Alcohol misuse often manifests in two different patterns of drinking; Binge Drinking (BD; ≥4 (women) or ≥ 5 (men) drinks/day, ≤12 days/month) or Heavy Drinking (HD; ≥3 (women) or ≥4 (men) drinks/day, ≥16 days/month). Although direct comparisons have not been made, structural MRI studies indicate that the two types of drinking behaviors might be associated with different neuromorphometric characteristics.

Osteosarcoma (OS) is the most frequent primary malignant tumour of bone and metastases occur in 30% of cases, the 5-year survival rate is 25-30%. Although pre- and post-operative chemotherapy has improved prognosis in osteosarcoma (OS), high toxicity and natural and acquired drug-resistance are the first cause of treatment failure. The identification of new predictive and therapeutic biomarkers may increase drug sensitivity and better control localized and metastatic disease. By the evidence that CXCR4 receptor by binding its ligand CXCL12 activates downstream critical endpoints for tumour malignancy, we first studied human OS progression correlating CXCR4 expression in OS biopsy with patient clinical data. By Real-time PCR and immunoistochemistry we found that high levels of CXCR4 gene and protein expression significantly correlated with OS progression, emphasizing the role of CXCR4/CXCL12 axis in tumour prognosis. This was supported by univariate analyses that showed a higher probability of local and/or systemic relapse in OS patients with a high CXCR4 gene expression and a significant increase of metastasis risk associated with an increasing score of CXCR4 protein staining intensity. Secondarily, to study the role of CXCR4 as a target for new therapeutic strategies, we evaluated the response of OS cells to the fully human CXCR4 antibody, MDX1338. In the study we also included AMD3100, the most studied CXCR4 antagonist. In CXCR4-positive OS cells cultured in CXCL12-rich BM-MCS-CM (bone marrow-derived mesenchymal stem conditioned medium), a decrease of cell proliferation up to 30%-40% of control was seen after drug exposure. However, an increase of apoptosis was seen in p53-positive U2OS and 143B after CXCR4 inhibitor incubation, while no changes were seen in treated SAOS-2 cells which also present a different labeling profile. The role of p53 in apoptotic response to CXCR4 inhibitors was confirmed by p53 silencing in U2OS cell line. Our data suggest that the response to anti-CXCR4 agents could be influenced by the genetic background and labeling profile which induces a different cross-talk between tumour cells and environment. The delay in cell cycle progression associated with increased apoptosis could sensitize p53-positive cells to conventional therapy and in vivo preclinical experiments are on going with the aim to suggest new combined target therapies in human OS.

Time and cost constraints abound in the Emergency Centre (EC). These resource-constraints are further magnified in low- and middle-income countries (LMIC). Almost half of all patients presenting to the EC require laboratory tests. Unfortunately, access to laboratory services in LMIC is commonly inadequate. Point-of-Care (POC) tests may assist to avert this shortcoming. The aims of this study were to evaluate the cost effectiveness of upfront POC blood tests performed prior to doctor assessment compared to the standard EC workflow.

Research at the intersection of substance use and protective factors among transgender youth is scarce; emerging evidence suggests high risk for substance use for transgender youth. We analyzed data from 323 transgender youth aged 14-18 (M age = 16.67) to investigate the extent that risk (enacted stigma) and protective factors (support from family, school, friends) were related to substance use (i.e., cannabis and tobacco use, binge drinking). Enacted stigma was linked to higher odds of substance use behaviors, family connectedness was related to lower levels of tobacco and cannabis use, and more than one protective factor significantly lowered the probability of engaging in substance use behaviors. Support from multiple sources may be differentially protective against substance use for transgender youth.

The aim of the present study is to firstly study the effect of the extraction solvents (ethanol, acetonitrile, distilled water), pH, temperature, and the extraction method (maceration, sonication, maceration in two steps) on the flavonoid and phenolic contents of olive leaves. Furthermore, qualitative and quantitative analyzes of phenolic compounds by (HPLC) were performed. Results showed that the extract macerated in two steps by ethanol followed by distilled water of dried leaves showed high contents of phenolic compounds and flavonoids compared to the extracts obtained by the other studied techniques and solvents. On the other hand, the macerated extracts were studied for their antibacterial activity against five pathogenic bacteria (Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Listeria monocytogenes). The results showed a strong antibacterial activity of the same macerated extract in two steps for dried leaves, which could be attributed to its richness in bioactive compounds such as oleuropein.

