O2 Consumption Rate Research Articles

Effects of hypoxia on the respiratory and metabolic responses to progressive cooling in newborn rodents that range in heterothermic expression.

What is the central question of this study? Adult homeotherms and heterotherms differ in cold and hypoxia tolerance and in how they match O2 supply and demand in response to these stressors. It has never been ascertained whether these differences reflect different developmental trajectories or whether they are already present at birth. What is the main finding and its importance? When exposed to cold and hypoxia, newborn rodents differed in how they matched O2 supply and demand, with responses reflecting the degree of heterothermic expression and tolerance. Our findings indicate that elements of the adult phenotype are already present at birth. There are physiological differences in how adult rodents regulate O2 supply and O2 demand when exposed to hypoxia in the cold. We examined whether these differences reflect divergent developmental trajectories of homeotherms and heterotherms or whether the differences are already present at birth. We exposed newborn rodents (0-4 days old) that ranged in heterothermic expression [a homeotherm, the rat (Rattus norvegicus); two facultative heterotherms, the mouse (Mus musculus) and the hamster (Mesocricetus auratus); and an obligate heterotherm, the ground squirrel (Ictidomys tridecemlineatus)] to either normoxia (21% O2 ) or hypoxia (7% O2 ) and measured their metabolic, thermoregulatory and ventilatory responses while progressively reducing the ambient temperature from 33 to 15°C. All newborns reduced their body temperature, O2 consumption rate and ventilation during progressive cooling, both in normoxia and in hypoxia. When progressively cooled in hypoxia, however, the homeothermic rats exhibited the greatest thermogenic response, depressed their O2 consumption rate the least and increased ventilation the most. In contrast, the obligate heterotherm, the ground squirrel, did not mount a thermogenic response, exhibited the greatest reduction in O2 consumption rate and increased O2 uptake not by increasing ventilation like the rat, but by extracting ≤80% of the O2 from each breath. Facultative heterotherms (mice and hamsters) exhibited responses in between these two extreme phenotypes. We conclude that even as newborns, homeotherms and heterotherms diverge in how they match O2 supply and O2 demand when progressively cooled in hypoxia, with responses reflecting the degree of heterothermic expression, in addition to reported hypoxia and cold tolerance.

Involvement of purinergic system and electron transport chain in two species of cichlids from the Amazon basin exposed to hypoxia

The Amazonian aquatic ecosystem undergoes seasonal variations and daily changes that directly affect the availability of oxygen. During the day the levels of oxygen can reach supersaturation, and at night can drop to zero. In this way, aquatic organisms are exposed daily to physiological challenges regarding the availability of oxygen. The present study revealed significant differences in the physiology and performance of two cichlids: Geophagus proximus (black water cichlid – from Negro River) and Chaetobranchopsis orbicularis (white water cichlid – from Amazon River), exposed to hypoxia. The white water cichlid showed lower value (1.99 ± 0.79 pKa) of critical pressure of oxygen (Pcrit) and a longer time (68.00 ± 14.11 min) for total loss of balance (LOE); however, this species showed 50% mortality during exposure to hypoxia, while the black water cichlid did not show mortality. Both cichlids presented a decrease in O2 consumption rate (OCR) during hypoxia.. In this sense, it was observed that the black water cichlid presented several physiological strategies during hypoxia, such as, a significant increase in plasma cortisol levels, nucleoside triphosphate diphosphohydrolase activity (for adenosine diphosphate (ADP) as a substrate) in the gills, and the activity of adenosine deaminase (ADA) in gills and liver, in addition to a significant increase in the activity of complexes (II-III) in the transporter chain of electrons in both analyzed tissues and succinate dehydrogenase activity of gills' mitochondria. On the other hand, the only physiological change observed in the white water cichlid was a significant reduction in the activity of complexes II-III in gills and liver. Based on our findings, we can hypothesize that the white water cichlid specie has less tolerant to hypoxia when compared to the black water cichlid.

α-Tocopherol Acetate Attenuates Mitochondrial Oxygen Consumption and Maintains Primitive Cells within Mesenchymal Stromal Cell Population.

