Characterizing Focused-Ultrasound Mediated Drug Delivery to the Heterogeneous Primate Brain In Vivo with Acoustic Monitoring

Focused ultrasound with microbubbles has been used to noninvasively and selectively deliver pharmacological agents across the blood-brain barrier (BBB) for treating brain diseases. Acoustic cavitation monitoring could serve as an on-line tool to assess and control the treatment. While it demonstrated a strong correlation in small animals, its translation to primates remains in question due to the anatomically different and highly heterogeneous brain structures with gray and white matteras well as dense vasculature. In addition, the drug delivery efficiency and the BBB opening volume have never been shown to be predictable through cavitation monitoring in primates. This study aimed at determining how cavitation activity is correlated with the amount and concentration of gadolinium delivered through the BBB and its associated delivery efficiency as well as the BBB opening volume in non-human primates. Another important finding entails the effect of heterogeneous brain anatomy and vasculature of a primate brain, i.e., presence of large cerebral vessels, gray and white matter that will also affect the cavitation activity associated with variation of BBB opening in different tissue types, which is not typically observed in small animals. Both these new findings are critical in the primate brain and provide essential information for clinical applications

Sweet cherry fruit cracking: follow-up testing methods and cultivar-metabolic screening

Background Rain-induced fruit cracking is a major physiological problem in most sweet cherry cultivars. For an in vivo cracking assay, the ‘Christensen method’ (cracking evaluation following fruit immersion in water) is commonly used; however, this test does not adequately simulate environmental conditions. Herein, we have designed and evaluated a cracking protocol, named ‘Waterfall method’, in which fruits are continuously wetted under controlled conditions. Results The application of this method alone, or in combination with ‘Christensen method, was shown to be a reliable approach to characterize sweet cherry cracking behavior. Seventeen cherry cultivars were tested for their cracking behavior using both protocols, and primary as well as secondary metabolites identification was performed in skin tissue using a combined GC–MS and UPLC-MS/MS platform. Significant variations of some of the detected metabolites were discovered and important cracking index–metabolite correlations were identified. Conclusions We have established an alternative/complementary method of cherry cracking characterization alongside to Christiansen assay

Combinatory annotation of cell membrane receptors and signalling pathways of Bombyx mori prothoracic glands

The cells of prothoracic glands (PG) are the main site of synthesis and secretion of ecdysteroids, the biochemical products of cholesterol conversion to steroids that shape the morphogenic development of insects. Despite the availability of genome sequences from several insect species and the extensive knowledge of certain signalling pathways that underpin ecdysteroidogenesis, the spectrum of signalling molecules and ecdysteroidogenic cascades is still not fully comprehensive. To fill this gap and obtain the complete list of cell membrane receptors expressed in PG cells, we used combinatory bioinformatic, proteomic and transcriptomic analysis and quantitative PCR to annotate and determine the expression profiles of genes identified as putative cell membrane receptors of the model insect species, Bombyx mori, and subsequently enrich the repertoire of signalling pathways that are present in its PG cells. The genome annotation dataset we report here highlights modules and pathways that may be directly involved in ecdysteroidogenesis and aims to disseminate data and assist other researchers in the discovery of the role of such receptors and their ligands. © The Author(s) 2016

Mortalin-mediated and ERK-controlled targeting of HIF-1α to mitochondria confers resistance to apoptosis under hypoxia

Hypoxia inducible factor-1 (HIF-1) is the main transcriptional activator of the cellular response to hypoxia and an important target of anticancer therapy. Phosphorylation by ERK1 and/or ERK2 (MAPK3 and MAPK1, respectively; hereafter ERK) stimulates the transcriptional activity of HIF-1α by inhibiting its CRM1 (XPO1)- dependent nuclear export. Here, we demonstrate that phosphorylation by ERK also regulates the association of HIF-1α with a so-far-unknown interaction partner identified as mortalin (also known as GRP75 and HSPA9), which mediates non-genomic involvement of HIF-1α in apoptosis. Mortalin binds specifically to HIF-1α that lacks modification by ERK, and the HIF-1α-mortalin complex is localized outside the nucleus. Under hypoxia, mortalin mediates targeting of unmodified HIF-1α to the outer mitochondrial membrane, as well as association with VDAC1 and hexokinase II, which promotes production of a C-terminally truncated active form of VDAC1, denoted VDAC1-ΔC, and protection from apoptosis when ERK is inactivated. Under normoxia, transcriptionally inactive forms of unmodified HIF-1α or its C-terminal domain alone are also targeted to mitochondria, stimulate production of VDAC1-ΔC and increase resistance to etoposide- or doxorubicin-induced apoptosis. These findings reveal an ERK-controlled, unconventional and anti-apoptotic function of HIF-1α that might serve as an early protective mechanism upon oxygen limitation and promote cancer cell resistance to chemotherapy. © 2017. Published by The Company of Biologists Ltd

