Explore the vast range of titles available.

Most mycolic acid-containing actinobacteria and some proteobacteria use steroids as growth substrates, but the catabolism of the last two steroid rings has yet to be elucidated. In Mycobacterium tuberculosis , this pathway includes virulence determinants and has been proposed to be encoded by the KstR2-regulated genes, which include a predicted coenzyme A (CoA) transferase gene ( ipdAB ) and an acyl-CoA reductase gene ( ipdC ). In the presence of cholesterol, Δ ipdC and Δ ipdAB mutants of either M. tuberculosis or Rhodococcus jostii strain RHA1 accumulated previously undescribed metabolites: 3aα- H -4α(carboxyl-CoA)-5-hydroxy-7aβ-methylhexahydro-1-indanone (5-OH HIC-CoA) and ( R )-2-(2-carboxyethyl)-3-methyl-6-oxocyclohex-1-ene-1-carboxyl-CoA (COCHEA-CoA), respectively. A Δ fadE32 mutant of Mycobacterium smegmatis accumulated 4-methyl-5-oxo-octanedioic acid (MOODA). Incubation of synthetic 5-OH HIC-CoA with purified IpdF, IpdC, and enoyl-CoA hydratase 20 (EchA20), a crotonase superfamily member, yielded COCHEA-CoA and, upon further incubation with IpdAB and a CoA thiolase, yielded MOODA-CoA. Based on these studies, we propose a pathway for the final steps of steroid catabolism in which the 5-member ring is hydrolyzed by EchA20, followed by hydrolysis of the 6-member ring by IpdAB. Metabolites accumulated by Δ ipdF and Δ echA20 mutants support the model. The conservation of these genes in known steroid-degrading bacteria suggests that the pathway is shared. This pathway further predicts that cholesterol catabolism yields four propionyl-CoAs, four acetyl-CoAs, one pyruvate, and one succinyl-CoA. Finally, a Δ ipdAB M. tuberculosis mutant did not survive in macrophages and displayed severely depleted CoASH levels that correlated with a cholesterol-dependent toxicity. Our results together with the developed tools provide a basis for further elucidating bacterial steroid catabolism and virulence determinants in M. tuberculosis. IMPORTANCE Bacteria are the only known steroid degraders, but the pathway responsible for degrading the last two steroid rings has yet to be elucidated. In Mycobacterium tuberculosis , this pathway includes virulence determinants. Using a series of mutants in M. tuberculosis and related bacteria, we identified a number of novel CoA thioesters as pathway intermediates. Analysis of the metabolites combined with enzymological studies establishes how the last two steroid rings are hydrolytically opened by enzymes encoded by the KstR2 regulon. Our results provide experimental evidence for novel ring-degrading enzymes, significantly advance our understanding of bacterial steroid catabolism, and identify a previously uncharacterized cholesterol-dependent toxicity that may facilitate the development of novel tuberculosis therapeutics. Bacteria are the only known steroid degraders, but the pathway responsible for degrading the last two steroid rings has yet to be elucidated. In Mycobacterium tuberculosis , this pathway includes virulence determinants. Using a series of mutants in M. tuberculosis and related bacteria, we identified a number of novel CoA thioesters as pathway intermediates. Analysis of the metabolites combined with enzymological studies establishes how the last two steroid rings are hydrolytically opened by enzymes encoded by the KstR2 regulon. Our results provide experimental evidence for novel ring-degrading enzymes, significantly advance our understanding of bacterial steroid catabolism, and identify a previously uncharacterized cholesterol-dependent toxicity that may facilitate the development of novel tuberculosis therapeutics.

The wMel strain of Wolbachia can reduce the permissiveness of Aedes aegypti mosquitoes to disseminated arboviral infections. Here, we report that wMel-infected Ae. aegypti (Ho Chi Minh City background), when directly blood-fed on 141 viremic dengue patients, have lower dengue virus (DENV) transmission potential and have a longer extrinsic incubation period than their wild-type counterparts. The wMel-infected mosquitoes that are field-reared have even greater relative resistance to DENV infection when fed on patient-derived viremic blood meals. This is explained by an increased susceptibility of field-reared wild-type mosquitoes to infection than laboratory-reared counterparts. Collectively, these field- and clinically relevant findings support the continued careful field-testing of wMel introgression for the biocontrol of Ae. aegypti-born arboviruses.

