The Red Seaweed Grateloupia turuturu Prevents Epidermal Dysplasia in HPV16-Transgenic Mice.

The role of dietary profiles in promoting or reducing the risk of multiple types of cancer is increasingly clear, driving the search for balanced foods and nutraceuticals. The red seaweed Grateloupia turuturu has been used as human food showing a balanced nutritional profile. This study aims to test in vivo chemopreventive effects of G. turuturu against cutaneous pre-malignant lesions in transgenic mice for the human papillomavirus type 16 (HPV16). Forty-four female HPV+/- or HPV-/- mice received a standard diet or were supplemented with 10% G. turuturu for 22 consecutive days. Cutaneous lesions (ear and chest skin) were identified histologically. Complementarily, the weights and histology of internal organs as well as blood biochemical and DNA integrity parameters were also assessed. G. turuturu consistently reduced the incidence of epidermal dysplasia induced by HPV16 on both cutaneous sites. Moreover, biochemical, DNA integrity and histological analyses confirmed G. turuturu edibility as no signs of toxicity were found. Dietary supplementation with G. turuturu is an effective and safe chemopreventive strategy in this model.

Domesticated Populations of Codium tomentosum Display Lipid Extracts with Lower Seasonal Shifts than Conspecifics from the Wild-Relevance for Biotechnological Applications of this Green Seaweed.

In the last decades, the use of algae in biotechnology and food industries has experienced an exponential growth. Codium tomentosum is a green macroalgae with high biotechnological potential, due to its rich lipidome, although few studies have addressed it. This study aimed to investigate the seasonal changes in lipid and pigment profiles of C. tomentosum, as well as to screen its antioxidant activity, in order to evaluate its natural plasticity. Samples of C. tomentosum were collected in two different seasons, early-autumn (September/October) and spring (May), in the Portuguese coast (wild samples), and in a land-based integrated multitrophic aquaculture (IMTA) system (IMTA samples). Total lipid extracts were analysed by LC-MS, GC-MS, and HPLC, and antioxidant activity was screened through free radical scavenging potential against DPPH and 2,20-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals. Wild samples showed a high seasonal variability, modifying their lipidome and pigment profiles according to environmental shifts, while IMTA samples showed a relatively stable composition due to early-stage culturing in controlled conditions. The lipids that contributed the most to seasonal discrimination were glycolipids (monogalactosyl diacylglycerol - MGDG and digalactosyl diacylglycerol - DGDG) and the lyso forms of phospholipids and glycolipids. Lipid extracts showed antioxidant activity ranging from 61 ± 2 to 115 ± 35 µmol Trolox g-1 of lipid extract in DPPH assay and from 532 ± 73 to 927 ± 92 µmol Trolox g-1 of lipid extract in ABTS assay, with a more intense antioxidant activity in wild spring samples. This study revealed that wild specimens of C. tomentosum presented a higher plasticity to cope with seasonal environmental changes, adjusting their lipid, pigment, and bioactivity profiles, while IMTA samples, cultured under controlled conditions, displayed more stable lipidome and pigment compositions.

Dietary Supplementation with the Red Seaweed Porphyra umbilicalis Protects against DNA Damage and Pre-Malignant Dysplastic Skin Lesions in HPV-Transgenic Mice.

Some diet profiles are associated with the risk of developing cancer; however, some nutrients show protective effects. Porphyra umbilicalis is widely consumed, having a balanced nutritional profile; however, its potential for cancer chemoprevention still needs comprehensive studies. In this study, we incorporated P. umbilicalis into the diet of mice transgenic for the human papillomavirus type 16 (HPV16), which spontaneously develop pre-malignant and malignant lesions, and determined whether this seaweed was able to block lesion development. Forty-four 20-week-old HPV+/- and HPV-/- mice were fed either a base diet or a diet supplemented with 10% seaweed. At the end of the study, skin samples were examined to classify HPV16-induced lesions. The liver was also screened for potential toxic effects of the seaweed. Blood was used to study toxicological parameters and to perform comet and micronucleus genotoxicity tests. P. umbilicalis significantly reduced the incidence of pre-malignant dysplastic lesions, completely abrogating them in the chest skin. These results suggest that P. umbilicalis dietary supplementation has the potential to block the development of pre-malignant skin lesions and indicate its antigenotoxic activity against HPV-induced DNA damage. Further studies are needed to establish the seaweed as a functional food and clarify the mechanisms whereby this seaweed blocks multistep carcinogenesis induced by HPV.

