An organotypic slice culture model of chronic white matter injury with maturation arrest of oligodendrocyte progenitors

<p>Abstract</p> <p>Background</p> <p>CNS myelination disturbances commonly occur in chronic white matter lesions in neurodevelopmental and adult neurological disorders. Recent studies support that myelination failure can involve a disrupted cellular repair mechanism where oligodendrocyte (OL) progenitor cells (OPCs) proliferate in lesions with diffuse astrogliosis, but fail to fully differentiate to mature myelinating OLs. There are no <it>in vitro </it>models that reproduce these features of myelination failure.</p> <p>Results</p> <p>Forebrain coronal slices from postnatal day (P) 0.5/1 rat pups were cultured for 1, 5, or 9 days <it>in vitro </it>(DIV). Slices rapidly exhibited diffuse astrogliosis and accumulation of the extracellular matrix glycosaminoglycan hyaluronan (HA), an inhibitor of OPC differentiation and re-myelination. At 1 DIV ~1.5% of Olig2⁺OLs displayed caspase-3 activation, which increased to ~11.5% by 9 DIV. At 1 DIV the density of PDGFRα⁺and PDGFRα⁺/Ki67⁺OPCs were significantly elevated compared to 0 DIV (<it>P </it>< 0.01). Despite this proliferative response, at 9 DIV ~60% of white matter OLs were late progenitors (preOLs), compared to ~7% in the postnatal day 10 rat (<it>P </it>< 0.0001), consistent with preOL maturation arrest. Addition of HA to slices significantly decreased the density of MBP⁺OLs at 9 DIV compared to controls (217 ± 16 <it>vs. </it>328 ± 17 cells/mm², respectively; <it>P </it>= 0.0003), supporting an inhibitory role of HA in OL lineage progression in chronic lesions.</p> <p>Conclusions</p> <p>Diffuse white matter astrogliosis and early OPC proliferation with impaired OL maturation were reproduced in this model of myelination failure. This system may be used to define mechanisms of OPC maturation arrest and myelination failure related to astrogliosis and HA accumulation.</p

Hyaluronan Synthesis, Catabolism, and Signaling in Neurodegenerative Diseases

The glycosaminoglycan hyaluronan (HA), a component of the extracellular matrix, has been implicated in regulating neural differentiation, survival, proliferation, migration, and cell signaling in the mammalian central nervous system (CNS). HA is found throughout the CNS as a constituent of proteoglycans, especially within perineuronal nets that have been implicated in regulating neuronal activity. HA is also found in the white matter where it is diffusely distributed around astrocytes and oligodendrocytes. Insults to the CNS lead to long-term elevation of HA within damaged tissues, which is linked at least in part to increased transcription of HA synthases. HA accumulation is often accompanied by elevated expression of at least some transmembrane HA receptors including CD44. Hyaluronidases that digest high molecular weight HA into smaller fragments are also elevated following CNS insults and can generate HA digestion products that have unique biological activities. A number of studies, for example, suggest that both the removal of high molecular weight HA and the accumulation of hyaluronidase-generated HA digestion products can impact CNS injuries through mechanisms that include the regulation of progenitor cell differentiation and proliferation. These studies, reviewed here, suggest that targeting HA synthesis, catabolism, and signaling are all potential strategies to promote CNS repair

Low-level dissolved organic carbon subsidies drive a trophic upsurge in a boreal stream