To assess the rate of postanesthesia respiratory depression (RD) and test for potential associations with clinical characteristics of patients undergoing urogynecologic procedures at ambulatory surgical centers (ASCs). Postanesthesia RD is poorly characterized for patients undergoing urogynecologic procedures in ASCs.

Obesity is associated with high mortality and morbidity rates and low levels of quality of life among adults globally. It is critical to examine evidence-based practices for developing lifestyle behavioral changes such as physical movement and structured exercise training. The DoIT protocol, a high-intensity interval exercise training (HIIT) program, effectively reduces body mass, alters energy balance, and improves performance of obese adults with a high adherence rate. This study aims to determine the dose-response effects of the DoIT protocol on body composition, health, performance and quality of life in sedentary obese adults. This study will recruit 88 sedentary, obese males and females (BMI 25.0-34.9; 30-50 years) who will be randomly assigned to one of four groups: (i) control (n = 22), (ii) one session/week (n = 22), (iii) two sessions/week (n = 22) or (iv) three sessions/week (n = 22). DoIT will use a supervised, circuit-type (1-3 rounds), functional/neuromotor and progressive exercise program for 12 months. DoIT incorporates 8-12 multi-planar, fundamental and complex, whole body movements and uses bodyweight and alternative exercise modes as a resistance. DoIT utilizes prescribed work-to-rest ratios which will be varied every four weeks. Each session will last less than 30 min. DoIT will be implemented for a year and its effects on body mass and body composition, physical fitness, functional capacity, bone health, leptin, adiponectin, blood lipids, glycemic control, inflammation, oxidative stress and quality of life will be assessed. The outcomes of the proposed study will provide insight on optimal exercise prescription guidelines for such HIIT-type exercise protocols for overweight or obese individuals.

The effects of kafirins on protein and amino acid ileal digestibility have not been evaluated in vivo in pigs. The aim of this study was to determine the effects of protein profile on apparent ileal digestibility (AID) of amino acids. We used a sorghum hybrid with low tannin content (<0.5%). The same hybrid was harvested from 2 different plots with different kafirin profile. Sorghum with greater content of total kafirins had less content of γ- and α1-kafirins and higher content of β- and α2-kafirins than that with lower content of total kafirins. Two sorghum-soybean meal (SBM) diets were formulated: 1) low kafirin (LK) content (32.2 g/kg) and 2) high kafirin (HK) content (48.1 g/kg). A control diet (maize-SBM) and a reference SBM-diet were also prepared. The reference diet was fed to all pigs following the experimental period and was used to estimate the AID of cereals by the difference method. "T" cannulas were fixed in the distal ileum of 18 barrows (6 by treatment), divided into 2 groups of 9 pigs. The pigs were fed 2.5 times their maintenance requirement of digestible energy (110 kcal/kg BW0.75). The AID of dry matter, protein, amino acids, and energy of the experimental diets was measured; the AID of cereals (maize, LK sorghum and HK sorghum) was estimated by the difference method. The maize-SBM diet was more digestible than the sorghum-SBM diets, only with respect to valine (P < 0.05). The AID of valine in the maize-SBM diet was higher than that in sorghum-SBM diets. The changes in kafirin profile between the diets only affected the AID of threonine (P < 0.01), which decreased by 9.5 percentage units in LK diet compared with HK diet. Regarding the AID of cereals, maize exhibited greater AID than sorghum, with respect to valine (P < 0.01) and serine (P < 0.10). A comparison of sorghum with LK and HK content showed that the AID of threonine and serine increased by 50.5 (P < 0.001) and 19.2 percentage units (P < 0.05) in the latter, respectively. The higher content of γ-kafirins in LK sorghum negatively affected threonine and serine digestibility, implying that the AID of amino acids is affected more by the profile than the content of kafirins.

