TiO2-MWCNT nanohybrid: Cytotoxicity, protein corona formation and cellular internalisation in RTG-2 fish cell line.

Titanium dioxide nanoparticles-multiwalled carbon nanotubes (TiO2-MWCNT) nanohydrid has an enhanced photocatalytic activity across the visible light with promising applications in environmental remediation, solar energy devices and antimicrobial technologies. However, it is necessary to evaluate the toxicological effects of TiO2-MWCNT towards safe and sustainable development of nanohybrids. In this work, we studied the cytotoxicity, protein corona formation and cellular internalisation of TiO2-MWCNT on fibroblasts derived from gonadal rainbow trout tissue (RTG-2) for the first time. This nanohydrid did not show any toxicity effect on RTG-2 cells up to 100 mg L-1 after 24 h of exposure as monitored by alamar blue, neutral red and trypan blue assays (in presence or absence of foetal bovine serum, FBS). Futhermore, cryo-transmission electron microscopy analysis demonstrated that TiO2 particles is attached on nanotube surface after FBS-protein corona formation in cell culture medium. Raman spectroscopy imaging showed that TiO2-MWCNT can be internalised by RTG-2 cells. This work is a novel contribution towards better understanding the nanobiointeractions of nanohydrids linked to their in vitro effects on fish cells in aquatic nanoecotoxicology.

Cellular mechanisms linking to outdoor and indoor air pollution damage during pregnancy.

Pregnancies are a critical window period for environmental influences over the mother and the offspring. There is a growing body of evidence associating indoor and outdoor air pollution exposure to adverse pregnancy outcomes such as preterm birth and hypertensive disorders of pregnancy. Particulate matter (PM) could trigger oxi-inflammation and could also reach the placenta leading to placental damage with fetal consequences. The combination of strategies such as risk assessment, advise about risks of environmental exposures to pregnant women, together with nutritional strategies and digital solutions to monitor air quality can be effective in mitigating the effects of air pollution during pregnancy.

Functional consequences of SARS-CoV-2 infection in pregnant women, fetoplacental unit, and neonate.

The SARS-CoV-2 infection causes COVID-19 disease, characterized by acute respiratory distress syndrome, bilateral pneumonia, and organ failure. The consequences of maternal SARS-CoV-2 infection for the pregnant woman, fetus, and neonate are controversial. Thus, it is required to determine whether there is viral and non-viral vertical transmission in COVID-19. The disease caused by SARS-CoV-2 leads to functional alterations in asymptomatic and symptomatic pregnant women, the fetoplacental unit and the neonate. Several diseases of pregnancy, including COVID-19, affect the fetoplacental function, which causes in utero programming for young and adult diseases. A generalized inflammatory state and a higher risk of infection are seen in pregnant women with COVID-19. Obesity, diabetes mellitus, and hypertension may increase the vulnerability of pregnant women to infection by SARS-CoV-2. Alpha, Delta, and Omicron variants of SARS-CoV-2 show specific mutations that seem to increase the capacity of the virus to infect the pregnant woman, likely due to increasing its interaction via the virus S protein and angiotensin-converting enzyme 2 receptors. This review shows the literature addressing to what extent COVID-19 in pregnancy affects the pregnant woman, fetoplacental unit, and neonate. Prospective studies that are key in managing SARS-CoV-2 infection in pregnancy are discussed.

Mechanism of high-mannose N-glycan breakdown and metabolism by Bifidobacterium longum.

