Detoxification, scavenging, and repair systems embody the archetypical antioxidant defenses of prokaryotic and eukaryotic cells. Metabolic rewiring also aids with the adaptation of bacteria to ...oxidative stress. Evolutionarily diverse bacteria combat the toxicity of reactive oxygen species (ROS) by actively engaging the stringent response, a stress program that controls many metabolic pathways at the level of transcription initiation via guanosine tetraphosphate and the α-helical DksA protein. Studies herein with Salmonella demonstrate that the interactions of structurally related, but functionally unique, α-helical Gre factors with the secondary channel of RNA polymerase elicit the expression of metabolic signatures that are associated with resistance to oxidative killing. Gre proteins both improve transcriptional fidelity of metabolic genes and resolve pauses in ternary elongation complexes of Embden-Meyerhof-Parnas (EMP) glycolysis and aerobic respiration genes. The Gre-directed utilization of glucose in overflow and aerobic metabolism satisfies the energetic and redox demands of Salmonella, while preventing the occurrence of amino acid bradytrophies. The resolution of transcriptional pauses in EMP glycolysis and aerobic respiration genes by Gre factors safeguards Salmonella from the cytotoxicity of phagocyte NADPH oxidase in the innate host response. In particular, the activation of cytochrome bd protects Salmonella from phagocyte NADPH oxidase-dependent killing by promoting glucose utilization, redox balancing, and energy production. Control of transcription fidelity and elongation by Gre factors represent important points in the regulation of metabolic programs supporting bacterial pathogenesis.
RNA polymerase is the only known protein partner of the transcriptional regulator DksA. Herein, we demonstrate that the chaperone DnaJ establishes direct, redox-based interactions with oxidized DksA. ...Cysteine residues in the zinc finger of DksA become oxidized in Salmonella exposed to low concentrations of hydrogen peroxide (H₂O₂). The resulting disulfide bonds unfold the globular domain of DksA, signaling high-affinity interaction of the C-terminal α-helix to DnaJ. Oxidoreductase and chaperone activities of DnaJ reduce the disulfide bonds of its client and promote productive interactions between DksA and RNA polymerase. Simultaneously, guanosine tetraphosphate (ppGpp), which is synthesized by RelA in response to low concentrations of H₂O₂, binds at site 2 formed at the interface of DksA and RNA polymerase and synergizes with the DksA/DnaJ redox couple, thus activating the transcription of genes involved in amino acid biosynthesis and transport. However, the high concentrations of ppGpp produced by Salmonella experiencing oxidative stress oppose DksA/DnaJ-dependent transcription. Cumulatively, the interplay of DksA, DnaJ, and ppGpp on RNA polymerase protects Salmonella from the antimicrobial activity of the NADPH phagocyte oxidase. Our research has identified redox-based signaling that activates the transcriptional activity of the RNA polymerase regulator DksA.
Salmonella invades host cells and replicates inside acidified, remodeled vacuoles that are exposed to reactive oxygen species (ROS) generated by the innate immune response. Oxidative products of the ...phagocyte NADPH oxidase mediate antimicrobial activity, in part, by collapsing the ΔpH of intracellular Salmonella. Given the role of arginine in bacterial resistance to acidic pH, we screened a library of 54 single-gene mutants in Salmonella that are each involved in, but do not entirely block, arginine metabolism. We identified several mutants that affected Salmonella virulence in mice. The triple mutant Δ
, which is deficient in arginine biosynthesis, was attenuated in immunocompetent mice, but recovered virulence in phagocyte NADPH oxidase deficient
mice. Furthermore, Δ
Salmonella was profoundly susceptible to the bacteriostatic and bactericidal effects of hydrogen peroxide. Peroxide stress led to a larger collapse of the ΔpH in Δ
mutants than occurred in wild-type Salmonella. The addition of exogenous arginine rescued Δ
Salmonella from peroxide-induced ΔpH collapse and killing. Combined, these observations suggest that arginine metabolism is a hitherto unknown determinant of virulence that contributes to the antioxidant defenses of Salmonella by preserving pH homeostasis. In the absence of phagocyte NADPH oxidase-produced ROS, host cell-derived l-arginine appears to satisfy the needs of intracellular Salmonella. However, under oxidative stress, Salmonella must additionally rely on
biosynthesis to maintain full virulence.
