Efficacy and Safety of Lidocam Topical Gel (4% Lidocaine-0.3% Meloxicam) for Pain and Inflammation Management during Castration and Tail Docking in Piglets.

(1) Background: It has been well established that castration and tail docking are both painful during and following the procedure, yet there are limited convenient and effective products to address both short-term and long-term pain. Lidocam Topical Gel (LTG) (4% lidocaine and 0.3% meloxicam) was developed to address industry needs for an effective and safe product to address animal welfare concerns regarding castration and tail docking in piglets. (2) Methods: Study 1: Male piglets aged 4-8 days of age were treated with LTG (n = 30) or a control gel (n = 30). Approximately 30 min after application of the gel, the piglets were surgically castrated and tail docked. The efficacy of pain control during the surgical procedures and post-procedure (24 h) pain and inflammation control were evaluated using both behavioral and physiological measurements. Study 2: Meloxicam residue depletion following LTG treatment was followed for 28 days. Study 3: Clinical and pathological safety were evaluated in five groups of eight piglets receiving LTG with: (1) no treatment, (2) nominal topical dose, (3) two times the nominal topical dose, (4) three times the nominal topical dose, and 5) one times the nominal topical dose and 2 mL of LTG by oral gavage daily for 3 days. (3) Results: LTG-treated piglets had a significant reduction in electrocutaneous stimulation response before the procedures and 4 and 24 h post-procedures. Stress vocalization intensity and duration were less in piglets receiving LTG during the surgical procedures. Plasma cortisol and substance P were significantly lower in LTG-treated piglets 3 h after castration and tail docking. The weight and average daily gain were significantly increased in piglets receiving LTG. LTG did not interfere with wound healing or cause irritation at the application sites. There were no abnormal clinical or pathological findings associated with the use of LTG at three times the nominal dose given daily for three days. As meloxicam persisted in the application site tissue, a slaughter withdrawal time of 24 days was determined. (4) Conclusions: When applied to the skin 30 min before castration and tail docking, LTG is effective in surgical pain control and provides post-surgical pain control for up to 24 h. LTG is safe for use in piglets and provides an acceptable withdrawal time for commercial use. LTG is a potentially effective product for commercial use for piglet castration and tail docking.

Assessment of the Effective Tissue Concentrations of Injectable Lidocaine and a Lidocaine-Impregnated Latex Band for Castration in Calves.

This study aimed to assess the effective tissue concentrations of the current standard of care for pain mitigation in calves during castration (injectable lidocaine) and to assess the ability of a lidocaine-loaded elastration band (LLB) to deliver effective concentrations into the scrotal tissue over time. This study comprised two different trials: (1) effective concentrations of injectable lidocaine in the scrotal tissue; and (2) the in vivo delivery of effective concentrations of lidocaine from LLBs placed on the calf scrotums. Sensation in the scrotal tissue was assessed by electrocutaneous stimulation. Injectable lidocaine allowed for short-term anesthesia for up to 60 min, highlighting the importance of finding additional strategies to mitigate long-term pain. An elastomeric ligation band impregnated with lidocaine could provide a suitable alternative, as it yielded tissue levels of lidocaine that approached EC50 and exceeded EC95 at 2 and 72 h following application, respectively, and remained above those levels for at least 28 days after application. Further studies are warranted to compare the use of LLBs to injectable local anesthetics.

Assessment of the Pharmacokinetics and Pharmacodynamics of Injectable Lidocaine and a Lidocaine-Impregnated Latex Band for Castration and Tail Docking in Lambs.

The objectives of this study were to assess the pharmacokinetics and pharmacodynamics of the current standard-of-care for pain mitigation in lambs during castration and tail docking (injectable lidocaine) and assess the ability of Lidocaine-Loaded Bands (LLBs) to deliver therapeutic concentrations into the contacted tissues over time. The study was comprised of four different trials: (1) investigation of in vitro release of lidocaine from LLBs; (2) pharmacokinetics and pharmacodynamics of injectable lidocaine in scrotal and tail tissue; (3) pharmacokinetics and pharmacodynamics of in vivo delivery of lidocaine with LLBs placed on the tail and scrotum of lambs; and (4) a "proof-of-concept" study comparing the sensation of control- versus LLB-banded tail tissue over time. The use of injectable lidocaine provides effective short-term anesthesia for 120 to 180 min following the injection; however, additional strategies are needed to manage long-term pain. The use of an LLB could provide an alternative where tissue lidocaine concentrations meet or exceed the EC50 for at least 21-28 days and, based on electrostimulation data, provides local anesthesia for at least 3 days when compared to a control band. Further studies are needed to compare the use of an injectable local anesthetic to the LLBs.