We present here the data showing, in standard cultures exposed to atmospheric O2 concentration, that alpha-tocopherol acetate (α-TOA) has a positive impact on primitive cells inside mesenchymal stromal cell (MstroC) population, by maintaining their proliferative capacity. α-TOA decreases the O2 consumption rate of MStroC probably by impacting respiratory chain complex II activity. This action, however, is not associated with a compensatory increase in glycolysis activity, in spite of the fact that the degradation of HIF-1α was decreased in presence of α-TOA. This is in line with a moderate enhancement of mtROS upon α-TOA treatment. However, the absence of glycolysis stimulation implies the inactivity of HIF-1α which might - if it were active - be related to the maintenance of stemness. It should be stressed that α-TOA might act directly on the gene expression as well as the mtROS themselves, which remains to be elucidated. Alpha-tocopherol acetate (α-TOA), a synthetic vitamin E ester, attenuates electron flow through electron transport chain (ETC) which is probably associated with a moderate increase in mtROS in Mesenchymal Stromal Cells. α-TOA action results in enhancement of the proliferative capacity and maintenance of the differentiation potential of the mesenchymal stem and progenitor cells.

Effects of Intertidal Position on Metabolism and Behavior in the Acorn Barnacle, Balanus glandula.

The intertidal zone is characterized by persistent, tidally-driven fluctuations in both abiotic (e.g., temperature, oxygen, and salinity) and biotic (e.g., food availability and predation) factors, which make this a physiologically challenging habitat for resident organisms. The relative magnitude and degree of variability of environmental stress differ between intertidal zones, with the most extreme physiological stress often being experienced by organisms in the high intertidal. Given that so many of the constantly shifting parameters in this habitat are primary drivers of metabolic rate (e.g., temperature, [O2], and food availability), we hypothesized that sessile conspecifics residing in different tidal zones would exhibit distinct “metabolic phenotypes,” a term we use to collectively describe the organisms’ baseline metabolic performance and capacity. To investigate this hypothesis, we collected acorn barnacles (Balanus glandula) from low, mid, and high intertidal positions in San Luis Obispo Bay, CA, and measured a suite of biochemical (whole-animal citrate synthase (CS) and lactate dehydrogenase (LDH) activity, and aerial [D-lactate]), physiological (O2 consumption rates), morphological (body size), and behavioral (e.g., cirri beat frequency and percentage of time operculum open) indices of metabolism. We found tidal zone-dependent differences in B. glandula metabolism that primarily related to anaerobic capacity, cirral activity patterns, and body size. Barnacles from the low intertidal tended to have a greater capacity for anaerobic metabolism (i.e., increased LDH activity and increased baseline [D-lactate]), have reduced cirral beating activity—and presumably reduced feeding—when submerged, and be smaller in size compared to conspecifics in the high intertidal. We did not, however, see any D-lactate accumulation in barnacles from any tidal height throughout 96 h of air exposure. This trend indicates that the enhanced capacity of low intertidal barnacles for anaerobic metabolism may have evolved to support metabolism during more prolonged episodes of emersion or during events other than emersion (e.g., coastal hypoxia and predation). There were also no significant differences in CS activity or baseline O2 consumption rates (in air or seawater at 14°C) across tidal heights, which imply that aerobic metabolic capacity may not be as sensitive to tidal position as anaerobic processes. Understanding how individuals occupying different shore heights differ in their metabolic capacity becomes increasingly interesting in the context of global climate change, given that the intertidal zone is predicted to experience even greater extremes in abiotic stress.

A two-compartment model of oxygen transport in skeletal muscle using continuously distributed capillaries

For future application to studying regulation of microvascular oxygen delivery, a model is developed for O2 transport within an idealized volume of tissue, that is perfused by a continuous distribution of capillaries. Considering oxygen diffusion, convection, and consumption, an O2-dependent transfer term between the capillaries and tissue is used to extend previous single-compartment approaches to include separate tissue and capillary compartments. The coupled tissue–capillary PDE system is considered for unidirectional capillary flow in z, as a simplified model of O2 transport in skeletal muscle, and steady-state 2D solutions are obtained using boundary conditions in x that are consistent with two experimental situations of interest. To validate the continuous capillary model, comparisons are made of an exact nonlinear solution (for no flux at x=0) to results of an established discrete capillary model (solved via finite differences) for varying capillary density, O2 consumption rate, and red blood cell velocity. In addition, comparisons of an approximate linearized solution (for fixed PO2 at x=0) are made to the corresponding discrete capillary solution. Results of the continuous capillary model are presented for varying inlet O2 saturation, showing the utility of the new model for studying physiological problems. Numerical solution of the new model for problems with time dependence and complex geometry is expected to be substantially more efficient than for the corresponding discrete capillary problems.

