Creating functional nanostructures: Encapsulation of caffeine into α-lactalbumin nanotubes

This work evaluated the stability and functionality of nanotubes obtained from α-lactalbumin (α-LA). α-LA nanotubes' structure was highly stable during a freeze-drying process but not after grinding. The ability of α-LA nanotubes to encapsulate caffeine, used as a model molecule, was evaluated. α-La nanotubes were highly effective for this purpose as encapsulation efficiency (%EE) was near 100% and loading capacity (%LC) near 10% at 1.5/20 and 2/20 ratios (caffeine/α-LA, w/w). α-LA nanotubes' structure was not affected by the presence of caffeine. Also, in general, refrigeration temperatures and neutral or alkaline conditions, under which the adverse effect of chelating agents was prevented, helped to stabilise α-LA nanotubes' structure and maintain caffeine encapsulated. At 8 °C and pH 7.5, in the presence of 75 μg mL− 1 of EDTA, > 50% of the caffeine remained encapsulated into α-LA nanotubes.Clara Fuciños gratefully acknowledge her Post-Doctoral grant (I2C 2014) from Consellería de Cultura, Educación e Ordenación Universitaria (Xunta de Galicia, Spain). Pablo Fuciños gratefully acknowledges his Marie Curie COFUND Postdoctoral Research Fellowship (Project No: 600375. NanoTRAINforGrowth - INL Fellowship programme in nanotechnologies for biomedical, environment and food applications). This study was supported by the Fundação para a Ciência e a Tecnologia (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462)

Cardiovascular events in Systemic Lupus Erythematosus: a nationwide study in Spain from the RELESSER Registry

This article estimates the frequency of cardiovascular (CV) events that occurred after diagnosis in a large Spanish cohort of patients with systemic lupus erythematosus (SLE) and investigates the main risk factors for atherosclerosis. RELESSER is a nationwide multicenter, hospital-based registry of SLE patients. This is a cross-sectional study. Demographic and clinical variables, the presence of traditional risk factors, and CV events were collected. A CV event was defined as a myocardial infarction, angina, stroke, and/or peripheral artery disease. Multiple logistic regression analysis was performed to investigate the possible risk factors for atherosclerosis. From 2011 to 2012, 3658 SLE patients were enrolled. Of these, 374 (10.9%) patients suffered at least a CV event. In 269 (7.4%) patients, the CV events occurred after SLE diagnosis (86.2% women, median [interquartile range] age 54.9 years [43.2-66.1], and SLE duration of 212.0 months [120.8-289.0]). Strokes (5.7%) were the most frequent CV event, followed by ischemic heart disease (3.8%) and peripheral artery disease (2.2%). Multivariate analysis identified age (odds ratio [95% confidence interval], 1.03 [1.02-1.04]), hypertension (1.71 [1.20-2.44]), smoking (1.48 [1.06-2.07]), diabetes (2.2 [1.32-3.74]), dyslipidemia (2.18 [1.54-3.09]), neurolupus (2.42 [1.56-3.75]), valvulopathy (2.44 [1.34-4.26]), serositis (1.54 [1.09-2.18]), antiphospholipid antibodies (1.57 [1.13-2.17]), low complement (1.81 [1.12-2.93]), and azathioprine (1.47 [1.04-2.07]) as risk factors for CV events. We have confirmed that SLE patients suffer a high prevalence of premature CV disease. Both traditional and nontraditional risk factors contribute to this higher prevalence. Although it needs to be verified with future studies, our study also shows-for the first time-an association between diabetes and CV events in SLE patients

Vigilancia virológica de Aedes (Stegomyia) aegypti y Aedes (Stegomyia) albopictus como apoyo para la adopción de decisiones en el control del dengue en Medellín