Identification of Protective Amino Acid Metabolism Events in Nursery Pigs Fed Thermally Oxidized Corn Oil.

Feeding thermally oxidized lipids to pigs has been shown to compromise growth and health, reduce energy digestibility, and disrupt lipid metabolism. However, the effects of feeding oxidized lipids on amino acid metabolism in pigs have not been well defined even though amino acids are indispensable for the subsistence of energy metabolism, protein synthesis, the antioxidant system, and many other functions essential for pig growth and health. In this study, oxidized corn oil (OCO)-elicited changes in amino acid homeostasis of nursery pigs were examined by metabolomics-based biochemical analysis. The results showed that serum and hepatic free amino acids and metabolites, including tryptophan, threonine, alanine, glutamate, and glutathione, as well as associated metabolic pathways, were selectively altered by feeding OCO, and more importantly, many of these metabolic events possess protective functions. Specifically, OCO activated tryptophan-nicotinamide adenosine dinucleotide (NAD+) synthesis by the transcriptional upregulation of the kynurenine pathway in tryptophan catabolism and promoted adenine nucleotide biosynthesis. Feeding OCO induced oxidative stress, causing decreases in glutathione (GSH)/oxidized glutathione (GSSG) ratio, carnosine, and ascorbic acid in the liver but simultaneously promoted antioxidant responses as shown by the increases in hepatic GSH and GSSG as well as the transcriptional upregulation of GSH metabolism-related enzymes. Moreover, OCO reduced the catabolism of threonine to α-ketobutyrate in the liver by inhibiting the threonine dehydratase (TDH) route. Overall, these protective metabolic events indicate that below a certain threshold of OCO consumption, nursery pigs are capable of overcoming the oxidative stress and metabolic challenges posed by the consumption of oxidized lipids by adjusting antioxidant, nutrient, and energy metabolism, partially through the transcriptional regulation of amino acid metabolism.

Inhibiting myostatin signaling partially mitigates structural and functional adaptations to hindlimb suspension in mice.

Novel treatments for muscle wasting are of significant value to patients with disease states that result in muscle weakness, injury recovery after immobilization and bed rest, and for astronauts participating in long-duration spaceflight. We utilized an anti-myostatin peptibody to evaluate how myostatin signaling contributes to muscle loss in hindlimb suspension. Male C57BL/6 mice were left non-suspended (NS) or were hindlimb suspended (HS) for 14 days and treated with a placebo vehicle (P) or anti-myostatin peptibody (D). Hindlimb suspension (HS-P) resulted in rapid and significantly decreased body mass (-5.6% by day 13) with hindlimb skeletal muscle mass losses between -11.2% and -22.5% and treatment with myostatin inhibitor (HS-D) partially attenuated these losses. Myostatin inhibition increased hindlimb strength with no effect on soleus tetanic strength. Soleus mass and fiber CSA were reduced with suspension and did not increase with myostatin inhibition. In contrast, the gastrocnemius showed histological evidence of wasting with suspension that was partially mitigated with myostatin inhibition. While expression of genes related to protein degradation (Atrogin-1 and Murf-1) in the tibialis anterior increased with suspension, these atrogenes were not significantly reduced by myostatin inhibition despite a modest activation of the Akt/mTOR pathway. Taken together, these findings suggest that myostatin is important in hindlimb suspension but also motivates the study of other factors that contribute to disuse muscle wasting. Myostatin inhibition benefitted skeletal muscle size and function, which suggests therapeutic potential for both spaceflight and terrestrial applications.

Environmental estrogens inhibit insulin-like growth factor (IGF) receptor mRNA expression, IGF binding, and IGF signaling ex vivo in rainbow trout (Oncorhynchus mykiss).

