Computation-guided scaffold exploration of 2E,6E-1,10-trans/cis-eunicellanes.

Eunicellane diterpenoids are a unique family of natural products containing a foundational 6/10-bicyclic framework and can be divided into two main classes, cis and trans, based on the configurations of their ring fusion at C1 and C10. Previous studies on two bacterial diterpene synthases, Bnd4 and AlbS, revealed that these enzymes form cis- and trans-eunicellane skeletons, respectively. Although the structures of these diterpenes only differed in their configuration at a single position, C1, they displayed distinct chemical and thermal reactivities. Here, we used a combination of quantum chemical calculations and chemical transformations to probe their intrinsic properties, which result in protonation-initiated cyclization, Cope rearrangement, and atropisomerism. Finally, we exploited the reactivity of the trans-eunicellane skeleton to generate a series of 6/6/6 gersemiane-type diterpenes via electrophilic cyclization.

First trans-eunicellane terpene synthase in bacteria.

Terpenoids are the largest family of natural products, but prokaryotes are vastly underrepresented in this chemical space. However, genomics supports vast untapped biosynthetic potential for terpenoids in bacteria. We discovered the first trans-eunicellane terpene synthase (TS), AlbS from Streptomyces albireticuli NRRL B-1670, in nature. Mutagenesis, deuterium labeling studies, and quantum chemical calculations provided extensive support for its cyclization mechanism. In addition, parallel stereospecific labeling studies with Bnd4, a cis-eunicellane TS, revealed a key mechanistic distinction between these two enzymes. AlbS highlights bacteria as a valuable source of novel terpenoids, expands our understanding of the eunicellane family of natural products and the enzymes that biosynthesize them, and provides a model system to address fundamental questions about the chemistry of 6,10-bicyclic ring systems.

Phytotherapeutic alternatives for neurodegenerative dementias: Scientific review, discussion and therapeutic proposal.

The incidence and prevalence of age-related neurodegenerative dementias have been increasing. There is no curative therapy and conventional drug treatment can cause problems for patients. Medicinal plants traditionally used for problems associated with ageing are emerging as a therapeutic resource. The main aim is to give a proposal for use and future research based on scientific knowledge and tradition. A literature search was conducted in several searchable databases. The keywords used were related to neurodegenerative dementias, ageing and medicinal plants. Boolean operators and filters were used to focus the search. As a result, there is current clinical and preclinical scientific information on 49 species used in traditional medicine for ageing-related problems, including neurodegenerative dementias. There are preclinical and clinical scientific evidences on their properties against protein aggregates in the central nervous system and their effects on neuroinflammation, apoptosis dysregulation, mitochondrial dysfunction, gabaergic, glutamatergic and dopaminergic systems alterations, monoamine oxidase alterations, serotonin depletion and oestrogenic protection. In conclusion, the potential therapeutic effect of the different medicinal plants depends on the type of neurodegenerative dementia and its stage of development, but more clinical and preclinical research is needed to find better, safer and more effective treatments.

Investigation of Acid-Base Catalysis in Halimadienyl Diphosphate Synthase Involved in Mycobacterium tuberculosis Virulence.

The devastating human pathogenMycobacterium tuberculosis (Mtb) is able to parasitize phagosomal compartments within alveolar macrophage cells due, in part, to the activity of its cell-surface lipids. Prominent among these is 1-tuberculosinyl-adenosine (1-TbAd), a derivative of the diterpenoid tuberculosinyl (halima-5,13-dienyl) diphosphate produced by the class II diterpene cyclase encoded by Rv3377c, termed here MtHPS. Given the demonstrated ability of 1-TbAd to act as a virulence factor for Mtb and the necessity for Rv3377c for its production, there is significant interest in MtHPS activity. Class II diterpene cyclases catalyze a general acid-base-mediated carbocation cascade reaction initiated by protonation of the terminal alkene in the general diterpenoid precursor (E,E,E)-geranylgeranyl diphosphate and terminated by deprotonation of the final cyclized (and sometimes also rearranged) intermediate. Here, structure-guided mutagenesis was applied to characterize the various residues contributing to activation of the enzymatic acid, as well as identify the enzymatic base in MtHPS. Particularly given the ability of conservative substitution for the enzymatic base (Y479F) to generate an alternative product (labda-7,13-dienyl diphosphate) via deprotonation of an earlier unrearranged intermediate, further mutational analysis was carried out to introduce potential alternative catalytic bases. The results were combined with mechanistic molecular modeling to elucidate how these mutations affect the catalytic activity of this important enzyme. This not only provided detailed structure-function insight into MtHPS but also further emphasized the inert nature of the active site of MtHPS and class II diterpene cyclases more generally.

