MicroRNA-25-5p (miR-25-5p) may be involved in the pathogenesis and processes of vascular diseases. The aim of this study was to investigate the role of miR-25-5p in oxidized low-density lipoprotein (ox-LDL)-treated human brain microvessel endothelial cells (HBMECs) and the underlying mechanisms. RT-qPCR and/or Western blot were used to detect the expression levels of miR-25-5p and neuronal growth regulator 1 (NEGR1). The effect of miR-25-5p overexpression and NEGR1 silencing on cell proliferation, migration, apoptosis and reactive oxygen species (ROS) production of HBMECs were measured by using CCK-8 assay, transwell assay and flow cytometry, respectively. The expression levels of apoptosis-related protein (cleaved caspase-3 and pro-caspase-3) were detected using Western blot, and the nitric oxide (NO) production was measured by a nitric oxide assay kit. The expression level of miR-25-5p was decreased in HBMECs treated with ox-LDL. Compared with the control group, miR-25-5p overexpression significantly promoted the proliferation and migration of HBMECs treated with ox-LDL (p < 0.01). Overexpression of miR-25-5p significantly suppressed cell apoptosis, ROS production and NO reduction of ox-LDL-induced HBMECs (p < 0.01). In addition, the target gene of miR-25-5p was predicted to be NEGR1 through Targetscan online analysis. The effect of NEGR1 silencing on cell proliferation, migration, apoptosis, ROS and NO production of ox-LDL-induced HBMECs was similar to that of miR-25-5p overexpression. Furthermore, miR-25-5p overexpression and NEGR1 silencing significantly downregulated the protein expression levels of JAK2 and STAT3. Thus, miR-25-5p neutralizes the effects of ox-LDL on multiple functions of HBMECs through suppressing the expression of NEGR1 via regulating the JAK/STA signaling pathway.

While reporting human immunodeficiency virus (HIV) viral load (VL) using dried blood spot (DBS) in the BioMerieux NucliSENS platform, application of the hematocrit correction factor has been suggested. In this cross-sectional study from the National Microbiology Reference Laboratory of Zimbabwe, we assessed whether hematocrit correction (individual and/or mean) in DBS results improved the correlation with plasma VL and prediction of VL non-suppression (≥1000 copies per ml in plasma). Of 517 specimens during August-December 2018, 65(12.6%) had non-suppressed plasma VL results. The hematocrit correction factor ranged from 1.3 to 2.0 with a mean of 1.6, standard deviation (SD: 1.5, 1.7). The intraclass correlation (ICC) for mean (0.859, 95% CI: 0.834, 0.880) and individual (0.809, 95% CI: 0.777, 0.837) hematocrit corrected DBS results were not significantly different. The uncorrected DBS results had a significantly lower ICC (0.640, 95% CI: 0.586, 0.688) when compared to corrected DBS results. There were no significant differences in validity, predictive values, and areas under the receiver operating characteristics curves for all three DBS results when predicting VL non-suppression. To conclude, hematocrit correction of DBS VL results improved agreement with the plasma results but did not improve prediction of VL non-suppression. The results were not significantly different for individual and mean corrected results.

Obesity-induced inflammation is frequently associated with higher oxidative stress. In vitro and experimental studies have considered baru almonds (Dipteryx alata Vog) as a legume seed with high antioxidant capacity. The aim of this study was to evaluate whether baru almonds are capable of improving the inflammatory and antioxidant status in overweight and obese women.

Decision-making is an important everyday function that deteriorates during normal aging. Here, we asked whether value-based decision-making can be improved in the elderly by cognitive training. We compared the effects of two training regimens on the performance in the Iowa Gambling Task (IGT), a real-life decision-making simulation task. Elderly participants (age 62-75 years) were randomized into three matched groups. The filter training (FT) group performed a selective attention task and the memory training (MT) group performed a memory storage task on five consecutive days. The control group (CG) did not perform another task besides the IGT. Only the FT group showed an improvement in IGT performance over the five days-the overall gain rose and the prominent deck B phenomenon decreased. The latter refers to the selection of cards associated with high gains and rare losses, which are nevertheless a disadvantageous choice as the frequent losses lead to a negative net outcome. As the deck B phenomenon has been associated with impaired cognitive abilities in aging, the positive effect of FT here is of special importance. In sum, attention training seems superior in improving decision-making in the elderly.