Bifidobacteria are early colonizers of the human gut and play central roles in human health and metabolism. To thrive in this competitive niche, these bacteria evolved the capacity to use complex carbohydrates, including mammalian N-glycans. Herein, we elucidated pivotal biochemical steps involved in high-mannose N-glycan utilization by Bifidobacterium longum. After N-glycan release by an endo-ß-N-acetylglucosaminidase, the mannosyl arms are trimmed by the cooperative action of three functionally distinct glycoside hydrolase 38 (GH38) α-mannosidases and a specific GH125 α-1,6-mannosidase. High-resolution cryo-electron microscopy structures revealed that bifidobacterial GH38 α-mannosidases form homotetramers, with the N-terminal jelly roll domain contributing to substrate selectivity. Additionally, an α-glucosidase enables the processing of monoglucosylated N-glycans. Notably, the main degradation product, mannose, is isomerized into fructose before phosphorylation, an unconventional metabolic route connecting it to the bifid shunt pathway. These findings shed light on key molecular mechanisms used by bifidobacteria to use high-mannose N-glycans, a perennial carbon and energy source in the intestinal lumen.

PEGylated versus Non-PEGylated pH-Sensitive Liposomes: New Insights from a Comparative Antitumor Activity Study.

PEGylated liposomes are largely studied as long-circulating drug delivery systems. Nevertheless, the addition of PEG can result in reduced interactions between liposomes and cells, hindering liposomal internalization into target cells. The presence of PEG on the surface of pH-sensitive liposomes is not advantageous in terms of biodistribution and tumor uptake, raising the question of whether the indiscriminate use of PEG benefits the formulation. In this study, two doxorubicin-loaded pH-sensitive liposomal formulations, PEGylated (Lip2000-DOX) or non-PEGylated (Lip-DOX), were prepared and characterized. Overall, the PEGylated and non-PEGylated liposomes showed no differences in size or morphology in Cryo-TEM image analysis. Specifically, DLS analysis showed a mean diameter of 140 nm, PDI lower than 0.2, and zeta potential close to neutrality. Both formulations showed an EP higher than 90%. With respect to drug delivery, Lip-DOX had better cellular uptake than Lip2000-DOX, suggesting that the presence of PEG reduced the amount of intracellular DOX accumulation. The antitumor activities of free-DOX and both liposomal formulations were evaluated in 4T1 breast tumor-bearing BALB/c mice. The results showed that Lip-DOX was more effective in controlling tumor growth than other groups, inhibiting tumor growth by 60.4%. Histological lung analysis confirmed that none of the animals in the Lip-DOX group had metastatic foci. These results support that pH-sensitive liposomes have interesting antitumor properties and may produce important outcomes without PEG.

Comparison of Three Gestational Weight Gain Guidelines Under Use in Latin America.

Presently, three guidelines are used in Latin America to assess adequacy of maternal body mass index (BMI) during pregnancy: (1) the chart proposed by the Institute of Medicine of the United States (IOM), (2) the Rosso-Mardones Chart (RM), and (3) a modified RM chart proposed by Atalah et al. (AEA). The aim of the present review was to explore available information on the sensitivity, specificity, and both positive (PPV) and negative predictive values (NPV) of these charts to detect women at risk of delivering babies with the following signs of abnormal fetal growth: (a) length at birth (BL) <50 cm; (b) birth weight (BW) <3,000 g; and (c) BW ≥ 4,000 or 4,250 g. Data from studies conducted in large samples of Chilean and Uruguayan women indicate that the RM chart has the greatest sensitivity to identify at risk cases. However, predictive values were similar for the three charts. Thus, the use of the RM chart should be preferred. The main limitation for using the IOM weight gain recommendations in Latin American women stems from the fact that their average height is approximately 20 cm lower than US women.

An atomic model for the human septin hexamer by cryo-EM.

In order to form functional filaments, human septins must assemble into hetero-oligomeric rod-like particles which polymerize end-to-end. The rules governing the assembly of these particles and the subsequent filaments are incompletely understood. Although crystallographic approaches have been successful in studying the separate components of the system, there has been difficulty in obtaining high resolution structures of the full particle. Here we report a first cryo-EM structure for a hexameric rod composed of human septins 2, 6 and 7 with a global resolution of ~3.6 Å and a local resolution of between ~3.0 Å and ~5.0 Å. By fitting the previously determined high-resolution crystal structures of the component subunits into the cryo-EM map, we are able to provide an essentially complete model for the particle. This exposes SEPT2 NC-interfaces at the termini of the hexamer and leaves internal cavities between the SEPT6-SEPT7 pairs. The floor of the cavity is formed by the two α0 helices including their polybasic regions. These are locked into place between the two subunits by interactions made with the α5 and α6 helices of the neighbouring monomer together with its polyacidic region. The cavity may serve to provide space allowing the subunits to move with respect to one another. The elongated particle shows a tendency to bend at its centre where two copies of SEPT7 form a homodimeric G-interface. Such bending is almost certainly related to the ability of septin filaments to recognize and even induce membrane curvature.