Medicinal plants and natural products as neuroprotective agents in age-related macular degeneration.

The retina may suffer neurodegenerative damages, as other tissues of the central nervous system do, and serious eye diseases may develop. One of them is age-related macular degeneration, which causes progressive loss of vision due to retina degeneration. Treatment of age-related macular degeneration focuses on antioxidant agents and anti-vascular endothelial growth factor compounds, among others, that prevent/diminish oxidative stress and reduce neovascularisation respectively. The phytochemicals, medicinal plants and/or plant-diet supplements might be a useful adjunct in prevention or treatment of age-related macular degeneration owing to their antioxidant and anti-vascular endothelial growth factor properties. This review article presents the most investigated plants and natural products in relation to age-related macular degeneration, such as saffron, ginkgo, bilberry and blueberry, curcuma or turmeric, carotenoids, polyphenols, and vitamins C and E. This study provides up-to-date information on the effects, treatments, safety and efficiency of these phytotherapy products.

Singlet oxygenation of triquinacene, barrelene, and homobarrelene.

The singlet oxygenation of three polycyclic hydrocarbons, triquinacene, barrelene and homobarrelene was studied. Triquinacene reacted by way of a perepoxide intermediate, transferring an oxygen atom to another triquinacene molecule to give exclusively the mono epoxide. Barrelene, on the other hand, underwent a rare homo-Diels-Alder reaction with 1O2 to give the decomposition product from the initial tetracyclic 1,2-dioxolane leading to benzofuran. The latter reacted with 1O2 in a [2+2] cycloaddition to give an unstable 1,2-dioxetane which collapsed to 2-formylphenyl formate. The latter was independently synthesize via singlet oxygenation of authentic benzofuran. Homobarrelene reacted in a similar fashion to give a homoDiels product, decomposition of which led to a keto aldehyde which was characterized spectroscopically. Computational work confirms the barrelene and homobarrelene reactions with 1O2 as concerted [π2s+π2s+π2s] cycloadditions.

Neuroprotective Potential of Ginkgo biloba in Retinal Diseases.

Like other tissues of the central nervous system, the retina is susceptible to damage by oxidative processes that result in several neurodegenerative disease such as age-related macular degeneration, diabetic retinopathy, glaucoma, ischaemic retinal disease, retinal disease produced by light oxidation, and detached retina, among other diseases. The use of antioxidant substances is a solution to some health problems caused by oxidative stress, because they regulate redox homeostasis and reduce oxidative stress. This is important for neurodegeneration linked to oxidation processes. In line with this, Ginkgo biloba is a medicinal plant with excellent antioxidant properties whose effects have been demonstrated in several degenerative processes, including retinal diseases associated with neurodegeneration. This review describes the current literature on the role of ginkgo in retinal diseases associated with neurodegeneration. The information leads to the conclusion that G. biloba extracts might be a good option to improve certain neurodegenerative retinal diseases, but more research is needed to determine the safety and efficacy of G. biloba in these retinal degenerative processes.

Improving university students' web savvy: An intervention study.

