Development of siRNA-loaded lipid nanoparticles targeting long non-coding RNA LINC01257 as a novel and safe therapeutic approach for t(8;21) pediatric acute myeloid leukemia

Standard of care therapies for children with acute myeloid leukemia (AML) cause potent off-target toxicity to healthy cells, highlighting the need to develop new therapeutic approaches that are safe and specific for leukemia cells. Long non-coding RNAs (lncRNAs) are an emerging and highly attractive therapeutic target in the treatment of cancer due to their oncogenic functions and selective expression in cancer cells. However, lncRNAs have historically been considered ‘undruggable’ targets because they do not encode for a protein product. Here, we describe the development of a new siRNA-loaded lipid nanoparticle for the therapeutic silencing of the novel oncogenic lncRNA LINC01257. Transcriptomic analysis of children with AML identified LINC01257 as specifically expressed in t(8;21) AML and absent in healthy patients. Using NxGen microfluidic technology, we efficiently and reproducibly packaged anti-LINC01257 siRNA (LNP-si-LINC01257) into lipid nanoparticles based on the FDA-approved Patisiran (Onpattro®) formulation. LNP-si-LINC01257 size and ζ-potential were determined by dynamic light scattering using a Malvern Zetasizer Ultra. LNP-si-LINC01257 internalization and siRNA delivery were verified by fluorescence microscopy and flow cytometry analysis. lncRNA knockdown was determined by RT-qPCR and cell viability was characterized by flow cytometry-based apoptosis assay. LNP-siRNA production yielded a mean LNP size of ~65 nm with PDI ≤ 0.22 along with a >85% siRNA encapsulation rate. LNP-siRNAs were efficiently taken up by Kasumi-1 cells (>95% of cells) and LNP-si-LINC01257 treatment was able to successfully ablate LINC01257 expression which was accompanied by a significant 55% reduction in total cell count following 48 h of treatment. In contrast, healthy peripheral blood mononuclear cells (PBMCs), which do not express LINC01257, were unaffected by LNP-si-LINC01257 treatment despite comparable levels of LNP-siRNA uptake. This is the first report demonstrating the use of LNP-assisted RNA interference modalities for the silencing of cancer-driving lncRNAs as a therapeutically viable and non-toxic approach in the management of AML

Delivery of PEGylated liposomal doxorubicin by bispecific antibodies improves treatment in models of high-risk childhood leukemia

High-risk childhood leukemia has a poor prognosis because of treatment failure and toxic side effects of therapy. Drug encapsulation into liposomal nanocarriers has shown clinical success at improving biodistribution and tolerability of chemotherapy. However, enhancements in drug efficacy have been limited because of a lack of selectivity of the liposomal formulations for the cancer cells. Here, we report on the generation of bispecific antibodies (BsAbs) with dual binding to a leukemic cell receptor, such as CD19, CD20, CD22, or CD38, and methoxy polyethylene glycol (PEG) for the targeted delivery of PEGylated liposomal drugs to leukemia cells. This liposome targeting system follows a "mix-and-match" principle where BsAbs were selected on the specific receptors expressed on leukemia cells. BsAbs improved the targeting and cytotoxic activity of a clinically approved and low-toxic PEGylated liposomal formulation of doxorubicin (Caelyx) toward leukemia cell lines and patient-derived samples that are immunophenotypically heterogeneous and representative of high-risk subtypes of childhood leukemia. BsAb-assisted improvements in leukemia cell targeting and cytotoxic potency of Caelyx correlated with receptor expression and were minimally detrimental in vitro and in vivo toward expansion and functionality of normal peripheral blood mononuclear cells and hematopoietic progenitors. Targeted delivery of Caelyx using BsAbs further enhanced leukemia suppression while reducing drug accumulation in the heart and kidneys and extended overall survival in patient-derived xenograft models of high-risk childhood leukemia. Our methodology using BsAbs therefore represents an attractive targeting platform to potentiate the therapeutic efficacy and safety of liposomal drugs for improved treatment of high-risk leukemia

The snow must go on: Ground ice encasement, snow compaction and absence of snow differently cause soil hypoxia, CO2 accumulation and tree seedling damage in boreal forest

