Chemotherapy-induced gut microbiome disruption, inflammation, and cognitive decline in female patients with breast cancer.

Chemotherapy is notorious for causing behavioral side effects (e.g., cognitive decline). Notably, the gut microbiome has recently been reported to communicate with the brain to affect behavior, including cognition. Thus, the aim of this clinical longitudinal observational study was to determine whether chemotherapy-induced disruption of the gut microbial community structure relates to cognitive decline and circulating inflammatory signals. Fecal samples, blood, and cognitive measures were collected from 77 patients with breast cancer before, during, and after chemotherapy. Chemotherapy altered the gut microbiome community structure and increased circulating TNF-α. Both the chemotherapy-induced changes in microbial relative abundance and decreased microbial diversity were related to elevated circulating pro-inflammatory cytokines TNF-α and IL-6. Participants reported subjective cognitive decline during chemotherapy, which was not related to changes in the gut microbiome or inflammatory markers. In contrast, a decrease in overall objective cognition was related to a decrease in microbial diversity, independent of circulating cytokines. Stratification of subjects, via a reliable change index based on 4 objective cognitive tests, identified objective cognitive decline in 35% of the subjects. Based on a differential microbial abundance analysis, those characterized by cognitive decline had unique taxonomic shifts (Faecalibacterium, Bacteroides, Fusicatenibacter, Erysipelotrichaceae UCG-003, and Subdoligranulum) over chemotherapy treatment compared to those without cognitive decline. Taken together, gut microbiome change was associated with cognitive decline during chemotherapy, independent of chemotherapy-induced inflammation. These results suggest that microbiome-related strategies may be useful for predicting and preventing behavioral side effects of chemotherapy.

EELS: Autonomous snake-like robot with task and motion planning capabilities for ice world exploration.

Ice worlds are at the forefront of astrobiological interest because of the evidence of subsurface oceans. Enceladus in particular is unique among the icy moons because there are known vent systems that are likely connected to a subsurface ocean, through which the ocean water is ejected to space. An existing study has shown that sending small robots into the vents and directly sampling the ocean water is likely possible. To enable such a mission, NASA's Jet Propulsion Laboratory is developing a snake-like robot called Exobiology Extant Life Surveyor (EELS) that can navigate Enceladus' extreme surface and descend an erupting vent to capture unaltered liquid samples and potentially reach the ocean. However, navigating to and through Enceladus' environment is challenging: Because of the limitations of existing orbital reconnaissance, there is substantial uncertainty with respect to its geometry and the physical properties of the surface/vents; communication is limited, which requires highly autonomous robots to execute the mission with limited human supervision. Here, we provide an overview of the EELS project and its development effort to create a risk-aware autonomous robot to navigate these extreme ice terrains/environments. We describe the robot's architecture and the technical challenges to navigate and sense the icy environment safely and effectively. We focus on the challenges related to surface mobility, task and motion planning under uncertainty, and risk quantification. We provide initial results on mobility and risk-aware task and motion planning from field tests and simulated scenarios.

Impulsivity and Treatment Outcomes in Individuals with Cocaine Use Disorder: Examining the Gap between Interest and Adherence.

Background: Impulsivity is implicated in the development and maintenance of Cocaine Use Disorder (CUD). Less work has examined impulsivity's role on interest in initiating treatment, treatment adherence, or treatment response. No pharmacotherapies are approved for CUD, so efforts to understand and bolster the effects of psychotherapy are important in guiding and refining treatment. The present study examined the impact of impulsivity on interest in treatment, treatment initiation, treatment adherence, and treatment outcomes in individuals with CUD. Methods: Following the completion of a larger study on impulsivity and CUD participants were offered 14 sessions of (12 weeks) Cognitive Behavioral Relapse Prevention (CBT-RP). Before starting treatment, participants completed seven self-report and four behavioral measures of impulsivity. Sixty-eight healthy adults (36% female) with CUD (aged 49.4 ± 7.9) expressed an interest in treatment. Results: Greater scores on several self-report measures of impulsivity, and fewer difficulties with delayed gratification were associated with increased interest in treatment in both males and females. 55 participants attended at least 1 treatment session, while 13 participants did attend a single session. Individuals who attended at least one treatment session scored lower on measures of lack of perseverance and procrastination. Still, measures of impulsivity did not reliably predict session attendance nor the frequency of cocaine-positive urine samples throughout treatment. Males attended nearly twice as many treatment sessions as females despite nonsignificant associations between impulsivity in males and the number of sessions attended. Conclusions: Greater impulsivity in individuals with CUD was associated with expressing an interest in treatment, but not treatment adherence or response.