The exquisite specificity between a sensor kinase and its cognate response regulator ensures faithful partner selectivity within two-component pairs concurrently firing in a single bacterium, ...minimizing crosstalk with other members of this conserved family of paralogous proteins. We show that conserved hydrophobic and charged residues on the surface of thioredoxin serve as a docking station for structurally diverse response regulators. Using the OmpR protein, we identify residues in the flexible linker and the C-terminal β-hairpin that enable associations of this archetypical response regulator with thioredoxin, but are dispensable for interactions of this transcription factor to its cognate sensor kinase EnvZ, DNA or RNA polymerase. Here we show that the promiscuous interactions of response regulators with thioredoxin foster the flow of information through otherwise highly dedicated two-component signaling systems, thereby enabling both the transcription of Salmonella pathogenicity island-2 genes as well as growth of this intracellular bacterium in macrophages and mice.
Stress fracture risk is elevated during initial military training (IMT), particularly in lower-extremity bones such as the tibia. Although the etiology of stress fractures is multifactorial, lower ...bone strength increases risk.
The objective of this study was to assess, through the use of peripheral quantitative computed tomography, whether adherence to a dietary pattern rich in calcium, potassium, and protein before IMT is positively associated with bone indexes in young adults entering IMT.
A cross-sectional analysis was performed with the use of baseline data from 3 randomized controlled trials in Army, Air Force, and Marine recruits (n = 401; 179 men, 222 women). Dietary intake was estimated from a food-frequency questionnaire. A dietary pattern characterized by calcium, potassium, and protein was derived via reduced rank regression and a pattern z score was computed for each volunteer, where higher scores indicated greater adherence to the pattern. At the 4% (metaphysis) and 14% (diaphysis) sites of the tibia, bone mineral content (BMC), volumetric bone mineral density, robustness, and strength indexes were evaluated. Associations between dietary pattern z score as the predictor variable and bone indexes as the response variables were evaluated by multiple linear regression.
Pattern z score was positively associated with BMC (P = 0.004) and strength (P = 0.01) at the metaphysis and with BMC (P = 0.0002), strength (P = 0.0006), and robustness (P = 0.02) at the diaphysis when controlling for age, sex, race, energy, smoking, education, and exercise. Further adjustment for BMI attenuated the associations, except with diaphyseal BMC (P = 0.005) and strength (P = 0.01). When height and weight were used in place of body mass index, the association with BMC remained (P = 0.046).
A dietary pattern rich in calcium, potassium, and protein is positively associated with measures of tibia BMC and strength in recruits entering IMT. Whether adherence to this dietary pattern before IMT affects injury susceptibility during training remains to be determined.
These trials were registered at clinicaltrials.gov as NCT01617109 and NCT02636348.
Courage. Endurance. Mateship. Sacrifice. These values, engraved in stone at the Isurava war memorial, have become synonymous with the Australian experience during the Kokoda campaign of 1942. The ...story of Kokoda and of the fighting in Papua has been told and retold in books, films and documentaries, but these popular narratives rarely explore beyond this one campaign. Kokoda: Beyond the Legend critically assesses not only the campaigns in Papua and their context in the wider lengthy Pacific war, but also the actions of senior Australian, American and Japanese military leaders. Moving beyond the legend, this book addresses the central question of why Kokoda holds such a significant place in Australian military history. In this book, Karl James brings together eminent military scholars to reassess the principal battles from both Allied and Japanese perspectives, providing readers with a more complete understanding of one of the major turning points in the Second World War.