Structure optimization of lipopeptide assemblies for aldol reactions in an aqueous medium.

Four amphiphilic peptides were synthesized, characterized, and evaluated regarding their efficiency in the catalysis of direct aldol reactions in water. The lipopeptides differ by having a double lipid chain and a guanidinium pyrrole group functionalizing one Lys side chain. All the samples are composed of the amino acids l-proline (P), l-arginine (R), or l-lysine (K) functionalized with the cationic guanidiniocarbonyl pyrrole unit (GCP), l-tryptophan (W), and l-glycine (G), covalently linked to one or two long aliphatic chains, leading to surfactant-like designs with controlled proline protonation state and different stereoselectivity. Critical aggregation concentrations (cac) were higher in the presence of the GCP group, suggesting that self-assembly depends on charge distribution along the peptide backbone. Cryogenic Transmission Electron Microscopy (Cryo-TEM) and Small Angle X-ray Scattering (SAXS) showed a rich polymorphism including spherical, cylindrical, and bilayer structures. Molecular dynamics simulations performed to assess the lipopeptide polymorphs revealed an excellent agreement with core-shell arrangements derived from SAXS data and provided an atomistic view of the changes incurred by modifying head groups and lipid chains. The resulting nanostructures behaved as excellent catalysts for aldol condensation reactions, in which superior conversions (>99%), high diastereoselectivities (ds = 94 : 6), and enantioselectivities (ee = 92%) were obtained. Our findings contribute to elucidate the effect of nanoscale organization of lipopeptide assemblies in the catalysis of aldol reactions in an aqueous environment.

Folate status in women of childbearing age in the Urban Metropolitan Region of Chile: results from the National Health Survey 2016-2017.

OBJECTIVE: To analyse serum folate levels in women of childbearing age in the Metropolitan Region (MR) of Chile. DESIGN: Cross-sectional design as part of the 2016-2017 National Health Survey (Encuesta Nacional de Salud, ENS 2016-2017), using a household-based multistage stratified random sample. Serum folate levels measured by electrochemiluminescence immunoassay in fasting venous blood samples were classified as deficient (<4·4 ng/ml), normal (4·4-20 ng/ml) or supraphysiological (>20 ng/ml). SETTING: The MR of Chile. PARTICIPANTS: Women of reproductive age (15-49 years, n 222) from the MR participated in the ENS 2016-2017. RESULTS: The mean, median and range of serum folate were 14·2 (se 0·4), 13·9 and 2·1-32·2 ng/ml, respectively. Folate deficiency was detected in 0·9 % of women, while 7·0 % had supraphysiological levels of the vitamin. No significant effects of age, educational level, marital status, parity, smoking status or nutritional status on serum folate levels were detected by univariate or multivariate analyses. Intake of folic acid supplements showed a significant association with serum folate levels, but only 1·2 % of women used supplements. CONCLUSIONS: Folate deficiency in women of reproductive age living in the MR of Chile is almost inexistent according to the ENS 2016-2017, suggesting that the current population-wide mandatory folic acid fortification of flour is an effective and equitable measure to prevent folate deficiency. These results support the option of maintaining current folic acid fortification in Chile, particularly based on the low adherence to supplementation regimes evidenced in other populations.