BACKGROUND: Young people increasingly turn to the Internet for information about social and political issues. However, they struggle to evaluate the trustworthiness of the information they encounter online. AIMS: This pilot study investigated whether a focused curricular intervention could improve university students' ability to make sound judgements of credibility. SAMPLE: Participants (n = 67) were students in four sections of a 'critical thinking and writing' course at a university on the West Coast of the United States. Course sections were randomly assigned to treatment (n = 29) and control conditions (n = 38). METHODS: We conducted a pre-and-posttest, treatment/control experiment using a 2 × 2 × 2 design (treatment condition × order × time) with repeated measures on the last factor. Students in the treatment group received two 75-min lessons on evaluating the credibility of online content. An assessment of online reasoning was administered to students 6 weeks prior to the intervention and again 5 weeks after. RESULTS: Students in the treatment group were significantly more likely than students in the control group to have shown gains from pretest to posttest. CONCLUSIONS: Results suggest that teaching students a small number of flexible heuristics that can be applied across digital contexts can improve their evaluation of online sources.

Antocianos: su potencial en la prevención de enfermedades neurodegenerativas / Antocianinas: potencialidades para a prevenção de doenças neurodegenerativas / Anthocyanins: potential for the prevention of neurodegenerative diseases

Los antocianos son compuestos polifenólicos, de origen vegetal, ampliamente distribuidos en la naturaleza, tanto en plantas medicinales como en alimentos, responsables de los colores rojizos, azulados y violáceos de diferentes frutos, hojas, raíces o bulbos. Numerosos estudios científicos han constatado sus propiedades saludables, principalmente relacionadas con sus actividades antiinflamatorias y antioxidantes. En base a ello han sido propuestos como candidatos para la prevención y tratamiento de enfermedades neurodegenerativas. En este artículo se revisan los últimos conocimientos científicos publicados sobre sus efectos beneficiosos sobre las funciones del SNC. Es interesante señalar que dichos estudios parecen confirmar, tanto en animales como en el hombre, su capacidad de atravesar la barrera hematoencefálica, cuestionada hasta hace poco (AU)

As antocianinas são compostos polifenólicos de origem vegetal, amplamente distribuídas na natureza, tanto em plantas medicinais como em alimentos, responsáveis pelas cores vermelha, azul e violácea de diversos frutos, fo-lhas, raízes ou bolbos. Numerosos estudos científicos evidenciaram as suas propriedades benéficas para a saúde, principalmente relacionadas com as suas atividades anti-inflamatórias e antioxidantes. Nesta base, foram propostos como candidatos para a prevenção e tratamento de doenças neurodegenerativas. Neste artigo fez-se a revisão dos mais recentes conhecimentos científicos publicados sobre os efeitos benéficos sobre as funções do SNC. É interessante assinalar que estes estudos parecem confirmar, tanto em animais como no homem, a sua capacidade de atravessar a barreira he-matoencefálica, questionada até recentemente

Anthocyans are polyphenolic compounds widely distributed in nature, found both in food and in medicinal plants. They are responsible for the red-blue colors of different fruits, leaves, roots or bulbs. Many scientific studies have confirmed its healthy properties, mainly related to their anti-inflammatory and antioxidant activities. For these reasons, they could be proposed as agents for the prevention and treatment of neurodegenerative diseases. In the present paper, the latest scientific knowledge on the effects of these compounds on central nervous system functions has been reviewed. Despite what was thought until recently, several studies, both in animal and in humans, seem to confirm their ability to pass the blood-brain barrier (AU)

Overexpression of IL-10 in C2D macrophages promotes a macrophage phenotypic switch in adipose tissue environments.

Adipose tissue macrophages are a heterogeneous collection of classically activated (M1) and alternatively activated (M2) macrophages. Interleukin 10 (IL-10) is an anti-inflammatory cytokine, secreted by a variety of cell types including M2 macrophages. We generated a macrophage cell line stably overexpressing IL-10 (C2D-IL10) and analyzed the C2D-IL10 cells for several macrophage markers after exposure to adipocytes compared to C2D cells transfected with an empty vector (C2D-vector). C2D-IL10 macrophage cells expressed more CD206 when co-cultured with adipocytes than C2D-vector cells; while the co-cultured cell mixture also expressed higher levels of Il4, Il10, Il1ß and Tnf. Since regular C2D cells traffic to adipose tissue after adoptive transfer, we explored the impact of constitutive IL-10 expression on C2D-IL10 macrophages in adipose tissue in vivo. Adipose tissue-isolated C2D-IL10 cells increased the percentage of CD206(+), CD301(+), CD11c(-)CD206(+) (M2) and CD11c(+)CD206(+) (M1b) on their cell surface, compared to isolated C2D-vector cells. These data suggest that the expression of IL-10 remains stable, alters the C2D-IL10 macrophage cell surface phenotype and may play a role in regulating macrophage interactions with the adipose tissue.

