Mycolyltransferase-mediated glycolipid exchange in Mycobacteria

Trehalose dimycolate (TDM), also known as cord factor, is a major surface glycolipid of the cell wall of mycobacteria. Because of its potent biological functions in models of infection, adjuvancy, and immunotherapy, it is important to determine how its biosynthesis is regulated. Here we show that glucose, a host-derived product that is not readily available in the environment, causes Mycobacterium avium to down-regulate TDM expression while up-regulating production of another major glycolipid with immunological roles in T cell activation, glucose monomycolate (GMM). In vitro, the mechanism of reciprocal regulation of TDM and GMM involves competitive substrate selection by antigen 85A. The switch from TDM to GMM biosynthesis occurs near the physiological concentration of glucose present in mammalian hosts. We further demonstrate that GMM is produced in vivo by mycobacteria growing in mouse lung. These results establish an enzymatic pathway for GMM production. More generally, these observations provide a specific enzymatic mechanism for dynamic alterations of cell wall glycolipid remodeling in response to the transition from noncellular to cellular growth environments, including factors that are monitored by the host immune system.     

The tuberculosis prodrug isoniazid bound to activating peroxidases

Isoniazid (INH, isonicotinic acid hydrazine) is one of only two therapeutic agents effective in treating tuberculosis. This prodrug is activated by the heme enzyme catalase peroxidase (KatG) endogenous to Mycobacterium tuberculosis but the mechanism of activation is poorly understood, in part because the binding interaction has not been properly established. The class I peroxidases ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP) have active site structures very similar to KatG and are also capable of activating isoniazid. We report here the first crystal structures of complexes of isoniazid bound to APX and CcP. These are the first structures of isoniazid bound to any activating enzymes. The structures show that isoniazid binds close to the delta-heme edge in both APX and CcP, although the precise binding orientation varies slightly in the two cases. A second binding site for INH is found in APX at the gamma-heme edge close to the established ascorbate binding site, indicating that the gamma-heme edge can also support the binding of aromatic substrates. We also show that in an active site mutant of soybean APX (W41A) INH can bind directly to the heme iron to become an inhibitor and in a different mode when the distal histidine is replaced by alanine (H42A). These structures provide the first unambiguous evidence for the location of the isoniazid binding site in the class I peroxidases and provide rationalization of isoniazid resistance in naturally occurring KatG mutant strains of M. tuberculosis.     

Landscape-scale patterns of shrub-species abundance in California chaparral – The role of topographically mediated resource gradients

We examine the degree to which landscape-scale spatial patterns of shrub-species abundance in California chaparral reflect topographically mediated environmental conditions, and evaluate whether these patterns correspond to known ecophysiological plant processes. Regression tree models are developed to predict spatial patterns in the abundance of 12 chaparral shrub and tree species in three watersheds of the Santa Ynez Mountains, California. The species response models are driven by five variables: average annual soil moisture, seasonal variability in soil moisture, average annual photosynthetically active radiation, maximum air temperature over the dry season (May–October), and substrate rockiness. The energy and moisture variables are derived by integrating high resolution (10 m) digital terrain data and daily climate observations with a process-based hydro-ecological model (RHESSys). Field-sampled data on species abundance are spatially integrated with the distributed environmental variables for developing and evaluating the species response models.The species considered are differentially distributed along topographically-mediated environmental gradients in ways that are consistent with known ecophysiological processes. Spatial patterns in shrub abundance are most strongly associated with annual soil moisture and solar radiation. Substrate rockiness is also closely associated with the establishment of certain species, such as Adenostoma fasciculatum and Arctostaphylos glauca. In general, species that depend on fire for seedling recruitment (e.g., Ceanothous megacarpus) occur at high abundance in xeric environments, whereas species that do not depend on fire (e.g., Heteromeles arbutifolia) occur at higher abundance in mesic environments. Model performance varies between species and is related to life history strategies for regeneration. The scale of our analysis may be less effective at capturing the processes that underlie the establishment of species that do not depend on fire for recruitment. Analysis of predication errors in relation to environmental conditions and the abundance of potentially competing species suggest factors not explicitly considered in the species response models.     

Restriction enzyme analysis of mitochondrial DNA of the Aspergillus flavus group: A. flavus, A. parasiticus, and A. nomius

Mitochondrial DNA restriction fragment length polymorphisms were identified that clearly distinguish Aspergillus flavus, A. parasiticus, and A. nomius. Mitochondrial DNAs of A. flavus and A. parasiticus were found to be circular, and their size was estimated size to be 32 kilobases. A restriction map was constructed for the mitochondrial genome of an A. parasiticus isolate by using four restriction endonucleases. Four genes tested were found to have the same order as in the mitochondrial genome of A. nidulans. The mitochondrial genome of A. nomius was estimated to be 33 kilobases.     

Elevated glutamate and NMDA disrupt production of the second messenger cyclic GMP in the early postnatal mouse cortex

The second messenger cyclic guanosine monophosphate (cGMP) plays many roles during nervous system development. Consequently, cGMP production shows complex patterns of regulation throughout early development. Elevated glutamate levels are known to increase cGMP levels in the mature nervous system. A number of clinical conditions including ischemia and perinatal asphyxia can result in elevated glutamate levels in the developing brain. To investigate the effects of elevated glutamate levels on cGMP in the developing cortex we exposed mouse brain slices to glutamate or N‐methyl D ‐aspartate (NMDA). We find that at early postnatal stages when the endogenous production of cGMP is high, glutamate or NMDA exposure results in a significant lowering of the overall production of cGMP in the cortex, unlike the situation in the mature brain. However, this response pattern is complex with regional and cell‐type specific exceptions to the overall lowered cGMP production. These data emphasize that the response of the developing brain to physiological disturbances can be different from that of the mature brain, and must be considered in the context of the developmental events occurring at the time of disturbance. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2009     

Inhibition of nicotinamide nucleotide transhydrogenase in rat liver submitochondrial particles by dicyclohexylcarbodi-imide and butanedione

Nicotinamide nucleotide transhydrogenase in rat liver submitochondrial particles is inhibited by treatment with NN'-dicyclohexylcarbodi-imide or butane-2,3-dione. Both inhibitions are pseudo-first-order with respect to enzyme activity. The reaction order with respect to inhibitor is close to unity for butanedione, but is significantly lower than unity for dicyclohexylcarbodi-imide.     

Use of leuprolide to treat endometriosis in a rhesus macaque.

Endometriosis was diagnosed in an aged dysmenorrheic rhesus monkey (Macaca mulatta) after biopsy of a 7 cm abdominal mass which could not be completely resected due to extensive adhesions. A 6-month course of treatment with leuprolide, a gonadotropin-releasing hormone agonist, resulted in cessation of menstrual cycles and marked clinical improvement. Dysmenorrhea and hypovolemic shock occurred 2 months after therapy was completed. Despite supportive treatment and resumption of leuprolide, the primate's clinical deterioration and abdominal mass enlargement necessitated euthanasia. To our knowledge, this is the first report of a case of endometriosis in a rhesus macaque treated with a gonadotropin-releasing hormone agonist. Although prolonged leuprolide therapy was clinically effective, its cost and the difficulty in early diagnosis of endometriosis may limit its use in nonhuman primate medicine.