Temperature distributions and gradients in laser-heated plasmas relevant to magnetized liner inertial fusion.

We present two-dimensional temperature measurements of magnetized and unmagnetized plasma experiments performed at Z relevant to the preheat stage in magnetized liner inertial fusion. The deuterium gas fill was doped with a trace amount of argon for spectroscopy purposes, and time-integrated spatially resolved spectra and narrow-band images were collected in both experiments. The spectrum and image data were included in two separate multiobjective analysis methods to extract the electron temperature spatial distribution T_{e}(r,z). The results indicate that the magnetic field increases T_{e}, the axial extent of the laser heating, and the magnitude of the radial temperature gradients. Comparisons with simulations reveal that the simulations overpredict the extent of the laser heating and underpredict the temperature. Temperature gradient scale lengths extracted from the measurements also permit an assessment of the importance of nonlocal heat transport.

Assessment of the relationship between a written measure of empathy and an independently rated interview of Motivational Interviewing.

Motivational Interviewing (MI) is an evidence-based practice shown to be effective when working with people in treatment for substance use disorders. However, MI is a complex treatment modality optimized by training with feedback. Feedback, assessment and monitoring of treatment fidelity require measurement, which is typically done using audiotaped sessions. The gold standard for such measurement of MI skill has been an audiotaped interview, scored by a rater with a detailed structured instrument such as the Motivational Interviewing Treatment Integrity 2.0 (MITI 2.0) Coding System (Moyers, et al., 2005). The Helpful Responses Questionnaire (HRQ) (Miller, Hedrick, & Orlofsky, 1991) is a pen-and-paper test of empathy (a foundational MI skill) that does not require an audiotaped session. A randomized trial of three different regimens for training counselors in MI (live supervision using Teleconferencing, Tape-based supervision and Workshop only) (Smith et al., 2012) offered the opportunity to evaluate the performance of the HRQ as a measure of MI ability, compared to the several MITI 2.0 global scores and subscales. Participants were counselors (N=97) working at community-based substance use treatment programs, whose MI proficiency was measured at four time points: baseline (before an initial 2-day MI workshop), post-workshop, 8weeks post-workshop (i.e., post-supervision), and 20weeks post-workshop with both MITI 2.0 and HRQ. HRQ total scores correlated significantly with the Reflection to Question Ratio from the MITI 2.0 at post-workshop (r=0.33), week 8 (r=0.34), and week 20 (r=0.38), and with the Spirit (r=0.32) and Empathy (r=0.32) global scores at week 20. Correlations of HRQ with other MITI 2.0 subscales and time points after workshop were small and not significant. As predicted, HRQ scores differed between training conditions (X2(2)=7.88, p=0.02), with counselors assigned to live supervision achieving better HRQ scores than those in Workshop only. In summary, HRQ is a modestly accurate measure, mainly of the Reflection to Question Ratio, considered a core marker of MI skill. It is sensitive to training effects and may help identify counselors needing more intensive supervision. Given its ease of administration and scoring, HRQ may be a useful marker of MI skill during training efforts.

Consumption of a high-fat breakfast on consecutive days alters preclinical biomarkers for atherosclerosis.

BACKGROUND/OBJECTIVES: Recent research has speculated that the risk of developing atherosclerosis is due to the accumulation of the effects of daily diet choices. The purpose of this study was to examine which of our previously identified preclinical disease risk biomarkers were further elevated when consuming a high-fat (644±50 kcal; 100% recommended dietary allowance for fat), high-calorie (1118±100 kcal; 70% daily caloric needs) breakfast on consecutive days. Young, normal weight females (N=7) participated in this study. SUBJECTS/METHODS: Blood samples were taken premeal and hourly for 5-h postprandial. Serum biomarkers (C-peptide, eotaxin, gastric inhibitory polypeptide, granulocyte colony-stimulating factor (G-CSF), granulocyte-monocyte colony-stimulating factor (GM-CSF), insulin, leptin, monocyte chemoattractant protein 1, pancreatic polypeptide (PPY) and tumor necrosis factor-α), monocyte concentration, and adhesion molecule expression (CD11a, CD18 and CD54) were measured. Area under the curve was calculated for each outcome variable as a function of day and data were analyzed for significance. RESULTS: We found significant (P<0.05) increases on Day 2 for: GM-CSF (+47%; P=0.041), G-CSF (+31%; P=0.012), PPY (+51%; P=0.049), total monocyte (+110%; P=0.043), pro-inflammatory (PI) monocyte (+60%; P=0.012), PI monocyte CD18 (+960%; P=0.003), PI monocyte CD11a (+230%; P=0.006), and PI monocyte CD54 (+208%; P=0.015). CONCLUSIONS: To our knowledge, the present study is the first to report changes in selected biomarkers and monocytes following eating a high-fat, high-calorie breakfast on consecutive days in humans. More research is needed to determine how transient the observed changes are and what the long-term implications for disease risk are.