Study of Ambient Temperature Oxidation in Low Metamorphic Coal and the Oxidation Mechanism

In view of the problem where there is abnormal continuous CO exceedance in the tailgate corner without any sign of spontaneous combustion associated with the mining of low metamorphic coal seam, both desorption experiments and ambient temperature oxidation experiments were carried out, and the ambient temperature oxidation characteristics and its mechanism were studied. The results demonstrate that the original coal seam from which the experimental coal sample was taken, does not contain CO and CO2, and the coal sample can oxidize at ambient temperature. The O2 consumption rate, CO and CO2 production rate is not directly related to the particle size. Low O2 concentration can inhibit but not eliminate the ambient temperature oxidation. In addition to the active sites of coal itself, the ambient temperature oxidation reaction rate is also affected by the inhibitory substances produced by the reaction itself. R1(△CO/△O2), R2(△CO2/△O2) and R3(△CO/△CO2) indexes are mostly not affected by the changes of particle size, O2 concentration and oxidation rates, and the ambient temperature oxidation of coal is dominated by a single mechanism. The ambient temperature oxidation mechanism is also theoretically deduced. The research will have important significance for controlling the abnormal exceedance of CO.

In Escherichia coli Ammonia Inhibits Cytochrome bo3 But Activates Cytochrome bd-I.

Interaction of two redox enzymes of Escherichia coli, cytochrome bo3 and cytochrome bd-I, with ammonium sulfate/ammonia at pH 7.0 and 8.3 was studied using high-resolution respirometry and absorption spectroscopy. At pH 7.0, the oxygen reductase activity of none of the enzymes is affected by the ligand. At pH 8.3, cytochrome bo3 is inhibited by the ligand, with 40% maximum inhibition at 100 mM (NH4)2SO4. In contrast, the activity of cytochrome bd-I at pH 8.3 increases with increasing the ligand concentration, the largest increase (140%) is observed at 100 mM (NH4)2SO4. In both cases, the effector molecule is apparently not NH4+ but NH3. The ligand induces changes in absorption spectra of both oxidized cytochromes at pH 8.3. The magnitude of these changes increases as ammonia concentration is increased, yielding apparent dissociation constants Kdapp of 24.3 ± 2.7 mM (NH4)2SO4 (4.9 ± 0.5 mM NH3) for the Soret region in cytochrome bo3, and 35.9 ± 7.1 and 24.6 ± 12.4 mM (NH4)2SO4 (7.2 ± 1.4 and 4.9 ± 2.5 mM NH3) for the Soret and visible regions, respectively, in cytochrome bd-I. Consistently, addition of (NH4)2SO4 to cells of the E. coli mutant containing cytochrome bd-I as the only terminal oxidase at pH 8.3 accelerates the O2 consumption rate, the highest one (140%) being at 27 mM (NH4)2SO4. We discuss possible molecular mechanisms and physiological significance of modulation of the enzymatic activities by ammonia present at high concentration in the intestines, a niche occupied by E. coli.

The impact of recently excavated dredge pits on coastal hypoxia in the northern Gulf of Mexico shelf

Large volumes of sand are needed in order to combat coastal land loss due to global sea-level rise for restoration of barrier island systems and beaches undergoing rapid erosion and submergence. The sediment required for such projects often originates from dredging of sand deposits on the adjacent shelf. Two dredge pits, with contrasting geology and located at varying distances from the Mississippi River Delta in the northern Gulf of Mexico shelf were sampled during spring and summer. Samples were also collected concurrently from surrounding continental shelf stations that are subject to seasonal hypoxia every summer. The bottom water dissolved O2 inside the dredge pits were found to be consistently hypoxic or near hypoxic throughout both seasons, with high sediment O2 consumption (SOC) rates of 23.7 to 51.8 mmol m−2 d−1 in spring and 34.3 to 51.3 mmol m−2 d−1 in summer. In contrast, control stations immediately outside the dredge pits showed lower SOC rates ranging between 6.3 and 35.9 mmol m−2 d−1. The SOC rates of the surrounding continental shelf subjected to annual seasonal hypoxia ranged between 25.7 and 59.6 mmol m−2 d−1 indicating that the dredge pits experienced similar high rates of SOC. Our results suggest that sluggish water circulation inside these topographic depressions coupled with higher SOC rates does result in persistent low bottom O2 conditions inside these dredge pits well beyond the duration of the seasonal hypoxia period in this region. This is the first study to provide insight on the impacts of dredge pits to surrounding hypoxia in this region which is critical as future dredging operations are expected to increase worldwide with projected sea-level rise.

