Identification of carboxymethyl (CM)-binding proteins derived from Lolium multiflorum pollen extract and antibody reactivity in Brazilian allergic patients.

Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

MR-guided HDR prostate brachytherapy with teleoperated steerable needles.

Conformity of tumour volumes and dose plans in prostate brachytherapy (BT) can be constrained by unwanted needle deflections, needle access restrictions and visualisation limitations. This work validates the feasibility of teleoperated robotic control of an active steerable needle using magnetic resonance (MR) for guidance. With this system, perturbations can be counteracted and critical structures can be circumvented to access currently inaccessible areas. The system comprises of (1) a novel steerable needle, (2) the minimally invasive robotics in an MR environment (MIRIAM) system, and (3) the daVinci Research Kit (dVRK). MR scans provide visual feedback to the operator controlling the dVRK. Needle steering is performed along curved trajectories to avoid the urethra towards targets (representing tumour tissue) in a prostate phantom with a targeting error of 1.2 ± 1.0 mm. This work shows the potential clinical applicability of active needle steering for prostate BT with a teleoperated robotic system in an MR environment.

Diet quality, asthma and airway inflammation in school-aged children.

Summary: Background. Asthma is a major public health problem, with increasing prevalence in most countries, particularly among children. Poor dietary quality is also increasing in children, and evidence of the overall quality of children's food patterns effects on asthma is scarce. Methods. This cross-sectional analysis (660 children: 49.1% females, aged 7-12years) evaluated the association between diet quality and asthma (n = 56) and airway inflammation among school-aged children according to body mass index (BMI). Diet quality was assessed through the Healthy Eating Index (HEI)-2015, and categorized by tertiles. Higher scores represent a healthier diet. A questionnaire was used to enquire about self-reported medical diagnosis of asthma and asthma under medication. Lung function and airway reversibility were measured, and airway inflammation assessed measuring exhaled fractional nitric oxide (eNO). Two categories of BMI were considered: non-overweight/obese (p less than 85th), (n = 491), and overweight/obese (p ≥ than 85th), (n = 169). The associations between diet quality and asthma and airway inflammation were estimated using logistic regression models. Results. Non-overweight/obese children in 2nd tertile of HEI-2015 score had decreased odds of having eNO ≥ 35ppb (OR 0.43, 95%CI 0.19-0.98), medical diagnosis of asthma (OR 0.18; 95%CI 0.04-0.84), and asthma treatment (OR 0.12; 95%CI 0.01-0.95), compared to children in the 1st tertile. Conclusions. Our findings suggest that a higher diet quality associates with lower levels of airway inflammation and reduced prevalence of asthma among non-overweight/obese school-aged children.

Identification of carboxymethyl (CM)-binding proteins derived from Lolium multiflorum pollen extract and antibody reactivity in Brazilian allergic patients

Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

Exosomal glypican-1 discriminates pancreatic ductal adenocarcinoma from chronic pancreatitis.

BACKGROUND AND AIMS: Pancreatic ductal adenocarcinoma (PDAC) diagnosis can be difficult in a chronic pancreatitis (CP) background, especially in its mass forming presentation. We aimed to assess the accuracy of glypican-1-positive circulating exosomes (GPC1+crExos) to distinguish PDAC from CP versus the state-of-the-art CA 19-9 biomarker. METHODS: This was a unicentric prospective cohort. Endoscopic ultrasound with fine-needle aspiration or biopsy and blood tests (GPC1+crExos and serum CA 19-9) were performed. RESULTS: The cohort comprised 60 PDAC and 29 CP (7 of which mass forming - MF) patients. Median levels of GPC1+crExos were significantly higher in PDAC (99.7%) versus CP (28.4%; p<0.0001) with an AUROC of 0.96 with 98.3% sensitivity and 86.2% specificity for a cut-off of 45.0% (p<0.0001); this outperforms CA 19-9 AUROC of 0.82 with 78.3% sensitivity and 65.5% specificity at a cut-off of 37 U/mL (p<0.0001). The superiority of% GPC1+crExos over CA 19-99 in differentiating PDAC from CP was observed in both early (stage I) and advanced tumors (stages II-IV). CONCLUSION: Levels of GPC1+crExos coupled to beads enable differential diagnosis between PDAC and CP including its mass-forming presentation.