AAV-encoded OTC activity persisting to adulthood following delivery to newborn spf(ash) mice is insufficient to prevent shRNA-induced hyperammonaemia.

Urea cycle defects presenting in the neonatal period with hyperammonaemia are associated with high morbidity and mortality, and necessitate liver transplantation for long-term management. Gene therapy is therefore an attractive possibility, with vectors based on adeno-associated virus (rAAV) currently showing exciting promise in liver-targeted clinical trials in adults. Successful use of rAAV vectors in infants, however, is more challenging as episomal rAAV genomes will be lost from proliferating hepatocytes during liver growth, leaving stable transgene expression dependent on the subset of vector genomes that undergo genomic integration. To explore this challenge, we exploited the partially ornithine transcarbamylase (OTC)-deficient spf(ash) mouse model and small hairpin RNA-mediated knockdown of residual endogenous OTC enzyme activity in adult mice that had received neonatal treatment with an OTC-encoding rAAV. This leaves mice reliant on vector-encoded OTC activity that has persisted from the newborn period. Despite stable transduction in approximately 8% of hepatocytes and residual vector-encoded OTC activity of up to 33% of wild-type, well above endogenous spf(ash) levels (5-7%), mice were not protected from hyperammonaemia. These data show that the distribution of OTC activity within the liver is critical and that rAAV vector re-delivery after early neonatal treatment is likely to be necessary for stable control of hyperammonaemia into adulthood.

Considerations to maximize fat mass gain in a mouse model of diet-induced weight gain.

Mouse experimental models of diet-induced weight gain are commonly used as analogs to human obesity; however, a wide variety of feeding methods have been used and the most effective way to maximize weight gain is not known. Maximizing weight gain may allow for a reduction in the number of animals required for a given experiment. The purpose of this study was how to cause the greatest amount of weight gain in CD-1 mice by modifying the composition and source of their diet. To accomplish this goal, we completed two experiments: (1) Effect of dietary macronutrient fat intake (60% (HF60), 45% (HF45), 30% (HF30), or 13.5% (CON) fat diet for 18 weeks); and (2) Effect of 1:1 mixed HF60 and CON diets. Outcome measures included food intake, body mass, and body composition, which were measured bi-weekly and statistically analyzed using a repeated measures analysis of variance (RM-ANOVA). In Experiment 1, the greatest increase in body and fat mass was observed in HF60 (36%) and HF45 (29%) compared with HF30 and CON (P < 0.05). In Experiment 2, HF + stock diet (SK) gained 25% more body mass and 70% more fat mass than HF (P < 0.05). Collectively, these findings suggest that using a high-fat based diet (>45% calories from fat), mixed with a stock diet, results in substantially more weight gain over a similar period, of time, which would allow an investigator to use ~40% fewer animals in their experimental model.

Baker's yeast ß-glucan supplementation increases monocytes and cytokines post-exercise: implications for infection risk?