Effect of cooking and germination on phenolic composition and biological properties of dark beans (Phaseolus vulgaris L.).

Legumes are the basés diet in several countries. They hold a high nutritional value, but other properties related to human health are nowadays being studied. The aim of this work was to study the influence of processes (boiling or germination) on the phenolic composition of dark beans (Phaseolus vulgaris L. c.v. Tolosana) and their effect on their antioxidant, neuroprotective and anticancer ability. Phenolic composition of raw and processed dark beans was analysed by HPLC-PAD and HPLC-ESI/MS. The antioxidant activity was evaluated by ORAC. Astrocytes cultures (U-373) have been used to test their neuroprotective effect. Anticancer activities were evaluated on three different cell lines (renal adenocarcinoma (TK-10), breast adenocarcinoma (MCF-7) and melanoma (UACC-62)) by sulphorhodamine B method. Qualitative and quantitative differences in phenolic composition have been observed between raw and processed dark beans that influence the antioxidant activity, mainly for germinated samples which show a decrease of antioxidant capacity. Although every assayed extracts decreased reactive oxygen species release and exhibited cytotoxicity activities on cancer cell lines, raw beans proved to be the most active in neuroprotective and antitumoral effects; this sample is especially rich in phenolic compounds, mainly anthocyanins. This study further demonstrated that phenolic composition of dark beans is related with cooking process and so with their neuroprotective and anticancer activity; cooking of dark beans improves their digestion and absorption at intestinal level, while maintaining its protective ability on oxidative process at cellular level.

Tratamiento de la sobrecarga de volumen en IC refractaria / Treatment of volume overload in refractory heart failure / Tratamento da sobrecarga de volume na insuficiência cardíaca refratária

La insuficiencia cardíaca es un problema de salud creciente debido al envejecimiento de la población y a la mejoría en el pronóstico de los pacientes con cardiopatía establecida. En los últimos años, han ido apareciendo nuevos tratamientos que intentan mejorar la situación clínica, la calidad de vida y la supervivencia de aquellos pacientes que desarrollan refractariedad a los diuréticos. El empleo de estos nuevos fármacos podría tener un impacto positivo en las situaciones agudas, pero no parece que aporten beneficios adicionales para los enfermos con insuficiencia cardíaca congestiva crónica. En estos casos, las técnicas de ultrafiltración extracorpórea y, sobre todo, la diálisis peritoneal parecen demostrar mejoría de la clase funcional, reducción en las tasas de hospitalización, mejoría en la calidad de vida y reducción de la mortalidad, siendo además coste-eficiente.

Heart failure is an increasing health problem due to the aging population and the improvement in the prognosis of patients with established heart disease. In recent years new treatments have emerged that seek to improve the clinical situation, the quality of life and survival of patients who develop refractoriness to diuretics. The use of these new drugs could have a positive impact on the acute but it would not provide additional benefits for patients with chronic congestive heart failure. In these cases, extracorporeal ultrafiltration techniques and especially peritoneal dialysis seem to show improvement in functional class, reduction in hospitalization rates, improved quality of life and reduction in mortality, besides being cost-efficient.

A insuficiência cardíaca é um problema crescente de saúde devido ao envelhecimento da população e a melhora do prognóstico dos pacientes com doença cardíaca estabelecida. Nos últimos anos, novos tratamentos surgiram que procuram melhorar a situação clínica, a qualidade de vida e sobrevida dos pacientes que desenvolvem refratariedade ao uso de diuréticos. O uso desses novos fármacos poderia ter um impacto positivo sobre os efeitos agudos, mas não oferecem benefícios adicionais para os pacientes com insuficiência cardíaca congestiva crônica. Nestes casos, as técnicas de ultrafiltração extracorpórea e, especialmente, a diálise peritoneal parecem mostrar melhoria da classe funcional, redução das taxas de hospitalização, melhoria da qualidade de vida e a redução da mortalidade, e tambémé custo-eficiente.

