Use of Di- and Tripropionate substrate analogs to probe the active site of human recombinant coproporphyrinogen oxidase

Background: Defects in the enzyme coproporphyrinogen oxidase result in accumulation of porphyrins which may affect the severity of a subset of porphyrias. Thus evaluation of this enzyme for substrate selectivity is of value. Kinetic evaluations of recombinant human coproporphyrinogen oxidase have been undertaken using six di- and tripropionate analogs of the natural substrate coproporphyrinogen-III. These Substrate analogs were modified by having alkyl groups in place of one or both of the ring 13- or 17-propionate moieties. Material/Methods: Cloned human enzyme was incubated with analogs under apparent first order conditions and with various substrate concentrations. The kinetic values, K-m and V-max, were determined. Results: Relative to the authentic substrate, the K-m values for the 13-ethyl, dimethyl and diethyl porphyrinogens were very comparable whereas the K-m values were much higher using dipropyl and dibutyl porphyrinogen and much lower for the 17-ethyl analog. For the dipropionate analogs, the V-max values were an apparent function of the carbon length of the substituent. on the C and D rings, with longer carbon length severely reducing product formation by some 4-5 orders of magnitude. Also, the two isomeric tripropionates that were tested indicated that it was more detrimental to have an ethyl group at the 13-position for both binding and catalysis. Conclusions: This work extends our understanding of porphyrin ring substituent effects reported by Cooper et al. (2005). The substituents on both the C and D rings have significant effects on both the substrate binding and catalysis by this important enzyme

Relationships Among Metabolic-Risk, Body Fatness, and Muscular Fitness in Young Obese Latino Children

International Journal of Exercise Science 13(3): 488-500, 2020. Given the high prevalence of obesity in Latino children and potential health risks, the purpose of this study was to: 1) evaluate relationships among metabolic-risk, fitness, and body fatness; 2) determine sex differences in cardio-metabolic risk factors and fitness of obese children of Latino descent. Sixty children (boys, n= 39, 7.8 ± 1.5 years; girls, n= 21, 7.2 ± 1.5 years; body mass index, 97.8 ± 2.5thpercentile) completed assessments of height, weight, and body fat, prior to fasted blood draws and a battery of fitness tests. Cardio-metabolic markers were analyzed, and a metabolic risk score created. Correlations and regression analyses evaluated the relationships among body fatness, metabolic-risk, and fitness. Independent samples t-tests determined sex differences (p \u3c 0.05). Body fat related negatively to lower body power (p \u3c 0.016), but positively to upper body power (p= 0.049). After controlling for age and sex, body fat (p\u3c 0.001) was a positive predictor of variance in metabolic-risk scores, (R2 = 0.39, p \u3c 0.001). Further, the association between body fat and metabolic-risk was not moderated by sex. Metabolic-risk scores and body fat were similar for both sexes, but boys performed better on muscular fitness tests, even after accounting for fat free mass (p \u3c 0.05). Higher body fatness in obese Latino children may result in greater metabolic-risk and difficulty performing weight-bearing tasks. Therefore, culturally adapted weight management programs should employ a multifaceted approach to improve metabolic-risk and fitness

General moments of the inverse real Wishart distribution and orthogonal Weingarten functions

Let $W$ be a random positive definite symmetric matrix distributed according to a real Wishart distribution and let $W^{-1}=(W^{ij})_{i,j}$ be its inverse matrix. We compute general moments $\mathbb{E} [W^{k_1 k_2} W^{k_3 k_4} ... W^{k_{2n-1}k_{2n}}]$ explicitly. To do so, we employ the orthogonal Weingarten function, which was recently introduced in the study for Haar-distributed orthogonal matrices. As applications, we give formulas for moments of traces of a Wishart matrix and its inverse.Comment: 29 pages. The last version differs from the published version, but it includes Appendi

Oceanids C2: An Integrated Command, Control, and Data Infrastructure for the Over-the-Horizon Operation of Marine Autonomous Systems

Long-range Marine Autonomous Systems (MAS), operating beyond the visual line-of-sight of a human pilot or research ship, are creating unprecedented opportunities for oceanographic data collection. Able to operate for up to months at a time, periodically communicating with a remote pilot via satellite, long-range MAS vehicles significantly reduce the need for an expensive research ship presence within the operating area. Heterogeneous fleets of MAS vehicles, operating simultaneously in an area for an extended period of time, are becoming increasingly popular due to their ability to provide an improved composite picture of the marine environment. However, at present, the expansion of the size and complexity of these multi-vehicle operations is limited by a number of factors: (1) custom control-interfaces require pilots to be trained in the use of each individual vehicle, with limited cross-platform standardization; (2) the data produced by each vehicle are typically in a custom vehicle-specific format, making the automated ingestion of observational data for near-real-time analysis and assimilation into operational ocean models very difficult; (3) the majority of MAS vehicles do not provide machine-to-machine interfaces, limiting the development and usage of common piloting tools, multi-vehicle operating strategies, autonomous control algorithms and automated data delivery. In this paper, we describe a novel piloting and data management system (C2) which provides a unified web-based infrastructure for the operation of long-range MAS vehicles within the UK's National Marine Equipment Pool. The system automates the archiving, standardization and delivery of near-real-time science data and associated metadata from the vehicles to end-users and Global Data Assembly Centers mid-mission. Through the use and promotion of standard data formats and machine interfaces throughout the C2 system, we seek to enable future opportunities to collaborate with both the marine science and robotics communities to maximize the delivery of high-quality oceanographic data for world-leading science

