A Drug Repurposing Approach Reveals Targetable Epigenetic Pathways in Plasmodium vivax Hypnozoites.

Radical cure of Plasmodium vivax malaria must include elimination of quiescent 'hypnozoite' forms in the liver; however, the only FDA-approved treatments are contraindicated in many vulnerable populations. To identify new drugs and drug targets for hypnozoites, we screened the Repurposing, Focused Rescue, and Accelerated Medchem (ReFRAME) library and a collection of epigenetic inhibitors against P. vivax liver stages. From both libraries, we identified inhibitors targeting epigenetics pathways as selectively active against P. vivax and P. cynomolgi hypnozoites. These include DNA methyltransferase (DNMT) inhibitors as well as several inhibitors targeting histone post-translational modifications. Immunofluorescence staining of Plasmodium liver forms showed strong nuclear 5-methylcystosine signal, indicating liver stage parasite DNA is methylated. Using bisulfite sequencing, we mapped genomic DNA methylation in sporozoites, revealing DNA methylation signals in most coding genes. We also demonstrated that methylation level in proximal promoter regions as well as in the first exon of the genes may affect, at least partially, gene expression in P. vivax. The importance of selective inhibitors targeting epigenetic features on hypnozoites was validated using MMV019721, an acetyl-CoA synthetase inhibitor that affects histone acetylation and was previously reported as active against P. falciparum blood stages. In summary, our data indicate that several epigenetic mechanisms are likely modulating hypnozoite formation or persistence and provide an avenue for the discovery and development of improved radical cure antimalarials.

Experimental evidence for trait utility of gill raker number in adaptive radiation of a north temperate fish.

North temperate fish in post-glacial lakes are textbook examples for rapid parallel adaptive radiation into multiple trophic specialists within individual lakes. Speciation repeatedly proceeded along the benthic-limnetic habitat axis, and benthic-limnetic sister species diverge in the number of gill rakers. Yet, the utility of different numbers of gill rakers for consuming benthic vs. limnetic food has only very rarely been experimentally demonstrated. We bred and raised families of a benthic-limnetic species pair of whitefish under common garden conditions to test whether these species (i) show heritable differentiation in feeding efficiency on zooplankton, and (ii) whether variation in feeding efficiency is predicted by variation in gill raker numbers. We used zooplankton of three different size classes to investigate prey size dependency of divergence in feeding efficiency and to investigate the effect strength of variation in the number of gill rakers. Our results show strong interspecific differences in feeding efficiency. These differences are largest when fish were tested with the smallest zooplankton. Importantly, feeding efficiency is significantly positively correlated with the number of gill rakers when using small zooplankton, also when species identity is statistically controlled for. Our results support the hypothesis that a larger number of gill rakers are of adaptive significance for feeding on zooplankton and provide one of the first experimental demonstrations of trait utility of gill raker number when fish feed on zooplankton. These results are consistent with the suggested importance of divergent selection driven feeding adaptation during adaptive radiation of fish in post-glacial lakes.

Psychosocial intervention in stroke: Families in Recovery from Stroke Trial (FIRST).

A family-focused psychosocial intervention for stroke survivors is described and illustrated with case studies. It is designed to improve functional recovery through four specific pathways: increased knowledge, efficacy, and control through stroke education; optimized social support; increased network cohesion; and improved problem-solving abilities. Rationales for these pathways are presented and methods of implementing them discussed.

Diethylstilbestrol on urinary catecholamines and on food intake in castrated male rats.

Moderate changes in food intake produced by diethylstilbestrol in rats were not well correlated with changes in urinary norepinephrine, vanillylmandelic acid or epinephrine. Apparently moderate dietary restrictions are not capable of decreasing adrenergic activity, and the decreased urinary norepinephrine produced by diethylstilbestrol is not associated with decreased availability of dietary precursors.

Management of a child with acute thyroxine ingestion.

We report a case of thyroxine overdose in a child. Despite extremely high thyroxine (T4RIA) levels on admission, the patient's only symptoms were mild hypertension and tachycardia. Both symptoms responded to propranolol, with a drop in pulse rate and a decrease in blood pressure to normal levels. After four days of cardiac monitoring, the patient was released and received propranolol for five additional days as an outpatient.

Dopamine reverses cardiovascular depression of toxic doses of pentobarbitone in dogs.

Pentobarbitone, 20 mg X kg-1 IV followed by infusion of 25 mg X kg-1 X hr-1, produced a progressive decrease in mean arterial pressure in dogs from 113 +/- 17 mmHg (SD) after one hour of infusion to 82 +/- 21 mmHg after 3.5 hours and to 49 +/- 22 mmHg after 5.5 hours. EEG silence occurred at 3.6 +/- 0.6 hours. In dogs similarly treated with pentobarbitone, a two hour infusion of dopamine 5 micrograms X kg-1 X min-1 beginning at the time of EEG silence prevented the further decrease in pressure and restored pressure to 87 +/- 18 mmHg. The mechanism for this effect of dopamine was an increase in cardiac output as systemic vascular resistance was unaffected by dopamine. The cardiac output increase was mainly the result of an increase in stroke volume as heart rate increased only slightly. Since reduced stroke volume was the main reason why pentobarbitone lowered blood pressure, the effect of dopamine on stroke volume and thus on blood pressure makes it an appropriate antagonist to the cardiovascular effects of toxic doses of pentobarbitone.

Comparison of cardiovascular effects of thiopental and pentobarbital at equivalent levels of CNS depression.

Thiopental and pentobarbital have been used in high doses to protect the brain from injury following hypoxia or to reduce intracranial pressure. This study was performed to determine whether these barbiturates differ in cardiovascular effects when present in plasma concentrations that produce equivalent CNS effects. The effects of thiopental and pentobarbital on heart rate, stroke volume/kg, cardiac output/kg, systemic vascular resistance, mean arterial pressure, and central venous pressure were statistically indistinguishable at plasma concentrations of each barbiturate ranging from 50% to 100% of their concentration producing EEG silence. Three of the seven dogs given thiopental developed ventricular bigeminy at plasma concentrations ranging from 45% to 65% of their concentration producing EEG silence. Lidocaine (1.4-2.0 mg/kg intravenously) reversed the bigeminy to sinus rhythm. When given more than the amount needed to produce a flat EEG, five of the seven dogs given thiopental died, but all dogs given pentobarbital survived. Pentobarbital may be a better choice than thiopental when large doses are indicated.

Contrasting effects of diethylstilbestrol and estradiol on contraction of isolated rat aorta.

Diethylstilbestrol (DES, 1 X 10(-6) M) inhibited norepinephrine (NE)-induced contractions of isolated rat aortic strips only at low doses of NE in contrast to estradiol-17 beta (1 X 10(-5) M) which depressed only the response to high doses. The degree of inhibition increased over a period of 90 min exposure to the estrogens. Microsomes accumulated process appears to be gradual requiring many minutes which may explain the gradual increase in intensity of the inhibitory effect with increased time of exposure to DES. The contrasting inhibitory effects of DES and estradiol in rat aorta may be due to non-homogeneous solution of these drugs in smooth muscle plasma membranes. DES, but not estradiol, caused contraction of isolated aortic strips prior to the onset of inhibition. This contraction showed fade and tachyphylaxis, was antagonized by alpha-adrenergic blockers, and was enhanced by prior treatment of the strips with NE. Thus, DES but not estradiol is capable of mobilizing NE from storage sites in rat aorta.