Microbial BMAA elicits mitochondrial dysfunction, innate immunity activation, and Alzheimer\u2019s disease features in cortical neurons

BackgroundAfter decades of research recognizing it as a complex multifactorial disorder, sporadic Alzheimer’s disease (sAD) still has no known etiology. Adding to the myriad of different pathways involved, bacterial neurotoxins are assuming greater importance in the etiology and/or progression of sAD. β-N-Methylamino-l-alanine (BMAA), a neurotoxin produced by some microorganisms namely cyanobacteria, was previously detected in the brains of AD patients. Indeed, the consumption of BMAA-enriched foods has been proposed to induce amyotrophic lateral sclerosis-parkinsonism-dementia complex (ALS-PDC), which implicated this microbial metabolite in neurodegeneration mechanisms.MethodsFreshly isolated mitochondria from C57BL/6 mice were treated with BMAA and O2 consumption rates were determined. O2 consumption and glycolysis rates were also measured in mouse primary cortical neuronal cultures. Further, mitochondrial membrane potential and ROS production were evaluated by fluorimetry and the integrity of mitochondrial network was examined by immunofluorescence. Finally, the ability of BMAA to activate neuronal innate immunity was quantified by addressing TLRs (Toll-like receptors) expression, p65 NF-κB translocation into the nucleus, increased expression of NLRP3 (Nod-like receptor 3), and pro-IL-1β. Caspase-1 activity was evaluated using a colorimetric substrate and mature IL-1β levels were also determined by ELISA.ResultsTreatment with BMAA reduced O2 consumption rates in both isolated mitochondria and in primary cortical cultures, with additional reduced glycolytic rates, decrease mitochondrial potential and increased ROS production. The mitochondrial network was found to be fragmented, which resulted in cardiolipin exposure that stimulated inflammasome NLRP3, reinforced by decreased mitochondrial turnover, as indicated by increased p62 levels. BMAA treatment also activated neuronal extracellular TLR4 and intracellular TLR3, inducing p65 NF-κB translocation into the nucleus and activating the transcription of NLRP3 and pro-IL-1β. Increased caspase-1 activity resulted in elevated levels of mature IL-1β. These alterations in mitochondrial metabolism and inflammation increased Tau phosphorylation and Aβ peptides production, two hallmarks of AD.ConclusionsHere we propose a unifying mechanism for AD neurodegeneration in which a microbial toxin can induce mitochondrial dysfunction and activate neuronal innate immunity, which ultimately results in Tau and Aβ pathology. Our data show that neurons, alone, can mount inflammatory responses, a role previously attributed exclusively to glial cells.

TNFα induces mitochondrial fragmentation and biogenesis in human airway smooth muscle.

