Detection of coca alkaloids in oral fluid from coca leaf (tea) consumers: using solid phase extraction to improve validation parameters and widen the detection window.

Hygrine and cuscohygrine, two coca leaf alkaloids, have been previously proposed as markers to differentiate legal and illegal cocaine consumption. This is a very common problem in some countries of South America, where the consumption of coca leaves has a long tradition. Analytical methods focusing on the assessment of coca leaf alkaloids, such as cuscohygrine, hygrine, tropacocaine and t-cinnamoylcocaine, in oral fluid are virtually non-existent in forensic toxicology laboratories worldwide due to their lack of application. However, the problem of differentiating legal and illegal cocaine use in criminal justice, DUID (drug-impaired driving) and WDT (workplace drug testing) programs is growing. Therefore, researchers are obliged to develop methods to measure coca leaf alkaloids (cuscohygrine, hygrine and t-cinnamoylcocaine) in biological matrices for further validation for routine analyses in forensic toxicology laboratories. This work aims to optimize a previously published separation method by protein precipitation in oral fluid by using solid-phase extraction (SPE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) operating in multiple reaction monitoring (MRM) mode. The use of SPE allowed the matrix effect and the background to be reduced in the chromatograms due to the obtained cleaner extracts. Consequently, improved detection and quantification limits were reached. Findings showed that the detection windows for coca leaf alkaloids were longer than three hours in real oral fluid samples from volunteers who drank a cup of coca tea. These detection windows are quite higher than those previously obtained when using the method based on separation by protein precipitation.

Understanding the hematopoietic microenvironment in chronic myeloid leukemia: A concise review.

Chronic myeloid leukemia (CML) is a myeloproliferative disease that results from the BCR-ABL gene-induced transformation of a primitive hematopoietic cell. This disease has been extensively studied, and, as a result, a very effective therapy has been developed: the tyrosine kinase inhibitors. Although, there is a significant knowledge about the intrinsic biology of CML cells, alterations in their bone marrow microenvironment are not yet completely understood. In this concise review, we summarized recent findings on the composition and function of the bone marrow microenvironment in CML, and their importance in the progression of the disease and treatment resistance.

Perfil de alcaloides de la hoja de coca en el fluido oral de un mascador de hoja de coca y un bebedor de té de coca: Estudio preliminar / Coca alkaloids profile in oral fluid from people chewing coca leaves and drinking coca tea: Preliminary study

Actualmente el fluido oral (FO) es aceptado como una matriz biológica alternativa para detectar drogas en toxicología clínica y forense. En países como Argentina donde el uso de hojas de coca (mascar hojas de coca o beber té de coca) es legal son necesarios procedimientos adecuados para logar una clara diferenciación entre los individuos que usan las hojas de coca de manera legal de aquéllos que usan cocaína en forma ilegal. Poca es la información que hay en la literatura sobre el perfil de los alcaloides de la hoja de coca en FO de personas que mascan hojas de coca o toman té de coca y hasta el presente trabajo no se hallaron datos sobre el perfil en FO de la higrina (HIG) y cuscohigrina (CUS). De este estudio preliminar participaron dos voluntarios. Los resultados mostraron que la CUS e HIG siguieron siendo positivas después que la cocaína (COC) y benzoilecgonina (BE) cayeron por debajo de los valores cut- off propuestos por las guías internacionales para FO en casos de screening (15 a 20 ng/ mL) y de confirmación (8 a 10 ng/mL) en el caso del mascador de coca. En el participante que tomó una taza de té de coca, en el último punto examinado (1 h) resultó ser positivo para la COC y BE y también para la CUS e HIG. El FO podría ser una muestra útil para confirmar el uso legal de la hoja de coca, aun cuando futuros estudios son necesarios para corroborar estos primeros datos.