Babesiosis and Theileriosis are important worldwide-distributed tick-borne diseases for human and animals. Their presence in a particular area depends on the presence of suitable tick-vector and host species as well as competent reservoirs such as roe deer, one of the most abundant wild cervids in Spain. Spleen samples from 174 roe deer hunted in Spain were analysed to determine the prevalence of Babesia and Theileria species. DNA of both piroplasms was firstly detected using a commercial qPCR. Then, positive samples were molecularly characterized at the 18S rRNA and ITS1 genes of Babesia spp. and Theileria spp. The possible influence of some factors such as ecological area, age and sex was also assessed. Overall, 89.7% of roe deer were positive to any of the two piroplasms. Theileria spp. was more prevalent (60.9%) than Babesia spp. (19.0%); species identification could not be achieved in 17.3% of positive samples. Babesia prevalence was significantly higher in young animals and in roe deer from Oceanic regions, in contrast to Theileria spp. Five species were identified: Theileria sp. OT3 (60.3%), Babesia capreoli (15.5%), Babesia venatorum (2.9%), Theileria sp. 3185/02 (0.6%) and Babesia bigemina (0.6%). The coinfection B. capreoli/T. sp. OT3 was the most common (4.6%) followed by B. venatorum/T. sp. OT3 (0.6%) and B. bigemina/T. sp. OT3 (0.6%). Our results reveal that Theileria spp. and Babesia spp. are prevalent piroplasms in roe deer from Spain. These cervids can act as reservoirs for several Babesia and Theileria species, including the zoonotic B. venatorum. This study represents the first description of B. venatorum and B. bigemina in roe deer from Spain.

Although cobalamin (vitamin B12) deficiency was described over a century ago, it is still difficult to establish the correct diagnosis and prescribe the right treatment. Symptoms related to vitamin B12 deficiency may be diverse and vary from neurologic to psychiatric. A number of individuals with vitamin B12 deficiency may present with the classic megaloblastic anemia. In clinical practice, many cases of vitamin B12 deficiency are overlooked or sometimes even misdiagnosed. In this review, we describe the heterogeneous disease spectrum of patients with vitamin B12 deficiency in whom the diagnosis was either based on low serum B12 levels, elevated biomarkers like methylmalonic acid and/or homocysteine, or the improvement of clinical symptoms after the institution of parenteral vitamin B12 therapy. We discuss the possible clinical signs and symptoms of patients with B12 deficiency and the various pitfalls of diagnosis and treatment.

Sudan is located in the heart of Africa and surrounded by eight countries with people of different ethnic origins. Historical records show that the population of the Sudan is a mixture of Arabic, West Asian Arabic and sub-Saharan African elements. The present survey provides data on allele lineages, and haplotype frequencies of Human Leukocyte Antigen (HLA) class I (HLA-A and HLA-B) and class II (HLA-DR and -HLA-DQ) loci in 11 Sudanese populations. The sampled individuals are all local transplant donors who provided informed consent for HLA analyses on their blood samples and were registered at National Cancer Institute, University of Gezira, Wad Medani. The HLA class I and II data reported here can be subjected for future analyses of genetic structure and health in Sudan. These include as reference datasets for identifying the association between HLA and diseases and for designing donor recruitment strategies.

Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1β), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α). This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types. This technique advances traditional ELISAs by: •FMIA provides insight into the profiles of multiple porcine cytokines in certain situations (e.g. disease, parturition).•Use of the Bio-Plex® 200 system to investigate novel sample types, including brain, placenta and synovial tissue.•Multiplexing utilises a fraction of the sample volume compared with multiple ELISAs.

A keloid (KD) is a benign dermal fibrotic tumor. Treatment of KDs is challenging and the recurrence rate is high; thus, there is an unmet need to explore new target sites and new treatment methods. As a tumor-associated cytokine, autocrine motility factor (AMF) can effectively stimulate the random and directional movement of cells. We first found that AMF was overexpressed in keloid fibroblasts (KFs) and the proliferation and migration of KFs were promoted by AMF stimulation. After treatment with Y-27632, RhoA kinase inhibitor, the proliferation and migration capacity of KFs declined significantly, and type I collagen protein, active RhoA and ROCK1 also were downregulated. In addition, a KD transplantation model was established under the skin of nude mice, with KD intramural injection AMF siRNA, we found that the weight of the KD was smaller than in the control group (P < 0.05), KD tissue sections stained by HE and Masson showed that fibers became loose and the blood vessels were visibly reduced. In conclusion, AMF siRNA is expected to be a novel strategy to treat KD by inhibiting signaling pathway of RhoA/ROCK1.

