Biomimetic Growth of Hydroxyapatite in Hybrid Polycaprolactone/Graphene Oxide Ultra-Porous Scaffolds.

This paper reports the preparation and characterization of hybrid scaffolds composed of polycaprolactone (PCL) and different graphene oxide (GO) amounts, intending to incorporate the intrinsic characteristics of their constituents, such as bioactivity and biocidal effect. These materials were fabricated by a solvent-casting/particulate leaching technique showing a bimodal porosity (macro and micro) that was around 90%. The highly interconnected scaffolds were immersed in a simulated body fluid, promoting the growth of a hydroxyapatite (HAp) layer, making them ideal candidates for bone tissue engineering. The growth kinetics of the HAp layer was influenced by the GO content, a remarkable result. Furthermore, as expected, the addition of GO neither significantly improves nor reduces the compressive modulus of PCL scaffolds. The thermal behavior of composites was investigated by differential scanning calorimetry, showing an increase in crystallinity as the addition of GO raised, which implies that GO nanosheets can act as seeds to induce the crystallization of PCL. The improved bioactivity was demonstrated by the deposition of an HAp layer on the surface of the scaffold with GO, especially with a 0.1% GO content.

A First Record of Organochlorine Pesticides in Barn Owls (Tyto alba) from Portugal: Assessing Trends from Variation in Feather and Liver Concentrations.

We evaluated feathers as a non-destructive biomonitoring tool documenting organochlorine pesticides (OCP) in liver and checked possible trends in pesticide use in two areas based on OCP concentrations in barn owls (Tyto alba). We measured the concentrations of 16 OCP in 15 primary feathers and 15 livers from barn owl carcasses collected on roadsides in Tagus Valley and Évora regions, south Portugal. Total OCP mean concentration was 8 120 ng g-1 in feathers and 178 ng g-1 in livers. All compounds were detected in feathers while in livers δ-HCH, endosulfan sulphate, p,p'-DDT and p,p'-DDD were not detected. The high ß-HCH and heptachlor concentrations in feathers most likely derived from external endogenous contamination. P,p'-DDE was the OCP with the highest hepatic concentration. Both matrices indicated an exposure to recently released heptachlor. The differing OCP concentrations between Tagus Valley and Évora seem to reflect differences in land-use and pesticide use histories of the two locations, and/or faster degradation of OCP in the Tagus area.

Presence of Anti-T. cruzi Antibodies in Inhabitants and Dogs of Two Rural Settlements in the Sierra de Los Tuxtlas, Veracruz, Mexico.

PURPOSE: The aim of this study was to identify the risk factors associated with house infestation by Triatoma dimidiata as well as with Trypanosoma cruzi infection in humans and owned dogs in two rural communities from the municipality of Catemaco, Veracruz, Mexico. METHODS: One hundred and 16 human blood samples and 34 dog blood samples were collected. The presence of anti-T. cruzi antibodies was determined using four different ELISA assays. Moreover, reactive ELISA sera from humans and dogs were processed by indirect immunofluorescence (IFI) to confirm the presence of anti-T. cruzi antibodies. RESULTS: Serologic tests for T. cruzi infection showed a prevalence of 5.1% (6/116) in humans and of 50% (17/34) in owned dogs. CONCLUSION: The presence of animals (dogs, chickens and wild animals), as well as some characteristics of house construction were identified as risk factors for infestation and infection. Complementary studies must be carried out to allow a better understanding of the transmission dynamics in the state of Veracruz, Mexico, and the implementation of adequate control programs.

High Levels of Heavy Metals detected in Feathers of an Avian Scavenger Warn of a High Pollution Risk in the Atacama Desert (Chile).