Nowadays oral fluid (OF) is accepted as an alternative biological sample for detecting drugs in clinical and forensic toxicology. In countries like Argentina, where the use of coca leaves (coca leaves chewing and coca tea drinking) is legal, adequate procedures are required to allow a clear differentiation between people who use coca leaves (legal practice) and those who use cocaine (illicit practice). There is scarce literature regarding coca leaf alkaloids profile in OF from people who chew coca leaves and drink coca tea. Until now, coca leaf alkaloids profile of hygrine (HYG) and cuscohygrine (CUS) in OF were not described in the literature. The current preliminary study was performed with two healthy volunteers. In this research CUS and HYG have been found to be positive (detectable) even when cocaine (COC) and benzoylecgonine (BE) are dropped below the cut-off values proposed by international guidelines for screening (15 to 20 ng/mL), and confirmation (8 to 10 ng/mL) in OF. In addition, CUS and HYG were also found to be positive at the same time of the last detection of COC and BE after coca tea consumption. The OF would be a useful sample to confirm the legal use of coca leaf, even when more researches are therefore needed.

La ley de salud mental en Inglaterra / The mental health act in England

Introduction: People can become extremely vulnerable during episodes of severe mental ill health. Their ability to protect themselves can become impaired. They can also pose significant risks to the safety of others through their erratic or hostile behaviour. It could be actually argued that it is precisely during a mental health crisis when explicit legal safeguards are of critical importance for the protection of people's rights, including their dignity and autonomy. Method: An outline is provided of how the Mental Health Act operates in England, with particular reference to involuntary admissions to hospital as well as the right of appeal through the Mental Health Review Tribunal. Results: The roles of medical professionals, Approved Mental Health Professionals, the Nearest Relative as well as the type of evidence that professionals must submit to the Mental Health Review Tribunal and how this evidence is challenged are discussed. Conclusions: The independent legal review of the involuntary detention of a person in hospital is discussed both regarding the protection of human rights it provides as well as in terms of its impact on promoting higher standards of care.

Introducción: Las personas afectadas de un episodio severo de enfermedad mental pueden encontrarse en un estado de alta vulnerabilidad, incluyendo el ser incapaces de protegerse. Durante estos episodios, dichas personas pueden también suponer un riesgo significativo para otros debido, por ejemplo, a una conducta errática u hostil. En este artículo se arguye que es durante estos períodos de crisis cuando se requiere de disposiciones legales explícitas que protejan los derechos, la dignidad, y la autonomía de dichas personas. Métodos: Se provee de un resumen de cómo funciona la Ley de Salud Mental en Inglaterra, con particular referencia a la hospitalización involuntaria así como al derecho de apelación que los pacientes tienen contra dicha hospitalización, la que es vista por el Tribunal de Salud Mental. Resultados: Se discuten los roles de los médicos, de los denominados Profesionales Aprobados de Salud Mental (AMHP) y del Familiar Más Cercano (Nearest Relative), así como el tipo de evidencia que todos los profesionales deben presentar ante el Tribunal de Salud Mental y cómo dicha evidencia es cuestionada en detalle tanto por el abogado del paciente como por los miembros del Tribunal de Salud Mental. Conclusión: Se discute la importancia del hecho de que la hospitalización involuntaria es sometida a una revisión legal independiente de la profesión médica. En particular, se hace referencia al efecto de dicho derecho de apelación en términos tanto de la protección de los derechos humanos de los pacientes así como de la mejoría de los estándares de la calidad de los servicios de psiquiatría.

La enseñanza de la educación para la salud: ¿una confrontación a la teoría y la práctica de la salud pública como disciplina? / Teaching education for health: ¿a confrontation with the theory and practice of public health as a discipline?

Objetivo: comprender la enseñanza de la educación para la salud (EpS) en la Facultad Nacional de Salud Pública (fnsp) de la Universidad de Antioquia. Metodología: entrevistas grupales e individuales a profesores, estudiantes y egresados, así como con la revisión de los planes de estudio de los programas académicos de la Facultad. Resultados: es predominante la enseñanza de la EpS como herramienta o estrategia, concebida desde una perspectiva biomédica y fundada en un modelo pedagógico tradicional/coactivo; además, que la enseñanza de la EpS en la fnsp no tiene la relevancia que se esperaría, lo que aparentemente supone una contradicción. Discusión: La reflexión teórica realizada propone una salida diferente: esta situación, lejos de ser una contradicción, corresponde a una consecuencia lógica para una perspectiva de salud pública imperante en la fnsp. El papel que cumple la educación en la salud pública depende de la corriente en la cual se inscribe. Se presentan otras versiones de salud pública en las cuales la educación tiene un sentido básico y fundante y se plantea desde modelos pedagógicos alternativos. Así, abordar la enseñanza de la EpS y definir el perfil del salubrista no es un asunto neutral ni técnico para incluir ciertos contenidos; corresponde al producto de un debate teórico y político sobre la salud pública que debe orientar los currículos de los programas académicos y, en general, las acciones de la Facultad.

