Uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily. Based upon its high homology with UCP1 and its restricted tissue distribution to skeletal muscle and brown adipose tissue, UCP3 has been suggested to play important roles in regulating energy expenditure, body weight, and thermoregulation. Other postulated roles for UCP3 include regulation of fatty acid metabolism, adaptive responses to acute exercise and starvation, and prevention of reactive oxygen species (ROS) formation. To address these questions, we have generated mice lacking UCP3 (UCP3 knockout (KO) mice). Here, we provide evidence that skeletal muscle mitochondria lacking UCP3 are more coupled (i.e. increased state 3/state 4 ratio), indicating that UCP3 has uncoupling activity. In addition, production of ROS is increased in mitochondria lacking UCP3. This study demonstrates that UCP3 has uncoupling activity and that its absence may lead to increased production of ROS. Despite these effects on mitochondrial function, UCP3 does not seem to be required for body weight regulation, exercise tolerance, fatty acid oxidation, or cold-induced thermogenesis. The absence of such phenotypes in UCP3 KO mice could not be attributed to up-regulation of other UCP mRNAs. However, alternative compensatory mechanisms cannot be excluded. The consequence of increased mitochondrial coupling in UCP3 KO mice on metabolism and the possible role of yet unidentified compensatory mechanisms, remains to be determined. Uncoupling protein 3 (UCP3)1 (1-3) is a member of the mitochondrial anion carrier superfamily with high homology (57%) to UCP1, a well characterized uncoupling protein (4, 5). UCP3 together with UCP1, UCP2 (6, 7), and possibly BMCP1 (brain mitochondrial carrier protein) (8) and UCP4 (9), form a family of uncoupling proteins located in the inner mitochondrial membrane. The evidence supporting the uncoupling activity of these proteins comes from studies where UCPs have been heterologously expressed in yeast or reconstituted into proteoliposomes. The expression of UCP2 and -3 decreases the mitochondrial membrane potential, as assessed by uptake of fluorescent membrane potential-sensitive dyes in whole yeast. They also increase state 4 respiration in isolated mitochondria, which serves as an indicator of inner membrane proton leak (3, 6, 10). More recently, reconstitution of UCPs into liposomes has shown that UCP2 and UCP3, like UCP1, mediate proton transport across bilipid layers (11). It is well established that UCP1 is exclusively expressed in brown fat, where it plays a key role in facultative thermogenesis in rodents. Although there is controversy about the molecular mechanisms involved (12-16), it is clear that activated UCP1 catalyzes a proton leak across the mitochondrial inner membrane leading to thermogenesis. The activity of UCP1 is highly regulated, facilitated by fatty acids and inhibited by purine ribose di-and trinucleotides (ATP, ADP, GTP, GDP) (17). UCP1 is also highly regulated at the transcriptional level (18) by cat...

Patterns of behavior exhibited by mice in their home cages reflect the function and interaction of numerous behavioral and physiological systems. Detailed assessment of these patterns thus has the potential to provide a powerful tool for understanding basic aspects of behavioral regulation and their perturbation by disease processes. However, the capacity to identify and examine these patterns in terms of their discrete levels of organization across diverse behaviors has been difficult to achieve and automate. Here, we describe an automated approach for the quantitative characterization of fundamental behavioral elements and their patterns in the freely behaving mouse. We demonstrate the utility of this approach by identifying unique features of home cage behavioral structure and changes in distinct levels of behavioral organization in mice with single gene mutations altering energy balance. The robust, automated, reproducible quantification of mouse home cage behavioral structure detailed here should have wide applicability for the study of mammalian physiology, behavior, and disease.circadian ͉ ingestion ͉ obesity ͉ phenotyping M olecular genetic approaches for manipulating gene expression and neural activity in mice, combined with the elucidation of the mouse genome, provide unprecedented opportunities for the investigation of diverse behavioral processes in the context of a mammalian system. While substantial insights have been gained through the application of existing behavioral assays, many of these examine behavior over a limited time window and focus on a single behavioral domain (1, 2). To complement such approaches, we developed an automated, readily-standardized quantitative approach for elucidating the complex organization of diverse behaviors exhibited by mice in their home cages.We focused on mice in their home cages because the organization of behavior in freely acting animals provides a window into the central integration of numerous behavioral and physiological systems (e.g., energy balance, thermal status, osmotic/volume status, sleep, reproduction, defense, and environmental entrainment). The functions and interactions of these systems result in the coordinated organization of multiple behaviors (3-5). Although several sophisticated approaches for automated behavioral data collection and home cage monitoring exist, they do not employ algorithms that quantitatively capture the rich temporal and spatial structure of diverse behaviors that occur over multiple time scales (1,(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19).As a first step in examining this structure, we made use of the observation that in natural environments animals typically alternate between two major discrete states, active and inactive (20)(21)(22). During active states (ASs), animals engage in behaviors such as foraging and patrolling within a regularly traversed home range. During inactive states (ISs), animals return to a refuge (nest, burrow, or home base) and engage in behaviors such as rest and sleep (23-26). These...

The relationships between cerebral blood flow (CBF), arterial oxygen content, whole blood viscosity and the transport of oxygen to the brain have been studied in 54 subjects with variations in arterial oxygen content resulting from alterations in haemoglobin concentration ranging from 5.7 to 19.1 g/dl. A highly significant relationship was found between CBF and arterial oxygen content, CBF altering to maintain the rate of transport of oxygen to the brain within certain limits. No significant effect of changes in blood viscosity was evident. In a multiple regression analysis arterial oxygen content emerged as the major determinant of CBF and after this effect had been taken into account there was no significant influence of blood viscosity, arterial pCO2, age or mean arterial blood pressure. The results imply that the alterations in CBF found in subjects with anaemia and polycythaemia are primarily physiological and not caused by the accompanying alterations in blood viscosity. It is proposed that the response of the cerebral circulation to arterial oxygen content is mediated by a local mechanism sensitive to alterations in local tissue oxygen tension.

Background:Patients with immune-mediated diseases on immunosuppressive therapies have more infectious episodes than healthy individuals, yet vaccination practices by physicians for this patient population remain suboptimal.Objectives:To evaluate the safety and efficacy of vaccines in individuals exposed to immunosuppressive therapies and provide evidence-based clinical practice recommendations.Methods:A literature search for vaccination safety and efficacy in patients on immunosuppressive therapies (2009-2017) was conducted. Results were assessed using the Grading of Recommendation, Assessment, Development, and Evaluation system.Results:Several immunosuppressive therapies attenuate vaccine response. Thus, vaccines should be administered before treatment whenever feasible. Inactivated vaccines can be administered without treatment discontinuation. Similarly, evidence suggests that the live zoster vaccine is safe and effective while on select immunosuppressive therapy, although use of the subunit vaccine is preferred. Caution regarding other live vaccines is warranted. Drug pharmacokinetics, duration of vaccine-induced viremia, and immune response kinetics should be considered to determine appropriate timing of vaccination and treatment (re)initiation. Infants exposed to immunosuppressive therapies through breastmilk can usually be immunized according to local guidelines. Intrauterine exposure to immunosuppressive agents is not a contraindication for inactivated vaccines. Live attenuated vaccines scheduled for infants and children ⩾12 months of age, including measles, mumps, rubella, and varicella, can be safely administered as sufficient time has elapsed for drug clearance.Conclusions:Immunosuppressive agents may attenuate vaccine responses, but protective benefit is generally maintained. While these recommendations are evidence based, they do not replace clinical judgment, and decisions regarding vaccination must carefully assess the risks, benefits, and circumstances of individual patients.