The Atacama Desert represents the highest levels of mining exploitation in Chile, which inexorably results in high levels of pollution. Raptors, and particularly scavengers, have shown their usefulness to evaluate exposure to environmental contaminants in polluted scenarios. In this study, we used primary feathers from a local avian scavenger, turkey vulture Cathartes aura, to evaluate the exposure to cadmium (Cd), lead (Pb), copper (Cu) and zinc (Zn) in two locations from the southern Atacama Desert (coastal and inland) and a third location from the bordering semiarid region, in northern Chile. All metals were detected in all analyzed samples (n = 54). Mean concentrations in Coastal Atacama were 0.68 ± 0.84 ppm for Cd; 1.97 ± 2.01 ppm for Pb; 59.11 ± 80.69 ppm for Cu; and 107.96 ± 51.00 ppm for Zn, while mean concentrations in Inland Atacama were 0.55 ± 0.42 ppm for Cd; 3.37 ± 2.61 ppm for Pb; 91.66 ± 77.74 ppm for Cu; and 214.03 ± 99.08 ppm for Zn. Mean concentrations in Coastal Coquimbo were 0.63 ± 0.69 ppm for Cd; 1.57 ± 0.92 ppm for Pb; 18.09 ± 6.12 ppm for Cu; and 149.37 ± 105.56 ppm for Zn. These differences could be explained by differences on abundance of mining settlements among areas. According to the literature, these values are very high for all metals, exceeding in some cases those values referred as responsible of health disorders in birds. We strongly recommend further research looking at potential adverse effects caused by heavy metal pollution on the health of human and wildlife populations in the southern Atacama Desert.

Hg and Se in Organs of Three Cetacean Species from the Murcia Coastline (Mediterranean Sea).

We determinated Hg and Se concentrations in liver, kidney, brain, lung and muscle of five bottlenose dolphin (Tursiops truncatus), four common dolphins (Delphinus delphis) and four Risso's dolphin (Grampus griseus) stranded along the Murcia coast, Southeast Spain, in order to evaluate the risk of Hg toxicity. Hg concentrations showed similar concentrations to other individuals in the Mediterranean Sea with the same length in the same period. We observed a positive correlation of Hg and Se in liver (r = 0.948, p < 0.001) and kidney (r = 0.939; p = 0.001) and ratio the Se/Hg molar was higher than 1 in most cases. Our results suggest that the protective effects of Se against Hg toxicity occur in cetaceans. However, we detected levels of Hg described as responsible liver damage and neurotoxicological effects so other tools, as biochemical markers, should be included. Besides, more studies are needed to evaluate the risk of Hg exposure in dolphins from Murcia coastline.

From Mexico to the Beagle Channel: A review of metal and metalloid pollution studies on wildlife species in Latin America.

Emissions of metals and metalloids (Hg; Cd; Cr; Cu; Pb; Ni; Zn; Fe; Mn; As; Se) generated by natural (e.g., geothermal activity) or anthropic causes (eg., industry or mining) represent a worldwide contamination problem, especially in developing countries. Exposure to high concentrations of these elements is harmful to living beings, including humans. Information on this type of contamination is scarce and fragmented, limiting research which could benefit from these data. To know the state of the research, we reviewed the studies of environmental pollution by metals and metalloids carried out on animal species in Latin America. The use of animals as biomonitors of contamination by metals and metalloids is a continuously expanding practice that allows for early detection of problems. With this work, we were able to identify the most studied areas in Latin America (Amazon, Gulf of California, coastal area between Rio de Janeiro and Florianopolis and River Plate Estuary). Moreover, we provide information on the most studied metals (Hg, Cd, Cu, Pb, Zn) and wild species, which evidence the use of endangered species. The data reviewed should help researchers to direct their efforts towards sparsely researched areas and facilitate bibliographic consultation of scientific information on exposure to metals and metalloids in Latin America.

Trace metals in striped dolphins (Stenella coeruleoalba) stranded along the Murcia coastline, Mediterranean Sea, during the period 2009-2015.

