CCL17 and CCL22 chemokines are upregulated in human obesity and play a role in vascular dysfunction.

Background/Aims: Chemokines are known to play critical roles mediating inflammation in many pathophysiological processes. The aim of this study was to investigate the role of chemokine receptor CCR4 and its ligands CCL17 and CCL22 in human morbid obesity. Methods: Circulating levels of CCL17 and CCL22 were measured in 60 morbidly obese patients (mean age, 45 ± 1 years; body mass index/BMI, 44 ± 1 kg/m2) who had undergone bariatric bypass surgery, and 20 control subjects. Paired subcutaneous (SCAT) and visceral adipose tissue (VCAT) from patients were analysed to measure expression of CCR4 and its ligands by RT-PCR, western blot and immunohistochemical analysis. The effects of CCR4 neutralization ex vivo on leukocyte-endothelial cells were also evaluated. Results: Compared with controls, morbidly obese patients presented higher circulating levels of CCL17 (p=0.029) and CCL22 (p<0.001) and this increase was positively correlated with BMI (p=0.013 and p=0.0016), and HOMA-IR Index (p=0.042 and p< 0.001). Upregulation of CCR4, CCL17 and CCL22 expression was detected in VCAT in comparison with SCAT (p<0.05). Using the parallel-plate flow chamber model, blockade of endothelial CCR4 function with the neutralizing antibody anti-CCR4 in morbidly obese patients significantly reduced leucocyte adhesiveness to dysfunctional endothelium, a key event in atherogenesis. Additionally, CCL17 and CCL22 increased activation of the ERK1/2 mitogen-activated protein kinase signalling pathway in human aortic endothelial cells, which was significantly reduced by CCR4 inhibition (p=0.016 and p<0.05). Conclusion: Based on these findings, pharmacological modulation of the CCR4 axis could represent a new therapeutic approach to prevent adipose tissue dysfunction in obesity.

Role of antiangiogenic VEGF-A165b in angiogenesis and systolic function after reperfused myocardial infarction.

INTRODUCTION AND OBJECTIVES: Angiogenesis helps to reestablish microcirculation after myocardial infarction (MI). In this study, we aimed to further understand the role of the antiangiogenic isoform vascular endothelial growth factor (VEGF)-A165b after MI and to explore its potential as a coadjuvant therapy to coronary reperfusion. METHODS: Two mice MI models were formed: a) permanent coronary ligation (nonreperfused MI); b) transient 45-minute coronary occlusion followed by reperfusion (reperfused MI); in both models, animals underwent echocardiography before euthanasia at day 21 after MI induction. We determined serum and myocardial VEGF-A165b levels. In both experimental MI models, we assessed the functional and structural role of VEGF-A165b blockade. In a cohort of 104 ST-segment elevation MI patients, circulating VEGF-A165b levels were correlated with cardiovascular magnetic resonance-derived left ventricular ejection fraction at 6 months and with the occurrence of adverse events (death, heart failure, and/or reinfarction). RESULTS: In both models, circulating and myocardial VEGF-A165b levels were increased 21 days after MI induction. Serum VEGF-A165b levels inversely correlated with systolic function evaluated by echocardiography. VEGF-A165b blockade increased capillary density, reduced infarct size, and enhanced left ventricular function in reperfused, but not in nonreperfused, MI experiments. In patients, higher VEGF-A165b levels correlated with depressed ejection fraction and worse outcomes. CONCLUSIONS: In experimental and clinical studies, higher serum VEGF-A165b levels are associated with worse systolic function. Their blockade enhances neoangiogenesis, reduces infarct size, and increases ejection fraction in reperfused, but not in nonreperfused, MI experiments. Therefore, VEGF-A165b neutralization represents a potential coadjuvant therapy to coronary reperfusion.

Modeling trade-offs across carbon sequestration, biodiversity conservation, and equity in the distribution of global REDD+ funds.