Introduction: Dengue represents an important public health problem in Colombia. No treatment is available and the vaccine has not been approved in all countries, hence, actions should be strengthened to mitigate its impact through the control of Aedes aegypti, the vector mosquito. In Colombia, surveillance is done using entomological indexes and case notification, which is usually informed late, leading to untimely interventions. Viral detection in urban mosquitoes using molecular techniques provides more accurate entomological information for decision-making.Objective: To report results of virological surveillance in Aedes specimens collected during routine entomological activities of the Secretaría de Salud de Medellín.Materials and methods: Specimens were collected during two periods in each of which we selected 18 dwellings around each one of the 250 larva traps arranged for mosquitoe surveillance, as well as 70 educational institutions and 30 health centers. Specimens were identified morphologically, and divided in pools for viral detection using reverse transcription polymerase chain reaction (RT-PCR). We calculated the minimum infection rate and the adult infestation index for each group.Results: We collected 1,507 adult mosquitoes, 10 of which were identified as A. albopictus. Out of the 407 pools, 132 (one of them Ae. albopictus) were positive, and 14.39% were A. aegypti males. The minimum infection rates for Ae. aegypti were 120.07 and 69,50 for the first and second periods, respectively, and the adult infestation index was higher in educational institutions (23.57%).Conclusions: Using RT-PCR we identified natural infectivity and vertical transmission of dengue virus in A. aegypti and A. albopictus. We suggest the use of molecular techniques in arbovirosis surveillance and control programs in Colombia.Introducción. El dengue en Colombia representa un grave problema de salud y, dado que no existe un tratamiento efectivo para la enfermedad y la vacuna no se ha aprobado en todos los países, se deben fortalecer acciones para mitigar su impacto mediante el control de Aedes aegypti, el mosquito vector. La vigilancia en el país se hace con base en los índices entomológicos y en la notificación de casos, la cual es frecuentemente tardía y por ello conduce a falta de oportunidad en las intervenciones. La detección viral en mosquitos urbanos mediante técnicas moleculares proporciona información entomológica más precisa para la adopción de decisiones.Objetivo. Reportar los resultados de la vigilancia virológica de especímenes de Aedes spp. recolectados durante actividades entomológicas rutinarias de la Secretaría de Salud de Medellín.Materiales y métodos. Los ejemplares se recolectaron durante dos periodos, en cada uno de los cuales se seleccionaron 18 viviendas alrededor de cada una de las 250 trampas para larvas dispuestas para la vigilancia entomológica, así como 70 instituciones educativas y 30 centros de salud. Los ejemplares se identificaron y se conformaron grupos para la detección viral mediante reacción en cadena de la polimerasa con transcripción inversa (Reverse Transcription Polymerase Chain Reaction, RT-PCR). Se calculó la tasa mínima de infección y el índice de infestación en adultos.Resultados. Se recolectaron 1.507 mosquitos, 10 de los cuales eran Ae. albopictus. De los 407 grupos conformados, 132 (uno de ellos de Ae. albopictus) fueron positivos, y 14,39 % correspondió a machos de Ae. aegypti. La tasa mínima de infección para Ae. aegypti fue de 120,07 y 69,50 en el primer y segundo períodos, respectivamente, y el índice de infestación en adultos fue mayor en las instituciones educativas (23,57 %).Conclusión. Mediante la RT-PCR se detectaron la infección natural y la transmisión vertical del virus del dengue en Ae. aegypti y en Ae. albopictus. Se propone considerar la incorporación de estas técnicas moleculares en los programas de vigilancia y control de las arbovirosis en el país

Incidence, associated factors and clinical impact of severe infections in a large, multicentric cohort of patients with systemic lupus erythematosus