In this study, we used juvenile rainbow trout to examine the direct effects of selected environmental estrogens (EE), specifically, 17 ß-estradiol (E2), ß-sitosterol (ßS), and 4-n-nonylphenol (NP), on target tissue sensitivity to insulin-like growth factor (IGF) as assessed by expression of IGF receptor type 1 (IGFR1) mRNAs and IGF-1 binding capacity, as well as on the cell signaling pathways through which EE exert their effects. E2 and NP inhibited IGFR1A and IGFR1B mRNA expression in a time- and concentration-related manner in gill and muscle; however, ßS had no effect on expression of IGFR1 mRNAs in either tissue. NP reduced 125I-IGF binding in gill and E2 and NP reduced 125I-IGF in white muscle; ßS had no effect on 125I-IGF binding in either gill or white muscle. Treatment of gill filaments with either E2 or NP rapidly deactivated (via reduced proportion of phosphorylation) JAK2, STAT5, Akt, and ERK; ßS had no effect on the activation state of any cell signaling elements tested. The effects of EE on IGFR mRNA expression in gill were estrogen receptor (ER) dependent as the inhibitory effects were rescued by the ER antagonist, ICI 182,780. All EE tested blocked growth hormone (GH)-stimulated IGFR mRNA expression in gill filaments. GH-stimulated activation of JAK2, STAT5, Akt, and ERK were blocked by E2, ßS, and NP. Lastly, E2 and NP stimulated suppressor of cytokine signaling 2 (SOCS-2) mRNA expression, an effect that also was ER dependent. These results indicate that EE directly reduce the sensitivity of peripheral tissues to IGF by reducing mRNA and functional expression of IGFRs. Such inhibitory actions of EE are mediated, at least in part, by ER-dependent mechanisms that deactivate JAK, STAT, Akt, and ERK and enhance expression of SOCS-2. These findings together with our previous results show that EE retard growth of post-embryonic rainbow trout through widespread direct effects on the GH-IGF system, specifically, by reducing tissue sensitivity to GH, inhibiting IGF production, reducing tissue sensitivity to IGF, and by deactivating post-receptor IGF cell signaling pathways.

Spatial Visualization of Human Anatomy through Art Using Technical Drawing Exercises.

Spatial visualization, the ability to mentally rotate three-dimensional (3D) images, plays a significant role in anatomy education. This study examines the impact of technical drawing exercises on the improvement of spatial visualization and anatomy education in a Neuroscience course. First-year medical students (n = 84) were randomly allocated into a control group (n = 41) or art-training group (n = 43). Variables including self-reported artistic drawing ability, previous technical drawing experience, or previous anatomy laboratory exposure were gathered. Participants who self-identified as artistic individuals were equally distributed between the two groups. Students in the art-training group attended four 1-hour sessions to solve technical drawing worksheets. All participants completed two Mental Rotations Tests (MRT), which were used to assess spatial visualization. Data were also collected from two neuroscience written examinations and an anatomical "tag test" practical examination. Participants in the art-training and control groups improved on the MRT. The mean of written examination two was significantly higher (P = 0.007) in the art-training group (12.95) than the control group (11.48), and higher (P = 0.027) in those without technical drawing experience (12.44) than those with (11.00). The mean of the anatomical practical was significantly higher (P = 0.010) in those without artistic ability (46.24) than those with (42.00). These results suggest that completing technical drawing worksheets may aid in solving anatomy-based written examination questions on complex brain regions, but further research is needed to determine its implication on anatomy practical scores. These results propose a simple method of improving spatial visualization in anatomy education.

Environmental estrogens inhibit the expression of insulin-like growth factor mRNAs in rainbow trout in vitro by altering activation of the JAK-STAT, AKT-PI3K, and ERK signaling pathways.