Cationic liposomes bearing IL-2 on their external surface induced mice leukocytes to kill human cervical cancer cells in vitro, and significantly reduced tumor burden in immunodepressed mice.

Tumor cells are known to modify their surroundings in order to escape immunologic detection, and IL-2, a killer cell activator, is one of the factors known to overcome this escape mechanism. In this regard, when we cocultured cells from the human cervical cancer cell line INBL with mice blood leukocytes, no inhibition of tumor cell growth was observed, but when a similar coculture was done in the presence of cationic liposomes bearing IL-2 on their external surface (CL-IL-2), all the INBL cells were killed. In order to evaluate whether this in vitro property of CL-IL-2 to overcome tumor cell detection by lymphocytes could also be reproduced in vivo, INBL cells were intraperitoneally (i.p.) inoculated into immunodepressed mice to produce solid tumors. We observed that the subsequent i.p. delivery of CL-IL-2 rendered the tumor masses significantly smaller. The presence of a large number of infiltrating lymphocytes on those tumors, and the fact that many had a cytotoxic CD8(+) phenotype suggests that these lymphocytes were responsible for the observed antitumor effect. Finally, the possible formation of a bridge between the IL-2R receptors on both, the lymphocytes and the INBL cells, mediated by the IL-2-bearing liposomes, and its possible effect on the activation of antitumor cytotoxic lymphocytes is discussed.

Papel de la diálisis peritoneal en el tratamiento de la insuficiencia cardíaca / Role of peritoneal dialysis in the treatment of heart failure / Papel da diálise peritoneal no tratamento da insuficiência cardíaca

Introducción. La insuficiencia cardíaca (IC) es una patología progresiva incluso con tratamiento adecuado. La eliminación de líquido ayuda en el manejo de estos pacientes. Evaluamos la eficacia de la diálisis peritoneal (DP) para tratar la IC refractaria. Pacientes y métodos. Estudio prospectivo no randomizado en pacientes con IC refractaria a dosis máximas toleradas de fármacos. Todos fueron tratados con DP. Se analizaron datos clínicos y estado funcional. Para determinar su eficacia se comparó el estado de salud percibido en los pacientes con DP respecto a los reportes de aquéllos que utilizaban tratamientos conservadores. Finalmente, se realizó un estudio de coste-utilidad. Resultados. Fueron incluidos 25 pacientes (68% varones, 66±10 años) y 16 siguieron en DP a la finalización del período de seguimiento (20±9 meses). Realizaron un solo intercambio nocturno 21 pacientes; el resto 2 ó 3 en relación al grado de insuficiencia renal. Todos mejoraron su estado funcional (4% tres grados, 64% dos y 32% uno; P<0,001); se redujo la presión sistólica en la arteria pulmonar (46±12 vs 25±9 mm Hg; P=0,007). Las tasas de hospitalización disminuyeron de forma importante (de 65±16 a 9±5 días/paciente/año; P=0,002). La expectativa de vida fue del 84% al año de tratamiento y del 72% y 58% tras 18 y 24 meses, respectivamente. La DP se asoció a mejor percepción del estado de salud que los tratamientos conservadores (0,6978 vs 0,4104; P<0,01). La DP es coste-efectiva comparada con los regímenes diuréticos. Conclusiones. La DP es una buena opción para el manejo del paciente con IC refractaria; mejora el grado funcional y la calidad de vida y reduce la hospitalización, mortalidad y los costes sanitarios.