In human airway smooth muscle (hASM), mitochondrial volume density is greater in asthmatic patients compared with normal controls. There is also an increase in mitochondrial fragmentation in hASM of moderate asthmatics associated with an increase in dynamin-related protein 1 (Drp1) and a decrease in mitofusin 2 (Mfn2) expression, mitochondrial fission, and fusion proteins, respectively. Proinflammatory cytokines such TNFα contribute to hASM hyperreactivity and cell proliferation associated with asthma. However, the involvement of proinflammatory cytokines in mitochondrial remodeling is not clearly established. In nonasthmatic hASM cells, mitochondria were labeled using MitoTracker Red and imaged in three dimensions using a confocal microscope. After 24-h TNFα exposure, mitochondria in hASM cells were more fragmented, evidenced by decreased form factor and aspect ratio and increased sphericity. Associated with increased mitochondrial fragmentation, Drp1 expression increased while Mfn2 expression was reduced. TNFα also increased mitochondrial biogenesis in hASM cells reflected by increased peroxisome proliferator-activated receptor-γ coactivator 1α expression and increased mitochondrial DNA copy number. Associated with mitochondrial biogenesis, TNFα exposure also increased mitochondrial volume density and porin expression, resulting in an increase in maximum O2 consumption rate. However, when normalized for mitochondrial volume density, O2 consumption rate per mitochondrion was reduced by TNFα exposure. Associated with mitochondrial fragmentation and biogenesis, TNFα also increased hASM cell proliferation, an effect mimicked by siRNA knockdown of Mfn2 expression and mitigated by Mfn2 overexpression. The results of this study support our hypothesis that in hASM cells exposed to TNFα mitochondria are more fragmented, with an increase in mitochondrial biogenesis and mitochondrial volume density resulting in reduced O2 consumption rate per mitochondrion.

Impacts of vesicular environment on Nox2 activity measurements in vitro

BackgroundThe production of superoxide anions (O2•-) by the phagocyte NADPH oxidase complex has a crucial role in the destruction of pathogens in innate immunity. Majority of in vitro studies on the functioning of NADPH oxidase indirectly follows the enzymatic reaction by the superoxide reduction of cytochrome c (cyt c). Only few reports mention the alternative approach consisting in measuring the NADPH consumption rate.When using membrane vesicles of human neutrophils, the enzyme specific activity is generally found twice higher by monitoring the NADPH oxidation than by measuring the cyt c reduction. Up to now, the literature provides only little explanations about such discrepancy despite the critical importance to quantify the exact enzyme activity. MethodsWe deciphered the reasons of this disparity in studying the role of key parameters, including.cyt c and arachidonic acid concentrations, in conjunction with an ionophore, a detergent and using Clark electrode to measure the O2 consumption rates. ResultsOur results show that the O2•- low permeability of the vesicle membrane as well as secondary reactions (O2•- and H2O2 disproportionations) are strong clues to shed light on this inconsistency. Conclusion and General SignificanceThese results altogether indicate that the cyt c reduction method underestimates the accurate Nox2 activity.

Knockout of VDAC1 in H9c2 Cells Promotes Oxidative Stress-Induced Cell Apoptosis through Decreased Mitochondrial Hexokinase II Binding and Enhanced Glycolytic Stress.

Background/Aims:The role of VDAC1, the most abundant mitochondrial outer membrane protein, in cell death depends on cell types and stimuli. Both silencing and upregulation of VDAC1 in various type of cancer cell lines can stimulate apoptosis. In contrast, in mouse embryonic stem (MES) cells and mouse embryonic fibroblasts (MEFs), the roles of VDAC1 knockout (VDAC1−/−) in apoptotic cell death are contradictory. The contribution and underlying mechanism of VDAC1−/− in oxidative stress-induced cell death in cardiac cells has not been established. We hypothesized that VDAC1 is an essential regulator of oxidative stress-induced cell death in H9c2 cells.Methods:We knocked out VDAC1 in this rat cardiomyoblast cell line with CRISPR-Cas9 genome editing technique to produce VDAC1−/− H9c2 cells, and determined if VDAC1 is critical in promoting cell death via oxidative stress induced by tert-butylhydroper-oxide (tBHP), an organic peroxide, or rotenone (ROT), an inhibitor of mitochondrial complex I by measuring cell viability with MTT assay, cell death with TUNEL stain and LDH release. The mitochondrial and glycolytic stress were examined by measuring O2 consumption rate (OCR) and extracellular acidification rate (ECAR) with a Seahorse XFp analyzer.Results:We found that under control conditions, VDAC1−/− did not affect H9c2 cell proliferation or mitochondrial respiration. However, compared to the wildtype (WT) cells, exposure to either tBHP or ROT enhanced the production of ROS, ECAR, and the proton (H+) production rate (PPR) from glycolysis, as well as promoted apoptotic cell death in VDAC1−/− H9c2 cells. VDAC1−/− H9c2 cells also exhibited markedly reduced mitochondria-bound hexokinase II (HKII) and Bax. Restoration of VDAC1 in VDAC1−/− H9c2 cells reinstated mitochondria-bound HKII and concomitantly decreased tBHP and ROT-induced ROS production and cell death. Interestingly, mitochondrial respiration remained the same after tBHP treatment in VDAC1−/− and WT H9c2 cells.Conclusion:Our results suggest that VDAC1−/− in H9c2 cells enhances oxidative stress-mediated cell apoptosis that is directly linked to the reduction of mitochondria-bound HKII and concomitantly associated with enhanced ROS production, ECAR, and PPR.