OBJECTIVES: To estimate the incidence of severe infection and investigate the associated factors and clinical impact in a large systemic lupus erythematosus (SLE) retrospective cohort. METHODS: All patients in the Spanish Rheumatology Society Lupus Registry (RELESSER) who meet ?4 ACR-97 SLE criteria were retrospectively investigated for severe infections. Patients with and without infections were compared in terms of SLE severity, damage, comorbidities, and demographic characteristics. A multivariable Cox regression model was built to calculate hazard ratios (HRs) for the first infection. RESULTS: A total of 3658 SLE patients were included: 90% female, median age 32.9 years (DQ 9.7), and mean follow-up (months) 120.2 (±87.6). A total of 705 (19.3%) patients suffered ?1 severe infection. Total severe infections recorded in these patients numbered 1227. The incidence rate was 29.2 (95% CI: 27.6-30.9) infections per 1000 patient years. Time from first infection to second infection was significantly shorter than time from diagnosis to first infection (p < 0.000). Although respiratory infections were the most common (35.5%), bloodstream infections were the most frequent cause of mortality by infection (42.0%). In the Cox regression analysis, the following were all associated with infection: age at diagnosis (HR = 1.016, 95% CI: 1.009-1.023), Latin-American (Amerindian-Mestizo) ethnicity (HR = 2.151, 95% CI: 1.539-3.005), corticosteroids (?10mg/day) (HR = 1.271, 95% CI: 1.034-1.561), immunosuppressors (HR = 1.348, 95% CI: 1.079-1.684), hospitalization by SLE (HR = 2.567, 95% CI: 1.905-3.459), Katz severity index (HR = 1.160, 95% CI: 1.105-1.217), SLICC/ACR damage index (HR = 1.069, 95% CI: 1.031-1.108), and smoking (HR = 1.332, 95% CI: 1.121-1.583). Duration of antimalarial use (months) proved protective (HR = 0.998, 95% CI: 0.997-0.999). CONCLUSIONS: Severe infection constitutes a predictor of poor prognosis in SLE patients, is more common in Latin-Americans and is associated with age, previous infection, and smoking. Antimalarials exerted a protective effect.Spanish Foundation of Rheumatology. FIS/ISCIII (grant number PI11/02857). Dr. Pego-Reigosa is supported by Grant 316265 (BIOCAPS) from the European Union 7th Framework Programme (FP7/REGPOT-2012–2013.1)

Comorbidities in patients with Primary Sjögren's Syndrome and Systemic Lupus Erythematosus: A comparative registries-based study

OBJECTIVE: To compare the prevalence of the main comorbidities in 2 large cohorts of patients with primary Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE), with a focus on cardiovascular (CV) diseases. METHODS: This was a cross-sectional multicenter study where the prevalence of more relevant comorbidities in 2 cohorts was compared. Patients under followup from SJOGRENSER (Spanish Rheumatology Society Registry of Primary SS) and RELESSER (Spanish Rheumatology Society Registry of SLE), and who fulfilled the 2002 American-European Consensus Group and 1997 American College of Rheumatology classification criteria, respectively, were included. A binomial logistic regression analysis was carried out to explore potential differences, making general adjustments for age, sex, and disease duration and specific adjustments for each variable, including CV risk factors and treatments, when appropriate. RESULTS: A total of 437 primary SS patients (95% female) and 2,926 SLE patients (89% female) were included. The mean age was 58.6 years (interquartile range [IQR] 50.0-69.9 years) for primary SS patients and 45.1 years (IQR 36.4-56.3 years) for SLE patients (P?<?0.001), and disease duration was 10.4 years (IQR 6.0-16.7 years) and 13.0 years (IQR 7.45-19.76 years), respectively (P?<?0.001). Smoking, dyslipidemia, and arterial hypertension were associated less frequently with primary SS (odds ratio [OR] 0.36 [95% confidence interval (95% CI) 0.28-0.48], 0.74 [95% CI 0.58-0.94], and 0.50 [95% CI 0.38-0.66], respectively) as were life-threatening CV events (i.e., stroke or myocardial infarction; OR 0.57 [95% CI 0.35-0.92]). Conversely, lymphoma was associated more frequently with primary SS (OR 4.41 [95% CI 1.35-14.43]). The prevalence of severe infection was lower in primary SS than in SLE (10.1% versus 16.9%; OR 0.54 [95% CI 0.39-0.76]; P?<?0.001). CONCLUSION: Primary SS patients have a consistently less serious CV comorbidity burden and a lower prevalence of severe infection than those with SLE. In contrast, their risk of lymphoma is greater

Functional characterisation and antimicrobial efficiency assessment of smart nanohydrogels containing natamycin incorporated into polysaccharide-based films