Valuation of angiogenesis in bovine xenografts implanted in intracorporal sites of rabbits as models of in-vivo bioreactors / Avaliação da angiogênese em xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos como modelos de biorreatores in vivo

The aim of this study was to evaluate neovascularization of bovine xenografts implanted in intracorporeal sites of rabbits (bioreactors). 30 rabbits were used, divided into 6 groups, according to the evaluation time (7, 15, 30, 45, and 60 days); each animal received xenogenic implants in 3 different intracorporeal sites (A1 - omentum bag; A2 - intermuscular space of quadriceps femoris; A3 - subperiosteal of ilium bone). Histological assessments graded the presence of angiogenesis, the number of inflammatory cells, newly formed bone tissue, and the presence of giant cells. Histological analyses showed intense angiogenesis in all implanted xenografts. Presence of inflammatory infiltrate and giant cells at the A1 implant site and presence of bone neoformation at the A3 implant site were noted. Degeneration of implants and formation of a fibrous capsule were noted. When comparing the interaction of the site with the days of evaluation, statistical analysis showed a significant difference (p≤0.05) in any time of neovascularization analysis. The vascular endothelial growth factor (VEGF) and inflammatory cells of the omentum in its structure, may have contributed to the greater presence of neovessels and inflammatory cells, a fact that may indicate functionality as a possible bone substitute.(AU)

O objetivo deste estudo foi avaliar a neovascularização de xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos (biorreatores). Foram utilizados 30 coelhos, os quais foram divididos em seis grupos, de acordo com o tempo de avaliação (sete, 15, 30, 45 e 60 dias); cada animal recebeu implantes xenogênicos em três diferentes sítios intracorpóreos (A1 - bolsa de omento; A2 - espaço intermuscular do quadríceps femoral; A3 - subperiosteal do osso ílio). Avaliações histológicas classificaram a presença de angiogênese, o número de células inflamatórias, de tecido ósseo neoformado e a presença de células gigantes. As análises histológicas mostraram intensa angiogênese em todos os xenoenxertos implantados. Observou-se presença de infiltrado inflamatório e células gigantes no local do implante A1 e presença de neoformação óssea no local do implante A3. Ao mesmo tempo, a degeneração dos implantes e a formação de uma cápsula fibrosa foram observadas. Ao comparar a interação do local com os dias de avaliação, a análise estatística mostrou diferença significativa (P≤0,05) em qualquer momento da análise de neovascularização. O fator de crescimento endotelial vascular (VEGF) e as células inflamatórias do omento em sua estrutura podem ter contribuído para a maior presença de neovasos e células inflamatórias, fato que pode indicar funcionalidade como possível substituto ósseo.(AU)

Subjective Wellbeing in Gypsy Students

Gypsy often experience substandard social conditions, poorer health, and discrimination. However, little is known about Gypsies students’ subjective wellbeing. This study aimed to compare Gypsy (n= 42) and non-Gypsy (n= 76) young-students (mean age= 13 years) in terms of composite Subjective Wellbeing (combining indicators of satisfaction with life, positive and negative affect, satisfaction with social support, and quality of life). A series of t-tests indicated that Gypsy and non-Gypsy students did not differ significantly in Subjective Wellbeing. A novelty and strength of the study were that we complemented mean difference tests with procedures for describing group similarities. Understanding similarity is important because it can lead to more accurate and positive perceptions of outgroups, including Gypsies. Across multiple dimensions of Subjective Wellbeing, the similarity between Gypsy and non-Gypsy students was greater than 92%. Although students gypsies experience several cultural specificities, they seem to register levels of composite wellbeing similar to non-gypsy peers. These results are relevant for inclusion practices and policies, as they suggest that research in quality of life is an important means to promote inclusion and equity and to reduce prejudice against ethnic minoritie (AU)

High fidelity defines the temporal consistency of host-parasite interactions in a tropical coastal ecosystem.

Host-parasite interactions represent a selective force that may reduce hosts' lifespan, their reproductive success and survival. Environmental conditions can affect host-parasite communities, leading to distinct patterns of interactions with divergent ecological and evolutionary consequences for their persistence. Here, we tested whether climatic oscillation shapes the temporal dynamics of bird-haemosporidian associations, assessing the main mechanisms involved in the temporal dissimilarity of their interactions' networks. For two years, we monthly sampled birds in a tropical coastal ecosystem to avian malaria molecular diagnosis. The studied networks exhibited high specialization, medium modularity, with low niche overlap among parasites lineages. Moreover, alpha and ß-diversity of hosts, parasites and their interactions, as well as the structure of their networks were temporally consistent, i.e., stable under fluctuations in temperature or precipitation over seasons. The structure and temporal consistency of the studied antagonistic networks suggest a high fidelity between partners, which is likely relevant for their evolutionary persistence.