The program on Reducing Emissions from Deforestation and Forest Degradation (REDD+) is one of the major attempts to tackle climate change mitigation in developing countries. REDD+ seeks to provide result-based incentives to promote emission reductions and increase carbon sinks in forest land while promoting other cobenefits, such as the conservation of biodiversity. We model different scenarios of international REDD+ funds distribution toward potential recipient countries using 2 carbon emission reduction targets (20% and 50% compared to the baseline scenario, i.e., deforestation and forest degradation without REDD+) by 2030. The model combines the prioritization of environmental outcomes in terms of carbon sequestration and biodiversity conservation and social equity, accounting for the equitable distribution of international REDD+ funds. Results highlight the synergy between carbon sequestration and biodiversity conservation under alternative fund allocation criteria, especially for scenarios of low carbon emission reduction. Trade-offs increase when distributional equity is considered as an additional criterion, especially under higher equity requirements. The analysis helps to better understand the inherent trade-offs between enhancing distributional equity and meeting environmental targets under alternative REDD+ fund allocation options.

Disruption of the CCL1-CCR8 axis inhibits vascular Treg recruitment and function and promotes atherosclerosis in mice.

The CC chemokine 1 (CCL1, also called I-309 or TCA3) is a potent chemoattractant for leukocytes that plays an important role in inflammatory processes and diseases through binding to its receptor CCR8. Here, we investigated the role of the CCL1-CCR8 axis in atherosclerosis. We found increased expression of CCL1 in the aortas of atherosclerosis-prone fat-fed apolipoprotein E (Apoe)-null mice; moreover, in vitro flow chamber assays and in vivo intravital microscopy demonstrated an essential role for CCL1 in leukocyte recruitment. Mice doubly deficient for CCL1 and Apoe exhibited enhanced atherosclerosis in aorta, which was associated with reduced plasma levels of the anti-inflammatory interleukin 10, an increased splenocyte Th1/Th2 ratio, and a reduced regulatory T cell (Treg) content in aorta and spleen. Reduced Treg recruitment and aggravated atherosclerosis were also detected in the aortas of fat-fed low-density lipoprotein receptor-null mice treated with CCR8 blocking antibodies. These findings demonstrate that disruption of the CCL1-CCR8 axis promotes atherosclerosis by inhibiting interleukin 10 production and Treg recruitment and function.

Synthesis and biological evaluation of sphingosine kinase 2 inhibitors with anti-inflammatory activity.

The synthesis of inhibitors of SphK2 with novel structural scaffolds is reported. These compounds were designed from a molecular modeling study, in which the molecular interactions stabilizing the different complexes were taken into account. Particularly interesting is that 7-bromo-2-(2-phenylethyl)-2,3,4,5-tetrahydro-1,4-epoxynaphtho[1,2-b]azepine, which is a selective inhibitor of SphK2, does not exert any cytotoxic effects and has a potent anti-inflammatory effect. It was found to inhibit mononuclear cell adhesion to the dysfunctional endothelium with minimal impact on neutrophil-endothelial cell interactions. The information obtained from our theoretical and experimental study can be useful in the search for inhibitors of SphK2 that play a prominent role in different diseases, especially in inflammatory and cardiovascular disorders.

Intravital Microscopy in the Cremaster Muscle Microcirculation for Endothelial Dysfunction Studies.

The intravital microscopy in the mouse cremaster muscle microcirculation is a method widely used to visualize in vivo blood cells interacting with the endothelium and within the vessels. Therefore, it is a suitable technique to study leukocyte-endothelial cell interactions along every stage of the canonical leukocyte recruitment cascade: rolling, adhesion, intravascular crawling, and migration both in postcapillary venules and arterioles of the mouse cremasteric microcirculation. This technique also enables to assess vessel functionality, since hemodynamic parameters such as shear stress, flow rate, and vasodilatation/vasoconstriction, among other vascular events, can be additionally determined. Furthermore, response to multiple drugs and mechanisms underlying blood cells interactions within the vascular system can be studied in a real scenario. This chapter describes a protocol for intravital microscopy in the mouse cremaster muscle microcirculation.