Protective effects of seaweed supplemented diet on antioxidant and immune responses in European seabass (Dicentrarchus labrax) subjected to bacterial infection.

European seabass (Dicentrarchus labrax) production is often hampered by bacterial infections such as photobacteriosis caused by Photobacterium damselae subsp. piscicida (Phdp). Since diet can impact fish immunity, this work investigated the effect of dietary supplementation of 5% Gracilaria sp. aqueous extract (GRA) on seabass antioxidant capacity and resistance against Phdp. After infection, mortality was delayed in fish fed GRA, which also revealed increased lysozyme activity levels, as well as decreased lipid peroxidation, suggesting higher antioxidant capacity than in fish fed a control diet. Dietary GRA induced a down-regulation of hepatic stress-responsive heat shock proteins (grp-78, grp-170, grp-94, grp-75), while bacterial infection caused a down-regulation in antioxidant genes (prdx4 and mn-sod). Diet and infection interaction down-regulated the transcription levels of genes associated with oxidative stress response (prdx5 and gpx4) in liver. In head-kidney, GRA led to an up-regulation of genes associated with inflammation (il34, ccr9, cd33) and a down-regulation of genes related to cytokine signalling (mif, il1b, defb, a2m, myd88). Additionally, bacterial infection up-regulated immunoglobulins production (IgMs) and down-regulated the transcription of the antimicrobial peptide leap2 in head kidney. Overall, we found that GRA supplementation modulated seabass resistance to Phdp infection.

A New Look for the Red Macroalga Palmaria palmata: A Seafood with Polar Lipids Rich in EPA and with Antioxidant Properties.

Palmaria palmata is an edible red macroalga widely used for human consumption and valued for its high protein value. Despite its low total lipid content, it is rich in eicosapentaenoic acid (EPA). This seaweed has been scarcely explored with regard to its lipid composition. The polar lipids of seaweeds are nowadays recognized as important phytochemicals contributing to their add value valorization and providing support for claims of potential health benefits. The present study aimed to disclose the polar lipid profile of P. palmata, farmed in an integrated multi-trophic aquaculture (IMTA) through modern lipidomic approaches using high-resolution LC-MS and MS/MS and to screen for the antioxidant properties of this red macroalga. A total of 143 molecular species of lipids were identified, belonging to several classes of polar lipids, such as glycolipids, phospholipids, and betaine lipids. It is noteworthy that the most abundant lipid species in each class were esterified with eicosapentaenoic acid (EPA), accounting for more than 50% of the lipid content. The polar lipid extract rich in EPA showed antioxidant activity with an inhibition concentration (IC) of IC30 = 171 ± 19.8 µg/mL for α,α-diphenyl-ß-picrylhydrazyl radical (DPPH●) and IC50 = 26.2 ± 0.1 µg/mL for 2,20-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical cation (ABTS●+). Overall, this study highlights that P. palmata farmed in an IMTA framework can be a sustainable source of beneficial lipids with antioxidant activity. Moreover, this red macroalga can be exploited for future applications as a source of lipids rich in EPA for food and feed, nutraceuticals, and cosmetics.

Leaf-Inspired Authentically Complex Microvascular Networks for Deciphering Biological Transport Process.