Environmental estrogens (EE) have been found to disrupt a host of developmental, reproductive, metabolic, and osmoregulatory process in a wide-range of animals, particularly those in aquatic ecosystems where such compounds concentrate. Previously, we showed that EE inhibited post-embryonic organismal growth of rainbow trout in vivo, but the precise mechanism(s) through which EE exert their growth inhibiting effects remain unknown. In this study, we used rainbow trout (Oncorhynchus mykiss) as a model to investigate the direct effects of 17ß-estradiol (E2), ß-sitosterol (ßS), and 4-n-nonylphenol (NP) on the synthesis of insulin-like growth factors (IGFs) and to elucidate the mechanism(s) by which EEs exert such effects. E2, ßS, and NP significantly inhibited the expression of both IGF-1 and IGF-2 mRNAs in liver and gill in a time- and concentration-related manner. Although the response evoked by each EEs on the expression of IGF mRNAs was similar, the potency and efficacy varied with EE; the rank order potency/efficacy was as follows: E2 > NP > ßS. The effects of EEs on the expression of IGF mRNAs was blocked by the estrogen receptor (ER) antagonist, ICI 182780. The mechanism(s) through which EEs inhibit IGF mRNA expression were investigated in isolated liver cells in vitro. EE treatment deactivated JAK, STAT, ERK, and AKT. Moreover, blockade of growth hormone (GH)-stimulated IGF expression by EE was accompanied by deactivation of JAK, STAT, ERK, and AKT. EEs also increased the expression of suppressor of cytokine signaling 2 (SOCS-2), a known inhibitor of JAK-2--an action that also was blocked by ICI 182780. These results indicate that EEs directly inhibit the expression of IGF mRNAs by disrupting GH post-receptor signaling pathways (e.g., JAK, STAT, ERK, and AKT) in an ER-dependent manner.

Hypogonadal hypertension in male Sprague-Dawley rats is renin-angiotensin system-dependent: role of endogenous androgens.

BACKGROUND: Acutely, testosterone (TES) and other androgens are efficacious vasodilators, both in vitro and in vivo; however, their long-term effects on arterial blood pressure (BP) remain unclear. It was hypothesized that endogenous androgens exert long-term anti-hypertensive effects on systemic BP through a combination of genomic and nongenomic effects to enhance vasodilation of the systemic vasculature. METHODS: The long-term effects of endogenous TES and exogenous TES replacement therapy (TRT) on BP were studied in intact (InT) and castrated (CsX) male Sprague-Dawley (SD) and testicular-feminized male (Tfm, androgen receptor defective) rats (12 weeks old). Systolic BP (tail-cuff plethysmography) was determined weekly for 15 weeks in InT-control and CsX rats. Some CsX-SD rats received androgen replacement therapy at 10-15 weeks with TES-enanthate (TRT; 1.75 mg/kg, 2x/week) or DHT-enanthate (DRT; 1.00 mg/kg. 2x/week) and a separate group of CsX-SD rats received losartan-potassium in drinking water (LST, 250 mg/L) for the entire 15 week period. Expression of renin, angiotensinogen (Agt), angiotensin converting enzyme (ACE), and angiotensin II type I receptor (AT1R) mRNA in kidney and aorta were determined by real-time PCR (rt-PCR) and plasma renin levels were determined by radioimmunoassay. RESULTS: There was a progressive rise in BP over 10 weeks in CsX (109 ± 3.3 vs. 143 ± 3.5 mmHg), while BP remained stable in InT-control (109 ± 3.0 vs. 113 ± 0.3). BP gradually declined to normal in CsX-TRT rats (113 ± 1.3), while BP remained elevated in CsX (140 ± 1.2) and normal in InT-control (113 ± 0.3). LST prevented the development of hypertension in CsX at 10 weeks (100 ± 1.5 in CsX + LST vs. 143 ± 3.5 in CsX). During the next 5 weeks with TES-RT, BP declined in CsX-TRT (113 ± 1.3) and remained lower in CsX + LST (99 ± 0.4). DHT-RT reduced BP in CxS to a similar extent. In Tfm, CsX resulted in a similar rise in BP (109 ± 0.7 vs. 139 ± 0.4 mmHg), but TRT reduced BP more rapidly and to a greater extent (106 ± 2.8). rt-PCR of the kidney revealed that CsX increased expression of mRNA for renin (92%), ACE (58%), and AT1R (80%) compared to InT, while TES RT normalized expression of renin, AT1R, and ACE mRNA to levels of InT rats. Plasma renin levels exhibited changes similar to those observed for renin mRNA expression. CONCLUSIONS: This is the first study to examine the long-term effects of endogenous and exogenous androgens on BP in male SD and Tfm rats. These data reveal that endogenous androgens (TES) exert anti-hypertensive effects that appear to involve non-genomic and possibly genomic mechanism(s), resulting in reductions in RAS expression in the kidney and enhanced systemic vasodilation.

Cooking and Meal Planning as Predictors of Fruit and Vegetable Intake and BMI in First-Year College Students.