Novel HIF-2α interaction with Reptin52 impairs HIF-2 transcriptional activity and EPO secretion

Hypoxia-inducible factor 2 (HIF-2), is essential for cellular response to hypoxia and holds an important role in erythropoiesis, angiogenesis, tissue invasion and metastasis, thus, constituting an important therapeutic target. Maximal HIF-2 transcriptional activation requires HIF-2α phosphorylation by ERK1/2 that impairs its CRM1-mediated nuclear export. Herein, we reveal a novel interaction of HIF-2α with Reptin52, a multifunctional protein involved in cellular functions orchestrated both in the nucleus and the cytoplasm. HIF-2α and Reptin52 interact both in nuclear and cytoplasmic fractions, however, ERK1/2 pathway inactivation seems to favour their association in the cytoplasm. Notably, we demonstrate that Reptin52 reduces HIF-2 transcriptional activity, which results in decreased EPO secretion under hypoxia, by impairing HIF-2α stability via a non-canonical PHD-VHL-proteasome independent mechanism. This interaction represents a novel HIF-2 fine tuning mechanism that allows for distinct HIF1/2 isoforms regulation. © 2021 Elsevier Inc

PAF-metabolic enzymes and PAF-like activity in L. infantum and L. major promastigotes

Leishmania spp are obligate intracellular protozoan parasites of the mononuclear phagocyte system that cause a spectrum of diseases known as leishmaniasis. Platelet Activating Factor (PAF) and some PAF-antagonists seem to play a key role in Leishmania infection. In this article we detected for the first time the specific activities of PAF basic metabolic enzymes, PAF cholinephosphotransferase (PAF-CPT) and Lyso-PAF-acetylotransferase (Lyso-PAF-AT), in two species of Leishmania, namely Leishmania major (L. major) and Leishmania infantum (L. infantum). Specific activity of PAF-CPT of L. major homogenates was substantially higher than that of L. infantum homogenates, while Lyso-PAF-AT specific activities of the homogenates of both species were detected in the same levels. In addition, PAF-like molecules were detected in cells and their culture medium of both species. These results show that PAF-biosynthesis exists and varies between Leishmania species, findings that are in correlation with the general idea that the presence of PAF in several Leishmania species is implicated in Leishmania infection. Copyright © by BIOLIFE, s.a.s

Paf-Metabolic Enzymes and Paf-like Activity in and Promastigotes

Leishmania spp are obligate intracellular protozoan parasites of the mononuclear phagocyte system that cause a spectrum of diseases known as leishmaniasis. Platelet Activating Factor (PAF) and some PAF-antagonists seem to play a key role in Leishmania infection. In this article we detected for the first time the specific activities of PAF basic metabolic enzymes, PAF cholinephosphotransferase (PAF-CPT) and Lyso-PAF-acetylotransferase (Lyso-PAF-AT), in two species of Leishmania , namely Leishmania major (L. major) and Leishmania infantum (L. infantum) . Specific activity of PAF-CPT of L. major homogenates was substantially higher than that of L. infantum homogenates, while Lyso-PAF-AT specific activities of the homogenates of both species were detected in the same levels. In addition, PAF-like molecules were detected in cells and their culture medium of both species. These results show that PAF-biosynthesis exists and varies between Leishmania species, findings that are in correlation with the general idea that the presence of PAF in several Leishmania species is implicated in Leishmania infection