The vascular transport of molecules, cells, and nanoconstructs is a fundamental biophysical process impacting tissue regeneration, delivery of nutrients and therapeutic agents, and the response of the immune system to external pathogens. This process is often studied in single-channel microfluidic devices lacking the complex tridimensional organization of vascular networks. Here, soft lithography is employed to replicate the vein system of a Hedera elix leaf on a polydimethilsiloxane (PDMS) template. The replica is then sealed and connected to an external pumping system to realize an authentically complex microvascular network. This satisfies energy minimization criteria by Murray's law and comprises a network of channels ranging in size from capillaries (∼50 µm) to large arterioles and venules (∼400 µm). Micro-PIV (micro-particle image velocimetry) analysis is employed to characterize flow conditions in terms of streamlines, fluid velocity, and flow rates. To demonstrate the ability to reproduce physiologically relevant transport processes, two different applications are demonstrated: vascular deposition of tumor cells and lysis of blood clots. To this end, conditions are identified to culture cells within the microvasculature and realize a confluent endothelial monolayer. Then, the vascular deposition of circulating breast (MDA-MB 231) cancer cells is documented throughout the network under physiologically relevant flow conditions. Firm cell adhesion mostly occurs in channels with low mean blood velocity. As a second application, blood clots are formed within the chip by mixing whole blood with a thrombin solution. After demonstrating the blood clot stability, tissue plasminogen activator (tPA) and tPA-carrying nanoconstructs (tPA-DPNs) are employed as thrombolytics. In agreement with previous data, clot dissolution is equally induced by tPA and tPA-DPNs. The proposed leaf-inspired chip can be efficiently used to study a variety of vascular transport processes in complex microvascular networks, where geometry and flow conditions can be modulated and monitored throughout the experimental campaign.

Searching for antigenotoxic properties of marine macroalgae dietary supplementation against endogenous and exogenous challenges.

The functional characterization of marine macroalgae toward their potential to strength genome protection is still scarce. Hence, the aim of this study was to assess the antigenotoxic potential of Ulva rigida, Fucus vesiculosus, and Gracilaria species in Drosophila melanogaster following dietary exposure and adopting the somatic mutation and recombination test (SMART). All macroalgae displayed a genoprotection activity, namely against an exogenous challenge (streptonigrin). The action against subtler endogenous pressures was also noted indicating that supplementation level is a critical factor. Gracilaria species provided ambivalent indications, since 10% of G. vermiculophylla inhibited the egg laying and/or larvae development, while 10% of G. gracilis promoted spontaneous genotoxicity. The effects of U. rigida were modulated (in intensity) by the growing conditions, demonstrating higher genoprotection against streptonigrin-induced damage when grown in an aquaculture-controlled system, while the effectiveness against spontaneous genotoxicity was more apparent in specimens grown under wild conditions. In contrast, F. vesiculosus did not produce significant differences in its potential under varying growing conditions. Overall, these findings shed some light on the macroalgae ability toward genome protection, contributing to the development of algaculture industry, and reinforcing the concept of functional food and its benefits.

High dosage folic acid supplementation, oral cleft recurrence and fetal growth.

OBJECTIVES: To evaluate the effects of folic acid supplementation on isolated oral cleft recurrence and fetal growth. PATIENTS AND METHODS: The study included 2,508 women who were at-risk for oral cleft recurrence and randomized into two folic acid supplementation groups: 0.4 and 4 mg per day before pregnancy and throughout the first trimester. The infant outcome data were based on 234 live births. In addition to oral cleft recurrence, several secondary outcomes were compared between the two folic acid groups. Cleft recurrence rates were also compared to historic recurrence rates. RESULTS: The oral cleft recurrence rates were 2.9% and 2.5% in the 0.4 and 4 mg groups, respectively. The recurrence rates in the two folic acid groups both separately and combined were significantly different from the 6.3% historic recurrence rate post the folic acid fortification program for this population (p = 0.0009 when combining the two folic acid groups). The rate of cleft lip with palate recurrence was 2.9% in the 0.4 mg group and 0.8% in the 4 mg group. There were no elevated fetal growth complications in the 4 mg group compared to the 0.4 mg group. CONCLUSIONS: The study is the first double-blinded randomized clinical trial (RCT) to study the effect of high dosage folic acid supplementation on isolated oral cleft recurrence. The recurrence rates were similar between the two folic acid groups. However, the results are suggestive of a decrease in oral cleft recurrence compared to the historic recurrence rate. A RCT is still needed to identify the effect of folic acid on oral cleft recurrence given these suggestive results and the supportive results from previous interventional and observational studies, and the study offers suggestions for such future studies. The results also suggest that high dosage folic acid does not compromise fetal growth.