Strenuous aerobic exercise is known to weaken the immune system, and while many nutritional supplements have been proposed to boost post-exercise immunity, few are known to be effective. The purpose of the present study was to evaluate whether 10 d of supplementation with a defined source of baker's yeast ß-glucan (BG, Wellmune WGP®) could minimise post-exercise immunosuppression. Recreationally active men and women (n 60) completed two 10 d trial conditions using a cross-over design with a 7 d washout period: placebo (rice flour) and baker's yeast BG (250 mg/d of ß-1,3/1,6-glucans derived from Saccharomyces cerevisiae) before a bout of cycling (49 ± 6 min) in a hot (38 ± 2°C), humid (45 ± 2 % relative humidity) environment. Blood was collected at baseline (before supplement), pre- (PRE), post- (POST) and 2 h (2H) post-exercise. Total and subset monocyte concentration was measured by four-colour flow cytometry. Plasma cytokine levels and lipopolysaccharide (LPS)-stimulated cytokine production were measured using separate multiplex assays. Total (CD14⁺) and pro-inflammatory monocyte concentrations (CD14⁺/CD16⁺) were significantly greater at POST and 2H (P<0·05) with BG supplementation. BG supplementation boosted LPS-stimulated production of IL-2, IL-4, IL-5 and interferon-γ (IFN-γ) at PRE and POST (P<0·05). Plasma IL-4, IL-5 and IFN-γ concentrations were greater at 2H following BG supplementation. It appears that 10 d of supplementation with BG increased the potential of blood leucocytes for the production of IL-2, IL-4, IL-5 and IFN-γ. The key findings of the present study demonstrate that BG may have potential to alter immunity following a strenuous exercise session.

Mouse blood monocytes: standardizing their identification and analysis using CD115.

Monocytes have been used to assess immune dysfunction and disease. While mouse models are a useful longitudinal analog, few researchers have assessed changes in mouse monocytes. The purpose of this study was to provide recommendations for the sample processing and flow cytometric analysis of mouse blood monocytes. Blood was drawn in a non-lethal manner from CD-1 male mice to be used in three experiments. Experiment 1 compared commonly used mouse monocyte markers. Experiment 2 compared the stability of CD115 expression after immediate (0h) and delayed (2 and 4h) processing following blood collection under various experimental conditions (laser strength, anticoagulant, and storage temp.). Experiment 3 compared the consistency of CD115(+) monocyte and subset concentrations using decreasing (40, 20, 10 and 5µL) volumes of blood. In experiment 1, >95% of CD115(+) events co-expressed CD11b; >85% co-expressed CD14. 70% of CD14(+) and 50% of CD11b(+) events co-expressed CD115. In experiment 2, CD115 expression decreased by 33% between 0 and 4h when stored at room temperature. Blood treated with EDTA and refrigerated maintained CD115 stability. In experiment 3, calculated concentrations for total monocyte events varied by <10% when 40, 20 and 10µL of blood were stained. While CD115 staining provides the most distinct monocyte population, it is important to treat blood with EDTA and refrigerate if sample processing will be delayed over 2h. Collectively, the findings of the present study outline important considerations that must be addressed when examining mouse monocytes in small, non-lethal blood samples.

Interpersonal competence and sex risk behaviours among Jamaican adolescents.

OBJECTIVE: Of particular public health concern to the Jamaican authorities is the consistently high numbers of new HIV infections among adolescents and young adults. The thrust in HIV/AIDS prevention campaigns has largely been toward an increase in knowledge and attitudes as opposed to personality variables. However, it is widely believed that persons with high interpersonal skills may be less likely to engage in sex risk behaviours. This study investigated interpersonal competence as a personality characteristic associated with sexual risk-taking among Jamaican adolescents. METHODS: A cross-sectional survey of 500 adolescents, ages 13-18 years (250 males and 250 females) from nine randomly selected secondary government schools within Kingston and St Andrew was used. The sample ensured maximum variation in age groups. The BarOn EQ-i:YV(S) was utilized to provide a measure of interpersonal competence and the Sex Risk Scale from the Adolescent Risk Inventory acted as a measure of sex risk behaviours. The Spearman's rho correlational statistic was used to investigate the hypothesis. RESULTS: Of the students surveyed, 58.6% reported that they were sexually active; 31.8% reported having multiple sexual partners and 28.2% reporting inconsistent condom use. A significant, inverse relationship was observed between interpersonal competence and sex risk behaviours (p < 0.05). CONCLUSION: This research provided evidence that adolescents with high interpersonal skills are less likely to participate in risky sexual behaviours. Therefore, interventions aimed at reducing risky adolescent sexual practices might benefit from the inclusion of strategies to build interpersonal skills.