Objective: to understand the manner in which health education (HE) is taught at the National Faculty of Public Health (NFPH) of the University of Antioquia in Medellín, Colombia. Methodology: group and individual interviews with teachers, students and graduates. Likewise, the curricula of the Faculty's academic programs were reviewed. Results: The predominant approach is to teach HE as a tool or strategy conceived from the biomedical standpoint and grounded on a traditional/coercive pedagogical model. Furthermore, the teaching OF HE at the NFPH does not have the relevance that should be expected, which is a contradiction in itself. Discussion: the theoretical reflection developed proposes a different explanation: it is not a contradiction; rather this situation pertains to a logical consequence of a public health perspective prevailing at the NFPH. The role of health education depends on the public health approach with which it is aligned. Other public health approaches are presented in which education has a basic and foundational sense and it is conceived from the point of view of alternative pedagogical models. So addressing HE teaching and defining the profile of public health professionals is not a neutral and technical issue for including certain topics. It is rather a consequence of a theoretical and political debate about the type of public health that should guide the curricula of academic programs and actions at the Faculty.

Relationship between SNPs and expression level for candidate genes in rheumatoid arthritis.

OBJECTIVES: The study of polymorphisms of genes differentially expressed may lead to the identification of putative causal genetic variants in multifactorial diseases such as rheumatoid arthritis (RA). Based on preceding transcriptomic results, we genotyped 10 single nucleotide polymorphisms (SNPs) belonging to six genes (S100A8, RNASE2, PGLYRP1, RUNX3, IL2RB, and LY96) showing the highest fold change (> 1.9) when level of expression was compared between RA patients and controls. These SNPs were then analysed to evaluate their role in RA. METHOD: The relationship between gene expression and genotypes of SNPs was first investigated by Kruskal-Wallis and Mann-Whitney tests in RA patients and controls. The genetic association of these SNPs with RA were then analysed using family-based association tests in trio families. RESULTS: We found that RNASE2 gene expression was related to rs2013109 genotypes in 14 RA patients (p = 0.030). The association study in a discovery sample of 200 French trio families revealed a significant association with RA for one SNP, PGLYRP1-rs2041992 (p = 0.019); this association was stronger in trios where RA patients carried the HLA-DRB1 shared epitope (SE) (p = 0.003). However, this association was not found in a replication sample of 240 European trio families (p = 0.6). CONCLUSIONS: Family-based association tests did not reveal an association between RA and any SNP of the candidate genes tested. However, RNASE2 gene expression was differentially expressed in RA patients considering a sequence polymorphism. This result led us to highlight the potential disease-specific regulation for this candidate gene in RA.

Speciation of iodine-containing proteins in Nori seaweed by gel electrophoresis laser ablation ICP-MS.

An analytical approach providing an insight into speciation of iodine in water insoluble fraction of edible seaweed (Nori) was developed. The seaweed, harvested in the Galician coast (Northwestern Spain), contained 67.7±1.3 µg g(-1) iodine of which 25% was water soluble and could be identifies as iodide. Extraction conditions of water insoluble residue using urea, NaOH, SDS and Triton X-100 were investigated. The protein pellets obtained in optimized conditions (after precipitation of urea extracts with acetone), were digested with trypsin and protease XIV. Size exclusion chromatography-ICP-MS of both enzymatic digests demonstrated the occurrence of iodoaminoacids putatively present in proteins. Intact proteins could be separated by gel electrophoresis after an additional extraction of the protein extract with phenol. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) with laser ablation ICP-MS detection of (127)I indicated the presence of iodine in protein bands corresponding to molecular masses of 110 kDa, 40 kDa, 27 kDa, 20 kDa and 10 kDa. 2D IEF-SDS PAGE with laser ablation ICP-MS (127)I imaging allowed the detection of 5 iodine containing protein spots in the alkaline pI range.