Mercury (Hg), cadmium (Cd), lead (Pb), selenium (Se) and arsenic (As) concentrations in internal tissues of 72 striped dolphins (Stenella coeruleoalba) from Murcia Region (Mediterranean coastline) have been investigated for the first time. Hg showed the highest concentration, followed by Se, Cd, As and Pb. In general, the levels of metal found in this study were similar to those described in similar studies in the Mediterranean Sea. However, in some adult specimens, Hg liver concentrations were related with toxic effects in cetacean. A significant correlation was observed with age, likewise between Se and Hg and Cd in tissues, which agree with detoxify effect attributed to Se through inert complex formation. Molar ratio Hg:Se in liver was close to 1:1 in some specimens, which would indicate overload of the Hg-Se detoxify function and compromised health. These results could contribute to a better knowledge of the distribution of these persistent pollutants in the Mediterranean Sea.

Utility of percutaneous catheters for local anaesthetics infusion for postoperative pain control in lumbar arthrodesis. A prospective cohort study. / Utilidad de los catéteres percutáneos para perfusión continua de anestésico local en el manejo del dolor postoperatorio en pacientes sometidos a artrodesis lumbar instrumentada. Estudio de cohortes prospectivas.

OBJECTIVE: To evaluate whether postoperative continuous wound infiltration of levobupivacaine through two submuscular catheters connected to two elastomeric pumps after lumbar instrumented arthrodesis is more effective than intravenous patient-controlled analgesia. MATERIAL AND METHODS: An observational, prospective cohorts study was carried out. The visual analogue scale, the need for additional rescue analgesia and the onset of adverse effects were recorded. RESULTS: Pain records measured with visual analogue scale scale were significantly lower in the 48hours postoperative record at rest (p=.032). The other records of visual analogue scale showed a clear tendency to lower levels of pain in the group treated with the catheters. No statistically significant differences were found in the rescue analgesia demands of the patients. The adverse effects were lower in the catheter group (6 cases versus 11 cases) but without statistical differences. CONCLUSIONS: A trend to lower pain records was found in the group treated with catheters, although differences were not statistically significant.

Development of a QuEChERS method for simultaneous analysis of antibiotics in carcasses for supplementary feeding of endangered vultures.

Antibiotics have been beneficial for human and animal health. However, an excessive use in livestock and a deficient management of the carcasses can lead to adverse effects in the scavengers that ingest them, especially in "supplementary feeding sites" (SFS). The aim of this study was to assess the potential risk of exposure to antibiotics for an endangered population of Cinereous vultures (Aegypius monachus) from southeastern Portugal. Hence, a multi-residue method based on QuEChERs was adapted and validated to analyse, in small volumes of tissues, the most frequent antibiotics used in livestock. The method was applied to 87 samples of liver, muscle and kidney from 7 goats and 25 sheep disposed in SFS. According to questionnaires to farmers, the animals had not been treated with antibiotics, but analyses showed residues in 29% of the samples. Antibiotics were more frequent in goats (42.9%) than in sheep (24.2%), and oxytetracycline and trimethoprim were the most common (both 13.8%). Oxytetracycline, the most common antibiotic for livestock in Portugal, showed the highest concentration (1452.68 ng g-1). To our knowledge, this is the first study of presence of antibiotics in carrion from SFS. The concentrations of antibiotics in carrion do not seem to pose a risk of acute intoxication for adult Cinereous vultures. However, subtle and likely chronic exposure with unknown health consequences may occur, which requires more research. Moreover, the results of this first study can be used in future studies to assess the risk for avian scavengers.

Influence of a Former Mining Area in the Heavy Metals Concentrations in Blood of Free-Living Mediterranean Pond Turtles (Mauremys leprosa).