Mental workload (MWL) can affect human performance and is considered critical in the design and evaluation of complex human-machine systems. While numerous physiological measures are used to assess MWL, there appears no consensus on their validity as effective agents of MWL. This study was conducted to provide a comprehensive understanding of the use of physiological measures of MWL and to synthesize empirical evidence on the validity of the measures to discriminate changes in MWL. A systematical literature search was conducted with four electronic databases for empirical studies measuring MWL with physiological measures. Ninety-one studies were included for analysis. We identified 78 physiological measures, which were distributed in cardiovascular, eye movement, electroencephalogram (EEG), respiration, electromyogram (EMG) and skin categories. Cardiovascular, eye movement and EEG measures were the most widely used across varied research domains, with 76%, 66%, and 71% of times reported a significant association with MWL, respectively. While most physiological measures were found to be able to discriminate changes in MWL, they were not universally valid in all task scenarios. The use of physiological measures and their validity for MWL assessment also varied across different research domains. Our study offers insights into the understanding and selection of appropriate physiological measures for MWL assessment in varied human-machine systems.

(1) Background: Parental feeding practices are related to child body mass index (BMI, kg/m2) and seem to be a consequence rather than cause of child BMI, but research so far is limited. Stability and continuity of feeding practices, probably explaining changes in food intake and child BMI, remain to be poorly examined. (2) Methods: Feeding practices (i.e., restriction, food as reward, pressure to eat, monitoring) assessed via the Child Feeding Questionnaire, child age, standardized BMI (zBMI), and socio-economic status were measured annually at multiple visits (range 2-8) in a population-based longitudinal cohort study of 1512 parents with their children aged 2 to 12 years. Stability, continuity, and bi-directionality of feeding practices and child zBMI were calculated using correlation coefficients, paired t tests, and cross-lagged panels, respectively. (3) Results: Feeding practices and child zBMI showed moderate to high stability. While continuity was high for restriction, minor temporal changes were observed for other feeding practices and child zBMI. Cross-lags indicated that child zBMI predicted restriction, pressure to eat, and monitoring, while food-rewarding predicted child zBMI only minorly. (4) Conclusions: Parents seem to adapt feeding practices to child zBMI with the exception of food-rewarding.

The threat imposed by the bacterial pathogen Xylella fastidiosa to crops of utter importance to European agriculture such as olive, stone fruit and grapevine calls for immediate research against the meadow spittlebug, Philaenus spumarius (L.), the main European vector. Management tools should consider reducing juveniles of vector populations growing on weeds or cover crops during spring as nymphs have limited movement and do not contribute to disease spread. We examined a wide range of insecticides with different modes of action against P. spumarius nymphs in laboratory and semi-field glasshouse conditions. Pyrethroids (delthamethrin and λ-cyhalothrin) and natural pyrethrin (Pirecris®) + piperonyl butoxide (PBO) efficacy surpassed 86% after 24 h of exposure, without significant differences in the PBO amount tested. The inclusion of PBO caused a 3-fold increase in the mortality of P. spumarius nymphs compared to pyrethrin alone. Sulfoxaflor (Closer®) exhibited similar efficacy at 48 and 72 h but it was slow acting and mortality only reached 60% at 24 h. The LC90 was 34 ppm at 72 h. Pymetrozine, spirotetramat, azadirachtin and kaolin were not effective against nymphs (mortality <33%) although in azadirachtin-treated plants, mortality had a 3-fold increase from 24 to 72 h. Our results will help decision-making policy bodies to set up a sustainable integrated pest management of P. spumarius in areas where X. fastidiosa becomes a problem.