The objective was to determine if cooking skills and meal planning behaviors are associated with greater fruit and vegetable intake and lower body mass index (BMI) in first-year college students who are at risk for excessive weight gain. A cross-sectional analysis was conducted using baseline data from a multi-state research project aimed at preventing weight gain in first-year college students. Cooking type, frequency and confidence, self-instruction for healthful mealtime behavior intention, self-regulation of healthful mealtime behavior, and cup equivalents of fruits and vegetables (FV) were measured using validated surveys. BMI was calculated from measured height and weight. First-year students (n = 1108) considered at risk for weight gain from eight universities completed baseline assessments within the first month of entering college. Multiple linear regression was used to determine associations among independent variables of cooking patterns, meal planning behaviors, and dependent variables of fruit and vegetable intake and BMI, after controlling for the influence of sex. Cooking more frequently, cooking with greater skills, and practicing meal planning behaviors are associated with greater fruit and vegetable intake and lower BMI in first-year college students. Interventions aimed at improving health in college students may be enhanced by incorporating cooking and meal planning components.

Addition of tert-butylhydroquinone (TBHQ) to maize oil reduces lipid oxidation but does not prevent reductions in serum vitamin E in nursery pigs.

BACKGROUND: Maize oil is abundantly used in foods and feeds and is highly susceptible to oxidation. Consequently, commercially available antioxidants should be evaluated for effectiveness against lipid oxidation in swine diets. Our study was conducted to evaluate growth performance of nursery pigs fed oxidized maize oil and to determine effects of using antioxidants on oxidative status in a 2 × 2 factorial design. Two hundred eight weaned pigs were blocked by initial BW into 13 blocks, resulting in 4 pigs per pen and 13 pens per treatment. Dietary treatments included 6% unoxidized or oxidized maize oil, and 0 or 60 mg/kg of tert-butylhydroquinone (TBHQ), which was added after lipid oxidation. Data for growth performance were collected from 5 time periods of a two-phase feeding program (Phase 1 = d 0 to 12 and Phase 2 = d 13 to 34). Serum and liver samples were collected from one pig per pen, which had initial BW closest to average BW to determine oxidative status on d 34. RESULTS: Oxidized maize oil was heated for 12 h at 185 °C with 12 L/min of air, yielding a peroxide value (PV) of 5.98 mEq O2/kg and TBARS of 0.11 mg MDA eq/g. Addition of TBHQ to diets containing oxidized maize oil decreased PV by 37% and increased the oil stability index by 69%. Final BW, ADG, ADFI, and G:F of pigs were not different among the four dietary treatments. However, pigs fed oxidized maize oil tended (P <  0.08) to increase hepatosomatic index by 5% compared with those fed unoxidized oil, and this was not affected by adding TBHQ. The serum vitamin E concentration of pigs fed oxidized maize oil was less (P < 0.03) than pigs fed unoxidized oil, but this reduction was not reversed by adding TBHQ. Finally, the serum and liver selenium concentration were not different among the treatments. CONCLUSIONS: The addition of TBHQ did not affect growth performance and vitamin E status in pigs fed moderately oxidized maize oil, but TBHQ reduced lipid oxidation, enhanced the oil stability, and appeared to reduce oxidative stress.

In situ transformation of ethoxylate and glycol surfactants by shale-colonizing microorganisms during hydraulic fracturing.

In the last decade, extensive application of hydraulic fracturing technologies to unconventional low-permeability hydrocarbon-rich formations has significantly increased natural-gas production in the United States and abroad. The injection of surface-sourced fluids to generate fractures in the deep subsurface introduces microbial cells and substrates to low-permeability rock. A subset of injected organic additives has been investigated for their ability to support biological growth in shale microbial community members; however, to date, little is known on how complex xenobiotic organic compounds undergo biotransformations in this deep rock ecosystem. Here, high-resolution chemical, metagenomic, and proteomic analyses reveal that widely-used surfactants are degraded by the shale-associated taxa Halanaerobium, both in situ and under laboratory conditions. These halotolerant bacteria exhibit surfactant substrate specificities, preferring polymeric propoxylated glycols (PPGs) and longer alkyl polyethoxylates (AEOs) over polyethylene glycols (PEGs) and shorter AEOs. Enzymatic transformation occurs through repeated terminal-end polyglycol chain shortening during co-metabolic growth through the methylglyoxal bypass. This work provides the first evidence that shale microorganisms can transform xenobiotic surfactants in fracture fluid formulations, potentially affecting the efficiency of hydrocarbon recovery, and demonstrating an important association between injected substrates and microbial growth in an engineered subsurface ecosystem.

