The turnover and distribution of labelled corticosterone during post-natal development of the duckling (Anas platyrhynchos)

During 6 months of post-natal development in the laboratory, the weight of the adrenal gland relative to body weight decreases exponentially. In the 3 day-old duckling a single intravenous dose of labelled corticosterone becomes distributed in a very large apparent volume and an "extracellular" pool that is greater than the extracellular fluid volume and the pattern of disappearance of labelled hormone from plasma is biphasic. Later during development the volumes of distribution decrease and the biphasic pattern of disappearance becomes less distinct until at 6 months only one phase of disappearance can be detected with confidence. No significant change in plasma corticosterone concentration occurs during this period of development in the laboratory. Estimations of the corticosterone secretory rates, however, indicate that whereas the adrenal weight-specific rate os secretion increases during the first 3 weeks and declines therafter, the bodyf weight-specific secretory rate continues to decline throughtout the period of increasing body weight.     

Fecal Coliform Removal within a Marshland Upwelling System Consisting of Scatlake Soils

The marshland upwelling system (MUS) was installed on private camps in the Grand Bay National Estuarine Research Reserve, Moss Point, Mississippi. The system was evaluated for its effectiveness in removing fecal coliforms from settled, raw wastewater. A suite of studies was performed at flow rates of 1.9, 2.8, and 5.5 L/min and an injection frequency of 30 min every 3 h to investigate fecal coliform removal. An additional study was performed at a flow rate of 2.8 L/min and an injection frequency of 15 min every hour. Overall, the MUS consistently maintained fecal concentrations below effluent regulatory standards for shellfish harvesting waters (14 most probable number of colonies per 100 mL). Mean influent concentrations of 55,269±2,218,016 colony forming units (CFU)/100 mL were reduced to effluent counts of 2.7±14.07 CFU/100 mL (observed in the 1.5 m wells). Three- to four-log reductions in influent counts were observed over the initial 1.4 vector?m from the injection well. The overall removal followed a first-order decay relationship with respect to vector distance, resulting in removal rate constants ranging from 5.6 to 6.6/m and predicted surface concentrations approaching 0 CFU/100 mL. The 2.8 L/min for 30 min every 3 h treatment provided the best effluent quality.     

Rough Terrain Autonomous Mobility—Part 1: A Theoretical Analysis of Requirements

A basic requirement of autonomous vehicles is that of guaranteeing the safety of the vehicle by avoiding hazardous situations. This paper analyses this requirement in general terms of real-time response, throughput, and the resolution and accuracy of sensors and computations. Several nondimensional expressions emerge which characterize requirements in canonical form.The automatic generation of dense geometric models for autonomously navigating vehicles is a computationally expensive process. Using first principles, it is possible to quantify the relationship between the raw throughput required of the perception system and the maximum safely achievable speed of the vehicle. We derive several useful expressions for the complexity of terrain mapping perception under various assumptions. All of them can be reduced to polynomials in the response distance.The significant time consumed by geometric perception degrades real-time response characteristics. Using our results, several strategies of active geometric perception arise which are practical for autonomous vehicles and increasingly important at higher speeds.     

Calorimetric and thermodynamic analysis of palladium-deuterium electrochemical cells

An analysis is made of the chemical heats liberated from a palladium-deuterium electrochemical cell operating inside a calorimeter. It is important, in such an analysis, to carefully identify the chemical and electrochemical sources of heat, before any “excess heats” can be ascribed to non-chemical reactions.⁽¹⁾ The calorimeter measures the enthalpy (ΔH _r ) of the reaction; whereas, the electrochemical voltage of the cell reflects the free energy (ΔG _r ) of the reaction within the Pd-D electrolysis cell. The heat energy from the calorimeter cell therefore doesnot equal the electrical energy supplied to the cell, as might initially be expected. The magnitudes of the differing calorimetric and electrochemical energies were found to be related through the “thermoneutral potential” (ξ_H) of the electrochemical reaction. The chemical heat theoretically expected from the calorimeter is given by (1) I(ξ_L-ξ_H), the cell current (I), multiplied by the difference between the operating cell voltage (ξ_L) and the thermoneutral potential (ξ_H), rather than (2) Iξ_L, the electrical input power. This was verified empirically using a freon vaporization calorimeter, which operates on the principle of accurate measurement of the vaporization rate of liquid freon which completely surrounds the electrochemical cell. The calorimetrically-measured heats observed from a Pt/D₂O, 0.1M LiOD/Pd electrochemical cell were within 2% of the thermoneutral potential predicted value, I(ξ_L-ξ_H); but were found to be 15–30% less than the electrical work supplied to the cell, Iξ_L. Measurements of D₂O consumed by the cell reactions also verified that essentially no significant recombination of D₂ and O₂ gases occurred within the cell. No “excess heats” were observed from this Pd cell during the 36 days of its electrolytic operation. Likewise, no increase in the neutron flux around the cell was found, using three³He radiation detectors.     

Evaluation of Slip Feed System for Vapor-Phase Bioreactors

This study evaluated the ability of a slip feed system to maintain the pollutant-degrading activity of biomass in a vapor phase bioreactor during periods of little or no contaminant feed. Three slip feed fractions (1, 5, and 9% by mass) were investigated in a bioreactor treating toluene-contaminated air. Results indicate that the slip feed system improved the contaminant degradation capacity in the carbon-deprived bioreactor zones by approximately 30% regardless of the slip feed fraction supplied. This yielded a 10–20% improvement in toluene removal efficiency when the entire bioreactor column was subjected to a sudden threefold spike in toluene loading. When a 12% slip feed was supplied to the bioreactor during a short-term (3 days) or longer-term (1 week) shutdown period, it reduced the reacclimation time required following restart by as much as 70%. These results indicate that as long as the biomass receives at least a trace of the pollutant by means of a slip feed system, it can maintain sufficient pollutant degrading capacity to respond quickly to pollutant loadings upon restart.     