The objective of this study was to assess metal exposure in the Mediterranean Pond Turtle (Mauremys leprosa) inhabiting a watercourse in an ancient mining district polluted by different metals ("Rambla de Las Moreras", southeastern Spain) and included in the Ramsar Convention. For this purpose, mercury (Hg), lead (Pb), copper (Cu), zinc (Zn) and cadmium (Cd) were analysed in blood samples from 42 M. leprosa healthy adults (27 males and 13 females). The highest median concentrations were found for Zn, followed by Cu, Pb, Hg and Cd (366, 33, 9, 0.83 and 0.14 µg/dL, respectively). Although the literature regarding toxic metals in freshwater turtles is relatively scarce, Pb may cause deleterious effects in our population. In general, males presented higher levels than females, which could be due to maternal transfer during egg formation. The significant correlations between Cu-Cd and Cu-Hg suggest the implication of an efficient mechanism of detoxification involving metallothioneins.

Per- and polyfluoroalkyl substances in plasma and feathers of nestling birds of prey from northern Norway.

Plasma samples from nestlings of two top predators, White-tailed eagle (Haliaeetus albicilla) and Northern goshawk (Accipiter gentilis) from northern Norway were analysed for a wide range of per- and polyfluoroalkyl substances (PFASs). Body feathers from the White-tailed eagles were also analysed and significant associations between specific PFASs in blood plasma and body feathers were found (0.36

Validation of a QuECheRS method for analysis of neonicotinoids in small volumes of blood and assessment of exposure in Eurasian eagle owl (Bubo bubo) nestlings.

Neonicotinoid pesticides have gained great interest in the last years both for agricultural and domestic use. Since the information on their environmental distribution or the effects derived from exposure to ecosystems and biota is scarce, new analytical techniques are being developed for monitoring studies. In this sense, two extraction techniques based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) methodology to detect the neonicotinoids authorised in Spain (acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, nitenpyram and thiamethoxam) were adapted and compared: a) using acetate buffer (AB); and b) using citrate buffer (CB). For detection and quantification, high performance liquid chromatography coupled with time of flight mass spectrometry (HPLC/TOF-MS) was used. The CB method provided a wide range of recoveries (68-134%) and accuracy (4-9%). The AB method provided good recoveries (59-76%, 59% corresponded to clothianidin) precision (4-11%) linearity (0.987-0.998%) and limit of quantification (2-10ng/mL) for all the compounds. To test the effectiveness of the technique, we analysed 30 blood samples of free-ranging nestlings of Eurasian eagle owl (Bubo bubo). The only compound detected, in one nestling from a dry land farming area, was imidacloprid, with a concentration of 3.28ng/mL. To our knowledge, this is the first study of neonicotinoid pesticides in free-ranging birds of prey using non-destructive samples, providing the first insight for biomonitoring studies. Further studies, including toxicokinetics and toxicodynamics, are recommended to assess the risk for these species.

Combined in situ effects of metals and nutrients on marine biofilms: Shifts in the diatom assemblage structure and biological traits.

The effects of multiple stressors on marine diatom assemblages are still poorly understood. The interactive effects of metals and nutrients were assessed in two coastal biofilms grown at a reference site and a historically contaminated site. The biofilms were exposed in situ to pulse exposures of metals (Zn and Pb) and nutrients (N and P) individually and in combination to mimic patterns of discharge in the study area. The reference community's structure (composition and abundance of taxa) was modified after metals and/or nutrients exposure, but each stressor acted in different way. Irrespective of the stressors or scenario, the abundance of the dominant species Opephora krumbeinii declined, and it is proposed as sensitive species. Nutrient supply favoured the proliferation of certain species with high nutrient tolerances (Fragilaria famelica, Tabularia ktenoeides), whereas metals promoted the colonisation of metal-tolerant species, e.g., Berkeleya fennica, Opephora marina. Simultaneous exposure induced an amplification of levels of accumulated metals, chlorophyll a and EPS contents and triggered the succession of species towards tolerant species with specific growth. Metals seemed to act as a selective factor of metal-tolerant species, and nutrients favoured the proliferation of those species forming zig-zag colonies (Neosynedra provincialis), mucous tubes (Berkeleya spp.) and motile diatoms (Navicula salinicola, Nitzschia incognita), resulting in biofilms with a more complex architecture. The diatom communities from the historically contaminated site were more resistant to pulse exposure, but metals or nutrients loads induced overproduction of mucilage. We propose that growth forms may complement taxonomic approaches and provide a quick and easy way to detect community changes related to metal and nutrient pollution.