High total dissolved solids in shale gas wastewater inhibit biodegradation of alkyl and nonylphenol ethoxylate surfactants.

Hydraulic fracturing fluids are injected into unconventional oil and gas systems to stimulate hydrocarbon production, returning to the surface in flowback and produced waters containing a complex mixture of xenobiotic additives and geogenic compounds. Nonionic polyethoxylates are commonly added surfactants that act as weatherizers, emulsifiers, wetting agents, and corrosion inhibitors in hydraulic fracturing fluid formulations. Understanding the biodegradability of these ubiquitous additives is critical for produced water pre-treatment prior to reuse and for improving treatment trains for external beneficial reuse. The objective of this study was to determine the effect of produced water total dissolved solids (TDS) from an unconventional natural gas well on the aerobic biodegradation of alkyl ethoxylate and nonylphenol ethoxylate surfactants. Changes in surfactant concentrations, speciation and metabolites, as well as microbial community composition and activity were quantified over a 75-day aerobic incubation period. Alkyl ethoxylates (AEOs) were degraded faster than nonylphenol ethoxylates (NPEOs), and both compound classes and bulk organic carbon biodegraded slower in TDS treatments (10 g L-1, 40 g L-1) as compared to controls. Short-chain ethoxylates were more rapidly biodegraded than longer-chain ethoxylates, and changes in the relative abundance of metabolites including acetone, alcohols, and carboxylate and aldehyde intermediates of alkyl units indicated metabolic pathways may shift in the presence of higher produced water TDS. Our key finding that polyethoxylated alcohol surfactant additives are less labile at high TDS has important implications for produced water management, as these fluids are increasingly recycled for beneficial reuse in hydraulic fracturing fluids and other purposes.

Viruses control dominant bacteria colonizing the terrestrial deep biosphere after hydraulic fracturing.

The deep terrestrial biosphere harbours a substantial fraction of Earth's biomass and remains understudied compared with other ecosystems. Deep biosphere life primarily consists of bacteria and archaea, yet knowledge of their co-occurring viruses is poor. Here, we temporally catalogued viral diversity from five deep terrestrial subsurface locations (hydraulically fractured wells), examined virus-host interaction dynamics and experimentally assessed metabolites from cell lysis to better understand viral roles in this ecosystem. We uncovered high viral diversity, rivalling that of peatland soil ecosystems, despite low host diversity. Many viral operational taxonomic units were predicted to infect Halanaerobium, the dominant microorganism in these ecosystems. Examination of clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins (CRISPR-Cas) spacers elucidated lineage-specific virus-host dynamics suggesting active in situ viral predation of Halanaerobium. These dynamics indicate repeated viral encounters and changing viral host range across temporally and geographically distinct shale formations. Laboratory experiments showed that prophage-induced Halanaerobium lysis releases intracellular metabolites that can sustain key fermentative metabolisms, supporting the persistence of microorganisms in this ecosystem. Together, these findings suggest that diverse and active viral populations play critical roles in driving strain-level microbial community development and resource turnover within this deep terrestrial subsurface ecosystem.

Members of Marinobacter and Arcobacter Influence System Biogeochemistry During Early Production of Hydraulically Fractured Natural Gas Wells in the Appalachian Basin.

Hydraulic fracturing is the prevailing method for enhancing recovery of hydrocarbon resources from unconventional shale formations, yet little is understood regarding the microbial impact on biogeochemical cycling in natural-gas wells. Although the metabolisms of certain fermentative bacteria and methanogenic archaea that dominate in later produced fluids have been well studied, few details have been reported on microorganisms prevelant during the early flowback period, when oxygen and other surface-derived oxyanions and nutrients become depleted. Here, we report the isolation, genomic and phenotypic characterization of Marinobacter and Arcobacter bacterial species from natural-gas wells in the Utica-Point Pleasant and Marcellus Formations coupled to supporting geochemical and metagenomic analyses of produced fluid samples. These unconventional hydrocarbon system-derived Marinobacter sp. are capable of utilizing a diversity of organic carbon sources including aliphatic and aromatic hydrocarbons, amino acids, and carboxylic acids. Marinobacter and Arcobacter can metabolize organic nitrogen sources and have the capacity for denitrification and dissimilatory nitrate reduction to ammonia (DNRA) respectively; with DNRA and ammonification processes partially explaining high concentrations of ammonia measured in produced fluids. Arcobacter is capable of chemosynthetic sulfur oxidation, which could fuel metabolic processes for other heterotrophic, fermentative, or sulfate-reducing community members. Our analysis revealed mechanisms for growth of these taxa across a broad range of salinities (up to 15% salt), which explains their enrichment during early natural-gas production. These results demonstrate the prevalence of Marinobacter and Arcobacter during a key maturation phase of hydraulically fractured natural-gas wells, and highlight the significant role these genera play in biogeochemical cycling for this economically important energy system.