Circular RNA NOX4 promotes the development of colorectal cancer via the microRNA‑485‑5p/CKS1B axis.

Colorectal cancer (CRC) is a common malignancy globally. The aim of the present study was to explore the role and the working mechanism of circular RNA NADPH oxidase 4 (circNOX4; circBase ID, hsa_circ_0023990) in CRC. Reverse transcription-quantitative (RT-q)PCR was used to examine the expression of circNOX4, NOX4 mRNA and microRNA (miR)-485-5p in CRC tissues and cell lines. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assays were performed to analyze CRC cell viability and motility. The glycolytic ability of CRC cells was assessed by measuring glucose consumption, lactate production, extracellular acidification and O2 consumption rates using commercial kits. The starBase database was used to predict the targets of circNOX4 and miR-485-5p, and the interaction was confirmed by dual-luciferase reporter and RNA immunoprecipitation assays. A murine xenograft model was established to verify the role of circNOX4 in CRC in vivo. The results demonstrated that the expression of circNOX4 was upregulated in CRC tissues and cell lines compared with that in adjacent normal tissues and a normal colon epithelial cell line, respectively. The expression of circNOX4 was negatively associated with the prognosis of patients with CRC. CircNOX4 silencing suppressed the proliferation, migration, invasion and glycolysis of CRC cells. miR-485-5p was identified as a target of circNOX4. CircNOX4 promoted CRC progression by sponging miR-485-5p. miR-485-5p was demonstrated to bind to the 3′ untranslated region (UTR) of CDC28 protein kinase regulatory subunit 1B (CKS1B). miR-485-5p overexpression-mediated malignant properties of CRC cells were partly reversed by the transfection with the CKS1B overexpression plasmid. CircNOX4 silencing restrained the CRC xenograft growth in vivo. Collectively, the results of the present study demonstrated that circNOX4 may serve an oncogenic role in CRC by promoting the proliferation, migration, invasion and glycolysis of CRC cells via the miR-485-5p/CKS1B axis.

IP3 receptor-mediated Ca2+ release from acidocalcisomes regulates mitochondrial bioenergetics and prevents autophagy in Trypanosoma cruzi

In contrast to animal cells, the inositol 1,4,5-trisphosphate receptor of Trypanosoma cruzi (TcIP3R) localizes to acidocalcisomes instead of the endoplasmic reticulum. Here, we present evidence that TcIP3R is a Ca2+ release channel gated by IP3 when expressed in DT40 cells knockout for all vertebrate IP3 receptors, and is required for Ca2+ uptake by T. cruzi mitochondria, regulating pyruvate dehydrogenase dephosphorylation and mitochondrial O2 consumption, and preventing autophagy. Localization studies revealed its co-localization with an acidocalcisome marker in all life cycle stages of the parasite. Ablation of TcIP3R by CRISPR/Cas9 genome editing caused: a) a reduction in O2 consumption rate and citrate synthase activity; b) decreased mitochondrial Ca2+ transport without affecting the membrane potential; c) increased ammonia production and AMP/ATP ratio; d) stimulation of autophagosome formation, and e) marked defects in growth of culture forms (epimastigotes) and invasion of host cells by infective stages (trypomastigotes). Moreover, TcIP3R overexpressing parasites showed decreased metacyclogenesis, trypomastigote host cell invasion and intracellular amastigote replication. In conclusion, the results suggest a modulatory activity of TcIP3R-mediated acidocalcisome Ca2+ release on cell bioenergetics in T. cruzi.

Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure.

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

Tumor vascular status controls oxygen delivery facilitated by infused polymerized hemoglobins with varying oxygen affinity.