[GEITDAH consensus on conduct disorders in children and adolescents]. / Consenso del GEITDAH sobre los trastornos de conducta en niños y adolescentes.

In this paper, the Special Interest Group on Attention Deficit Hyperactivity Disorder (GEITDAH, from its name in Spanish) presents a consensus reached by experts from all over Spain on conduct disorders in children and adolescents. Following the initial work by the team at the Pedopsychiatry Unit at the Quiron-Teknon Hospital in Barcelona, agreements have been reached on a number of basic aspects that could be the starting point for future consensuses. A top priority aim of the work was also to update the criteria in the Diagnostic and statistical manual of mental disorders, fifth edition, for conduct disorders in children and adolescents, together with their comorbidity with attention deficit hyperactivity disorder.

TITLE: Consenso del GEITDAH sobre los trastornos de conducta en niños y adolescentes.El Grupo de Especial Interes en el Trastorno por Deficit de Atencion/Hiperactividad (GEITDAH) presenta en este articulo un consenso de expertos de toda España sobre los trastornos de conducta en niños y adolescentes. A partir del trabajo inicial del equipo de la Unidad de Paidopsiquiatria del Hospital Quiron-Teknon de Barcelona, se han consensuado aspectos basicos que podrian ser el punto de partida para futuros consensos. Ha sido tambien objetivo prioritario del trabajo actualizar en los trastornos de conducta en niños y adolescentes los criterios del Manual diagnostico y estadistico de los trastornos mentales, quinta edicion, y su comorbilidad con el trastorno por deficit de atencion/hiperactividad.

Novel applications of motif-directed profiling to identify disease resistance genes in plants.

BACKGROUND: Molecular profiling of gene families is a versatile tool to study diversity between individual genomes in sexual crosses and germplasm. Nucleotide binding site (NBS) profiling, in particular, targets conserved nucleotide binding site-encoding sequences of resistance gene analogs (RGAs), and is widely used to identify molecular markers for disease resistance (R) genes. RESULTS: In this study, we used NBS profiling to identify genome-wide locations of RGA clusters in the genome of potato clone RH. Positions of RGAs in the potato RH and DM genomes that were generated using profiling and genome sequencing, respectively, were compared. Largely overlapping results, but also interesting discrepancies, were found. Due to the clustering of RGAs, several parts of the genome are overexposed while others remain underexposed using NBS profiling. It is shown how the profiling of other gene families, i.e. protein kinases and different protein domain-coding sequences (i.e., TIR), can be used to achieve a better marker distribution. The power of profiling techniques is further illustrated using RGA cluster-directed profiling in a population of Solanum berthaultii. Multiple different paralogous RGAs within the Rpi-ber cluster could be genetically distinguished. Finally, an adaptation of the profiling protocol was made that allowed the parallel sequencing of profiling fragments using next generation sequencing. The types of RGAs that were tagged in this next-generation profiling approach largely overlapped with classical gel-based profiling. As a potential application of next-generation profiling, we showed how the R gene family associated with late blight resistance in the SH*RH population could be identified using a bulked segregant approach. CONCLUSIONS: In this study, we provide a comprehensive overview of previously described and novel profiling primers and their genomic targets in potato through genetic mapping and comparative genomics. Furthermore, it is shown how genome-wide or fine mapping can be pursued by choosing different sets of profiling primers. A protocol for next-generation profiling is provided and will form the basis for novel applications. Using the current overview of genomic targets, a rational choice can be made for profiling primers to be employed.

[Clinical use of 30:70 controlled-release methylphenidate in the treatment of attention deficit hyperactivity disorder]. / Uso clínico de metilfenidato 30:70 de liberación modificada en el tratamiento del trastorno por déficit de atención/hiperactividad.