Coupled laboratory and field investigations resolve microbial interactions that underpin persistence in hydraulically fractured shales.

Hydraulic fracturing is one of the industrial processes behind the surging natural gas output in the United States. This technology inadvertently creates an engineered microbial ecosystem thousands of meters below Earth's surface. Here, we used laboratory reactors to perform manipulations of persisting shale microbial communities that are currently not feasible in field scenarios. Metaproteomic and metabolite findings from the laboratory were then corroborated using regression-based modeling performed on metagenomic and metabolite data from more than 40 produced fluids from five hydraulically fractured shale wells. Collectively, our findings show that Halanaerobium, Geotoga, and Methanohalophilus strain abundances predict a significant fraction of nitrogen and carbon metabolites in the field. Our laboratory findings also exposed cryptic predatory, cooperative, and competitive interactions that impact microorganisms across fractured shales. Scaling these results from the laboratory to the field identified mechanisms underpinning biogeochemical reactions, yielding knowledge that can be harnessed to potentially increase energy yields and inform management practices in hydraulically fractured shales.

Identification of activation of tryptophan-NAD+ pathway as a prominent metabolic response to thermally oxidized oil through metabolomics-guided biochemical analysis.

Consumption of thermally oxidized oil is associated with metabolic disorders, but oxidized oil-elicited changes in the metabolome are not well defined. In this study, C57BL/6 mice were fed the diets containing either control soybean oil or heated soybean oil (HSO) for 4 weeks. HSO-responsive metabolic events were examined through untargeted metabolomics-guided biochemical analysis. HSO directly contributed to the presence of new HSO-derived metabolites in urine and the decrease of polyunsaturated fatty acid-containing phospholipids in serum and the liver. HSO disrupted redox balance by decreasing hepatic glutathione and ascorbic acid. HSO also activated peroxisome proliferator-activated receptors, leading to the decrease of serum triacylglycerols and the changes of cofactors and products in fatty acid oxidation pathways. Most importantly, multiple metabolic changes, including the decrease of tryptophan in serum; the increase of NAD+ in the liver; the increases of kynurenic acid, nicotinamide and nicotinamide N-oxide in urine; and the decreases of the metabolites from pyridine nucleotide degradation in the liver indicated that HSO activated tryptophan-NAD+ metabolic pathway, which was further confirmed by the upregulation of gene expression in this pathway. Because NAD+ and its metabolites are essential cofactors in many HSO-induced metabolic events, the activation of tryptophan-NAD+ pathway should be considered as a central metabolic response to the exposure of HSO.

Isometric Midthigh Pull Reliability and Relationship to Deadlift One Repetition Maximum.

De Witt, JK, English, KL, Crowell, JB, Kalogera, KL, Guilliams, ME, Nieschwitz, BE, Hanson, AM, and Ploutz-Snyder, LL. Isometric midthigh pull reliability and relationship to deadlift one repetition maximum. J Strength Cond Res 32(2): 528-533, 2018-The purpose of this investigation was to examine the reliability of the isometric midthigh pull (IMTP) and the relationship between IMTP peak force and deadlift 1 repetition maximum (1RM). Nine subjects (5 men and 4 women; 40.6 ± 8.0 years; 1.72 ± 0.10 m; 75.6 ± 13.4 kg) participated in this study. Isometric midthigh pull and deadlift 1RM were both performed during 2 testing sessions. For IMTP, peak force and peak rate of force development (RFD) were determined, in addition to RFD at 30 ms, 50 ms, 90 ms, 150 ms, 200 ms, and 250 ms after initiation of the pull. Intraclass correlation coefficients (ICCs) were calculated to evaluate the reliability of IMTP measures. Pearson product-moment correlations and linear regression were used to determine associations between IMTP and deadlift 1RM. Isometric midthigh pull peak force was reproducible both within (ICC = 0.98 and 0.97) and between sessions (ICC = 0.89) and significantly correlated with deadlift 1RM (r = 0.88, p ≤ 0.05), but intermediate force outputs and RFD were not. Lack of associations between RFD and deadlift 1RM indicate that the ability to create explosive force may be independent of the ability to create maximal force. The strong relationship between IMTP peak force and deadlift 1RM was present regardless of which IMTP repetition across the 2 sessions was examined. Peak force generated during IMTP is a reliable method to assess full body maximal strength. A single IMTP repetition, provided adequate familiarization and warm-up, correlates strongly with deadlift 1RM. Practitioners can use the IMTP test as a method to estimate maximal deadlift strength in a quick and potentially less provocative manner than traditional 1RM testing.