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Ecosystem resistance in the face of climate change: a case study from the freshwater marshes of the Florida Everglades

Shaped by the hydrology of the Kissimmee-Okeechobee-Everglades watershed, the Florida Everglades is composed of a conglomerate of wetland ecosystems that have varying capacities to sequester and store carbon. Hydrology, which is a product of the region’s precipitation and temperature patterns combined with water management policy, drives community composition and productivity. As shifts in both precipitation and air temperature are expected over the next 100 years as a consequence of climate change, CO2 dynamics in the greater Everglades are expected to change. To reduce uncertainties associated with climate change and to explore how projected changes in atmospheric CO2 concentration and climate can alter current CO2 exchange rates in Everglades freshwater marsh ecosystems, we simulated fluxes of carbon among the atmosphere, vegetation, and soil using the DAYCENT model. We explored the effects of low, moderate, and high scenarios for atmospheric CO2 (550, 850, and 950 ppm), mean annual air temperature (þ1, þ2.5, and þ4.28C) and precipitation (2, þ7, and þ14%), as predicted by the IPCC for the year 2100 for the region, on CO2 exchange rates in short- and long-hydroperiod wetland ecosystems. Under 100 years of current climate and atmospheric CO2 concentration, Everglades freshwater marsh ecosystems were estimated to be CO2-neutral. As atmospheric CO2 concentration increased and under climate change projections, there were slight shifts in the start and length of the wet season (1 to þ7 days) and a small enhancement in the sink capacity (by 169 to 573 g C m2 century1 ) occurred at both short- and longhydroperiod ecosystems compared to CO2 dynamics under the current climate regime. Over 100 years, rising temperatures increased net CO2 exchange rates (þ1 to 13 g C m2 century1 ) and shifts in precipitation patterns altered cumulative net carbon uptake by þ13 to 46 g C m2 century1 . While changes in ecosystem structure, species composition, and disturbance regimes were beyond the scope of this research, results do indicate that climate change will produce small changes in CO2 dynamics in Everglades freshwater marsh ecosystems and suggest that the hydrologic regime and oligotrophic conditions of Everglades freshwater marshes lowers the ecosystem sensitivity to climate change. Key word

COVID Isolation Eating Scale (CIES): Analysis of the impact of confinement in eating disorders and obesity-A collaborative international study

Confinement during the COVID-19 pandemic is expected to have a serious and complex impact on the mental health of patients with an eating disorder (ED) and of patients with obesity. The present manuscript has the following aims: (1) to analyse the psychometric properties of the COVID Isolation Eating Scale (CIES), (2) to explore changes that occurred due to confinement in eating symptomatology; and (3) to explore the general acceptation of the use of telemedicine during confinement. The sample comprised 121 participants (87 ED patients and 34 patients with obesity) recruited from six different centres. Confirmatory Factor Analyses (CFA) tested the rational-theoretical structure of the CIES. Adequate goodness-of-fit was obtained for the confirmatory factor analysis, and Cronbach alpha values ranged from good to excellent. Regarding the effects of confinement, positive and negative impacts of the confinement depends of the eating disorder subtype. Patients with anorexia nervosa (AN) and with obesity endorsed a positive response to treatment during confinement, no significant changes were found in bulimia nervosa (BN) patients, whereas Other Specified Feeding or Eating Disorder (OSFED) patients endorsed an increase in eating symptomatology and in psychopathology. Furthermore, AN patients expressed the greatest dissatisfaction and accommodation difficulty with remote therapy when compared with the previously provided face-to-face therapy. The present study provides empirical evidence on the psychometric robustness of the CIES tool and shows that a negative confinement impact was associated with ED subtype, whereas OSFED patients showed the highest impairment in eating symptomatology and in psychopathology.This manuscript and research was supported by grants from the Ministeriode Economía y Competitividad (PSI2015-68701-R), Instituto de Salud Carlos III (ISCIII) (FIS PI14/00290/ INT19/00046nd PI17/01167) and co-funded by FEDER funds/European Regional Development Fund (ERDF), a way to build Europe. CIBERobn, CIBERsam and CIBERDEM are all initiatives of ISCIII. GMB is supported by a postdoctoral grant from FUNCIVA. This initiative is supported by Generalitat de Catalunya. LM is supported by a postdoctoral grant of the mexican institution Consejo Nacional de Ciencia y Tecnología (CONACYT). PPM was supported, in part, by a Portuguese Foundation for Science and Technology grant (POCI-01-0145-FEDER-028145). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript

A review of methods of utilizing crude glycerine phase from biodiesel production

Zagospodarowanie fazy glicerynowej - produktu odpadowego, powstającego podczas wytwarzania estrów metylowych oleju rzepakowego (RME - Rapeseed Methyl Esters), jest ważnym elementem wspierającym produkcję biopaliw zarówno w aspekcie ekonomicznym, jak i ekologicznym. Rafinerie wytwarzające biodiesla na skalę przemysłową stosują najczęściej technologie oczyszczania fazy glicerynowej do postaci czystej gliceryny, możliwej do zastosowania w przemyśle kosmetycznym, spożywczym czy farmaceutycznym. Oczekiwana skala produkcji biodiesla będzie oznaczać jednak bardzo wysoki poziom wytwarzania fazy glicerynowej, której nie będą w stanie zagospodarować wspomniane sektory gospodarki. Niezmiernie ważne jest więc zaproponowanie efektywnych metod wykorzystania tego odpadu, w tym zagospodarowania na cele energetyczne, przy jak najmniejszych nakładach związanych z jego przetwarzaniem. Jeszcze trudniejsze, a jednocześnie jeszcze bardziej znaczące w aspekcie ekonomicznym jest efektywne zagospodarowanie fazy glicerynowej w sektorze produkcji indywidualnej biodiesla. Wykorzystanie wartościowych energetycznie odpadów z przetwarzania rzepaku w olej, a następnie w biodiesel jest często warunkiem rentowności całego procesu. W niniejszym artykule przedstawiono najczęściej stosowane rozwiązania oraz dokonano rozważań na temat potencjalnych metod zagospodarowania fazy glicerynowej zwłaszcza w indywidualnej produkcji RME.The utilization of glycerłne phase - a waste substance from RME (Rapeseed Methyl Esters) manufacturing process -is a vital element improving the ecological and economical worthwhileness of tnę whole biodiesel technology. Industrial refmeries most commonly purify and prepare glycerine for pharmaceutical, cosmetic or food production purposes. However, the expected future ratę of biodiesel production will cause a great increase of amount of crude glycerine produced, and the mentioned industries' demand will not be enough to consume this amount of waste. Therefore, it is crucial to further develop different methods of effective glycerine phase utilization, including energetic use of glycerine, requiring as little preparation efforts as possible. Also, it is difficult and, in the same time, very important from an economical point of view, to research methods of glycerine utilization in a household biodiesel production facility. Energetically efficient use of glycerine waste in individual production may have a great impact on the economical balance of the implemented technology. A review of most popular crude glycerine utilization methods and a study on perspectives of glycerine use in a household biodiesel installation have been presented in this paper

Evidence of Ash Tree (Fraxinus spp.) Specific Associations with Soil Bacterial Community Structure and Functional Capacity

The spread of the invasive emerald ash borer (EAB) across North America has had enormous impacts on temperate forest ecosystems. The selective removal of ash trees (Fraxinus spp.) has resulted in abnormally large inputs of coarse woody debris and altered forest tree community composition, ultimately affecting a variety of ecosystem processes. The goal of this study was to determine if the presence of ash trees influences soil bacterial communities and/or functions to better understand the impacts of EAB on forest successional dynamics and biogeochemical cycling. Using 16S rRNA amplicon sequencing of soil DNA collected from ash and non-ash plots in central Ohio during the early stages of EAB infestation, we found that bacterial communities in plots with ash differed from those without ash. These differences were largely driven by Acidobacteria, which had a greater relative abundance in non-ash plots. Functional genes required for sulfur cycling, phosphorus cycling, and carbohydrate metabolism (specifically those which breakdown complex sugars to glucose) were estimated to be more abundant in non-ash plots, while nitrogen cycling gene abundance did not differ. This ash-soil microbiome association implies that EAB-induced ash decline may promote belowground successional shifts, altering carbon and nutrient cycling and changing soil properties beyond the effects of litter additions caused by ash mortality