Recent data on erosive tooth wear (ETW) in Belgium have associated a vocational/technical type of education with ETW risk. Since the role of schools is essential to the promotion of healthy diets, this study aimed to investigate school food policies (SFP) related to soft drink and fruit juice consumption and to detect differences among schools in Flanders, Belgium (BE-F). An online questionnaire related to the control of acidic beverages and promotion of healthy drinking habits was sent to all Flemish secondary schools. For analysis, schools (n = 275) were grouped by type of education (vocational secondary education (VSE) and general secondary education (GSE)), and by socioeconomic status. Multiple factor analyses (MFA) were performed to identify schools with a similar SFP profile. Additionally, descriptive analyses were performed to determine other associations. Overall, 44% of schools in BE-F claimed to have written SFP related to the consumption of soft drinks. SFP expressly prohibiting or limiting acidic beverages were significantly more frequent in GSE schools (p < 0.05), where a higher economic status was present. This study shows that a considerable group of schools in BE-F have no or incomplete rules concerning acidic beverage consumption. Such rules differ between types of education, with VSE schools reporting less control regarding the consumption of drinks.

Lanthanide-based nanophosphors (NPhs) are herein developed as contrast agents for spectral X-ray imaging, highlighting the chemical, macromolecular and structural differences derived from ligand exchange on computed tomography (CT) and solvent dispersibility. Taking advantage of the ability of spectral X-ray imaging with photon-counting detectors to perform image acquisition, analysis, and processing at different energy windows (bins), enhanced signal of our K-edge materials was derived, improving sensitivity of CT imaging, and differentiation between water, tumor-mimic phantoms, and contrast materials. Our results indicate that the most effective of our oleic acid-stabilized K-edge nanoparticles can achieve 2-4x higher contrast than the examined iodinated molecules, making them suitable for deep tissue imaging of tissues or tumors. On the other hand, ligand exchange yielding poly(acrylic acid)-stabilized K-edge nanoparticles allows for high dispersibility and homogeneity in water, but with a lower contrast due to the high density of the polymer grafted, unless further engineering is probed. This is the first well-defined study that manages to correlate NPh grafting density with CT numbers and water dispersibility, laying the groundwork for the development of the next generation CT-guided diagnostic and/or theranostic materials.

Duration of receptor antagonism, measured as the recovery of agonist responsiveness, is gaining attention as a method to evaluate the 'effective' target-residence for antagonists. These functional assays might be a good alternative for kinetic binding assays in competition with radiolabeled or fluorescent ligands, as they are performed on intact cells and better reflect consequences of dynamic cellular processes on duration of receptor antagonism. Here, we used a bioluminescence resonance energy transfer (BRET)-based assay that monitors heterotrimeric G protein activation via scavenging of released Venus-Gβ1γ2 by NanoLuc (Nluc)-tagged membrane-associated-C-terminal fragment of G protein-coupled receptor kinase 3 (masGRK3ct-Nluc) as a tool to probe duration of G protein-coupled receptor (GPCR) antagonism. The Gαi-coupled histamine H3 receptor (H3R) was used in this study as prolonged antagonism is associated with adverse events (e.g., insomnia) and consequently, short-residence time ligands might be preferred. Due to its fast and prolonged response, this assay can be used to determine the duration of functional antagonism by measuring the recovery of agonist responsiveness upon washout of pre-bound antagonist, and to assess antagonist re-equilibration time via Schild-plot analysis. Re-equilibration of pre-incubated antagonist with agonist and receptor could be followed in time to monitor the transition from insurmountable to surmountable antagonism. The BRET-based G protein activation assay can detect differences in the recovery of H3R responsiveness and re-equilibration of pre-bound antagonists between the tested H3R antagonists. Fast dissociation kinetics were observed for marketed drug pitolisant (Wakix®) in this assay, which suggests that short residence time might be beneficial for therapeutic targeting of the H3R.

In the silkworm, the sex-determination primary signal Fem controls sex differentiation by specific binding of Fem-derived piRNA to the cleavage site in Masc mRNA, thus inhibiting Masc protein production in the female. In this study, we identified a novel splicing isoform of Masc, named Masc-S, which lacks the intact sequence of the cleavage site, encoding a C-terminal truncated protein. Results of RT-PCR showed that Masc-S was expressed in both sexes. Over-expression of Masc-S and Masc in female-specific cell lines showed that Masc-S could be translated against the Fem-piRNA cut. By RNA-protein pull-down, LC/MS/MS, and EMSA, we identified a protein BmEXU that specifically binds to an exclusive RNA sequence in Masc compared to Masc-S. Knockdown of Masc-S resulted in abnormal morphology in female external genital and increased expression of the Hox gene Abd-B, which similarly occurred by Bmexu RNAi. These results suggest that the splice variant Masc-S against Fem-piRNA plays an important role in female external genital development, of which function is opposite to that of full-length Masc. Our study provides new insights into the regulatory mechanism of sex determination in the silkworm.