Background. Heart failure (HF) is a progressive disorder even with adequate treatment. Fluid removal may aid in the management of these patients. We evaluated the efficacy of peritoneal dialysis (PD) in the treatment of refractory HF. Patients and methods. Prospective, non-randomized study involving patients with congestive HF refractory to maximum tolerable drug treatment. All of them were treated with PD. We analysed clinical data and functional status. To determine efficacy we compared the perceived state of health to PD patients respect to those reported with conservative therapies. Finally, we carried out a cost-utility evaluation. Results. Twenty-five patients (68% men, 66±10 years) were included and 16 were still undergoing PD at the end of the follow-up period (20±9 months). Twenty-one patients underwent only one daily nocturnal exchange; the rest, 2 or 3 exchanges due to different degrees of renal failure. All of them improved their NYHA functional status, (4% three classes, 64% two, 32% one; P<0.001), with a reduction in their pulmonary artery systolic pressure (46±12 vs 25±9 mm Hg; P=0.007). Hospitalization rates underwent a dramatic reduction (from 65±16 to 9±5 days/patient/year; P=0.002). PD life expectancy was 84% after 12 months of treatment, and 72% and 58% after 18 and 24 months. PD was associated with a higher perception state of health than the conservative therapy, (0.6978 vs 0.4104; P<0.01). PD is cost-effective compared with the diuretic regimens. Conclusions. PD is a good option for patients with refractory HF; it improves the functional status and quality of life, reduces morbidity, mortality and health care costs.

Introdução. A insuficiência cardíaca (IC) é uma patologia progressiva, mesmo com tratamento adequado. A remoção do líquido pode auxiliar no tratamento desses pacientes. Nós avaliamos a eficácia da diálise peritoneal (PD) no tratamento da insuficiência cardíaca refratária. Pacientes e métodos. Estudo prospectivo, não randomizado envolvendo pacientes com insuficiência cardíaca congestiva refratária ao tratamento medicamentoso máximo tolerado. Todos foram tratados com DP. Nós analisamos dados clínicos e estado funcional. Nós determinamos a eficácia, comparando o estado de saúde em PD em comparação com relação aos relatados com tratamento conservador. Finalmente, foi realizada uma avaliação de custo-utilidade. Resultados. Foram incluídos 25 pacientes (68% homens, 66±10 anos) e 16 estavam ainda em fase de PD no final do período de follow-up (20±9 meses). Vinte e um pacientes foram submetidos a apenas uma troca diária noturna, o resto, as trocas de 2 ou 3 devido a diferentes graus de insuficiência renal. Todos melhoraram seu status funcional da NYHA (4% três graus, 64% dois y 32% um; P<0,001); com uma redução da sua pressão sistólica da artéria pulmonar (46±12 vs 25±9 mm Hg; P=0,007). As taxas de hospitalização decresceram significativamente (de 65±16 a 9±5 dias/paciente/ano; P=0,002). A expectativa de vida era de 84% por ano de tratamento e 72% e 58% após 18 e 24 meses, respectivamente. PD foi associada com um estado de maior percepção de saúde do que o tratamento conservador (0,6978 vs. 0,4104; P<0,01). O DP é custo-utilidade em comparação com os regimes de diuréticos. Conclusàµes. O DP é uma boa opção para o manejo de pacientes com IC refratária, que melhora o estado funcional e qualidade de vida, reduz a internação, mortalidade e custos de saúde.

Efficacy of peritoneal ultrafiltration in the treatment of refractory congestive heart failure.

