Effects of vagotomy on cardiovascular response to periodic apneas in sedated pigs.

There are few studies investigating the influence of vagally mediated reflexes on the cardiovascular response to apneas. In 12 sedated preinstrumented pigs, we studied the effects of vagotomy during apneas, controlling for apnea periodicity and thoracic mechanical effects. Nonobstructive apneas were produced by paralyzing and mechanically ventilating the animals, then turning the ventilator off and on every 30 s. Before vagotomy, relative to baseline, apnea caused increased mean arterial pressure (MAP; +19 +/- 25%, P < 0.05), systemic vascular resistance (SVR; +33 +/- 16%, P < 0.0005), and heart rate (HR; +5 +/- 6%, P < 0.05) and decreased cardiac output (CO) and stroke volume (SV; -16 +/- 10% P < 0.001). After vagotomy, no significant change occurred in MAP, SVR, and SV during apneas, but CO and HR increased relative to baseline. HR was always greater ( approximately 14%, P < 0.01) during the interapneic interval compared with during apnea. We conclude that vagally mediated reflexes are important mediators of the apneic pressor response. HR increases after apnea termination are related, at least in part, to nonvagally mediated reflexes.     

Toxicity and neurophysiological effects of fipronil and fipronil sulfone on the western corn rootworm (Coleoptera: Chrysomelidae)

Fipronil is a member of the relatively new phenylpyrazole insecticide class that is active at the neuro‐inhibitory gamma amino butyric acid (GABA)‐gated chloride channel/ionopore complex. The toxicity and neurophysiological effects of fipronil and its oxidative sulfone metabolite [5‐amino‐1‐(2,6‐dichloro‐4‐trifluoromethylphenyl)‐3‐cyano‐4‐trifluoromethylsulfonylpyrazole] were investigated by using an insecticide‐susceptible western corn rootworm population. In topical bioassays using adult rootworms, fipronil was toxic at very low doses (LD₅₀ = 0.07; LD₉₀ = 0.33 ng/mg). At the LD₉₀, pre‐treatment with the cytochrome P450 monooxygenase inhibitor piperonyl butoxide led to mild antagonism of fipronil toxicity (LD₉₀ = 0.42 ng/mg), while the sulfone analog had greater toxicity (LD₉₀ = 0.22 ng/mg). In neurophysiological studies of spontaneous electrical activity, adult and larval rootworms were equally affected by fipronil and the sulfone analog at 10 μM (in the presence of 5 mM GABA) in comparison to GABA‐treated baselines. Using larval rootworms, insensitivity of the GABA receptor to binding by picrotoxinin or dieldrin (10 μM ) was not apparent in the presence of 5 mM GABA. Further neurophysiological investigation using a range of concentrations (0.625–20.0 μM) on larval rootworms indicated concentration‐dependent effects on bursting activity for both fipronil and the sulfone analog; however, subtle differences were observed between these two compounds. Results indicate that both fipronil and its oxidative sulfone metabolite have similar toxicological and neurological effects on rootworms. Arch. Insect Biochem. Physiol. 40:150–156, 1999. © 1999 Wiley‐Liss, Inc.     

Structural basis for rifamycin resistance of bacterial RNA polymerase by the three most clinically important RpoB mutations found in Mycobacterium tuberculosis

Since 1967, Rifampin (RMP, a Rifamycin) has been used as a first line antibiotic treatment for tuberculosis (TB), and it remains the cornerstone of current short‐term TB treatment. Increased occurrence of Rifamycin‐resistant (RIF^R) TB, ～41% of which results from the RpoB S531L mutation in RNA polymerase (RNAP), has become a growing problem worldwide. In this study, we determined the X‐ray crystal structures of the Escherichia coli RNAPs containing the most clinically important S531L mutation and two other frequently observed RIF^R mutants, RpoB D516V and RpoB H526Y. The structures reveal that the S531L mutation imparts subtle if any structural or functional impact on RNAP in the absence of RIF. However, upon RMP binding, the S531L mutant exhibits a disordering of the RIF binding interface, which effectively reduces the RMP affinity. In contrast, the H526Y mutation reshapes the RIF binding pocket, generating significant steric conflicts that essentially prevent any RIF binding. While the D516V mutant does not exhibit any such gross structural changes, certainly the electrostatic surface of the RIF binding pocket is dramatically changed, likely resulting in the decreased affinity for RIFs. Analysis of interactions of RMP with three common RIF^R mutant RNAPs suggests that modifications to RMP may recover its efficacy against RIF^R TB.     

The plant sHSP superfamily: five new members in Arabidopsis thaliana with unexpected properties

The small heat shock proteins (sHsps), which are ubiquitous stress proteins proposed to act as chaperones, are encoded by an unusually complex gene family in plants. Plant sHsps are classified into different subfamilies according to amino acid sequence similarity and localization to distinct subcellular compartments. In the whole Arabidopsis thaliana genome, 19 genes were annotated to encode sHsps, of which 14 belong to previously defined plant sHsp families. In this paper, we report studies of the five additional sHsp genes in A. thaliana, which can now be shown to represent evolutionarily distinct sHsp subfamilies also found in other plant species. While two of these five sHsps show expression patterns typical of the other 14 genes, three have unusual tissue specific and developmental profiles and do not respond to heat induction. Analysis of intracellular targeting indicates that one sHsp represents a new class of mitochondrion-targeted sHsps, while the others are cytosolic/nuclear, some of which may cooperate with other sHsps in formation of heat stress granules. Three of the five new proteins were purified and tested for chaperone activity in vitro. Altogether, these studies complete our basic understanding of the sHsp chaperone family in plants. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.