Control de la concentración de metilmercurio en productos pesqueros para el consumo humano mediante el empleo de polímeros de impronta molecular y de nanopartículas derivatizadas / Control of the methylmercury concentration in fish and seafood products for human consumption using molecularly imprinted polymers (MIPs) and derivatized nanoparticles

Objetivo: Desarrollar un polímero de impronta molecular para la determinación de metilmercurio en muestras de alimentos de origen marino. Material y métodos: Se sintetizó un polímero de impronta molecular (MIP) que emplea fenobarbital como ligando no vinilado para unirse al metilmercurio. Se utilizó la técnica de polimerización por precipitación. El MIP sintetizado fue empleado para funcionalizar nanopartículas magnéticas con el fin de mejorar el proceso de extracción del metilmercurio de las muestras. Resultados: El MIP con fenobarbital fue aplicado para la determinación de la concentración de metilmercurio en dos materiales de referencia de concentración certificada para el análisis de muestras de alimentos marinos (BCR-463, tejido de atún, y TORT-2, hepatopáncreas de langosta). Los valores de concentración experimentales obtenidos fueron de 3,04±0,16 μg/g y 0,152±0,013 μg/g respectivamente, coincidiendo con los valores certificados de forma estadísticamente significativa (test t, 95% confianza). Conclusiones: Se ha sintetizado un MIP para la determinación de la concentración de metilmercurio a bajas concentraciones en muestras de alimentos de origen marino (AU)

Objective: The development of a molecularly imprinted polymer support for methylmercury determination in fish and seafood samples. Materials and methods: A molecularly imprinted polymer was synthesized using the precipitation technique and phenobarbital as a non-vinylated ligand for binding methylmercury. The synthesized MIP was used for the functionalization of magnetic nanoparticles in order to improve the process of methylmercury extraction from the samples. Results: The MIP prepared with phenobarbital was applied to the determination of methylmercury concentration in two certified reference materials used for fish and seafood samples analysis (BCR-463, tuna fish, and TORT-2, lobster hepatopancreas). The experimental values (3.04±0.16 μg/g and 0.152±0.013 μg/g respectively) and the certified values showed no statistically significant difference (t test, 95% confidence level). Conclusions: We have synthesized a MIP for determining methylmercury in fish and seafood samples at low concentrations (AU)

Functionalized gold nanoparticles for the detection of arsenic in water.

Gold nanoparticles are attractive as sensing materials because of their size and shape are related with their optical properties. The color change produced by the aggregation of functionalized AuNPs allows the detection of arsenic at low levels. A simple, cheap and fast analytical procedure to perform arsenic determination using functionalized gold nanoparticles (AuNPs) and VIS spectrometry as a detection technique is studied. Three different synthesis procedures to obtain the AuNPs and two different functionalization modes were studied. AuNPs functionalized with GSH-DTT-CYs-PDCA were selected as the most adequate. The correlation between the decrease in the absorbance signal and the arsenic concentration was good in the 2-20 µg l(-1)interval. Repeatability, expressed as average of RSD (%), obtained for the different arsenic concentrations studied was 0.6%. The average value of the analytical recovery was 99.7%. The detection and quantifications limits were 2.5 and 8.4 µg l(-1) respectively. These limits are sufficient to detect World Health Organization's guideline value of 10 µg l(-1).

Comparison of two lab-made spray chambers based on MSIS™ for simultaneous metal determination using vapor generation-inductively coupled plasma optical emission spectroscopy.