Preussin, a hydroxyl pyrrolidine derivative isolated from the marine sponge-associated fungus Aspergillus candidus KUFA 0062, displayed anticancer effects in some cancer cell lines, including MCF7. Preussin was investigated for its cytotoxic and antiproliferative effects in breast cancer cell lines (MCF7, SKBR3, and MDA-MB-231), representatives of major breast cancers subtypes, and in a non-tumor cell line (MCF12A). Preussin was first tested in 2D (monolayer), and then in 3D (multicellular aggregates), cultures, using a multi-endpoint approach for cytotoxicity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), resazurin and lactate dehydrogenase (LDH)) and proliferative (5-bromo-2'-deoxyuridine (BrdU)) assays, as well as the analysis of cell morphology by optical/electron microscopy and immunocytochemistry for caspase-3 and ki67. Preussin affected cell viability and proliferation in 2D and 3D cultures in all cell lines tested. The results in the 3D culture showed the same tendency as in the 2D culture, however, cells in the 3D culture were less responsive. The effects were observed at different concentrations of preussin, depending on the cell line and assay method. Morphological study of preussin-exposed cells revealed cell death, which was confirmed by caspase-3 immunostaining. In view of the data, we recommend a multi-endpoint approach, including histological evaluation, in future assays with the tested 3D models. Our data showed cytotoxic and antiproliferative activities of preussin in breast cancer cell lines in 2D and 3D cultures, warranting further studies for its anticancer potential.

Decarboxylative aldol reaction of aliphatic carboxylic acids is a useful method for C-C bond formation because carboxylic acids are an easily available class of compounds. In this study, we found that the decarboxylative aldol reaction of tertiary β-ketocarboxylic acids and trifluoropyruvates proceeded smoothly to yield the corresponding aldol products in high yields and with high diastereoselectivity in the presence of a tertiary amine catalyst. In this reaction, we efficiently constructed a quaternary carbon center and an adjacent trifluoromethylated carbon center. This protocol was also extended to an enantioselective reaction with a chiral amine catalyst, and the desired product was obtained with up to 73% enantioselectivity.

The immobilization of fluorescent proteins is a key technology enabling to fabricate a new generation of photoactive materials with potential technological applications. Herein we have exploited superfolder green (sGFP) and red (RFP) fluorescent proteins expressed with different polypeptide tags. We fused these fluorescent proteins to His-tags to immobilize them on graphene 3D hydrogels, and Cys-tags to immobilize them on porous microparticles activated with either epoxy or disulfide groups and with Lys-tags to immobilize them on upconverting nanoparticles functionalized with carboxylic groups. Genetically programming sGFP and RFP with Cys-tag and His-tag, respectively, allowed tuning the protein spatial organization either across the porous structure of two microbeads with different functional groups (agarose-based materials activated with metal chelates and epoxy-methacrylate materials) or across the surface of a single microbead functionalized with both metal-chelates and disulfide groups. By using different polypeptide tags, we can control the attachment chemistry but also the localization of the fluorescent proteins across the material surfaces. The resulting photoactive material formed by His-RFP immobilized on graphene hydrogels has been tested as pH indicator to measure pH changes in the alkaline region, although the immobilized fluorescent protein exhibited a narrower dynamic range to measure pH than the soluble fluorescent protein. Likewise, the immobilization of Lys-sGFP on alginate-coated upconverting nanoparticles enabled the infrared excitation of the fluorescent protein to be used as a green light emitter. These novel photoactive biomaterials open new avenues for innovative technological developments towards the fabrication of biosensors and photonic devices.