Mitochondria, endoplasmic reticulum and innate immune dysfunction in mood disorders: Do Mitochondria-Associated Membranes (MAMs) play a role?

Mood disorders like major depression and bipolar disorder (BD) are among the most prevalent forms of mental illness. Current knowledge of the neurobiology and pathophysiology of these disorders is still modest and clear biological markers are still missing. Thus, a better understanding of the underlying pathophysiological mechanisms to identify potential therapeutic targets is a prerequisite for the design of new drugs as well as to develop biomarkers that help in a more accurate and earlier diagnosis. Multiple pieces of evidence including genetic and neuro-imaging studies suggest that mood disorders are associated with abnormalities in endoplasmic-reticulum (ER)-related stress responses, mitochondrial function and calcium signalling. Furthermore, deregulation of the innate immune response has been described in patients diagnosed with mood disorders, including depression and BD. These disease-related events are associated with functions localized to a subdomain of the ER, known as Mitochondria-Associated Membranes (MAMs), which are lipid rafts-like domains that connect mitochondria and ER, both physically and biochemically. This review will outline the current understanding of the role of mitochondria and ER dysfunction under pathological brain conditions, particularly in major depressive disorder (MDD) and BD, that support the hypothesis that MAMs can act in these mood disorders as the link connecting ER-related stress response and mitochondrial impairment, as well as a mechanisms behind sterile inflammation arising from deregulation of innate immune responses. The role of MAMs in the pathophysiology of these pathologies and its potential relevance as a potential therapeutic target will be discussed.

Adherence to a Mediterranean Dietary Pattern and Functional Parameters: A Cross-Sectional Study in an Older Population.

OBJECTIVES: In older adults, diet and physical activity are among the most important behaviours that influence health. The Mediterranean Dietary Pattern has been related to longevity and can benefit physical function in older adults. The present study aims to quantify the association of adherence to a Mediterranean Dietary Pattern with physical functioning through by handgrip strength, as well as, with sedentary behaviour based on sitting time, in a sample of older adults. METHODS: A cross-sectional study was conducted in 1,491 older adults ≥ 65 years old. The adherence or non-adherence to a Mediterranean Dietary Pattern was assessed using the previously validated and translated 14 item questionnaire, the PREDIMED (Prevención com Dieta Mediterránea). Handgrip strength was measured with a Jamar Dynamometer. Sitting time per day was used as an indicator of sedentary behaviour. Multinomial logistic regressions were performed to quantify the association of adherence to a Mediterranean Dietary Pattern with handgrip strength and sitting time. RESULTS: Compared to participants who adhered to a Mediterranean Dietary Pattern, those who did not adhere to this diet have a significantly higher adjusted odds ratio for low handgrip strength [adjusted odds ratio: 1.50; 95% confidence interval: 1.09-2.05], as well as for longer sitting time [adjusted odds ratio: 1.43; 95% confidence interval: 1.04-1.96]. CONCLUSION: In older adults, the non-adherence to a Mediterranean Dietary Pattern was associated with both lower values of handgrip strength and longer sitting time.

Are hypohydrated older adults at increased risk of exhaustion?

BACKGROUND: Dehydration appears to affect muscle strength and weakness, although its influence on exhaustion remains unclear. The present study aimed to quantify the association between hydration status and exhaustion among older adults. METHODS: A cluster sampling approach was used, representing Portuguese older adults (≥65 years) according to age, sex, education level and region within the Nutrition UP65 cross-sectional study. A 24-h urine sample was collected to estimate free water reserve (FWR), which was categorised into tertiles according to sex. Subjects with incomplete 24-h urine and renal disease were excluded. From a sample size of 1500 subjects, 1143 were eligible. Exhaustion was self-reported according to the Center for Epidemiologic Studies Depression Scale. A logistic regression model was conducted to evaluate the association between FWR and exhaustion. Odds ratios and the respective 95% confidence intervals were calculated by sex and age. RESULTS: Free water reserve median (interquartile range) was 0.52 (0.68) L in women and 0.36 (0.77) L in men. Hypohydration affected 11.6% of women and 25.1% of men, whereas exhaustion was reported by 39.3% of women and 25.1% of men. After adjusting for confounders, women ≥80 years classified in the highest tertile of FWR showed a decreased risk of exhaustion (third tertile: odds ratio = 0.38; 95% confidence interval = 0.15-0.96) compared to women in the lowest FWR tertile. No such significant association was observed in women with <80 years and in men. CONCLUSIONS: These results show an association between worse hydration status and exhaustion in older women, highlighting the need to implement further studies clarifying this association.