Interpersonal competence and sex risk behaviours among Jamaican adolescents / Competencia interpersonal y comportamientos de riesgo sexual entre los adolescentes Jamaicanos

OBJECTIVE: Of particular public health concern to the Jamaican authorities is the consistently high numbers of new HIV infections among adolescents and young adults. The thrust in HIV/AIDS prevention campaigns has largely been toward an increase in knowledge and attitudes as opposed to personality variables. However, it is widely believed that persons with high interpersonal skills may be less likely to engage in sex risk behaviours. This study investigated interpersonal competence as a personality characteristic associated with sexual risk-taking among Jamaican adolescents. METHODS: A cross-sectional survey of 500 adolescents, ages 13−18 years (250 males and 250 females) from nine randomly selected secondary government schools within Kingston and St Andrew was used. The sample ensured maximum variation in age groups. The BarOn EQ-i:YV(S) was utilized to provide a measure of interpersonal competence and the Sex Risk Scale from the Adolescent Risk Inventory acted as a measure of sex risk behaviours. The Spearman's rho correlational statistic was used to investigate the hypothesis. RESULTS: Of the students surveyed, 58.6% reported that they were sexually active; 31.8% reported having multiple sexual partners and 28.2% reporting inconsistent condom use. A significant, inverse relationship was observed between interpersonal competence and sex risk behaviours (p < 0.05). CONCLUSION: This research provided evidence that adolescents with high interpersonal skills are less likely to participate in risky sexual behaviours. Therefore, interventions aimed at reducing risky adolescent sexual practices might benefit from the inclusion of strategies to build interpersonal skills.

OBJETIVO: De interés particular para las autoridades jamaicanas de la salud pública es el número consistentemente alto de nuevas infecciones de VIH entre adolescentes y adultos jóvenes. El desempeño principal de las campañas de prevención del VIH/SIDA ha estado dirigido en gran parte hacia el aumento de conocimientos y actitudes, en vez de a variables de la personalidad. Sin embargo, existe la creencia ampliamente extendida de que las personas con buen desarrollo de sus habilidades interpersonales, serán menos propensas a involucrarse en conductas de riesgo sexual. Este estudio investigó la competencia interpersonal como una característica de personalidad asociada con riesgos sexuales entre adolescentes jamaicanos. MÉTODOS: Se llevó a cabo un estudio transversal de 500 adolescentes, con edades entre 13−18 años (250 varones, y 250 hembras) de nueve escuelas secundarias del gobierno, seleccionadas al azar en las áreas de Kingston y Saint Andrew. La muestra garantizó una máxima variación en los grupos de edad. Se usó el test BarOn EQ-i:YV (S) para obtener una medida de la competencia interpersonal, en tanto que la Escala de Riesgo Sexual del Inventario de Riesgo Adolescente actuó como una medida de los comportamientos de riesgo sexual. Las estadísticas correlaciónales de Spearman fueron utilizadas para investigar la hipótesis. RESULTADOS: De los estudiantes encuestados, 58.6% informó estar sexualmente activos; 31.8% informó tener múltiples parejas sexuales, y el 28.2%, reportó un uso sistemático del condón. Se observó una relación inversa significativa entre la competencia interpersonal y las conductas de riesgo sexual (p < 0.05). CONCLUSIÓN: Esta investigación proporcionó evidencia de que los adolescentes con alto desarrollo de habilidades interpersonales son menos propensos a participar en conductas sexuales riesgosas. Por lo tanto, las intervenciones destinadas a reducir las prácticas sexuales adolescentes riesgosas, podrían beneficiarse con la inclusión de las estrategias para desarrollar las habilidades interpersonales.

An improved ultra performance liquid chromatography-tandem mass spectrometry method for the determination of alloisoleucine and branched chain amino acids in dried blood samples.

BACKGROUND: Branched chain amino acid (BCAA) analysis is needed for the diagnosis and management of patients with maple syrup urine disease (MSUD). We report an improved ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of BCAAs and Allo-Ile in dried blood spot (DBS) samples. METHODS: BCAAs were extracted from a 3 mm blood spot into methanol/water containing stable isotope internal standards. Eluents were dried and reconstituted in the mobile phase. Gradient elution was performed on an Acquity™ BEH C(18) (100 × 2.1 mm, 1.7 µm) column. BCAAs were detected and quantified in the multiple reaction monitoring mode in a five-minute analysis. RESULTS: The assay was calibrated to give best possible alignment with plasma results. Retrospective analysis of newborn DBSs from six classic MSUD patients showed elevated alloisoleucine (Allo-Ile) in all cases. Two of four patients with mild disease had normal values; the other two had significant elevations in Allo-Ile. CONCLUSIONS: Analysis of BCAA in DBS by UPLC-MS/MS is a useful second tier newborn screening test to identify classical MSUD and for monitoring of remote patients.