Trans-Reduction of Cerebral Small Vessel Disease Proteins by Notch-Derived EGF-like Sequences

Cysteine oxidation states of extracellular proteins participate in functional regulation and in disease pathophysiology. In the most common inherited dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), mutations in NOTCH3 that alter extracellular cysteine number have implicated NOTCH3 cysteine states as potential triggers of cerebral vascular smooth muscle cytopathology. In this report, we describe a novel property of the second EGF-like domain of NOTCH3: its capacity to alter the cysteine redox state of the NOTCH3 ectodomain. Synthetic peptides corresponding to this sequence (NOTCH3 N-terminal fragment 2, NTF2) readily reduce NOTCH3 N-terminal ectodomain polypeptides in a dose- and time-dependent fashion. Furthermore, NTF2 preferentially reduces regional domains of NOTCH3 with the highest intensity against EGF-like domains 12–15. This process requires cysteine residues of NTF2 and is also capable of targeting selected extracellular proteins that include TSP2 and CTSH. CADASIL mutations in NOTCH3 increase susceptibility to NTF2-facilitated reduction and to trans-reduction by NOTCH3 produced in cells. Moreover, NTF2 forms complexes with the NOTCH3 ectodomain, and cleaved NOTCH3 co-localizes with the NOTCH3 ectodomain in cerebral arteries of CADASIL patients. The potential for NTF2 to reduce vascular proteins and the enhanced preference for it to trans-reduce mutant NOTCH3 implicate a role for protein trans-reduction in cerebrovascular pathological states such as CADASIL.     

An Iron Refractory Phenotype in Obese Adipose Tissue Macrophages Leads to Adipocyte Iron Overload

Adipocyte iron overload is a maladaptation associated with obesity and insulin resistance. The objective of the current study was to determine whether and how adipose tissue macrophages (ATMs) regulate adipocyte iron concentrations and whether this is impacted by obesity. Using bone marrow-derived macrophages (BMDMs) polarized to M0, M1, M2, or metabolically activated (MMe) phenotypes, we showed that MMe BMDMs and ATMs from obese mice have reduced expression of several iron-related proteins. Furthermore, the bioenergetic response to iron in obese ATMs was hampered. ATMs from iron-injected lean mice increased their glycolytic and respiratory capacities, thus maintaining metabolic flexibility, while ATMs from obese mice did not. Using an isotope-based system, we found that iron exchange between BMDMs and adipocytes was regulated by macrophage phenotype. At the end of the co-culture, MMe macrophages transferred and received more iron from adipocytes than M0, M1, and M2 macrophages. This culminated in a decrease in total iron in MMe macrophages and an increase in total iron in adipocytes compared with M2 macrophages. Taken together, in the MMe condition, the redistribution of iron is biased toward macrophage iron deficiency and simultaneous adipocyte iron overload. These data suggest that obesity changes the communication of iron between adipocytes and macrophages and that rectifying this iron communication channel may be a novel therapeutic target to alleviate insulin resistance.     

Assessing the Morphological and Behavioral Toxicity of Catechol Using Larval Zebrafish

Catechol is a ubiquitous chemical used in the manufacturing of fragrances, pharmaceuticals and flavorants. Environmental exposure occurs in a variety of ways through industrial processes, during pyrolysis and in effluent, yet despite its prevalence, there is limited information regarding its toxicity. While the genotoxicity and gastric carcinogenicity of catechol have been described in depth, toxicological studies have potentially overlooked a number of other effects relevant to humans. Here, we have made use of a general and behavioral larval zebrafish toxicity assay to describe previously unknown catechol-based toxicological phenomena. Behavioral testing revealed catechol-induced hypoactivity at concentrations an order of magnitude lower than observable endpoints. Catechol exposure also resulted in punctate melanocytes with concomitant decreases in the expression of pigment production and regulation markers mitfa, mc1r and tyr. Because catechol is converted into a number of toxic metabolites by tyrosinase, an enzyme found almost exclusively in melanocytes, an evaluation of the effects of catechol on these cells is critical to evaluating the safety of this chemical. This work provides insights into the toxic nature of catechol and highlights the benefits of the zebrafish larval testing platform in being able to dissect multiple aspects of toxicity with one model.     

A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping

We recently described cell-projection pumping as a mechanism transferring cytoplasm between cells. The uptake of fibroblast cytoplasm by co-cultured SAOS-2 osteosarcoma cells changes SAOS-2 morphology and increases cell migration and proliferation, as seen by single-cell tracking and in FACS separated SAOS-2 from co-cultures. Morphological changes in SAOS-2 seen by single cell tracking are consistent with previous observations in fixed monolayers of SAOS-2 co-cultures. Notably, earlier studies with fixed co-cultures were limited by the absence of a quantitative method for identifying sub-populations of co-cultured cells, or for quantitating transfer relative to control populations of SAOS-2 or fibroblasts cultured alone. We now overcome that limitation by a novel Cartesian plot analysis that identifies individual co-cultured cells as belonging to one of five distinct cell populations, and also gives numerical measure of similarity to control cell populations. We verified the utility of the method by first confirming the previously established relationship between SAOS-2 morphology and uptake of fibroblast contents, and also demonstrated similar effects in other cancer cell lines including from melanomas, and cancers of the ovary and colon. The method was extended to examine global DNA methylation, and while there was no clear effect on SAOS-2 DNA methylation, co-cultured fibroblasts had greatly reduced DNA methylation, similar to cancer associated fibroblasts.