Riverbank filtration (RBF) is used worldwide to produce high quality drinking water. With river water often contaminated by micropollutants (MPs) from various sources, this study addresses the occurrence and fate of such MPs at three different RBF sites with oxic alluvial sediments and short travel times to the drinking water well down to hours. A broad range of MPs with various physico-chemical properties were analysed with detection limits in the low ng L-1 range using solid phase extraction followed by liquid chromatography coupled to tandem high resolution mass spectrometry. Out of the 526 MPs targeted, a total of 123 different MPs were detected above the limit of quantification at the three different RBF sites. Of the 75-96 MPs detected in each river, 43-59% were attenuated during RBF. The remaining total concentrations of the MPs in the raw drinking water accounted to 0.6-1.6 μgL-1 with only a few compounds exceeding 0.1 μgL-1, an often used threshold value. The attenuation was most pronounced in the first meters of infiltration with a full elimination of 17 compounds at all three sites. However, a mixing with groundwater related to regional groundwater flow complicated the characterisation of natural attenuation potentials along the transects. Additional non-target screening at one site revealed similar trends for further non-target components. Overall, a risk assessment of the target and estimated non-target compound concentrations finally indicated during the sampling period no health risk of the drinking water according to current guidelines. Our results demonstrate that monitoring of contamination sources within a catchment and the affected water quality remains important in such vulnerable systems with partially short residence times.

A protocol for determining the characteristic temperatures of thermomechanical transitions on gluten-free flour doughs is proposed. This protocol is based on the mathematical analysis of experimental curve of storage modulus (G') vs temperature obtained by means of Dynamic Mechanical Thermal Analysis (DMTA) technique. Doughs at constant consistency of chestnut flour with different levels (3, 6 and 9% flour basis, f.b.) of brown seaweed (Bifurcaria bifurcata, Fucus vesiculosus and Ascophyllum nodosum) powders addition, 2% f.b. of guar gum and 1.8% f.b. of salt with different water absorption were used to test the proposed protocol. The ranges of temperatures corresponding to starch gelatinization (59-97 °C), amylopectin crystallites melting (82-101 °C), reversible dissociation of lipid-amylose complexes (107-128 °C) and amylose melting (133-171 °C) showed a strong dependence with water absorption of samples. Doughs with the same water absorption submitted to starch gelatinization during mixing were also analysed to corroborate the protocol suitability. Total polyphenols content and radical scavenging activity of extracts from chestnut flour-seaweed powder blends and seaweed-enriched chestnut cookies baked at 180 °C were determined. Extraction assisted with ultrasounds was carried out employing acetone-water (70:30 v/v) solution as solvent during 4 min with a liquid/solid ratio of 30 w/w. Seaweed powder addition had a positive effect on antioxidant properties of doughs before baking. However, the seaweed powder addition effect on baked products (cookies) is not clear due to antioxidant activity is overlapped by Maillard's products generated during baking.

Flow cytomtery (FCM) has become a standard approach to enumerate viruses in water research. However, the nature of the fluorescent signal in flow cytometric analysis of water samples and the mechanism of its formation, have not been addressed for bacteriophages expected in wastewaters. Here we assess the behaviour of fluorescent DNA-staining dyes in aqueous solutions, as well as sensitivity and accuracy of FCM for enumeration of DNA-stained model bacteriophages λ, P1, and T4. We demonstrate that in aqueous systems fluorescent dyes form a self-stabilized (pseudolyophilic) emulsion of auto-fluorescing colloid particles. Sample shaking and addition of surfactants enhance auto-fluorescence due to increased dispersion and, in the presence of surfactants, stabilization of the dye emulsion. Bacteriophages with genome sizes <100 kbp (i.e. λ & P1) did not generate a distinct population signal to be detected by one of the most sensitive FCM instruments available (BD LSR Fortessa™ X-20), whereas the larger T4 bacteriophage was resolved as a distinct population of events. These results indicate that the use of fluorescent dyes for bacteriophage enumeration by flow cytometry can produce false positive signals and lead to wrong estimation of total virus counts by misreporting colloid particles as virions, depending on instrument sensitivity.