Candidate predisposing germline copy number variants in early onset colorectal cancer patients.

PURPOSE: A great proportion of the heritability of colorectal cancer (CRC) still remains unexplained, and rare variants, as well as copy number changes, have been proposed as potential candidates to explain the so-called 'missing heritability'. We aimed to identify rare high-to-moderately penetrant copy number variants (CNVs) in patients suspected of having hereditary CRC due to an early onset. METHODS/PATIENTS: We have selected for genome-wide copy number analysis, 27 MMR-proficient early onset CRC patients (<50 years) without identifiable germline mutations in Mendelian genes related to this phenotype. Rare CNVs were selected by removing all CNVs detected at MAF >1% in the in-house control CNV database (n = 629 healthy controls). Copy number assignment was checked by duplex real-time quantitative PCR or multiplex ligation probe amplification. Somatic mutation analysis in candidate genes included: loss of heterozygosity studies, point mutation screening, and methylation status of the promoter. RESULTS: We have identified two rare germline deletions involving the AK3 and SLIT2 genes in two patients. The search for a second somatic mutational event in the corresponding CRC tumors showed loss of heterozygosity in AK3, and promoter hypermethylation in SLIT2. Both genes have been previously related to colorectal carcinogenesis. CONCLUSIONS: These findings suggest that AK3 and SLIT2 may be potential candidates involved in genetic susceptibility to CRC.

Tracking pan-continental trends in environmental contamination using sentinel raptors-what types of samples should we use?

Biomonitoring using birds of prey as sentinel species has been mooted as a way to evaluate the success of European Union directives that are designed to protect people and the environment across Europe from industrial contaminants and pesticides. No such pan-European evaluation currently exists. Coordination of such large scale monitoring would require harmonisation across multiple countries of the types of samples collected and analysed-matrices vary in the ease with which they can be collected and the information they provide. We report the first ever pan-European assessment of which raptor samples are collected across Europe and review their suitability for biomonitoring. Currently, some 182 monitoring programmes across 33 European countries collect a variety of raptor samples, and we discuss the relative merits of each for monitoring current priority and emerging compounds. Of the matrices collected, blood and liver are used most extensively for quantifying trends in recent and longer-term contaminant exposure, respectively. These matrices are potentially the most effective for pan-European biomonitoring but are not so widely and frequently collected as others. We found that failed eggs and feathers are the most widely collected samples. Because of this ubiquity, they may provide the best opportunities for widescale biomonitoring, although neither is suitable for all compounds. We advocate piloting pan-European monitoring of selected priority compounds using these matrices and developing read-across approaches to accommodate any effects that trophic pathway and species differences in accumulation may have on our ability to track environmental trends in contaminants.

[SEIP-AEPAP-SEPEAP consensus document on the aetiology, diagnosis and treatment of bacterial skin infections in out-patients]. / Documento de consenso SEIP-AEPAP-SEPEAP sobre la etiología, el diagnóstico y el tratamiento de las infecciones cutáneas bacterianas de manejo ambulatorio.

Skin infections are a common cause for dermatological consultations in the paediatric setting. A review is presented of the clinical manifestations, diagnosis and treatment of the main bacterial skin infections, as well as the diagnosis and treatment of super-infected puncture and bite wounds. The most prevalent bacteria in skin infections are Staphylococcus aureus and Streptococcus pyogenes. Treatment is usually empirical, since microbiological studies are only recommended under certain circumstances or lack of improvement with common therapies. Superficial skin infections can be treated with local antiseptics or antibiotics (mupirocin or fusidic acid). Systemic treatment is usually reserved for patients with extensive or severe disease or with other risk factors. Systemic treatment depends on the suspected infecting bacteria, with penicillin, amoxicillin, amoxicillin-clavulanic acid and first or second generation cephalosporin being the most frequently used drugs. Due to the low incidence of community-acquired methicillin-resistant infection by S. aureus in Spain, the use of clindamycin or co-trimoxazole is only recommended after severe disease, relapses or a clear epidemiological background.