The objective of this study is to evaluate the performance of two lab-made systems based on the Multimode Introduction System (MSIS™) and the modified MSIS™, to generate and introduce vapors of Ag, Cu, Cd, Cu, Ni, Sn, Zn, and also Au in the ICP torch. An univariate procedure was used to select the optimized working conditions (Ar flow, sample, reductant and waste flows, and reagent concentrations). Optimum conditions for working with modified MSIS were: nitric acid concentration 0.35 M, 8-hydroxyquinoline concentration: 40 mg L(-1), sodium borohydride concentration: 1.75% (w/v)+0.4% (w/v) NaOH, argon purge flow to sweep the vapors to the torch: 1.2 L min(-1), sample flow and sodium borohydride flows: 2.3 L min(-1); waste flow: 7.7 mL min(-1). For the optimum working conditions for lab-made MSIS in dual mode the concentration of 8-hydroxyquinoline was 225 mg L(-1), the Ar purge flow was 0.75 L min(-1), and the conventional nebulization flow was 2.3 L min(-1). The sensitivity obtained was higher using the lab-made MSIS than using the lab-made modified MSIS or a forced outlet gas-liquid separator. The limits of detection were better for Au, Cd, Sn than those obtained using conventional nebulization; the measurements were precise (RSDs≤5% in dual mode) and a good accuracy was obtained in the determination of Cd, Cu, Ni and Zn in a wastewater reference material using aqueous calibration and the lab-made MSIS in dual mode.

Preliminary findings on the antimony levels of Quiroga river water in the vicinity of a long-abandoned stibnite mine.

A UV-Vis spectrophotometric method was developed as a preliminary approach to the determination of antimony in water samples from a river that flows very close to an abandoned mining site. The analyte is complexed with ammonium pyrrolidinedithiocarbamate and absorbance of the complex is measured at 291.06 nm. The standard additions method is mandatory in view of the matrix effect observed, and the response is linear at least up to 9.3 microg/mL of antimony. The sensitivity of the method is 2.609 x 10(-2) mL/microg, whereas the limits of detection and quantification are, respectively, 0.2 and 0.6 microg/mL. The repeatability, expressed as mean relative standard deviation of the measurements within the calibration range, is 2.0%, whereas the repeatability of the entire procedure is 0.3%. The mean analytical recovery within the calibration range was 102.6%. The method was successfully applied to river water samples.

Consumo de calcoo: evolución y situación actual / The evolution and current situation of calcium consumption

El calcio es un mineral esencial en la nutrición humana, contribuye a la formación de masa ósea, interviene en funciones metabólicas y en la contracción muscular. Debido a la importancia de este nutriente en la salud pública, se han realizado estudios en varios países, entre ellos Colombia, tendientes a determinar su consumo en diferentes grupos poblacionales, encontrando en la mayoría de ellos una ingesta insuficiente, que cubre menos del 50% de la recomendación, independientemente del desarrollo económico. Las Encuestas de la Situación Nutricional en Colombia (2005, 2010) reportaron déficit en el consumo de lácteos, especialmente en población de bajo estrato socioeconómico y en las regiones Pacífica, Amazonía y Orinoquía. Recientemente el Institute of Medicine (IOM) estableció la ingesta diaria recomendada de calcio para cubrir las necesidades fisiológicas de la población. Esta medida es necesaria teniendo en cuenta que la evidencia científica señala una fuerte asociación entre baja ingesta y disminución de la densidad mineral ósea con incremento de la osteoporosis y de las fracturas asociadas. El IOM, también estableció los niveles máximos tolerables (UL) para evitar problemas asociados al exceso con el uso de suplementos. En todos los países, las guías alimentarias tienen como objetivo promover una alimentación saludable, por lo tanto la divulgación de este instrumento es responsabilidad de todos los profesionales de la salud, especialmente de los nutricionistas. Se requiere fortalecer la investigación y desarrollar estrategias que involucren diferentes sectores para mejorar la ingesta de calcio y la salud ósea de la población.