INTRODUCTION: Attention deficit hyperactivity disorder (ADHD) is one of the most frequent reasons for visits in daily clinical practice, with a prevalence rate of 1-7% in Spain. The effectiveness of stimulants for the treatment of ADHD has been widely demonstrated and methylphenidate (MPD) is the most commonly used. There are currently several different immediate-release or extended-release formulations of MPD on the market. AIMS: To review the characteristics of the different formulations of MPD, with special attention paid to the studies on Equasym®, an extended-release preparation soon to be made available in Spain. The article also includes recommendations for clinical practice and the choice of drugs. DEVELOPMENT: Several studies have assessed the effectiveness of Equasym® versus placebo or in comparison to other MPD formulations. The extended-release preparations have a therapeutic action that is similar to that of the immediate-release versions, the difference between them being the plasma concentration profiles over time during the day, which are reflected in the pharmacodynamic effects. Equasym® is more effective in the morning, whereas other formulations, such as Concerta®, allow greater control of the symptoms in the afternoon. These differences are important when it comes to prescribing the treatment. CONCLUSIONS: One of the main advantages of having different formulations of MPD available is that it allows the professional to choose the drug that best suits the clinical features and needs of each patient. The individual response is the essential criterion in deciding on the most appropriate treatment.

[Inflammatory response during ischaemic processes: adhesion molecules and immunomodulation]. / Respuesta inflamatoria durante los procesos de isquemia: moléculas de adhesión e inmunomodulación.

INTRODUCTION: After cerebral ischemia, necrotic cell death occurs specially for neurons, mainly due to the deprivation of oxygen and glucose. Cell necrosis triggers the activation of the immune system followed by an inflammatory response. This reaction is characterized by the activation of astrocytes and microglia together with the infiltration of peripheral immune cells. DEVELOPMENT: Both, microglia and inflammatory cells, including circulating peripheral inflammatory cells, get activated and release a plethora of inflammatory mediators, cytokines, chemokines, etc. Such released factors induce the overexpression of adhesion molecules, increasing the blood brain barrier permeability, thus favoring even more inflammatory cell infiltration. In the end, this contributes to increase brain damage. Inflammatory response is nevertheless necessary in order to eliminate cellular debris from both apoptotic and necrotic cells. It seems to be also implicated in the initiation of certain mechanisms responsible for brain repair and plasticity. As a result, the inflammatory response is a coordinated effort. Activation of inflammation triggers an immunosuppressant and anti-inflammatory response. A high rate of infections in patients suffering from stroke, together with increased serum levels of anti-inflammatory molecules in these patients, support this statement. The anti-inflammatory response could be interpreted as the organism attempting to control the heightened inflammatory response that occurs after cerebral ischemia. On the other hand, following an ischemic event, there are several new cerebral epitopes that get exposed to the immune system, which would never have been exposed under normal physiological conditions. CONCLUSION: Therefore immunosuppression after an ischemic accident hinders the development of auto-immune responses.

Leukotriene B4-loaded microspheres: a new therapeutic strategy to modulate cell activation.

BACKGROUND: Leukotriene B4 (LTB4) is a potent inflammatory mediator that also stimulates the immune response. In addition, it promotes polymorphonuclear leukocyte phagocytosis, chemotaxis, chemokinesis and modulates cytokines release. Regarding chemical instability of the leukotriene molecule, in the present study we assessed the immunomodulatory activities conferred by LTB4 released from microspheres (MS). A previous oil-in-water emulsion solvent extraction-evaporation method was chosen to prepare LTB4-loaded MS. RESULTS: In the mice cremasteric microcirculation, intraescrotal injection of 0.1 ml of LTB4-loaded MS provoked significant increases in leukocyte rolling flux, adhesion and emigration besides significant decreases in the leukocyte rolling velocity. LTB4-loaded MS also increase peroxisome proliferator-activated receptor-alpha (PPARalpha) expression by murine peritoneal macrophages and stimulate them to generate nitrite levels. Monocyte chemoattractant protein-1 (MCP-1) and nitric oxide (NO) productions were also increased when human umbilical vein and artery endothelial cells (HUVECs and HUAECs, respectively) were stimulated with LTB4-loaded MS. CONCLUSION: LTB4-loaded MS preserve the biological activity of the encapsulated mediator indicating their use as a new strategy to modulate cell activation, especially in the innate immune response.