Effects of indoor endocrine-disrupting chemicals on childhood rhinitis

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Implementation of an in vitro methodology for phototoxicity evaluation in a human keratinocyte cell line.

To assess photoxicity, several in vitro methods using different cellular models have been developed for preclinical testing. Over prediction of the in vivo photosafety hazard has been however appointed. Herein, we describe the implementation and validation of an in vitro methodology for phototoxicity evaluation based on the 3T3 neutral red uptake phototoxicity test using the HaCaT human keratinocyte cell line, and UVA/UVB radiation. Known positive (5-methoxypsoralen, chlorpromazine, and quinine) and negative (acetyl salicylic acid, hexachlorophene, and sodium lauryl sulphate) controls were tested together with a set of chemical currently used in cosmetic/pharmaceutical formulations. Apart from the advantage of using a cell line of human origin, these cells were generally more resistant to the cytotoxic effects of the test substances relative to the 3T3 mouse fibroblasts when exposed to an UVA irradiation dose of 1.7 mW/cm2. Therefore, this HaCaT NRU assay provides a more realistic experimental model that overcomes the over/high sensitivity frequently noted with the 3T3 NRU assay and that is more consistent with the human in vivo situation. Using a more representative method can prevent time-consuming and expensive in vivo testing in both animal models and humans that can significantly delay the clinical development of new chemicals.

Fruit, vegetable consumption and blood pressure in healthy adolescents: A longitudinal analysis from the LabMed study.

BACKGROUND AND AIMS: The associations between fruit and vegetable consumption and high blood pressure among adults are well studied. Nonetheless, data on the influence of a low consumption of fruit and vegetables on cardiovascular disease risk, particularly blood pressure, among healthy adolescents are scarce. Therefore, we aim to analyse the associations between fruit and/or vegetable intake and blood pressure over a two-year period in healthy adolescents. METHODS AND RESULTS: As part of a cohort, 606 adolescents from the LabMed Physical Activity study were evaluated in 2011 (baseline) and 2013 (follow-up). Blood pressure was measured according to standardized procedures and fruit and vegetable consumption was assessed with a food frequency questionnaire. Anthropometric variables, socioeconomic status, pubertal stage and lifestyle determinants were gathered and used as confounders. Prospective associations between fruit and/or vegetable intake and blood pressure were examined using generalized linear models. Girls who consumed more fruit at baseline had a significant decrease in diastolic blood pressure at follow-up [unstandardized beta: -0.005 mmHg (95%CI: -0.01; -0.0002) (p = 0.038)]. CONCLUSION: In apparently healthy adolescents, fruit intake may already start to have an effect in blood pressure. Girls who consumed more fruit exhibited lower levels of diastolic blood pressure.

Vitamin D status and functional parameters: A cross-sectional study in an older population.