Symmetric and asymmetric action integration during cooperative object manipulation in virtual environments

Cooperation between multiple users in a virtual environment (VE) can take place at one of three levels. These are defined as where users can perceive each other (Level 1), individually change the scene (Level 2), or simultaneously act on and manipulate the same object (Level 3). Despite representing the highest level of cooperation, multi-user object manipulation has rarely been studied. This paper describes a behavioral experiment in which the piano movers' problem (maneuvering a large object through a restricted space) was used to investigate object manipulation by pairs of participants in a VE. Participants' interactions with the object were integrated together either symmetrically or asymmetrically. The former only allowed the common component of participants' actions to take place, but the latter used the mean. Symmetric action integration was superior for sections of the task when both participants had to perform similar actions, but if participants had to move in different ways (e.g., one maneuvering themselves through a narrow opening while the other traveled down a wide corridor) then asymmetric integration was superior. With both forms of integration, the extent to which participants coordinated their actions was poor and this led to a substantial cooperation overhead (the reduction in performance caused by having to cooperate with another person)

A structural and physical study of sol–gel methacrylate–silica hybrids: intermolecular spacing dictates the mechanical properties

Sol–gel hybrids are inorganic/organic co-networks with nanoscale interactions between the components leading to unique synergistic mechanical properties, which can be tailored, via a selection of the organic moiety. Methacrylate based polymers present several benefits for class II hybrids (which exhibit formal covalent bonding between the networks) as they introduce great versatility and can be designed with a variety of chemical side-groups, structures and morphologies. In this study, the effect of high cross-linking density polymers on the structure–property relationships of hybrids generated using poly(3-trimethoxysilylpropyl methacrylate) (pTMSPMA) and tetraethyl orthosilicate (TEOS) was investigated. The complexity and fine scale of the co-network interactions requires the development of new analytical methods to understand how network evolution dictates the wide-ranging mechanical properties. Within this work we developed data manipulation techniques of acoustic-AFM and solid state NMR output that provide new approaches to understand the influence of the network structure on the macroscopic elasticity. The concentration of pTMSPMA in the silica sol affected the gelation time, ranging from 2 h for a hybrid made with 75 wt% inorganic with pTMSPMA at 2.5 kDa, to 1 minute for pTMSPMA with molecular weight of 30 kDa without any TEOS. A new mechanism of gelation was proposed based on the different morphologies derived by AC-AFM observations. We established that the volumetric density of bridging oxygen bonds is an important parameter in structure/property relationships in SiO2 hybrids and developed a method for determining it from solid state NMR data. The variation in the elasticity of pTMSPMA/SiO2 hybrids originated from pTMSPMA acting as a molecular spacer, thus decreasing the volumetric density of bridging oxygen bonds as the inorganic to organic ratio decreased

Precision measurement of cis-regulatory energetics in living cells

Gene expression in all organisms is controlled by cooperative interactions between DNA-bound transcription factors (TFs). However, measuring TF-TF interactions that occur at individual cis-regulatory sequences remains difficult. Here we introduce a strategy for precisely measuring the Gibbs free energy of such interactions in living cells. Our strategy uses reporter assays performed on strategically designed cis-regulatory sequences, together with a biophysical modeling approach we call "expression manifolds". We applied this strategy in Escherichia coli to interactions between two paradigmatic TFs: CRP and RNA polymerase (RNAP). Doing so, we consistently obtain measurements precise to ~0.1 kcal/mol. Unexpectedly, CRP-RNAP interactions are seen to deviate in multiple ways from the prior literature. Moreover, the well-known RNAP binding motif is found to be a surprisingly unreliable predictor of RNAP-DNA binding energy. Our strategy is compatible with massively parallel reporter assays in both prokaryotes and eukaryotes, and should thus be highly scalable and broadly applicable

A non perturbative approach of the principal chiral model between two and four dimensions

We investigate the principal chiral model between two and four dimensions by means of a non perturbative Wilson-like renormalization group equation. We are thus able to follow the evolution of the effective coupling constants within this whole range of dimensions without having recourse to any kind of small parameter expansion. This allows us to identify its three dimensional critical physics and to solve the long-standing discrepancy between the different perturbative approaches that characterizes the class of models to which the principal chiral model belongs.Comment: 5 pages, 1 figure, Revte

Specific Subpopulations of Hypothalamic Leptin Receptor-Expressing Neurons Mediate the Effects of Early Developmental Leptin Receptor Deletion on Energy Balance

ACKNOWLEDGEMENTS We thank MedImmune, Inc. and James Trevaskis, PhD and Christopher Rhodes, PhD for the gift of leptin. We thank members of the Myers and Olson labs for helpful discussions. Research support was provided by the Michigan Diabetes Research Center (NIH P3 0 DK020572, including the Molecular Genetics, Animal Phenotyping, and Clinical Cores), the American Diabetes Association (MGM), the Marilyn H. Vincent Foundation (MGM), the NIH (MGM: D K05673 1; ACR:DK071212; MBA: DK097861), the BBSRC (LKH: BB/NO17838/1) and WellcomeTrust (LKH: 098012).Peer reviewedPublisher PD