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.

[Speech and general development in children receiving early cochlear implants]. / Sprachentwicklung und allgemeine Entwicklung bei Kindern mit früher Cochleaimplantation.

BACKGROUND: After cochlear implantation, most parents expect a normal speech and general development of their child. However, it remains unclear how quickly after early cochlear implantation these children can compensate for their deficits compared to normal-hearing children. METHODS: This study retrospectively analyzed ELFRA-1 questionnaire data from 40 children with borderline deafness or high-grade hearing loss (without other known impairments) who had undergone cochlear implantation at a university medical center before reaching 2 years of age. ELFRA-1 questionnaires were filled out parents assisted by specialists 12 months after implantation. Questions assessed the children's speech production and comprehension, as well as their use of gestures and fine motoric skills. RESULTS: At an average hearing-age of 12 months, the children achieved normal values in all of the subgroups that were comparable to those of 12-month-old children without hearing impairments. A significant correlation (p = 0.01) between the individual subgroups of the ELFRA-1 (speech production, speech comprehension, gestures and fine motor skills) was observed. Unilingual educated children performed significantly better overall. CONCLUSION: Within 12 months of receiving a cochlear implant, all children passed the four categories of the ELFRA-1. This demonstrates a rapid compensation of deficits in speech, motor skills and gesture development by children undergoing early cochlear implantation.

Malaria parasite liver stages render host hepatocytes susceptible to mitochondria-initiated apoptosis.

Intracellular eukaryotic parasites and their host cells constitute complex, coevolved cellular interaction systems that frequently cause disease. Among them, Plasmodium parasites cause a significant health burden in humans, killing up to one million people annually. To succeed in the mammalian host after transmission by mosquitoes, Plasmodium parasites must complete intracellular replication within hepatocytes and then release new infectious forms into the blood. Using Plasmodium yoelii rodent malaria parasites, we show that some liver stage (LS)-infected hepatocytes undergo apoptosis without external triggers, but the majority of infected cells do not, and can also resist Fas-mediated apoptosis. In contrast, apoptosis is dramatically increased in hepatocytes infected with attenuated parasites. Furthermore, we find that blocking total or mitochondria-initiated host cell apoptosis increases LS parasite burden in mice, suggesting that an anti-apoptotic host environment fosters parasite survival. Strikingly, although LS infection confers strong resistance to extrinsic host hepatocyte apoptosis, infected hepatocytes lose their ability to resist apoptosis when anti-apoptotic mitochondrial proteins are inhibited. This is demonstrated by our finding that B-cell lymphoma 2 family inhibitors preferentially induce apoptosis in LS-infected hepatocytes and significantly reduce LS parasite burden in mice. Thus, targeting critical points of susceptibility in the LS-infected host cell might provide new avenues for malaria prophylaxis.

[Characterizing the passive opening of the eustachian tube in a hypo-/hyperbaric pressure chamber]. / Charakterisierung der passiven Tubenöffnung in einer hypo-/hyperbaren Druckkammer.

BACKGROUND: Beside arbitrary and not arbitrary active pressure equalization systems there is a passive equalization system via the Eustachian tube (ET) at pressure difference between the epipharyngeal space and the middle ear. Aim of this study was to characterize this passive equalization system in a hypobaric/hyperbaric pressure chamber by continuously measuring the tympanic impedance. In contrast to other studies, which are measured only in a hypobaric pressure chamber it is possible to include participants with Eustachian tube dysfunction (ETD). MATERIAL AND METHODS: Following a fixed pressure profile 39 participants were exposed to phases of pressure rising and decompression. By continuously measuring the tympanic impedance in the pressure chamber it was possible to measure data of the Eustachian Tube opening Pressure (ETOP), Eustachian Tube closing pressure (ETCP) and Eustachian Tube opening duration (ETOD). In addition it was possible to characterize the gradient of pressure during decompression, while the ET was open. RESULTS: Beside the measurement of the arithmetic average of the ETOP (30.2 ± 15.1 mbar), ETCP (9.1 ± 7.7 mbar) and ETOD (0.65 ± 0.38 s) it was obvious that there are recurrent samples of pressure progression during the phase of tube opening. Generally it is possible to differentiate between the type of complete opening and partial opening. CONCLUSION: The fundamental characterization of the action of the passive tube opening, including the measurement of the ETOP, ETCP and ETOD, is a first step in understanding the physiological and pathophysiological function of the ET.

The impact of cidofovir treatment on viral loads in adult recurrent respiratory papillomatosis.

Cidofovir is an antiviral agent used in the therapy of recurrent respiratory papillomatosis (RRP). In this study, we hypothesized that cidofovir is effective in decreasing the viral load of human papillomavirus (HPV). We established a type specific real-time PCR and measured HPV DNA loads. The course of viral load of HPV types 6 and 11 after repeated applications of cidofovir intralesionally was compared to the clinical outcome using a modified Derkay score. In 6 of the 8 (75 %) patients, we detected HPV 6. In 2 (25 %) patients, we detected HPV 11. In all of the patients, the viral load and the modified Derkay score decreased significantly during the treatment. We conclude that viral load of HPV can be monitored using the technique described here. Cidofovir in combination with surgical debulking reduces the viral load in patients with RRP. Relapses of the symptoms cannot be avoided but might be delayed.