A review on exposure and effects of arsenic in passerine birds.

Arsenic (As) is a metalloid of high concern because of its toxic effects for plants and animals. However, it is hard to find information on this metalloid in passerines. This review presents a comprehensive overview of As exposure and effects in birds, and more particularly in passerines, as a result of an extensive search of the literature available. Internal tissues are the most frequently analyzed matrices for As determination in passerines (37.5% of the reviewed studies used internal tissues), followed by feathers and eggs (32.5% each), feces (27.5%), and finally blood (15%). A clear tendency is found in recent years to the use of non-destructive samples. Most studies on As concentrations in passerines have been done in great tit (Parus major; 50%), followed by pied flycatcher (Ficedula hypoleuca; 22.5%). Some factors such as diet and migratory status are crucial on the interspecific differences in As exposure. More studies are needed to elucidate if intraspecific factors like age or gender affect As concentrations in different tissues. The literature review shows that studies on As concentrations in passerines have been done mainly in the United States (30%), followed by Belgium (22.5%), and Finland (20%), making evident the scarce or even lack of information in some countries, so we recommend further research in order to overcome the data gap, particularly in the southern hemisphere. Studies on humans, laboratory animals and birds have found a wide range of effects on different organ systems when they are exposed to different forms of As. This review shows that few field studies on As exposure and effects in passerines have been done, and all of them are correlative so far. Arsenic manipulation experiments on passerines are recommended to explore the adverse effects of As in free-living populations at similar levels to those occurring in the environment. CAPSULE: This review summarizes the most interesting published studies on As exposure and effects in passerines.

The effect of orally administered probiotic Lactobacillus reuteri-containing tablets in peri-implant mucositis: a double-blind randomized controlled trial.

BACKGROUND AND OBJECTIVES: Probiotics create a biofilm and protect the oral tissues against the action of periodontal pathogenic bacteria. The aim of this study was to evaluate the effects of the oral probiotic Lactobacillus reuteri Prodentis upon the peri-implant health of edentulous patients with dental implants and peri-implant mucositis, establishing comparisons vs implants without peri-implant disease. MATERIAL AND METHODS: A double-blind, placebo-controlled, prospective cross-over study was made. The patients were all edentulous and were divided into two groups, (A) no peri-implant disease, and (B) peri-implant mucositis affecting one or more implants. Patients with peri-implantitis were excluded. The dosage was one tablet every 24 h over 30 d. All patients in both groups initially received the oral probiotic Lactobacillus reuteri Prodentis, followed by placebo. Patients started with probiotic treatment during 30 d, followed by a 6 mo washout period and the administration of placebo for the same period. The following parameters were studied: crevicular fluid volume, modified plaque index, probing depth, modified gingival index, and concentrations of interleukin 1ß, interleukin 6 and interleukin 8. RESULTS: A total of 77 implants were evaluated in 34 patients. Group A involved 22 patients with 54 implants without peri-implant alterations, and group B, 12 patients with mucositis affecting one or more implants (23 implants). After treatment with the probiotic, both the patients with mucositis and the patients without peri-implant disease showed improvements in the clinical parameters, with reductions in cytokine levels. In contrast, no such changes were observed with placebo. CONCLUSIONS: After treatment with the probiotic Lactobacillus reuteri in patients with implants presenting mucositis, the clinical parameters improved, and the cytokine levels decreased - in contraposition to the observations in the placebo group. Probiotic administration may be regarded as a good alternative for both the treatment of peri-implant mucositis and its prevention, as it also improved clinical parameters in the healthy individuals. Further studies involving larger patient series are needed regarding the effects of probiotics upon peri-implant health.