Transmission systems in agricultural tractors marketed in Brazil / Transmissões que equipam os tratores agrícolas comercializados no Brasil

ABSTRACT: In a tractor, the transmission system transmits the power generated in the engine to the driving wheels, hydraulic systems, and power take-off (PTO). There are several types of transmission systems available in the Brazilian market ranging from the simplest to the most advanced. Therefore, the objective of this study was to verify the different types of transmission systems present in 228 models of agricultural tractors produced and marketed in Brazil during 2019. The tractor models were grouped based on the engine power and the transmission systems were classified as sliding, partially synchronized, synchronized, semi powershift, full powershift, and continuously variable transmission (CVT). According to this classification, it was found that most tractors belonged to Class III (51.5-73.5 kW) with 66 models in total. However, even with continuous modernization in this field and with tractors becoming increasingly independent from the operator, 66% of transmission systems are still mechanical. However, CVT systems, which are available only in tractors with an engine power above 117.7 kW, are expected to be equipped on less powerful tractors in the coming decades.

RESUMO: Em um trator, o sistema de transmissão tem a função de transmitir a potência gerada no motor para as rodas motrizes, sistemas hidráulicos e tomada de potência (TDP). São diversos os tipos de transmissões disponíveis no mercado brasileiro, desde as mais simples até as de última geração. Neste sentido, o objetivo deste trabalho foi verificar os tipos de transmissões que equipam 228 modelos de tratores agrícolas, produzidos e comercializados no Brasil, durante o ano de 2019. Os modelos de tratores foram agrupados em classes de potência e as transmissões classificadas em: deslizante; parcialmente sincronizada; sincronizada; Semi Powershift; Full Powershift; e CVT. Como resultado, verificou-se que, na Classe III (51,5 a 73,5 kW) são ofertados o maior número de tratores, 66 modelos no total. Mesmo o trator acompanhando as constantes modernizações do campo e estando cada vez mais independente do operador, 66% das transmissões ainda são mecânicas. Porém, há o surgimento da transmissão CVT que, mesmo de modo incipiente, disponível apenas nos tratores com potência acima de 117,7 kW, tende a equipar tratores menos potentes nas próximas décadas.

Spatial variability of soil penetration resistance in the lowland area cultivated with soybean / Variabilidade espacial da resistência do solo à penetração em área de várzea cultivada com soja

ABSTRACT: This study determined the spatial variability of soil penetration resistance and yield of the soybean crop in lowland areas. The soil resistance to penetration at four different depths (0 to 0.10 m; 0.11 to 0.20 m; 0.21 to 0.30 m and 0.31 to 0.40 m), volumetric humidity of the soil at two depths (0 to 0.20 m and 0.21 to 0.40 m) and soybean yield were determined in an area of 1.13 hectares, using a sample mesh of 10 x 10 m. The corresponding data were subjected to descriptive statistical analysis. Pearson's simple linear correlation analysis (p≤0.05) was conducted and the spatial dependence was assessed by analyzing the isotropic semivariograms using spherical, exponential, linear, and Gaussian models. The results showed that the soil penetration resistance increased with depth, with restrictive values to root growth between 0.05 and 0.35 m. There was no correlation between yield and soil penetration resistance, and the semivariograms did not show a defined ascending region regarding the soil penetration resistance data. For the conditions under which the experiment was conducted, the sample 10 x 10 m mesh was suitable for assessing the spatial variability of soil resistance to penetration in depths exceeding 0.10 m.