Calcium is an essential mineral in human nutrition; it contributes towards bone formation, intervenes in metabolic functions and muscular contraction. Studies have been carried out in several countries, including Colombia, due to this nutrient's importance in public health; they have usually determined its consumption in different groups of the population, inadequate intake being found in most of them, covering less than 50% of the recommended intake, regardless of economic development. Nutritional situation surveys (Encuestas de la Situación Nutricional) in Colombia (2005, 2010) have reported a deficit in dairy product consumption, especially in low socioeconomic strata and people living in Colombia's Pacific, Amazonian and Orinoquia regions. The Institute of Medicine (IOM) has recently established the recommended daily calcium intake for covering the population's physiological needs; this measure is needed bearing in mind that the scientific evidence has revealed a strong association between low intake and reduced bone mineral density with increased osteoporosis and associated fractures. The IOM has also established maximum tolerable levels (UL) for avoiding excess- and supplement useassociated problems. Dietary guidelines in all countries are aimed at promoting healthy eating habits; disclosing/ disseminating such instrument is thus the responsibility of all health professionals, especially nutritionists. Research must be strengthened and strategies developed involving different sectors to improve the population's calcium intake and bone health.

[Epidemiology of injuries in a male college indoor soccer team]. / Epidemiología de las lesiones en un equipo varonil de fútbol rápido universitario.

BACKGROUND: So far there are no reports on the incidence of injuries in indoor soccer and their characteristics, so we decided to find out what happens in a college team of this sport. MATERIAL AND METHODS: The physician in a team recorded the injuries that occurred during the trainings and games from April to December 2009. The number of exposure hours of players was determined. The frequencies and incidence per exposure were calculated. RESULTS: A total of 62 injuries were recorded. The incidence during games was 8.97 injuries/100 hours, and during trainings 0.88 injuries/100 hours. Midfielders and defenders had the highest incidence of injuries, with 1.50 and 1.29 injuries per 100 hours of exposure, respectively. Eighty percent of the injuries involved the pelvic limbs and 11% the thoracic limbs. Sprains represented 25% of the injuries, followed by contusions and muscle injuries, with 23 and 18%, respectively. Ninety-two percent of the injuries were mild, 6.5% moderate and 1.5% severe. CONCLUSIONS: There are similarities with other studies; however, it is difficult to compare the studies reported on soccer and indoor soccer with this one due to the differences in collection methods, population and participants. The strength of this study is that a single physician made the diagnosis and follow-up, care was provided immediately and all the injuries that occurred in the team were recorded. This leads to continue conducting studies on the mechanisms and causes of injuries, which may be used to implement preventive measures.

Gasto energétic en reposo y composición corporal en adultos / Energy expenditure in repose related to body composition in adults

El requerimiento de energía de una persona está relacionado con su gasto energético (GE) y se define como la energía que consume un organismo, está representado por la tasa metabólica basal (TMB), la actividad física (AF) y la termogénesis inducida por la dieta (TID). La TMB es la mínima cantidad de energía que un organismo requiere para estar vivo y representa del 60-70% del total del gasto energético (TGE), en la mayoría de los adultos sedentarios. La AF representa entre el 25-75% del TGE y la TID representa cerca del 10% del TGE. Debido a su simplicidad, bajo costo y alta precisión el método comúnmente usado en el estudio de la composición corporal es la antropometría.

People's energy requirements are related to their energy expenditure (EE); this is defined as being the energy which an organism consumes. It is represented by the basal metabolic rate (BMR), physical activity (PA) and dietinduced thermogenesis (DIT). The BMR is the minimum amount of energy which an organism requires to stay alive and represents 60%-70% of total energy expenditure (TEE) in most sedentary adults. The PA represents 25%- 75% of TEE whilst DIT represents around 10% of TEE. Anthropometry is the method most frequently used in studying body composition due to its simplicity, low cost and great precision.

Direct LC-ES-MS/MS determination of phthalates in physiological saline solutions.

A method for determining a group of phthalic esters (PAEs) in physiological saline solutions has been developed. The PAEs studied were dimethyl phthalate, diethyl phthalate, butyl benzyl phthalate and dibutyl phthalate. These groups of phthalates were determined by liquid chromatography-electrospray ionization-tandem mass spectrometry, working in positive ion mode. The compounds were separated by liquid chromatography working in gradient mode with acetonitrile-ultrapure water as a mobile phase. The separation was performed starting with 5% of acetonitrile and increasing to 75% in 5 min, followed by isocratic elution for 8 min. The method was precise (with relative standard deviation (RSD) from 1.0 to 6.8%) and sensitive, with LODs of 0.05, 0.38, 0.05 and 0.82 µg L⁻¹ for DMP, DEP, BBP and DBP, respectively. The proposed analytical method has been applied to determine these compounds in different physiological saline solutions commercialized in plastic bottles.