The encapsulation of Ib-M6 antibacterial peptide in pellets of polyvinyl alcohol (PVA) and polyvinyl alcohol-alginate (PVA-Alg) matrices was carried out in order to explore its controlled release and activity against Escherichia coli K-12. The pellets were obtained by combined ice segregation induced self-assembly (ISISA) and freezing-thawing methods and their microstructure was studied by scanning electron microscopy. Bromothymol blue was used as a model compound to study the transport mechanisms and release from pellets. The results show that there is a significant effect of the total concentration of PVA precursor solutions, the mass ratio of PVA of different molecular weights and the addition of alginate on the microstructure and transport properties of pellets. The antibacterial activity of Ib-M6 against Escherichia coli K-12 was not affected by the encapsulation in PVA pellets. However, the release of Ib-M6 from PVA-Alg pellets was not possible, probably due to the electrostatic interaction of positively charged Ib-M6 and negatively alginate structure. Nonetheless, the controlled release of Ib-M6 from polymeric matrices can be fitting by modifying parameters such as the concentration and type of polymer precursors.

If touch is perceived as pleasant, it can counteract the experience of pain. However, its pain-inhibitory function might be disturbed in chronic pain and this could contribute to pain-related interference. We investigated the perception of pleasant touch and its brain correlates in chronic back pain patients (CBP) compared to subacute back pain patients (SABP) and healthy controls (HC) using soft brush strokes. CBP showed less positive evaluations of touch. We found the highest activation in somatosensory and insular cortices in CBP, ventral striatum (VS) in SABP, and the orbitofrontal cortex in HC. Brain responses were significantly positively correlated with pleasantness ratings in HC and SABP, but not CBP. Further, the insula responses in CBP were positively correlated with pain-related interference and the VS activation in SABP correlated negatively with affective distress. Brain and behavioral changes in the processing of touch and its pleasantness may be a marker of pain chronicity and raise questions about the therapeutic value of pleasant touch in pain prevention and treatment.

Concurrent with rising production of carbon-based engineered nanomaterials is a potential increase in respiratory and cardiovascular diseases due to exposure to nanomaterials in the workplace atmosphere. While single-cell models of pulmonary exposure are often used to determine the potential toxicity of nanomaterials in vitro, previous studies have shown that coculture cell models better represent the cellular response and crosstalk that occurs in vivo. This study identified differential gene regulation in human small airway epithelial cells (SAECs) grown either in monoculture or in coculture with human microvascular endothelial cells following exposure of the SAECs to multiwalled carbon nanotubes (MWCNTs). SAEC genes that either changed their regulation direction from upregulated in monoculture to downregulated in coculture (or vice versa) or had a more than a two-fold changed in the same regulation direction were identified. Genes that changed regulation direction were most often involved in the processes of cellular growth and proliferation and cellular immune response and inflammation. Genes that had a more than a two-fold change in regulation in the same direction were most often involved in the inflammatory response. The direction and fold-change of this differential gene regulation suggests that toxicity testing in monoculture may exaggerate cellular responses to MWCNTs, and coculture of cells may provide a more in-depth assessment of toxicological responses.

Small extracellular vesicles (EVs) are 50-200 nm sized mediators in intercellular communication that reflect both physiological and pathophysiological changes of their parental cells. Thus, EVs hold great potential for biomarker detection. However, reliable purification methods for the downstream screening of the microRNA (miRNA) cargo carried within urinary EVs by small RNA sequencing have yet to be established. To address this knowledge gap, RNA extracted from human urinary EVs obtained by five different urinary EV purification methods (spin column chromatography, immunoaffinity, membrane affinity, precipitation and ultracentrifugation combined with density gradient) was analyzed by small RNA sequencing. Urinary EVs were further characterized by nanoparticle tracking analysis, Western blot analysis and transmission electron microscopy. Comprehensive EV characterization established significant method-dependent differences in size and concentration as well as variances in protein composition of isolated vesicles. Even though all purification methods captured enough total RNA to allow small RNA sequencing, method-dependent differences were also observed with respect to library sizes, mapping distributions, number of miRNA reads and diversity of transcripts. Whereas EVs obtained by immunoaffinity yielded the purest subset of small EVs, highly comparable with results attained by ultracentrifugation combined with density gradient, precipitation and membrane affinity, sample purification by spin column chromatography indicated a tendency to isolate different subtypes of small EVs, which might also carry a distinct subset of miRNAs. Based on our results, different EV purification methods seem to preferentially isolate different subtypes of EVs with varying efficiencies. As a consequence, sequencing experiments and resulting miRNA profiles were also affected. Hence, the selection of a specific EV isolation method has to satisfy the respective research question and should be well considered. In strict adherence with the MISEV (minimal information for studies of extracellular vesicles) guidelines, the importance of a combined evaluation of biophysical and proteomic EV characteristics alongside transcriptomic results was clearly demonstrated in this present study.