Since its introduction in the 1970s, computed tomography(CT) has revolutionized diagnostic decision-making. One of the major concerns associated with the widespread use of CT is the associated ...increased radiation exposure incurred by patients. The link between ionizing radiation and the subsequent development of neoplasia has been largely based on extrapolating data from studies of survivors of the atomic bombs dropped in Japan in 1945 and on assessments of the increased relative risk of neoplasia in those occupationally exposed to radiation within the nuclear industry. However, the association between exposure to low-dose radiation from diagnostic imaging examinations and oncogenesis remains unclear. With improved technology, significant advances have already been achieved with regards to radiation dose reduction. There are several dose optimization strategies available that may be readily employed including omitting unnecessary images at the ends of acquired series, minimizing the number of phases acquired, and the use of automated exposure control as opposed to fixed tube current techniques. In addition, new image reconstruction techniques that reduce radiation dose have been developed in recent years with promising results. These techniques use iterative reconstruction algorithms to attain diagnostic quality images with reduced image noise at lower radiation doses.
Abstract
Contributions to election campaigns are tracked by the National Institute on Money in State Politics. In this paper, we use this data to examine patterns in political campaign contributions ...made by the insurance industry. We compile a state‐level data set that aggregates over 67,000 election campaign contributions to insurance regulators and governors; we show that candidate incumbency status is associated with a 20% to 30% increase in the share of contributions made to the incumbent insurance regulator's political party. Because incumbents are overwhelmingly favored to win elections, the industry's behavior suggests that insurers are more concerned with access to the elected official than with supporting a candidate that shares the industry's goals.
Load carriage (LC) exercise may exacerbate inflammation during training. Nutritional supplementation may mitigate this response by sparing endogenous carbohydrate stores, enhancing glycogen ...repletion, and attenuating negative energy balance. Two studies were conducted to assess inflammatory responses to acute LC and training, with or without nutritional supplementation. Study 1: 40 adults fed eucaloric diets performed 90‐min of either LC (treadmill, mean ± SD 24 ± 3 kg LC) or cycle ergometry (CE) matched for intensity (2.2 ± 0.1 VO2peak L min−1) during which combined 10 g protein/46 g carbohydrate (223 kcal) or non‐nutritive (22 kcal) control drinks were consumed. Study 2: 73 Soldiers received either combat rations alone or supplemented with 1000 kcal day−1 from 20 g protein‐ or 48 g carbohydrate‐based bars during a 4‐day, 51 km ski march (~45 kg LC, energy expenditure 6155 ± 515 kcal day−1 and intake 2866 ± 616 kcal day−1). IL‐6, hepcidin, and ferritin were measured at baseline, 3‐h post exercise (PE), 24‐h PE, 48‐h PE, and 72‐h PE in study 1, and before (PRE) and after (POST) the 4‐d ski march in study 2. Study 1: IL‐6 was higher 3‐h and 24‐h post exercise (PE) for CE only (mode × time, P < 0.05), hepcidin increased 3‐h PE and recovered by 48‐h, and ferritin peaked 24‐h and remained elevated 72‐h PE (P < 0.05), regardless of mode and diet. Study 2: IL‐6, hepcidin and ferritin were higher (P < 0.05) after training, regardless of group assignment. Energy expenditure (r = 0.40), intake (r = −0.26), and balance (r = −0.43) were associated (P < 0.05) with hepcidin after training. Inflammation after acute LC and CE was similar and not affected by supplemental nutrition during energy balance. The magnitude of hepcidin response was inversely related to energy balance suggesting that eating enough to balance energy expenditure might attenuate the inflammatory response to military training.
Unaccustomed military training may exacerbate inflammation and degrade nutritional status, particularly when performed in negative energy balance. However, during energy balance, we found that the inflammatory response and downstream effects on hepcidin after military‐like exercise (load carriage) were not different from those observed after traditional aerobic exercise (cycling), regardless of supplemental dietary intake. However, manipulating dietary intake during a multi‐day, metabolically challenging military training may attenuate inflammation and hepcidin release by limiting negative energy balance.
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