Oxygen (O2) delivery facilitated by hemoglobin (Hb)-based O2 carriers (HBOCs) is a promising strategy to increase the effectiveness of chemotherapeutics for treatment of solid tumors. However, the heterogeneous vascular structures present within tumors complicates evaluating the oxygenation potential of HBOCs within the tumor microenvironment. To account for spatial variations in the vasculature and tumor tissue that occur during tumor growth, we used a computational model to develop artificial tumor constructs. With these simulated tumors, we performed a polymerized human hemoglobin (hHb) (PolyhHb) enhanced oxygenation simulation accounting for differences in the physiologic characteristics of human and mouse blood. The results from this model were used to determine the potential effectiveness of different treatment options including a top load (low volume) and exchange (large volume) infusion of a tense quaternary state (T-State) PolyhHb, relaxed quaternary state (R-State) PolyhHb, and a non O2 carrying control. Principal component analysis (PCA) revealed correlations between the different regimes of effectiveness within the different simulated dosage options. In general, we found that infusion of T-State PolyhHb is more likely to decrease tissue hypoxia and modulate the metabolic rate of O2 consumption. Though the developed models are not a definitive descriptor of O2 carrier interaction in tumor capillary networks, we accounted for factors such as non-uniform vascular density and permeability that limit the applicability of O2 carriers during infusion. Finally, we have used these validated computational models to establish potential benchmarks to guide tumor treatment during translation of PolyhHb mediated therapies into clinical applications.

Effects of crude oil on the microstructure and spontaneous combustion characteristics of coal: a case study of weakly caking coal in the Huangling No. 2 mine, China

The effects of crude oil on the microstructure and spontaneous combustion characteristics of coal were assessed, using raw coal sample and coal samples containing 5%, 10% and 15% oil. The pore structures, oxygen adsorption capacity, surface functional groups and spontaneous combustion characteristics were characterized by the LP-N2A, TG, FTIR and temperature-programmed experiments. The LP-N2A and TG tests revealed that the presence of crude oil reduced the porosity, specific surface area and pore volume in the coal sample containing crude oil, which would suppress the transport capacity of oxygen in mesopores and micropores not to adsorb on the specific active sites of coal surfaces. FTIR analysis indicated that the amount of oxygen-containing functional groups and –OH groups decreased tremendously in the coals containing crude oil, but the content of aliphatic C–H groups showed an increasing trend. Moreover, the values of A(CH2)/A(CH3), Ahy/Aar, Aoxygen/Aar all decrease gradually with the rising crude oil contents, indicating that the crude oil reduced the average alkyl chain length and compressed the space between aromatic clusters. The results of temperature-programmed experiments showed that O2 consumption and CO production rates decreased gradually with the increase in crude oil content. The results were in agreement with the varying trend of characterization of microstructure and function groups obtained by LP-N2A, TG and FTIR tests. Furthermore, the Pearson correlation coefficient cleared that the breaking of long aliphatic chains of aromatic rings and reduction of hydrogen bonds content have a greater contribution to the suppression of spontaneous combustion than the attenuation of physical adsorption. This study concluded that crude oil lowers down the spontaneous combustion properties of coal, which could provide a reference for the prevention of spontaneous combustion of coal seam containing crude oil.

Atorvastatin Improves Mitochondrial Function and Prevents Oxidative Stress in Hippocampus Following Amyloid-β1-40 Intracerebroventricular Administration in Mice.

Amyloid-β (Aβ) peptides play a significant role in the pathogenesis of Alzheimer's disease (AD). Neurotoxic effects promoted by Aβ peptides involve glutamate transmission impairment, decrease of neurotrophic factors, mitochondrial dysfunction, oxidative stress, synaptotoxicity, and neuronal degeneration. Here, we assessed the early events evoked by Aβ1-40 on the hippocampus. Additionally, we sought to unravel the molecular mechanisms of atorvastatin preventive effect on Aβ-induced hippocampal damage. Mice were treated orally (p.o.) with atorvastatin 10mg/kg/day during 7 consecutive days before the intracerebroventricular (i.c.v.) infusion of Aβ1-40 (400pmol/site). Twenty-four hours after Aβ1-40 infusion, a reduced content of mature BDNF/proBDNF ratio was observed in Aβ-treated mice. However, there is no alteration in synaptophysin, PSD-95, and doublecortin immunocontent in the hippocampus. Aβ1-40 promoted an increase in reactive oxygen species (ROS) and nitric oxide (NO) generation in hippocampal slices, and atorvastatin prevented this oxidative burst. Mitochondrial OXPHOS was measured by high-resolution respirometry. At this time point, Aβ1-40 did not alter the O2 consumption rates (OCR) related to phosphorylating state associated with complexes I and II, and the maximal OCR. However, atorvastatin increased OCR of phosphorylating state associated with complex I and complexes I and II, maximal OCR of complexes I and II, and OCR associated with mitochondrial spare capacity. Atorvastatin treatment improved mitochondrial function in the rodent hippocampus, even after Aβ infusion, pointing to a promising effect of improving brain mitochondria bioenergetics. Therefore, atorvastatin could act as an adjuvant in battling the symptoms of AD to preventing or delaying the disease progression.