RESUMO: Este trabalho teve como objetivo identificar a variabilidade espacial da resistência do solo à penetração e na produtividade da cultura da soja, em área de várzea. Foram realizadas determinações de resistência do solo à penetração, em quatro profundidades (0 a 0,10 m; 0,11 a 0,20 m; 0,21 a 0,30 m e 0,31 a 0,40 m); umidade volumétrica do solo, em duas profundidades (0 a 0,20 m e 0,21 a 0,40 m); e produtividade da soja em uma área de 1,13 hectares, utilizando-se malha amostral de 10 x 10 m. Os dados foram submetidos à análise estatística descritiva. Realizou-se análise de correlação linear simples de Pearson (p≤0,05) e a dependência espacial foi avaliada pela análise de semivariogramas isotrópicos, utilizando os modelos: esférico, exponencial, linear e gaussiano. Os resultados indicaram que, a resistência do solo à penetração aumentou em profundidade, com valores restritivos ao crescimento radicular entre 0,05 e 0,35 m. Não se obteve correlação entre produtividade e resistência do solo à penetração sendo que, para os dados de resistência do solo à penetração, os semivariogramas não apresentaram uma região ascendente definida. Para as condições em que o experimento foi realizado, a malha amostral de 10 x 10 m utilizada foi adequada para avaliar a variabilidade espacial da resistência do solo à penetração em profundidades superiores a 0,10 m.

A prática corporal na disciplina natação nos cursos de formação: saber ou não saber nadar? 1 Este trabalho é um recorte de algumas fontes e categorias de análises da pesquisa de mestrado intitulada "Análise da prática corporal na disciplina Natação, nos cursos de Educação Física das universidades públicas da cidade do Recife-PE" que o pesquisador principal realizou através do Programa Associado de Pós-Graduação em Educação Física UPE/UFPB. / Body practice in the swimming subject in undergraduate courses: knowing or not knowing how to swim? / Práctica corporal en la disciplina de natación en cursos de pregrado: ¿saber o no nadar?

RESUMO Alguns estudantes passam pela disciplina Natação nos cursos de formação em Educação Física e não desenvolvem a habilidade do nadar. Este fato nos motivou a desenvolver este estudo que teve por objetivo analisar a prática corporal durante o saber fazer e o saber ensinar na disciplina Natação em três universidades públicas da cidade de Recife-PE. Os dados advindos da pesquisa de campo surgiram por meio de uma entrevista semiestruturada com três professores(as). A análise dos dados foi feita através do método da hermenêutica-dialética com a contribuição da análise de conteúdo do tipo categorial por temática. Como resultado, é visto que o saber nadar é uma condição essencial para os futuros profissionais de educação física e para o ensino da natação.

ABSTRACT Some students go through the Swimming subject in Physical Education courses and do not develop the ability to swim.This fact motivated us to develop this study, which aimed to analyze a body practice during the know-how and teach how to teach in the swimming discipline at three public universities in the city of Recife-PE. The data from the field emerged through a semi-structured interview with three professors. Data analysis was done with a view of Hermeneutics-Dialectics as a paradigm and with the contribution of categorical content analysis by theme. As a result, it is seen that knowing how to swim is an essential condition for future physical education professionals and for swimming teaching.

RESUMEN Algunos estudiantes pasan por la disciplina de Natación en cursos de formación de Educación Física y no desarrollan la habilidad de nadar. Este hecho nos motivó a desarrollar este estudio que tuvo como objetivo analizar la práctica corporal durante o saber hacer y saber enseñar en la disciplina de natación en tres universidades públicas de la ciudad de Recife-PE. Los datos de la investigación de campo surgieron a través de una entrevista semiestructurada con tres profesores y un cuestionario electrónico para estudiantes de las universidades mencionadas. El análisis de datos se realizó mediante el método hermenéutico dialéctico con el aporte del análisis de contenido categórico por tema. Como resultado, se ve que saber nadar es una condición fundamental para los futuros profesionales de la educación física y para la enseñanza de la natación

Visualization of supramolecular structure of Pluronic F127 micellar hydrogels using cryo-TEM.