Combination therapy for gonorrhoea: in vitro synergy testing.

OBJECTIVES: Antimicrobial resistance in Neisseria gonorrhoeae is an increasing problem worldwide and combinations of antimicrobial agents have been recommended to delay the onset of treatment failures. The objective of this study was to obtain in vitro data on the activity of current (ceftriaxone or cefixime plus azithromycin) and alternative (gentamicin plus azithromycin) regimens. METHODS: A panel of 64 gonococcal isolates displaying various cefixime MICs was selected for inclusion in the study. Determination of the activities of the antimicrobial combinations of ceftriaxone, cefixime or gentamicin with azithromycin was performed using the agar dilution method and subsequent calculation of the fractional inhibitory concentration index (FICI) values. RESULTS: No antagonism for any of the antimicrobial combinations was detected among the 64 gonococcal isolates. When cefixime or ceftriaxone was combined with azithromycin all isolates showed additivity/indifference with a mean FICI of 2.0. All gonococcal isolates also showed additivity/indifference with the antimicrobial combination of gentamicin with azithromycin, but with a lower mean FICI of 1.7. No significant difference in the mean FICI between isolates fully susceptible to cefixime and isolates with decreased susceptibility to cefixime was observed. CONCLUSIONS: The results obtained support the gonorrhoea treatment currently recommended in the UK national guidelines and suggest that gentamicin with azithromycin could be a future treatment option. The in vivo activity and efficacy of these combinations remain unknown and prospective clinical studies should be addressed.

Oral cleft prevention program (OCPP).

BACKGROUND: Oral clefts are one of the most common birth defects with significant medical, psychosocial, and economic ramifications. Oral clefts have a complex etiology with genetic and environmental risk factors. There are suggestive results for decreased risks of cleft occurrence and recurrence with folic acid supplements taken at preconception and during pregnancy with a stronger evidence for higher than lower doses in preventing recurrence. Yet previous studies have suffered from considerable design limitations particularly non-randomization into treatment. There is also well-documented effectiveness for folic acid in preventing neural tube defect occurrence at 0.4 mg and recurrence with 4 mg. Given the substantial burden of clefting on the individual and the family and the supportive data for the effectiveness of folic acid supplementation as well as its low cost, a randomized clinical trial of the effectiveness of high versus low dose folic acid for prevention of cleft recurrence is warranted. METHODS/DESIGN: This study will assess the effect of 4 mg and 0.4 mg doses of folic acid, taken on a daily basis during preconception and up to 3 months of pregnancy by women who are at risk of having a child with nonsyndromic cleft lip with/without palate (NSCL/P), on the recurrence of NSCL/P. The total sample will include about 6,000 women (that either have NSCL/P or that have at least one child with NSCL/P) randomly assigned to the 4 mg and the 0.4 mg folic acid study groups. The study will also compare the recurrence rates of NSCL/P in the total sample of subjects, as well as the two study groups (4 mg, 0.4 mg) to that of a historical control group. The study has been approved by IRBs (ethics committees) of all involved sites. Results will be disseminated through publications and presentations at scientific meetings. DISCUSSION: The costs related to oral clefts are high, including long term psychological and socio-economic effects. This study provides an opportunity for huge savings in not only money but the overall quality of life. This may help establish more specific clinical guidelines for oral cleft prevention so that the intervention can be better tailored for at-risk women. CLINICALTRIALS.GOV IDENTIFIER: NCT00397917.