1. Energy pathways in stream food webs are often driven by allochthonous basal resources. However, allochthonous dissolved organic carbon (DOC) is generally viewed as a minor if not insignificant basal resource because much of the DOC pool comprises high molecular weight, recalcitrant compounds and is inefficiently incorporated into biomass. Nevertheless, there is increasing evidence that the relatively small, labile fraction of DOC may indeed fuel microbial activity to a level that stimulates productivity across multiple trophic levels, resulting in a trophic upsurge. Here, we tested the trophic upsurge hypothesis by subsidising the labile DOC pool of an Alaskan boreal stream that had relatively high nutrient availability but low levels of naturally occurring DOC. 2. We continuously added ecologically relevant (0.250 mg C/L, c. 10% increase above ambient bulk DOC) concentrations of labile DOC (acetate-C) for 62 days to a treatment reach that was statistically indistinguishable in its channel form and chemistry from an upstream reference reach. We measured responses of pe-riphyton production and biomass, whole reach metabolism and nutrient uptake, benthic invertebrate abundances, and juvenile salmonid (Dolly Varden, Salvelinus malma) abundance and growth. 3. Measurements of basal ecosystem responses collectively indicated increased en-ergy mobilization at the base of the food web in response to labile DOC addition. Periphyton bacterial production in the treatment reach was generally >1.5× refer-ence reach values, and periphyton ash-free dry mass, chlorophyll-a, and chloro-phyll-a:ash-free dry mass were all greater in the treatment reach by the end of the study. Throughout dosing, ecosystem respiration was 1.3× greater in the treat-ment reach and dissolved inorganic nitrogen uptake was greater in the treatment reach on eight out of nine measurements. 4. Benthic invertebrate counts, dominated by Baetis spp. and Chironomidae, were c. 4× greater after 28 dosing days and c. 8× greater after 56 days in the upstream portion of the treatment reach. Abundance generally declined with increasing dis-tance from the dosing station. Dolly Varden fry and parr age classes were nearly 2× more abundant in the upstream portion of the treatment reach than in any section of the reference reach and also declined with increasing distance from the dosing station. Further, Dolly Varden tagged with passive integrated transponders prior to the experiment had significantly higher instantaneous growth rates in the treatment reach than those recaptured in the reference reach. 5. The strong consumer responses to small quantities of labile DOC mirrored sig-nificant treatment reach increases in basal ecosystem function and therefore demonstrated a response consistent with a trophic upsurge. Terrestrial DOC has historically been viewed as contributing little to metazoan consumers, instead modulating the influence of nutrients and being respired out of a disconnected microbial loop. Because we dosed the treatment reach with a relevant concentra-tion of labile DOC, based on measurements in nearby peatland-draining streams, we suggest that terrestrial DOC deserves more attention as a basal resource for whole food webs, akin to nutrients fuelling green (autochthonous) pathways.Alaska Sustainable Salmon Fund, Grant/Award Number: 4470

Nets, Spray or Both? The Effectiveness of Insecticide-Treated Nets and Indoor Residual Spraying in Reducing Malaria Morbidity and Child Mortality in sub-Saharan Africa.

Malaria control programmes currently face the challenge of maintaining, as well as accelerating, the progress made against malaria with fewer resources and uncertain funding. There is a critical need to determine what combination of malaria interventions confers the greatest protection against malaria morbidity and child mortality under routine conditions. This study assesses intervention effectiveness experienced by children under the age of five exposed to both insecticide-treated nets (ITNs) and indoor residual spraying (IRS), as compared to each intervention alone, based on nationally representative survey data collected from 17 countries in sub-Saharan Africa. Living in households with both ITNs and IRS was associated with a significant risk reduction against parasitaemia in medium and high transmission areas, 53% (95% CI 37% to 67%) and 31% (95% CI 11% to 47%) respectively. For medium transmission areas, an additional 36% (95% CI 7% to 53%) protection was garnered by having both interventions compared with exposure to only ITNs or only IRS. Having both ITNs and IRS was not significantly more protective against parasitaemia than either intervention alone in low and high malaria transmission areas. In rural and urban areas, exposure to both interventions provided significant protection against parasitaemia, 57% (95% CI 48% to 65%) and 39% (95% CI 10% to 61%) respectively; however, this effect was not significantly greater than having a singular intervention. Statistically, risk for all-cause child mortality was not significantly reduced by having both ITNs and IRS, and no additional protectiveness was detected for having dual intervention coverage over a singular intervention. These findings suggest that greater reductions in malaria morbidity and health gains for children may be achieved with ITNs and IRS combined beyond the protection offered by IRS or ITNs alone

A TLR/AKT/FoxO3 immune tolerance–like pathway disrupts the repair capacity of oligodendrocyte progenitors

Cerebral white matter injury (WMI) persistently disrupts myelin regeneration by oligodendrocyte progenitor cells (OPCs). We identified a specific bioactive hyaluronan fragment (bHAf) that downregulates myelin gene expression and chronically blocks OPC maturation and myelination via a tolerance-like mechanism that dysregulates pro-myelination signaling via AKT. Desensitization of AKT occurs via TLR4 but not TLR2 or CD44. OPC differentiation was selectively blocked by bHAf in a maturation-dependent fashion at the late OPC (preOL) stage by a noncanonical TLR4/TRIF pathway that induced persistent activation of the FoxO3 transcription factor downstream of AKT. Activated FoxO3 selectively localized to oligodendrocyte lineage cells in white matter lesions from human preterm neonates and adults with multiple sclerosis. FoxO3 constraint of OPC maturation was bHAf dependent, and involved interactions at the FoxO3 and MBP promoters with the chromatin remodeling factor Brg1 and the transcription factor Olig2, which regulate OPC differentiation. WMI has adapted an immune tolerance–like mechanism whereby persistent engagement of TLR4 by bHAf promotes an OPC niche at the expense of myelination by engaging a FoxO3 signaling pathway that chronically constrains OPC differentiation

A TLR/AKT/FoxO3 immune tolerance–like pathway disrupts the repair capacity of oligodendrocyte progenitors