Pluronic® F127 micellar hydrogels are of growing interest to the biomedical field due to their versatility as drug delivery systems. Pluronic® F127 is a symmetric and amphiphilic triblock copolymer which in aqueous medium self-assembles into micelles that pack togetherwith increasing temperature or concentration, leading to non-flowable hydrogels. The microstructure of these hydrogels is usually investigated by small-angle X-ray scattering, which is not a readily available technique. Conversely, cryo-TEM is a widespread technique used for investigating the morphology of aqueous systems. In the case of Pluronic® F127 micellar systems, the elevated viscosity poses a significant challenge for specimen preparation and, consequently, for cryo-TEM observation. Herein, we show a trustworthy, inexpensive and readily available methodology for preparing specimens of highly viscous micellar solutions and non-flowable hydrogels using an automated vitrification system. With this methodology we were able to visualize not only the morphology of individual Pluronic® F127 micelles -but also the supramolecular structure evolution as a function of concentration. This methodology opens up a wide range of opportunities for hydrogel characterization, although additional systematic studies might be required in order to optimize and replicate it for similar systems.

Early origins of allergy and asthma (ARIES): study protocol for a prospective prenatal birth cohort in Chile.

BACKGROUND: Growing evidence shows that atopic dermatitis (AD), food allergy (FA), allergic rhinitis, and asthma are largely determined during the first 1000 days (time elapsed from conception to the 2nd birthday). The ARIES birth cohort aims to determine prenatal and perinatal conditions, as well as genetic and epigenetic factors, that participate in the early setting of immune responses, and the role of these in the later determination of the risk of allergic diseases and asthma in the offspring. METHODS: We have designed a birth cohort of 250 families with prenatal recruitment (~ 14 weeks). We will genotype relevant allergy/asthma-associated variants in trios and will perform immunophenotyping and evaluation of allergy biomarkers in cord blood. At 1 and 2 years of age we will assess if infants have developed allergic sensitization, AD, FA, as well as biomarkers of asthma including the asthma predictive index. We will also evaluate how maternal conditions modify immune programming through epigenetic modifications and will then depict newborn epigenetic cues of allergy/asthma risk. Next, we will assess composition/diversity of maternal gut, placenta, breastmilk and infant gut microbiome and their association with immunophenotype and biomarkers at birth, and clinical outcomes at age 1 and 2. Finally, we plan to assess how environmental exposures (perinatal outdoor and indoor pollution, allergens and endotoxin) affect the incidence of allergic sensitization, AD, FA, and risk of asthma. DISCUSSION: The in-depth study of the ARIES birth cohort shall provide crucial information to understand the rising incidence of allergies and asthma in developing countries, and hopefully provide cues on how to prevent and treat these diseases. TRIAL REGISTRATION: clinicaltrials.gov NCT04186949, retrospectively registered on December 5, 2019.

Bacterioruberin from Haloarchaea plus dexamethasone in ultra-small macrophage-targeted nanoparticles as potential intestinal repairing agent.

Oral administration of antioxidant and anti-inflammatory drugs have the potential to improve the current therapy of inflammatory bowel disease. Success of oral treatments, however, depends on the capacity of drugs to remain structurally stable along the gastrointestinal tract, and on the feasibility of accessing the target cells. Delivering anti-inflammatory and antioxidant drugs to macrophages using targeted nanoparticles, could make treatments more efficient. In this work structural features and in vitro activity of macrophage-targeted nanostructured archaeolipid carriers (NAC) containing the high antioxidant dipolar C50 carotenoid bacterioruberin (BR) plus dexamethasone (Dex): NAC-Dex, are described. Ultra-small (66 nm), -32 mV ζ potential, 1200 µg Dex /ml NAC-Dex, consisted of a compritol and BR core, covered by a shell of sn 2,3 ether linked archaeolipids and Tween 80 (2: 2: 1.2: 3 % w/w) were obtained. NAC-Dex were extensively captured by macrophages and Caco-2 cells and displayed high anti-inflammatory and antioxidant activities on a gut inflammation model made of Caco-2 cells and lipopolysaccharide stimulated THP-1 derived macrophages reducing 65 % and 55 % TNF-α and IL-8 release, respectively and 60 % reactive oxygen species production. NAC-Dex also reversed the morphological changes induced by inflammation and increased the transepithelial electrical resistance, partly reconstituting the barrier function. Activity of BR and Dex in NAC-Dex was partially protected after simulated gastrointestinal digestion, improving the chances of BR-Dex joint activity. Results suggest that oral NAC-Dex deserve further exploration as intestinal barrier repairing agent.