The present study aims to clarify the association of vitamin D status with functionality, measured through gait speed and hand grip strength, in a sample of older adults, considering sex-specific characteristics and the potential confounding effect of lifestyle factors, disease, skin phenotype, season of blood collection and vitamin D supplementation. The Nutrition UP 65 cross-sectional study was conducted in a population-based sample of 1,425 older adults ≥ 65 years old. Serum levels of 25-hydroxyvitamin D were quantified through electrochemiluminescence immunoassay. Multinomial logistic regressions were carried out using quartiles of gait speed and of hand grip strength as dependent variables. Participants at risk of vitamin D inadequacy (30.0-49.9 nmol/L) and deficiency (< 30.0 nmol/L) presented higher adjusted odds ratio of the lowest values of gait speed and hand grip strength than those with adequate vitamin D levels (≥ 50.0 nmol/L). These associations were strongest among men at risk of vitamin D deficiency [adjusted odds ratio for the lowest quartile of gait speed = 3.24; 95% CI: 1.56-6.73 and for the lowest quartile of hand grip strength = 3.28; 95% CI: 1.47-7.31] than in women at risk of vitamin D deficiency [adjusted odds ratio for the lowest quartile of gait speed = 2.72; 95% CI: 1.37-5.41 and for the lowest quartile of hand grip strength = 1.56; 95% CI: 0.81-3.00]. In conclusion, in older adults, particularly in men, the risk of vitamin D deficiency was directly associated with the lowest values of gait speed and of hand grip strength. However, randomized controlled trials are needed to overcome the possibility of reverse causation and residual confounding. Present results emphasise the need for strategies to promote the reduction of the high prevalence of low vitamin D status among the Portuguese older adult population.

Emotional distress in haemophilia: Factors associated with the presence of anxiety and depression symptoms among adults.

INTRODUCTION: Haemophilia is related to several clinical and psychosocial challenges that have been associated with increased emotional distress. These may impact on disease adjustment and health outcomes, reinforcing the attention given to psychosocial health of people with haemophilia (PWH), in the scope of optimal comprehensive care. AIM: To identify potentially modifiable factors associated with the presence of anxiety and depression symptoms among adult PWH. METHODS: This was a cross-sectional observational study examining socio-demographic, clinical and psychosocial variables among 102 patients with haemophilia A or B who participated on a mail survey of haemophilia in Portugal. RESULTS: People with haemophilia revealing higher anxiety and depression symptoms were more likely to have had, in the previous year, more urgent hospital visits due to haemophilia, more bleeding episodes, more affected joints and pain, as well as worst levels of perceived functionality and quality of life. After controlling for demographic (age and education) and clinical (haemophilia severity and joint deterioration) variables in multivariate hierarchical logistic regression analyses, professional status (OR = 4.646, P = .004; OR = 3.333, P = .029) and pain interference (OR = 1.397, P = .011; OR = .1.347, P = .037) were significantly associated with both anxiety and depression symptoms. Additionally, physical activity (OR = 0.302, P = .024) and the perception of consequences underlying haemophilia (OR = 1.600, P = .012) also emerged as key factors significantly associated with depression symptoms. CONCLUSION: Current findings increased knowledge on factors associated with anxiety and depression among PWH. These highlight potential intervention targets, which are amenable to change through evidence-based tailored interventions aiming to decrease emotional distress, promote well-being and improving haemophilia-related health outcomes among these patients.

Brain insulin signalling, glucose metabolism and females' reproductive aging: A dangerous triad in Alzheimer's disease.

Alzheimer's disease (AD) constitutes a major socioeconomic challenge due to its disabling features and the rise in prevalence (especially among (peri)menopausal women and type 2 diabetes patients). The precise etiopathogenesis of AD remains poorly understood. Importantly, its neurodegenerative perspective has been challenged towards a more "systemic" view. Amyloid-ß (Aß) and hyperphosphorylated Tau protein (P-Tau) (the main AD neuropathological features) affect and are affected by peripheral and brain insulin signalling dysfunction, leading to glucose dysmetabolism, synaptic loss and AD-related cognitive deficits. This may be anticipated and exacerbated by the progressive loss of estrogen (and interactions, e.g., with insulin) during females' aging, increasing their risk for AD, especially during menopause. Under this perspective, we aimed to discuss the recent findings (and controversies) behind the peripheral view of AD, and the role for insulin deficits and brain glucose dysmetabolism in such diseased brain. We also focused on the metabolic shift and the putative effects of gender (especially during midlife/perimenopause) herein. We finally discussed AD as the potential "type 3 diabetes", and the therapeutic potential of restoring brain insulin levels or glucose energy metabolism via administration of intranasal insulin and use of ketogenic diets. In sum, AD appears to lie on an intricate crosstalk between age-related metabolic, hormonal and specific genetic changes that challenge its traditional view. Hence, clarification of AD risk factors (besides aging and gender) and pathophysiological mechanisms will allow to establish accurate preventive strategies, biomarkers and more efficient drugs - all urgent medical needs in our increasingly aged societies. This article is part of the Special Issue entitled 'Metabolic Impairment as Risk Factors for Neurodegenerative Disorders.'