Cerebral white matter injury (WMI) persistently disrupts myelin regeneration by oligodendrocyte progenitor cells (OPCs). We identified a specific bioactive hyaluronan fragment (bHAf) that downregulates myelin gene expression and chronically blocks OPC maturation and myelination via a tolerance-like mechanism that dysregulates pro-myelination signaling via AKT. Desensitization of AKT occurs via TLR4 but not TLR2 or CD44. OPC differentiation was selectively blocked by bHAf in a maturation-dependent fashion at the late OPC (preOL) stage by a noncanonical TLR4/TRIF pathway that induced persistent activation of the FoxO3 transcription factor downstream of AKT. Activated FoxO3 selectively localized to oligodendrocyte lineage cells in white matter lesions from human preterm neonates and adults with multiple sclerosis. FoxO3 constraint of OPC maturation was bHAf dependent, and involved interactions at the FoxO3 and MBP promoters with the chromatin remodeling factor Brg1 and the transcription factor Olig2, which regulate OPC differentiation. WMI has adapted an immune tolerance–like mechanism whereby persistent engagement of TLR4 by bHAf promotes an OPC niche at the expense of myelination by engaging a FoxO3 signaling pathway that chronically constrains OPC differentiation

Extreme sensitivity of myelinating optic nerve axons in a rodent model of perinatal ischemic injury

Cerebral white matter injury (WMI) is increasingly recognized as a common form of perinatal brain injury that predisposes to cerebral palsy as well as cognitive and learning disabilities. Despite the growing impact of WMI, it is unclear whether common cellular and molecular mechanisms define WMI pathogenesis in preterm and term neonates. Although numerous studies have defined a role for glial vulnerability in WMI, there has been limited study of the susceptibility of immature axons.peer-reviewe

Author Correction: A consensus-based transparency checklist.

An amendment to this paper has been published and can be accessed via a link at the top of the paper

Widespread use of herbal medicines by people living with human immunodeficiency virus and contamination of herbal medicines with antiretrovirals in Nigeria.

Herbal medication use amongst people living with human immunodeficiency virus (PLWH) is widespread and understudied. This study aimed to evaluate the prevalence of herbal medicine use amongst PLWH and possible contamination with antiretrovirals (ARVs). Countrywide collection of herbal samples sold by street vendors in Nigeria for the following indications: human immunodeficiency virus (HIV), acquired immune deficiency syndrome, fever and general weakness. Samples were screened using a validated liquid chromatography-mass spectrometry/mass spectrometry method for the presence of the following ARVs: efavirenz, nevirapine, lopinavir, darunavir, ritonavir, atazanavir, emtricitabine, tenofovir and lamivudine. A survey was conducted among 742 PLWH attending four HIV clinics in Nigeria. Data were collected using a structured questionnaire and analysed using IBM SPSS statistics version 22.0 (IBM Corp., 2013, Armond, NY). Of the 138 herbal medicines sampled, three (2%) contained detectable levels of tenofovir, emtricitabine and/or lamivudine. Additionally, of the 742 PLWH surveyed, 310 (41.8%) reported herbal medicine use. Among the users, 191 (61.6%) started taking herbals after commencing HIV therapy while herbal medicine use preceded ARVs treatment in 119 (38.4%) PLWH. We found herbal use to be widespread among PLWH in Nigeria, with increasing use after commencing ARV. Three herbal preparations were also found to contain detectable levels of ARVs. This is a concern and should be studied widely across the region and countries where herbal medicine use is prevalent and poorly regulated

Who sleeps under bednets in Ghana? A doer/non-doer analysis of malaria prevention behaviours

BACKGROUND: Malaria prevention programmes should be based in part on knowledge of why some individuals use bednets while others do not. This paper identifies factors and characteristics of women that affect bednet use among their children less than five years of age in Ghana. METHODS: Data come from the baseline component of an evaluation of Freedom from Hunger's malaria curriculum. A quasi-experimental design was used to select clients (n = 516) of Credit with Education (an integrated package of microfinance and health education) and non-clients (n = 535). Chi-squares, Fisher's Exact tests and logistic regression were used to compare the characteristics of mothers whose children use bednets (doers) with those whose children do not (non-doers) and to identify factors associated with bednet use among children less than five years of age. RESULTS: The following factors were most closely associated with bednet use: region of residence; greater food security; and caregivers' beliefs about symptoms, causation and groups most vulnerable to malaria. Most respondents knew mosquitoes caused malaria; however, 20.6% of doers and 12.3% of non-doers (p = .0228) thought overworking oneself caused malaria. Ninety percent of doers and 77.0% of non-doers felt that sleeping under a net was protective against malaria (p = .0040). In addition, 16.5% of doers and 7.5% of non-doers (p = .0025) identified adult males as most vulnerable to malaria. CONCLUSION: Greater knowledge about malaria does not always translate into improved bednet use. Though culturally-based ideas about malaria may vary between communities, integrating them into traditional health education messages may enhance the effectiveness of public health efforts