BACKGROUND: Heart failure (HF) is a major health problem in developed countries. HF is a progressive, lethal disorder, even with adequate treatment. There exists a vicious circle in the pathophysiology of HF that perpetuates and magnifies the problem. Concomitant fluid accumulation may worsen the congestive HF, it is responsible for numerous hospitalizations and it is an important cause of mortality. In this situation, any means of fluid removal may aid in the management of these patients. The objective of this study was to evaluate the efficacy of peritoneal dialysis (PD) in the treatment of refractory HF in terms of functional status, hospitalization and mortality. We also determined the improvement in health-related quality of life with the use of PD, and examined the economic consequences of its use. METHODS: We conducted a single centre, prospective, non-randomized study involving patients showing symptoms and signs of congestive HF refractory to maximum tolerable drug treatment. All of them were treated with PD. We analysed physical and biochemical determinations, functional status (according to the NYHA classification) and echocardiogram parameters. Also, to determine the efficacy of the technique we compared the perceived state of health (measured by the EQ5D) to PD patients respect to those reported with conservative therapies. Finally, we carried out a cost-utility evaluation measured by the incremental cost-utility ratio between these two options. RESULTS: Seventeen patients (65% men, 64 +/- 9 years) were included in the study, and 12 were still undergoing PD treatment at the end of the follow-up period (15 +/- 9 months). All patients improved their NYHA functional status (65% two classes; the rest, one; P < 0.001), with an important improvement in their pulmonary artery systolic pressure (44 +/- 12 versus 27 +/- 9 mmHg; P = 0.007), but no changes in left ventricular ejection fraction. Hospitalization rates underwent a dramatic reduction (from 62 +/- 16 to 11 +/- 5 days/patient/year; P = 0.003) before and after PD treatment. PD treatment raised life expectancy of 82% after 12 months of treatment, and 70% and 56% after 18 and 24 months, respectively, much better outcomes than those reported about conservative therapies, which only use diverse diuretic regimens. PD was associated with a higher perception state of health than the conservative therapy (0.6727 versus 0.4305; P < 0.01). Finally, we found that PD is cost-effective compared with the conservative therapy. CONCLUSIONS: We demonstrate that congestive HF programmes should consider offering PD in hope of seeing better functional status, reduced morbidity and mortality, better quality of life as well as reduced health care costs.

Study of red wine neuroprotection on astrocytes.

Phenolic composition of wine depends not only on the grape variety from which it is made, but on some external factors such as winemaking technology. Red wine possesses the most antioxidant effect because of its high polyphenolic content. The aim of this work is to study for the first time, the neuroprotective activity of four monovarietal Spanish red wines (Merlot (ME), Tempranillo (T), Garnacha (G) and Cabernet-Sauvignon (CS)) through its antioxidant ability, and to relate this neuroprotection to its polyphenolic composition, if possible. The wine effect on neuroprotection was studied through its effect as free radical scavenger against FeSO4, H2O2 and FeSO4 + H2O2. Effect on cell survival was determined by 3(4,5-dimethyltiazol-2-il)-2,5-diphenyltetrazolium reduction assay (MTT) and lactate dehydrogenase (LDH) release assay on astrocytes cultures. Results showed that most of the studied wine varieties induced neuroprotection through their antioxidant ability in astrocytes, Merlot being the most active; this variety is especially rich in phenolic compounds, mainly catechins and oligomeric proanthocyanidins. Our results show that red wine exerts a protection against oxidative stress generated by different toxic agents and that the observed neuroprotective activity is related to their polyphenolic content.

Identification of epitopes and immunodominant regions on the MICA protein defined by alloantibodies from kidney transplant patients.

BACKGROUND: Several reports showed a contribution of anti-MICA (major histocompatibility complex class I chain-related molecule A) antibodies (Abs) to the development of acute and chronic rejection. Identification of the epitopes to which the Abs bind may help to determine immunoreactive regions essential for the major histocompatibility complex compatibility between donor and recipients, leading to the best outcome of the transplant. METHODS: Sera from 284 kidney transplant patients were screened for anti-MICA Abs by Luminex assay. MICA allele typing of the recipients was determined. The epitopes of MICA were mapped by screening a synthesized library of overlapping peptides from the extracellular domains of the protein against the sera from kidney transplant patients with anti-MICA Abs. RESULTS: Antibodies against MICA were detected in 50 of 284 patients (17.6%) and correlated with the development of acute rejection. Nine antigenic regions were immunoreactive with anti-MICA Abs in the sera samples. Four of these continuous epitopes mapped to polymorphic amino acids (aa). Five antigenic regions were shared epitopes found in all the MICA alleles. The polymorphic residues, 173 (E/K), 175 (S/G), and 181 (R/T), had determined allele-specific epitopes (reactivity patterns 1 and 2). In contrast, the aa 208Y and 213T were implicated in the cross-reactivity among alleles. CONCLUSIONS: The presence of anti-MICA Abs could be an important marker for diagnosis because of their contribution to the outcome of the graft, regardless of presence of anti-HLA Abs. Additionally, the identification of epitopes revealed the in vivo antigens of the transplant and is spurring the development of new matching strategies to reduce the incidence of acute and chronic rejection.