Impact of salinity changes on growth, oxygen consumption and expression pattern of selected candidate genes in the orange mud crab ( Scylla olivacea )

Change in environmental salinity level is a major limiting factor for the aquaculture productivity because it imposes severe stress on organisms that in turn retards growth. The orange mud crab (Scylla olivacea) is an important coastal aquaculture species (farming is practised in 10‰–20‰ salinity levels) in Bangladesh. The present study was conducted to investigate the changes in growth, O2 consumption and mRNA expression levels of five selected genes in the orange mud crab (S. olivacea) exposed to three different experimental salinity levels (0‰, 10‰ and 20‰) for three months. Crabs reared at 10‰ and 20‰, showed significantly higher (p < .05) growth performance and expression of growth regulatory genes (Actin and α-amylase). The highest levels (p < .05) of O2 consumption and expression of ion regulatory genes (Na+-K+-ATPase, V-type H+-ATPase and Diuretic Hormone) were obtained at 0‰. Moderate levels of growth and expression of selected candidate genes were observed at 10‰ treatment while the highest levels of growth and gene expression were obtained at 20‰ (control salinity). Strong interactions were observed between growth performance and expression of growth genes (R2 = 0.81–0.91), and rate of O2 consumption and expression of ion regulatory genes (R2 = 0.83–0.93), implying that the selected genes are important candidates for growth and ionic balance in S. olivacea. Growth performance was found to be very low at 0‰ initially, after 30 days crabs showed better growth performance at this salinity level. It is thus inferred that orange mud crab individuals might require 3–5 days for acclimation to salinity stress but it can take at least 30 days for acclimation to regular growth. Results indicate that with proper acclimation, the orange mud crab (Scylla olivacea) can be farmed at low salinity conditions and possibly in freshwater condition.

Tidal Freshwater Zones as Hotspots for Biogeochemical Cycling: Sediment Organic Matter Decomposition in the Lower Reaches of Two South Texas Rivers

While organic and inorganic nutrient inputs from land are recognized as a major driver of primary production in estuaries, remarkably little is known about how processes within the tidal freshwater zones (TFZs) of rivers modify these inputs. This study quantifies organic matter (OM) decomposition rates in surface sediment layers in the lower reaches of two south Texas river channels and identifies key parameters that influence sediment decomposition rates. Sediment cores were collected from non-tidal and tidal freshwater sites in the Mission and Aransas rivers during two summers (June 2015 and June 2016) and two winters (February 2016, January 2017). We measured oxygen consumption rates, organic carbon and nitrogen content, stable isotope ratios (δ13C and δ15N of OM), and sediment porosity. O2 consumption rates in TFZ sediments were 385 ± 88 μmol O2 m−2 h−1 (summer) and 349 ± 87 μmol O2 m−2 h−1 (winter) in the Aransas River and 767 ± 153 μmol O2 m−2 h−1 (summer) and 691 ± 95 μmol O2 m−2 h−1 (winter) in the Mission River. These rates in TFZs were similar to rates in estuaries and higher than rates at non-tidal riverine sites. Rates of sediment O2 consumption were primarily controlled by OM content and temperature. Sediment OM was dominated by algal biomass from in situ production in both TFZs. We hypothesize that algal production and sinking within TFZs is a major pathway for translocation of watershed-derived nutrients from the water column to the sediments within TFZs. Further work is needed to quantify linkages between decomposition, nutrient remineralization, and potential removal through processes such as denitrification.