Gestational Diabetes Mellitus Treatment Schemes Modify Maternal Plasma Cholesterol Levels Dependent to Women´s Weight: Possible Impact on Feto-Placental Vascular Function.

: Gestational diabetes mellitus (GDM) associates with fetal endothelial dysfunction (ED), which occurs independently of adequate glycemic control. Scarce information exists about the impact of different GDM therapeutic schemes on maternal dyslipidemia and obesity and their contribution to the development of fetal-ED. The aim of this study was to evaluate the effect of GDM-treatments on lipid levels in nonobese (N) and obese (O) pregnant women and the effect of maternal cholesterol levels in GDM-associated ED in the umbilical vein (UV). O-GDM women treated with diet showed decreased total cholesterol (TC) and low-density lipoproteins (LDL) levels with respect to N-GDM ones. Moreover, O-GDM women treated with diet in addition to insulin showed higher TC and LDL levels than N-GDM women. The maximum relaxation to calcitonin gene-related peptide of the UV rings was lower in the N-GDM group compared to the N one, and increased maternal levels of TC were associated with even lower dilation in the N-GDM group. We conclude that GDM-treatments modulate the TC and LDL levels depending on maternal weight. Additionally, increased TC levels worsen the GDM-associated ED of UV rings. This study suggests that it could be relevant to consider a specific GDM-treatment according to weight in order to prevent fetal-ED, as well as to consider the possible effects of maternal lipids during pregnancy.

Pair Distribution Function from Electron Diffraction in Cryogenic Electron Microscopy: Revealing Glassy Water Structure.

In recent years, cryogenic electron microscopy (Cryo-EM) has revolutionized the structure determination of wet samples and especially that of biological macromolecules. The glassy-water medium in which the molecules are embedded is considered an almost in vivo environment for biological samples. The local structure of amorphous ice is known from neutron- and X-ray-diffraction studies, techniques appropriate for much larger volumes than those used in cryo-EM. We here present a first study of the pair-distribution function g(r) of glassy water under cryo-EM conditions using electron diffraction data. We found g(r) to be between that of low-density amorphous ice and that of supercooled water. Under electron exposure, cubic-ice regions were found to nucleate in thicker glassy-water samples. Our work enables to obtain quantitative structural information using g(r) from cryo-EM.

Nanoparticles containing ß-cyclodextrin potentially useful for the treatment of Niemann-Pick C.

ß-Cyclodextrin (ß-CD) is being considered a promising therapy for Niemann-Pick C (NPC) disease because of its ability to mobilise the entrapped cholesterol from lysosomes, however, a major limitation is its inability to cross the blood-brain barrier (BBB) and address the central nervous system (CNS) manifestations of the disease. Considering this, we aimed to design nanoparticles able to cross the BBB and deliver ß-CD into the CNS lysosomes. The physicochemical characteristics of ß-CD-loaded nanoparticles were evaluated by dynamic light scattering, small-angle X-ray scattering, and cryogenic transmission electron microscopy. The in vitro analyses were performed with NPC dermal fibroblasts and the ß-CD-loaded nanoparticles were tracked in vivo. The nanoparticles showed a mean diameter around 120 nm with a disordered bicontinuous inner structure. The nanoparticles did not cause decrease in cell viability, impairment in the antioxidant enzymes activity, damage to biomolecules or release of reactive species in NPC dermal fibroblasts; also, they did not induce genotoxicity or alter the mitochondrial function in healthy fibroblasts. The ß-CD-loaded nanoparticles were taken up by lysosomes reducing the cholesterol accumulated in NPC fibroblasts and reached the CNS of mice more intensely than other organs, demonstrating advantages compared to the free ß-CD. The results demonstrated the potential of the ß-CD-loaded nanoparticles in reducing the brain impairment of NPC.