CD127(low) expression in CD4+CD25(high) T cells as immune biomarker of renal function in transplant patients.

BACKGROUND: Noninvasive tests measuring cellular immunity could help predict immunologic risk and subsequent allograft dysfunction in transplant patients. CD25 is a promising marker of activation. Recent descriptions of CD127 expression as a discriminating factor between regulatory and activated T cells suggest its potential utility. METHODS: Expression of CD127 in CD4+CD25 T cells was analyzed by flow cytometry in peripheral blood from 62 renal transplanted patients and 30 healthy controls. Forty patients presented stable graft function and 22 suffered renal failure. RESULTS: Renal transplant patients showed higher levels of CD127(high) and a lower frequency of CD127(low) than healthy controls (0.63% vs. 0.29% [P<0.001] and 1.4% vs. 2.4% [P<0.001], respectively). However, high frequencies of not only CD127(high) but also CD127(low) showed a significant correlation with serum creatinine levels (P=0.012 and P=0.003, respectively). Allogenic stimulation in vitro increased the frequency of CD127(low) subset in a dose dependent manner. Furthermore, in patients with a high frequency of CD127(low) subset, this consisted mostly of FoxP3 negative cells, discarding their regulatory origin. Median frequency of CD127(low), but not CD127(high), cells showed significant differences between patients with stable function and with renal failure (P<0.005), with 16.7% and 53.1% of individuals above the median CD127(low) value (1.4%), respectively. CONCLUSION: Quantification of CD127(low) subset through staining of CD4+ T cells with the combined markers CD127/CD25/CD45RO has been demonstrated to be a significant tool for monitoring the outcome course of renal transplant patients.

Characterization of cationic liposomes having IL-2 expressed on their external surface, and their affinity to cervical cancer cells expressing the IL-2 receptor.

Anionic, cationic, and neutral liposomes were constructed to contain IL-2 in order to evaluate their affinity to a cervical cancer cell line (INBL) and to determine whether they can present IL-2 on their external surface. When these liposomes were co-cultured with INBL, the anionic liposomes were the only ones found to be cytotoxic. When neutral and cationic liposomes were constructed in the presence of IL-2, IL-2 was detected only on the surface of cationic liposomes by using a fluorescent anti-IL-2 antibody. By co-culturing INBL with IL-2-containing cationic liposomes, and by using fluorescent anti-IL-2 antibody, we found a strong IL-2 presence on the cell membranes thus suggesting a high affinity of the liposomes to the INBL cells.

The utility of a specific measure for heart transplant patients: reliability and validity of the Kansas City Cardiomyopathy Questionnaire.

BACKGROUND: Health-Related Quality of Life of patients with heart transplantation is an important variable; however, it has received little attention so far, and only two Spanish validated measurement instruments are available. The aim of our study was to validate the Spanish version of the Kansas City Cardiomyopathy Questionnaire (KCCQ) in heart transplant patients. METHODS: A prospective study was performed in 186 patients awaiting heart transplantation in nine transplant hospitals. Hundred transplant recipients filled out the KCCQ, the Euroqol 5-D (EQ5D), and the Short Form-36 (SF-36) Health Survey at pretransplant, after 3 months, 6 months, and 1 year of follow-up. A complete set of sociodemographic and clinical data were also collected. The validity, reliability, sensitivity to change, and effect size were studied. Two questionnaires, the SF-36 and EQ5D, were used to evaluate the validity. RESULTS: Mean age of patients was 56.0 years, and 80.5% were men. Twenty-six percent had acute rejection. A five-dimensional factorial structure could be discerned. The questionnaire presented a Cronbach's alpha coefficient of more than 0.7. Correlations between the KCCQ and the other questionnaires and clinical variables were satisfactory. CONCLUSIONS: The KCCQ features adequate psychometric properties. The KCCQ offers several advantages over other questionnaires because it quantifies symptoms (frequency, severity, and stability) and it is much more sensitive to change, even when compared with the SF-36. The specific questionnaire for heart transplant patients is a useful and user-friendly instrument for measuring the Health-Related Quality of Life related to functional status, quality of life, and social limitation more accurately.