1. Many prey organisms exhibit adaptive phenotypic plasticity in life-history traits that facilitate a better chance of survival in the presence of predators. The evolution of such plastic traits requires that the defensive phenotype incurs a cost in the absence of prédation. 2. Model systems that are used to examine predator-induced defences are often organisms with complex life histories that only induce defences during the larval stage. While many studies have detected costs of inducible defences during the larval stage, detecting the costs of larval defences after metamorphosis is also important. 3. We examine the benefits and costs of inducible larval defences in the urban mosquito, Aedes notoscriptus, by rearing them in the presence and absence of predation cues. We compared survival of larvae inducing behavioural defences, when exposed to predation cues, in predation trials with predatory fish Hypseleotris galii to that of larvae reared in the absence predation cues. We also compared life-history traits of predator-exposed larvae to larvae reared in control conditions. 4. Larvae exposed to chemical predation cues limited activity and were able to avoid predation for longer in trials with H. galii. However, predator-exposed larvae suffered retarded larval growth and development, were smaller at metamorphosis and less resistant to starvation as adults. 5. While it is difficult to understand the 'fitness costs' that poorer starvation resistance might confer to adult mosquitoes, it is likely that smaller adult size of predator-exposed individuals would reduce fitness, particularly for females where body size limits the size of blood meal they could take to facilitate egg production. We suggest that the demonstrable costs of inducible defences in mosquito larvae make this a good system for testing theoretical models for the evolutionary maintenance of adaptive phenotypic plasticity.

The evolutionary maintenance of inducible defences is governed by the costs and benefits of the defensive traits. The defensive traits should increase the chances of survival in the presence of predators, but be costly in their absence. The costs and benefits of inducible defensive traits can be influenced by environmental conditions, which subsequently affect the ability of prey to induce those defensive traits. We examine how temperature affects the costs of behavioural defences in larval mosquitoes, Aedes notoscriptus , which reduce activity in the presence of predator cues to limit detectability. The costs of reducing activity could either be exacerbated at warmer temperatures via increased metabolic demand, or ameliorated at warmer temperatures via accelerated development reducing exposure time. We compared life history traits of A. notoscriptus reared in control conditions to those exposed to predation cues as larvae at 18, 23 and 28 °C. Larvae reared in predation cues reduced activity, grew and developed slower and emerged later and smaller. While the reduction in activity increased with temperature, the negative effects on life history of A. notoscriptus were greatest at the coolest temperature. Our results show that the costs of inducible defences in A. notoscriptus are temperature dependent. This work suggests that variation in the thermal environment may have a strong influence on the dynamics of predator–prey interactions and the evolutionary maintenance of plasticity of defensive traits in natural populations.

This study explored gender differences in how correctional officers react to severe occupational stress and addressed differences in coping processes and social support utilization. The subjects were 244 correctional officers (167 men and 77 women) employed in Kentucky medium security institutions for male offenders. The study employed a survey administered to the volunteering participants. The Ways of Coping Questionnaire was used to assess coping processes used to deal with a stressful work event, and the Maslach Burnout Inventory was used to assess severe occupational stress. Statistical analyses revealed that correctional officers experience high levels of occupational stress but did not indicate gender differences in emotional exhaustion or depersonalization. The results indicated that female officers more frequently than male officers processed stress by seeking social support, while male officers more frequently than female officers processed stress by “planful problem solving.” No differences were found in correctional officers’ relationships with oppositesex peers. The results suggested that male and female correctional officers follow traditional sex roles in coping with occupational stress, but they revealed no differences in how they are affected by occupational stress in terms of emotional exhaustion, depersonalization, and personal accomplishment.