(1) Background: Genetically based hyperinflammation may play a role in pathogen defense. We here questioned whether alterations in circulating monocytes/macrophages, inflammatory biomarkers and a functional SNP (single nucleotide polymorphisms) of the Interleukin-6 (IL-6) promotor might play a role in patients with persistent, and treatment resistant borreliosis. (2) Methods: Leukocyte subpopulations were studied by flow cytometry; plasma cytokines were determined by a chemiluminescence based ELISA (Immulite®), and genotypes of the IL-6 promotor SNP rs1800795 were determined by pyrosequencing. (3) Results: In a cohort of n = 107 Lyme borreliosis patients, who concomitantly manifested either malignant diseases (group 1), autoimmune disorders (group 2), neurological diseases (group 3), or morbidities caused by multiple other infectious complications (group 4), we found decreased numbers of anti-inflammatory CD163-positive macrophages, elevated concentrations of inflammatory cytokines, and an imbalance of IL-6 promotor SNP rs1800795 genotypes. The most prominently upregulated cytokines were IL-1β, and IL-8. (4) Conclusions: Increased pro-inflammatory phenotypes identified by monocyte/macrophage subtypes and concomitantly increased cytokines appear to be valid to monitor disease activity in patients with persistent Lyme borreliosis. Patterns may vary by additional co-morbidities. In patients with autoimmune diseases, increased frequencies of a heterozygous IL-6 promotor SNP rs1800795 were identified. This functional SNP may guide chronic inflammation, impacting other cytokines to trigger trigger chronicity and therapeutic resistance in Lyme borreliosis.

Overexpression of Bmi1 gene is an important feature of cancer stem cell in various human tumors. Therefore, Bmi1 gene can be a potential target for small interfering RNA (siRNA) mediated cancer therapy. Ursolic acid (UA) as a natural product plays a pivotal role in anti-tumor field, although its performance is limited by low bioavailability and poor hydrophilicity. A folate receptor-targeted cationic liposome system was designed for the purpose of investigating the relationship between Bmil siRNA and UA. The folate receptor-targeted cationic liposomes co-delivering UA and Bmi1 siRNA (FA-UA/siRNA-L) were fabricated by electrostatic interaction between folate UA liposome (FA-UA-L) and Bmi1 siRNA. Tumor growth is inhibited by FA-UA/siRNA-L in vitro and in vivo and this inhibition is contributed by a synergistic anti-tumor effect of UA and Bmi1 siRNA. The western blot measurement of apoptosis-protein and cancer stem cell (CSC) marked-protein demonstrated that UA led to activation-induced tumor cell death and Bmi1 siRNA resulted in inhibition of cancer stem cells. Overall, these results indicate that Bmi1 as a regulating gene for cancer stem cell is an effective target for cancer treatment using siRNA and co-delivery of UA and Bmi1 siRNA using folate-targeted liposomes is a promising strategy for improved anti-tumor effect.

Oyster is rich in glycogen and free amino acids and is called "the milk of sea". To understand the main genetic effects of these traits and the genetic networks underlying their correlation, we have conducted the whole genome resequencing with 427 oysters collected from the world-wide scale.

Despite significant advances in surgical techniques, immunosuppression protocols, follow up periods and antimicrobial stewardship in modern medicine; post-renal transplantation urinary tract infection remained a major public health problem globally. This multiple serious squeals includes asymptomatic bacteriuria, cystitis and pyelonephritis. Among these, the bacterial origin of infection complications accounts for the most significant clinical, socio-economic impacts in many countries of the world. Therefore, the aim of the study was to investigate the prevalence of bacterial isolates that cause urinary tract infections, assess antibiotic susceptibility pattern among symptomatic and asymptomatic renal transplant recipients attending at St. Paul's Hospital Millennium Medical College, Addis Ababa, Ethiopia.

Claudin-low breast cancer is a molecular subtype associated with poor prognosis and without targeted treatment options. The claudin-low subtype is defined by certain biological characteristics, some of which may be clinically actionable, such as high immunogenicity. In mice, the medroxyprogesterone acetate (MPA) and 7,12-dimethylbenzanthracene (DMBA)-induced mammary tumor model yields a heterogeneous set of tumors, a subset of which display claudin-low features. Neither the genomic characteristics of MPA/DMBA-induced claudin-low tumors nor those of human claudin-low breast tumors have been thoroughly explored.

Many low- and middle-income countries (LMIC) are moving towards enforcing prescription-only access to antibiotics. This systematic literature review aims to assess the interventions used to enforce existing legislation prohibiting over-the-counter (OTC) sales of antibiotics in LMICs, their impact and examine the methods chosen for impact measurement including their strengths and weaknesses.