Sensitivity analysis of a parameterization of the stomatal component of the DO3SE model for Quercus ilex to estimate ozone fluxes.

A sensitivity analysis of a proposed parameterization of the stomatal conductance (g(s)) module of the European ozone deposition model (DO(3)SE) for Quercus ilex was performed. The performance of the model was tested against measured g(s) in the field at three sites in Spain. The best fit of the model was found for those sites, or during those periods, facing no or mild stress conditions, but a worse performance was found under severe drought or temperature stress, mostly occurring at continental sites. The best performance was obtained when both f(phen) and f(SWP) were included. A local parameterization accounting for the lower temperatures recorded in winter and the higher water shortage at the continental sites resulted in a better performance of the model. The overall results indicate that two different parameterizations of the model are needed, one for marine-influenced sites and another one for continental sites.

Tibouchina pulchra (Cham.) Cogn., a native Atlantic Forest species, as a bio-indicator of ozone: visible injury.

Tibouchina pulchra saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF) and ambient non-filtered air+40 ppb ozone (NF+O3) 8 h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12,895 ppb h(-1), respectively, for the three treatments. After 25 days of exposure (AOT40=3871 ppb h(-1)), interveinal red stippling appeared in plants in the NF+O3 chamber. In the NF chamber, symptoms were observed only after 60 days of exposure (AOT40=910 ppb h(-1)). After 60 days, injured leaves per plant corresponded to 19% in NF+O3 and 1% in the NF treatment; and the average leaf area injured was 7% within the NF+O3 and 0.2% within the NF treatment. The extent of leaf area injured (leaf injury index) was mostly explained by the accumulated exposure of ozone (r2=0.89; p<0.05).

Ozone in Spain's national parks and protected forests.

In general, it is difficult to measure air pollutant concentrations in remote areas, as they are mostly national parks and protected areas. Passive samplers provide an accurate and inexpensive method for measuring cumulative exposures of different air pollutants. They have been used to collect ozone data in both laboratory and field at different geographical scales. The objective of the present study is to fill the knowledge gap regarding air quality in remote areas of Spain, such as national parks and protected areas. Because there were no systematic data sets on the main air pollutants that could affect these areas, an air quality measurement network was established between 2001 and 2004 on 19 locations inside Spanish national parks and protected areas. The data collected suggest that ozone levels in mountainous areas are high enough to affect sensitive vegetation. Most of the locations registered moderate-to-high ozone levels, with important interannual variability. Altitudinal ozone gradients were observed in most of the parks with complex topography due to the establishment of local circulations that incorporate polluted air masses from polluted airsheds or even long-range transport (i.e., Canary Islands). Different latitude-dependent, yearly cycles were also observed, showing two, one, or no clear peaks depending on the region. These findings extend to the most southerly locations, except in the Canary Islands, where pollution transported from other regions in the upper transport layers probably led to the high concentrations observed.

Parameterization of the stomatal component of the DO3SE model for Mediterranean evergreen broadleaf species.

An ozone (O3) deposition model (DO3SE) is currently used in Europe to define the areas where O3 concentrations lead to absorbed O3 doses that exceed the flux-based critical levels above which phytotoxic effects would be likely recorded. This mapping exercise relies mostly on the accurate estimation of O3 flux through plant stomata. However, the present parameterization of the modulation of stomatal conductance (g(s)) behavior by different environmental variables needs further adjustment if O3 phytotoxicity is to be assessed accurately at regional or continental scales. A new parameterization of the model is proposed for Holm oak (Quercus ilex), a tree species that has been selected as a surrogate for all Mediterranean evergreen broadleaf species. This parameterization was based on a literature review, and was calibrated and validated using experimentally measured data of g(s) and several atmospheric and soil parameters recorded at three sites of the Iberian Peninsula experiencing long summer drought, and very cold and dry winter air (El Pardo and Miraflores) or milder conditions (Tietar). A fairly good agreement was found between modeled and measured data (R2 = 0.64) at Tietar. However, a reasonable performance (R2 = 0.47-0.62) of the model was only achieved at the most continental sites when g(s) and soil moisture deficit relationships were considered. The influence of root depth on g(s) estimation is discussed and recommendations are made to build up separate parameterizations for continental and marine-influenced Holm oak sites in the future.