Antihypertensive responses of vasoactive androgens in an in vivo experimental model of preeclampsia.

Dehydroepiandrosterone (DHEA), testosterone (TES) and its 5-reduced metabolites induce a nongenomic vasorelaxation in several vascular beds of mammals; similarly these hormones produce systemic hypotensive and antihypertensive responses in normotensive and hypertensive male rats. Thus, it was hypothesized that the antihypertensive response of androgens, whose levels are elevated during gestation, protect against gestational hypertension. An animal model of preeclampsia was induced in female Wistar rats using DOCA-salt-treated pregnant (PT) and normal pregnant (NP) rats. In vivo experiments in conscious rats revealed that bolus intravenous injections of DHEA, TES, 5α- or 5ß-dihydrotestosterone (-DHT) log -1.0 to 2.0µmolk-1min-1, produced substantial transient reductions in arterial blood pressure (BP), without significant changes in heart rate (HR). Mean arterial blood pressure (MAP) was reduced significantly in both groups. PT rats were more sensitive to the antihypertensive responses of androgens than NP. DHEA and 5ß-DHT were the most potent to reduce MAP: 66±07 and 69±2.0mmHg in PT but only 33±0.5 and 35±1.2mmHg in NP rats, respectively. In isolated aortas of PT and NP, the concentration-response curves to each androgen (0.1-100µM) indicated that KCl-induced pre-contraction is more sensitive to all androgens than phenylephrine (Phe) pre-contractions. Notably, 5ß-DHT is the greatest vasorelaxant with KCl-induced contraction than with Phe contraction of both groups, suggesting a preferential blockade on L-VOCCs. TES exhibited minor vasorelaxing effect of aortas pre-contracted with KCl, compared to its precursor DHEA and its 5-reduced metabolites. These data show that these androgens exert acute vasorelaxing effects in vitro and remarkably, reduce the BP in vivo in PT and NP at term pregnancy. Moreover, a deficit in feto-placental androgen production during pregnancy may trigger the development of preeclampsia or gestational hypertension.

Ground Reaction Forces During Reduced Gravity Running in Parabolic Flight.

BACKGROUND: Treadmills have been employed as both a form of exercise and a countermeasure to prevent changes in the musculoskeletal system on almost all NASA missions and many Russian missions since the early Space Shuttle flights. It is possible that treadmills may also be part of exercise programs on future Mars missions and that they may be a component of exercise facilities in lunar or Martian habitats. METHODS: In order to determine if the ambient gravity on these destinations will provide osteogenic effects while performing exercise on a treadmill, ground reactions forces (GRFs) were measured on eight subjects (six women and two men) running at 6 mph during parabolic flight in Martian and lunar gravity conditions. RESULTS: On average, stride length increased as gravity decreased. The first and second peaks of the GRFs decreased by 0.156 and 0.196 bodyweights, respectively, per 1/10 g change in ambient gravity. DISCUSSION: Based on comparisons with previously measured GRF during loaded treadmill running on the International Space Station, we conclude that unloaded treadmill running under lunar and Martian conditions during exploration missions is not likely to be an osteo-protective exercise.Cavanagh P, Rice A, Glauberman M, Sudduth A, Cherones A, Davis S, Lewis M, Hanson A, Wilt G. Ground reaction forces during reduced gravity running in parabolic flight. Aerosp Med Hum Perform. 2017; 88(8):730-736.