The potential application of polysaccharide-based films containing smart nanohydrogels for the controlled release of food preservatives is demonstrated here. Smart active packaging is the most promising alternative to traditional packaging as it provides a controlled antimicrobial effect, which allows reducing the amount of preservatives in the food bulk, releasing them only on demand. This work evaluates the usefulness of smart thermosensitive poly(N-isopropylacrylamide) (PNIPA) nanohydrogels with or without acrylic acid (AA) incorporated into polysaccharide-based films (GA) to transport natamycin and release it as a response to environmental triggers. Release kinetics in liquid medium from GA films containing PNIPA/AA nanohydrogels (GA-PNIPA(5) and GA-PNIPA-20AA(5)) presented a characteristic feature regarding the films without nanohydrogels that was the appearance of a lag time in natamycin release, able to reach values of around 35 h. Another important feature of natamycin release kinetics was the fact that the release from GA-PNIPA/AA films only occurred when temperature was increased, so that the natamycin release was restricted to when there is a risk of growth of microorganisms that cause food spoilage or the development of pathogenic microorganisms. Additionally, it could be observed that the relative fraction of natamycin released from GA-PNIPA/AA films was significantly (p<0.05) higher than that released from GA films loaded with the same amount of free natamycin. It can be hypothesised that the encapsulation of natamycin into nanohydrogels helped it to be released from GA films, creating reservoirs of natamycin into the films and, therefore, facilitating its diffusion through the film matrix when the nanohydrogel collapses. In a solid medium, the low water availability limited natamycin release from GA-PNIPA/AA films restricting the on/off release mechanism of PNIPA/AA nanohydrogels and favouring the hydrophobic interactions between natamycin and polymer chains at high temperatures. Despite the low natamycin release in solid media, antimicrobial efficiency of GA-PNIPA(5) films containing natamycin in acidified agar plates was higher than that obtained with GA films without natamycin and GA films with free natamycin, probably due to the protecting effect against degradation when natamycin was included in the nanohydrogels, allowing its release only when the temperature increased.Clara Fucinos and Miguel A. Cerqueira are recipients of a fellowship (SFRH/BPD/87910/2012 and SFRH/BPD/72753/2010, respectively) from the Fundacao para a Ciencia e Tecnologia (FCT, POPH-QREN, and FSE Portugal). The authors thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and the project "BioInd - Biotechnology and Bioengineering for improved Industrial and Agro-Food processes", Ref. NORTE-07-0124-FEDER-000028 co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER and the project from the "Ministerio de Educacion y Ciencia" (Spain) "Nanohidrogeles inteligentes sensibles a cambios de pH y Temperatura: Diseno, sintesis y aplicacion en terapia del cancer y el envasado activo de alimentos", Ref. MAT2010-21509-C03-01

Evaluation of antimicrobial effectiveness of pimaricin-loaded thermosensitive nanohydrogels in grape juice

Pimaricin-loaded poly(N-isopropylacrylamide) nanohydrogels with and without acrylic acid, were evaluated as food-spoilage inhibitors in a model system and a real food product: grape juice. Pimaricin was proposed as a non-allergenic alternative to sulphites for protecting juices against recontamination. However, pimaricin may degrade under conditions and treatments (heating, acidification, lighting) commonly applied in producing fresh juices. Nanohydrogel encapsulation may be a feasible procedure to avoid pimaricin degradation, improving its antimicrobial activity. Pimaricin-free nanohydrogels did not affect the growth of the indicator yeast either in the food model system or in grape juice. Conversely, pimaricin-loaded nanohydrogels effectively inhibited the growth of the indicator yeast. In some cases, the inhibition was extended even further than using free pimaricin. For instance, in the food model system, pimaricin-loaded nanohydrogels with acrylic acid (NPPNIPA-20AA(5)) prevented the yeast growth for more than 81 h while free pimaricin was only effective for 12 h. Despite pimaricin-loaded nanohydrogels without acrylic acid (NPPNIPA(5)) were able to reduce maximum yeast growth, as in all treatments with pimaricin, the extent of the inhibitory effect was not significantly (p>0.05) different to that achieved with free pimaricin. In grape juice, both free pimaricin and NPPNIPA-20AA(5) treatment completely inhibited the growth of the indicator yeast until the end of the bioassay. However, the latter provided similar inhibition levels using a smaller amount of pimaricin due to PNIPA-20AA(5) protection and its controlled release from the nanohydrogel. Therefore, nanohydrogel encapsulation may help to optimise antifungal treatments and decrease the incidence of food allergies.Funded by grant (MAT 2006-11662-CO3-CO2-C01/MAT 2010-21509-C03-01/EUI 2008-00115) from the “Ministerio de Educación y Ciencia” (Spain). Grant (SFRH/BPD/87910/2012) from the Fundação para a Ciência e Tecnologia (FCT, Portugal). Marie Curie COFUND Postdoctoral Research Fellow

Relevance of gastrointestinal manifestations in a large Spanish cohort of patients with systemic lupus erythematosus: what do we know?