[Incidental prostatic cancer: T1a-T1b. Our experience after observation/radical surgery and literature review]. / Cáncer de próstata incidental: T1a-T1b. Nuestra experiencia tras observación/intervención radical y revisión de la literatura.

OBJECTIVES: To analyse the progress of T1a and T1b prostate cancer diagnosed in our hospital. MATERIAL AND METHODS: Retrospective study of 40 patients in T1a-T1b clinical stage diagnosed with prostate adenocarcinoma in our hospital, from 1986 to 1999. A restaging biopsy was performed on the 16 T1a patients after initial diagnosis and control. A radical prostatectomy was performed on the 24 T1b patients. They were all monitored every six months with rectal exam and PSA. We analysed biological and/or clinical progression, time to progression, mortality caused by the tumour and survival. RESULTS: None of the 16 patients with T1a clinical stage presented tumour progression, with a median follow-up of 90 months. 12,5% of the 24 T1b cases presented tumour progression, with a median follow-up of 70 months. Cancer-specific mortality was one patient (4,16 %) in the T1b group. CONCLUSIONS: Observation and follow-up with PSA and rectal exam appears to be a good option for T1a clinical stage, given the good prognosis. Our results show that patients with T1a clinical stage and good prognostic factors could be at a similar risk of suffering from a new prostate cancer as the normal population, although prospective studies are required to validate these results. T1b cases require active treatment and closer monitoring.

[Papillary carcinoma after 13 years of correct function of renal graft]. / Carcinoma papilar hallado en injerto renal tras 13 años de correcto funcionamiento.

It is known the greater probability appearance of malignancy injuries in patients with renal graft due to its inmunosupresión. We expose a case in which after thirteen years of correct operation of the renal transplant a tumor is diagnosed of accidental form by means of ultrasonography of graft's control. It was demonstrated by percutaneous biopsy that it was a carcinoma to papilar and later transplanctectomy was made. We raised a reflection about the novo tumors on renal graft given to the high number of patients with funcionante transplant during long years and the little evidence in Literature, proposing a possible registry of such valuing its behavior and comparing it with the well-known ones on native kidneys without inmunosupresión situation.

Physiological responses of saplings of Caesalpinia echinata Lam., a Brazilian tree species, under ozone fumigation.

The effects induced by long-term (30 day) and short-term (6h) exposures to ozone on the physiological parameters in young plants of Caesalpinia echinata Lam., a Brazilian tree species, were determined. Potted plants were maintained in open-top chambers in Valencia, Spain, under charcoal filtered air (mean O3 level: 29 microg m3), nonfiltered air (NF; 43 microg m3), and nonfiltered air plus O3 (NF + O3; 68 microg m3), simulating prevailing concentrations observed in the city of São Paulo, Brazil, during spring months (50 microg m3 in 2002). In the plants kept in NF + O3 for 30 days, although no foliar visible injuries were observed, the net carbon assimilation rate was reduced to 50%, stomatal conductance 42%, and transpiration 40%, when compared to the results for the NF plants. No changes in antioxidants, in leaf, stem, and root biomass, and in the root/shoot ratio were observed. Significant reductions were observed in gas exchange and in PSII photochemical efficiency (Fv/Fm) after 6 h of exposure to an O3 peak. The species was shown to be sensitive to ambient O3 concentrations measured in São Paulo.

[Renal teratoma, case report and review of the literature]. / Teratoma renal, aportación de un caso y revisión de la literatura.

We report a case of intrarenal teratoma in a 39-year-old female patient. The clinical course after three years of follow-up has been satisfactory, finding the patient totally asymptomatic. Extragonadal teratoma occurs predominantly along the median line of the body. Intrarenal teratoma is extremely rare; however, it should be distinguished from other cystic lesions.