Estrategias de optimización del trasplante hepático / No disponible

No disponible

Type I interferons might form the link between Toll-like receptor (TLR) 3/7 and TLR4-mediated synovial inflammation in rheumatoid arthritis (RA).

BACKGROUND: Rheumatoid arthritis (RA) has been associated with an increased risk of infections, but the underlying pathways have not yet been identified. Toll-like receptors (TLR) probably play a role in synovial inflammation and may also contribute to the understanding of the role of infections in RA. OBJECTIVES: To investigate if the synovial expression of TLR3 and TLR7 in RA correlates with that of inflammatory cytokines, and to assess whether this has functional consequences for local cytokine production and to study potential links between the TLR3/7 axis and TLR4 in RA synovium. METHODS: Immunohistochemistry was used to study the expression of TLR3, TLR7, interferon alpha (IFNalpha), tumour necrosis factor alpha (TNFalpha) and interleukins IL1beta, IL12, IL17 and IL18 in RA synovium obtained by arthroscopy from 34 patients with RA. Monocytes, monocyte-derived dendritic cells (MoDCs) and RA synovial fibroblasts were stimulated via TLR3 (poly-IC) and TLR7 (loxorubin), after which IL1beta, IL6 and TNFalpha were measured by Luminex bead array technology. Following preincubation with IFNalpha, IL1beta and IL18, TLR3 and TLR7 mRNA expression was assessed using real-time PCR. Cytokine production after preincubation with IFNalpha and subsequent TLR stimulation was measured. RESULTS: Synovial TLR3/7 expression was co-expressed with IFNalpha, IL1beta and IL18, but not with TNFalpha, IL12 and IL17. Stimulation of TLR3/TLR7 on monocytes, MoDCs or synovial fibroblasts led to secretion of type I IFN but no biologically active IL1beta or IL18 could be detected. Type I IFNalpha increased TLR3/7 mRNA expression whereas IL1beta and IL18 did not. In spite of the fact that the mRNA level of TLR4 remained unchanged, IFNalpha enhanced the response to TLR4 agonists, a phenomenon that was clearly more marked in patients with RA. CONCLUSION: Type I interferons are highly co-expressed with TLR3/TLR7 in RA synovium. They enhance TLR3/TLR7-mediated cytokine production and also TLR4-mediated responses.

Anti-inflammatory therapy with tumour necrosis factor alpha inhibitors improves high-density lipoprotein cholesterol antioxidative capacity in rheumatoid arthritis patients.

OBJECTIVE: High-density lipoprotein (HDL) antiatherogenic functions seem to be diminished during inflammatory conditions such as rheumatoid arthritis (RA). The aim of this study was to investigate the effects of tumour necrosis factor (TNF) inhibition on the antioxidative capacity of HDL in RA. METHODS: Plasma lipids and paraoxonase (PON-1) activity were investigated in 45 RA patients, before and during 6 months of anti-TNF therapy. In addition, HDL was isolated and tested for its ability to inhibit copper-induced oxidation of low-density lipoprotein in vitro. RESULTS: Plasma HDL concentrations did not change considerably after 6 months of therapy. However, stable increases of PON-1 activities were observed throughout the same period (p<0.03). The increases were more obvious when related to HDL or apolipoprotein AI concentrations. HDL total antioxidative capacity significantly improved 6 months after the initiation of anti-TNF therapy (p = 0.015). The initial improvement of PON-1 activity paralleled a decrease in the inflammatory status, whereas specific TNF blockade was likely to be responsible for the long-term effects. CONCLUSIONS: Anti-TNF therapy with infliximab has beneficial effects on lipids through changes in HDL antioxidative capacity, which might be clinically relevant and contribute to the reported protective effect of anti-TNF on cardiovascular morbidity in RA. This emphasises the importance of HDL antiatherogenic capacity for cardiovascular risk in chronic inflammatory conditions.