial, sustainable, positive financially, and transferrable to other specialties.