Structural, physical, and chemical modifications induced by microwave heating on native agar-like galactans.

Native agars from Gracilaria vermiculophylla produced in sustainable aquaculture systems (IMTA) were extracted under conventional (TWE) and microwave (MAE) heating. The optimal extracts from both processes were compared in terms of their properties. The agars' structure was further investigated through Fourier transform infrared and NMR spectroscopy. Both samples showed a regular structure with an identical backbone, ß-d-galactose (G) and 3,6-anhydro-α-l-galactose (LA) units; a considerable degree of methylation was found at C6 of the G units and, to a lesser extent, at C2 of the LA residues. The methylation degree in the G units was lower for MAE(opt) agar; the sulfate content was also reduced. MAE led to higher agar recoveries with drastic extraction time and solvent volume reductions. Two times lower values of [η] and M(v) obtained for the MAE(opt) sample indicate substantial depolymerization of the polysaccharide backbone; this was reflected in its gelling properties; yet it was clearly appropriate for commercial application in soft-texture food products.

Patterns of pharmacogenetic diversity in African populations: role of ancient and recent history.

AIMS: The knowledge that genetic variation influencing drug response is clearly structured among human populations has prompted many studies aimed at obtaining pharmacogenetic profiles in specific populations. While large amounts of data are being produced for populations from developed countries, the African continent still remains very poorly studied. To help to fill this gap, this work characterized three previously uncharacterized African populations for a set of pharmacogenetically relevant polymorphisms. MATERIALS & METHODS: Seven polymorphic variations in four genes that encode enzymes from phase I (CYP2C9, CYP3A4 and CYP3A5) or phase III (ABCB1) of drug metabolism were analyzed in population samples from Cabinda (n = 107), Mozambique (n = 109) and São Tomé e Príncipe (n = 126). All three populations shared strong recent historical links with Portugal. RESULTS: The majority of the screened variations displayed large contrasts in allele frequencies between European and African populations, and this study identified a substantial higher European influence in São Tomé e Príncipe than in Cabinda or Mozambique. Admixture analysis demonstrated that the European contribution to the population of São Tomé e Príncipe amounted to 13.3 +/- 3.3%. Furthermore, the proportion of African or European pharmacogenetic ancestry varied widely across each São Tomean individual. DISCUSSION & CONCLUSION: This implies that genetic association studies conducted in São Tomé e Príncipe should take into account the confounding factor of admixture to avoid spurious positive or negative results. Our findings also indicate that drug dosage requirements may be different in São Tomé e Príncipe than in other African populations. Thus, the search for pharmacogenetic risk factors should be assessed at an individual level, therefore fulfilling the perspective of personalized medicine. This study further indicates that the common notion of 'African population' might hide a pattern of pharmacogenetic heterogeneity whose real extent still needs to be evaluated by means of a refined sampling of the entire continent.

Avaliação da qualidade de vida em pessoas com feridas crónicas: o caso dos doentes com úlcera de perna de etiologia venosa / Quality of life assessment in persons with chronic wounds: the case of patients with venous leg ulcer