SLE can affect any part of the gastrointestinal (GI) tract. GI symptoms are reported to occur in >50% of SLE patients. To describe the GI manifestations of SLE in the RELESSER (Registry of SLE Patients of the Spanish Society of Rheumatology) cohort and to determine whether these are associated with a more severe disease, damage accrual and a worse prognosis. METHODS: We conducted a nationwide, retrospective, multicentre, cross-sectional cohort study of 3658 SLE patients who fulfil =4 ACR-97 criteria. Data on demographics, disease characteristics, activity (SLEDAI-2K or BILAG), damage (SLICC/ACR/DI) and therapies were collected. Demographic and clinical characteristics were compared between lupus patients with and without GI damage to establish whether GI damage is associated with a more severe disease. RESULTS: From 3654 lupus patients, 3.7% developed GI damage. Patients in this group (group 1) were older, they had longer disease duration, and were more likely to have vasculitis, renal disease and serositis than patients without GI damage (group 2). Hospitalizations and mortality were significantly higher in group 1. Patients in group 1 had higher modified SDI (SLICC Damage Index). The presence of oral ulcers reduced the risk of developing damage in 33% of patients. CONCLUSION: Having GI damage is associated with a worse prognosis. Patients on a high dose of glucocorticoids are at higher risk of developing GI damage which reinforces the strategy of minimizing glucocorticoids. Oral ulcers appear to decrease the risk of GI damage. © The Author(s) 2021. Published by Oxford University Press on behalf of the British Society for Rheumatology

Increased Expression of Fatty-Acid and Calcium Metabolism Genes in Failing Human Heart

Heart failure (HF) involves alterations in metabolism, but little is known about cardiomyopathy-(CM)-specific or diabetes-independent alterations in gene expression of proteins involved in fatty-acid (FA) uptake and oxidation or in calcium-(Ca(2+))-handling in the human heart.RT-qPCR was used to quantify mRNA expression and immunoblotting to confirm protein expression in left-ventricular myocardium from patients with HF (n = 36) without diabetes mellitus of ischaemic (ICM, n = 16) or dilated (DCM, n = 20) cardiomyopathy aetiology, and non-diseased donors (CTL, n = 6).Significant increases in mRNA of genes regulating FA uptake (CD36) and intracellular transport (Heart-FA-Binding Protein (HFABP)) were observed in HF patients vs CTL. Significance was maintained in DCM and confirmed at protein level, but not in ICM. mRNA was higher in DCM than ICM for peroxisome-proliferator-activated-receptor-alpha (PPARA), PPAR-gamma coactivator-1-alpha (PGC1A) and CD36, and confirmed at the protein level for PPARA and CD36. Transcript and protein expression of Ca(2+)-handling genes (Two-Pore-Channel 1 (TPCN1), Two-Pore-Channel 2 (TPCN2), and Inositol 1,4,5-triphosphate Receptor type-1 (IP3R1)) increased in HF patients relative to CTL. Increases remained significant for TPCN2 in all groups but for TPCN1 only in DCM. There were correlations between FA metabolism and Ca(2+)-handling genes expression. In ICM there were six correlations, all distinct from those found in CTL. In DCM there were also six (all also different from those found in CTL): three were common to and three distinct from ICM.DCM-specific increases were found in expression of several genes that regulate FA metabolism, which might help in the design of aetiology-specific metabolic therapies in HF. Ca(2+)-handling genes TPCN1 and TPCN2 also showed increased expression in HF, while HF- and CM-specific positive correlations were found among several FA and Ca(2+)-handling genes