Weight gain in response to high-fat feeding in CD-1 male mice.

The purpose of this study was to compare weight gain and food intake during high-fat feeding in outbred CD-1 male mice while considering several different experimental designs. This study was completed using data from three separate experiments and was designed to address different experimental design issues. Experiment 1 compared mice housed in groups or singly. Experiment 2 compared adolescent and young adult mice. Experiment 3 examined mice that had been previously exercise-trained prior to diet-induced weight gain. Data from each experiment were analysed using repeated measures analysis of variance and linear regression. While housing and age did not significantly affect weight gain, mice that were previously exercise-trained consumed significantly more kilocalories than sedentary mice while maintaining comparable body weights. We generated a linear prediction model using data from Experiments 1 and 2 that will allow investigators to calculate the weeks of high-fat feeding needed to reach a target body weight. Our key findings characterize the issues related to and affecting experimental design when utilizing an outbred mouse diet-induced weight gain model and will serve as a guide for future researchers.

Measures of patient safety in developing and emerging countries: a review of the literature.

CONTEXT: The World Alliance for Patient Safety was formed to accelerate worldwide research progress towards measurably improving patient safety. Although rates of adverse events have been studied in industrialised countries, little is known about the rates of adverse events in developing and emerging countries. PURPOSE: To review the literature on patient safety issues in developing and emerging countries, to identify patient safety measures presently used in these countries and to propose a method of measurably improving patient safety measurement in these countries. METHODS: Using the Medline database for 1998 to 2007, we identified and reviewed 23 English-language articles that examined patient safety measurement in developing and emerging countries. Results Our review included 12 studies that prospectively measured patient safety and 11 studies that retrospectively measured safety. Two studies used measures of structure and the remaining used process measures, outcome measures or both. Whereas a few studies used surveys or direct observation, most studies used chart audits to measure patient safety. Most studies addressed safety at a single facility. CONCLUSIONS: Investigation of patient safety in developing and emerging countries has been infrequent and limited in scope. Establishing fundamental safe patient practices, integrating those processes into routine health services delivery and developing patients' expectations that such processes be present are necessary prerequisites to measuring and monitoring progress towards safe patient care in emerging and developing countries.

Ipilimumab-induced hypophysitis: MR imaging findings.

Ipilimumab is a promising new immunotherapeutic antineoplastic agent with clinical activity in the treatment of metastatic melanoma and renal cell carcinoma. With advances in immunotherapy, however, a host of new side effects related to the mechanism of action of these drugs has appeared. At our institution, 3 patients presented with hypophysitis, which was attributed to an autoimmune process based on the documented relationship of the drug to other autoimmune phenomena and significant and rapid improvement with discontinuation of the drug and addition of steroids. We present the imaging findings in 3 patients with presumed ipilimumab-induced hypophysitis.

In vivo assessment of mutations in OTC for dominant-negative effects following rAAV2/8-mediated gene delivery to the mouse liver.

Mutant proteins have the potential to exert dominant-negative effects that might limit the therapeutic efficacy of their wild-type counterparts after gene transfer. For ornithine transcarbamylase (OTC) deficiency, in vitro studies have suggested the presence of dominant-negative effects, however, supporting in vivo studies have not been conducted. In this study, we exploited the capacity of recombinant adeno-associated virus (rAAV) 2/8 vectors to deliver transgenes to the mouse liver with high efficiency to determine whether expression of selected OTC mutant proteins exert inhibitory effects on endogenous wild-type OTC enzymatic activity. Using site-directed mutagenesis we constructed three OTC mutants with a theoretical or reported in vitro capacity to exert dominant-negative effects, and delivered these to the liver using rAAV2/8. Each mutation had been earlier identified in patients with OTC deficiency. Treated mice showed no increase in urinary orotic acid levels or reduction in OTC activity despite supra-physiological expression of the mutant proteins, consistent with an absence of dominant-negative effects. These data have important implications for the development of gene therapy strategies for OTC deficiency and validate a model system in which potential dominant-negative effects of specific mutations in prospective patients can be examined empirically before gene therapy.