As investigações desenvolvidas sobre as pessoas com úlcera crónica de perna, apesar de ainda incipientes, descortinam uma forte relação entre as condições sociais e modos de vida e a prevalência desta afeção que tem também em Portugal uma relevância significativa. Por outro lado, apesar de todos os sintomas associados à presença de úlcera crónica de perna, muitos dos estudos realizados não determinam o impacto que estes produzem nas pessoas aos mais diferentes níveis, bem como na sua perceção e avaliação do próprio estado de saúde e da qualidade de vida. Deste modo, foi desenvolvido um estudo de cariz transversal, descritivo e exploratório consubstanciado num estudo de caso, tendo por base uma metodologia quantitativa. A amostra é constituída por 78 doentes que apresentavam um ou mais registos de diagnóstico de úlcera venosa ativo há pelo menos três meses, tendo a colheita de dados ocorrido em 2008. Os objetivos deste estudo passam por avaliar em que medida as condições sociais influenciam a qualidade de vida destes doentes; identificar quais dimensões, de acordo com a perspetiva dos próprios, se encontravam mais afetadas pela ocorrência do fenómeno de úlcera de perna e determinar qual o nível de qualidade de vida das pessoas com feridas crónicas, nomeadamente dos doentes com úlcera venosa. Para este efeito utilizou-se um questionário composto por duas partes: uma primeira que visava a caracterização inicial sociodemográfica, económica e clínica. A segunda, recorrendo à versão Portuguesa do CWIS© (Esquema Cardiff de Impacto de Ferida), pretendeu medir a ocorrência e impacto associado à existência de úlcera venosa e as dimensões «Bem-estar¼, «Sintomas físicos e vida diária¼ e «Vida social¼. Em complementaridade, foi avaliada a perceção de qualidade de vida e o respetivo grau de satisfação com a mesma por parte dos inquiridos. Em termos dos principais resultados, verificou-se uma relação entre os fatores sociodemográficos e económicos desfavoráveis e a existência de úlcera de perna crónica. No entanto, não se constatou idêntica relação linear entre estes mesmos fatores e avaliação global do estado de saúde em geral, bem como da qualidade de vida e da satisfação com a mesma de um modo particular. Em paralelo, observou-se que existe uma ocorrência e impacto significativo entre esta afeção crónica e sintomas físicos e atividades de vida diária, bem-estar e interação social. Como principal conclusão e na ótica da otimização da qualidade de vida destas pessoas, no âmbito de um paradigma holístico do cuidar, os enfermeiros na sua prática clínica deverão atender e considerar a influência destas dimensões, procurando com os doentes e famílias as medidas de remediação adequadas. Neste sentido, será igualmente determinante a valorização e monitorização contínua das condições sociais e modos de vida destas pessoas e os seus efeitos na saúde.

Despite all the symptoms associated with the presence of chronic leg ulcers, many studies do not determine the impact these have on people at many different levels, as well as their perception and evaluation of quality of life. Moreover, the investigations in this area, although still incomplete, reveal a strong relationship between social conditions and lifestyles and the prevalence of this disease that in Portugal has a significant importance. Therefore was developed a cross-sectional descriptive and exploratory study based in a study of cases using a quantitative methodology. The sample his constituted by 78 patients from the council of Matosinhos and who had one or more records of diagnosis of venous ulcers, active for at least three months, and the data collect occurred in 2008. This study objectives his to assess the extent to which social conditions affect the quality of life of these patients, identify which dimensions that, according to the perspective of themselves, were most affected by the occurrence of the phenomenon of the leg ulcer and to determinate what quality of life level of people with chronic wounds, particularly patients with venous ulcers. For this purpose it was used a questioner composed of two parts: the first aimed the socio-demographic, economic and clinical characterization. The second and using the Portuguese version of the CWIS© (Cardiff Wound Impact Schedule), sought to measure the occurrence and impact associated with the existence of venous ulcers and dimensions: «Welfare¼, «Physical symptoms and daily life¼ and «Social life¼. Complementarily was evaluated in the perception of quality of life and their satisfaction with it by the respondents. In terms of the main results, there was a relationship between the socio-demographic and economic existence of adversity and chronic leg ulcer. However there is not a linear relationship between these factors and overall assessment of general health status, and the quality of life in general and satisfaction with it in a particular way. In parallel, we observed that there is an occurrence and impact of this chronic condition and the dimensions: physical symptoms and daily life activities, welfare and social interaction. As a main conclusion and in the optical of optimizing the quality of life of these individuals and within a holistic paradigm of care, nurses in their clinical practice should address and consider the influence of these dimensions, looking for patients and families with the appropriate remedial action. In this sense is also crucial the continuous monitoring of social conditions and lifestyles of these people and their effects in health.