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The yield and composition of essential oils from leaves, stems, umbel rays and also from whole aerial parts of Bupleurum gibraltarium Lam. (Apiaceae) were determined during the pre-flowering, full flowering, late flowering, and fruiting vegetative periods. A fruit sample was also studied for comparative purposes. Leaves (0.8-1.8% yield) and stems (0.3-0.7%) reached their maximum oil content during the late flowering period, whereas umbel rays (2.2-3.7%) reached it in full flowering. Oil samples were analyzed by capillary GC and GC/MS: components which were found in amounts greater than 2% were selected to carry out a seasonal study. In the leaf oils, sabinene (12.0-33.9%) and limonene (7.8-23.4%) were the main components, the sabinene level being minimum in full flowering and maximum in fruiting. In stem oils, sabinene (4.7-21.6%) and 2,3,4-trimethylbenzaldehyde (9.3-13.6%) were the main components, the sabinene level being minimum in pre-flowering and maximum in full flowering. In umbel ray oils, sabinene (20.7-43.1%) was the first component in all the phenological periods, followed by a-pinene (7.3-28.2%). Both monoterpenes increased their levels in late flowering and reached minimum amounts in fruiting. [ABSTRACT FROM AUTHOR]

Mexico harbors great cultural and ethnic diversity, yet fine-scale patterns of human genome-wide variation from this region remain largely uncharacterized. We studied genomic variation within Mexico from over 1000 individuals representing 20 indigenous and 11 mestizo populations. We found striking genetic stratification among indigenous populations within Mexico at varying degrees of geographic isolation. Some groups were as differentiated as Europeans are from East Asians. Pre-Columbian genetic substructure is recapitulated in the indigenous ancestry of admixed mestizo individuals across the country. Furthermore, two independently phenotyped cohorts of Mexicans and Mexican Americans showed a significant association between subcontinental ancestry and lung function. Thus, accounting for fine-scale ancestry patterns is critical for medical and population genetic studies within Mexico, in Mexican-descent populations, and likely in many other populations worldwide. [ABSTRACT FROM AUTHOR]

Objective The mitochondrion is known to be important to chondrocyte survival. This study was undertaken to analyze protein expression profiles in chondrocyte mitochondria that are affected by interleukin-1β (IL-1β). Methods Normal human chondrocytes were isolated from knee cartilage obtained at autopsy from subjects with no history of joint disease. Cells were incubated for 48 hours with or without IL-1β (5 ng/ml). Proteins were separated by 2-dimensional electrophoresis and stained with Sypro Ruby, Coomassie brilliant blue, or silver. Qualitative and quantitative analyses were carried out using PDQuest software. Proteins were identified by mass spectrometry using matrix-assisted laser desorption ionization-time-of-flight/time-of-flight technology. The proteomic results were validated by real-time polymerase chain reaction, Western blotting, and microscopy. Nitric oxide (NO) was quantified using Griess reagent. Results Comparative analysis revealed differential expression of signal transduction proteins that regulate cytoskeleton, transcription, metabolic, and stress-related pathways. In total extracts, dimethylarginine dimethylaminohydrolase 2 (DDAH-2) did not show any change in expression after stimulation with IL-1β. However, in mitochondrial extracts, DDAH-2 expression was significantly increased after exposure to IL-1β. Conventional immunofluorescence and confocal microscopy revealed the presence of DDAH-2 in the mitochondria of IL-1β-stimulated chondrocytes. These results were reproducible in cartilage explants treated with IL-1β. In addition, we demonstrated that inhibition of the expression of DDAH-2, as well as interruption of its translocation to the mitochondria, reduced the NO production induced by IL-1β. DDAH-2 protein expression was higher in osteoarthritic (OA) cartilage than in normal cartilage. Conclusion In the present study, the presence of DDAH-2 in normal human chondrocytes and cartilage was identified for the first time. DDAH-2 could play an important role in IL-1β-induced NO production and in OA pathogenesis. [ABSTRACT FROM AUTHOR]

This article discusses a study on the contribution of mitochondrial DNA (mtDNA) haplogroups to the prevalence and severity of knee osteoarthritis (OA). The study used the single-base extension assay and the polymerase chain reaction-restriction fragment length polymorphism technique in the examination. It showed a decreased risk of knee OA in individuals carrying haplogroup J.

El uso de cohortes de osteoartritis bien caracterizadas es obligatorio para estudiar y profundizar en el conocimiento en esta enfermedad. En España no existe actualmente ninguna cohorte prospectiva en este ámbito; por ello, el objetivo de este trabajo es describir la primera cohorte de osteoartritis en España: la PROCOAC (PROspective COhort of A Coruña). El Servicio de Reumatología del Hospital Universitario de La Coruña inició un estudio de seguimiento prospectivo en el año 2006. Los criterios de inclusión fueron: a) pacientes mayores de 55 años a los que se les realizó una radiografía abdominal que permitiese estudiar ambas caderas; b) pacientes diagnosticados de osteoartritis radiográfica de mano según los criterios ACR; c) pacientes diagnosticados de osteoartritis radiográfica de rodilla o cadera según los criterios ACR. Se realizó seguimiento cada 2 años y se recogió información clínica, analítica, genética y radiográfica. La cohorte consta de 937 individuos; 873 tienen osteoartritis radiográfica de rodilla, 783 de cadera y 679 de mano. La edad media de la población es 63,9 ± 8,9 años y el IMC promedio de 29,6 ± 5,1. Más de la mitad de la población tiene hipertensión arterial y el 17%, diabetes. La osteoartritis predominante en la mano es la nodular (78,1%), seguida de la rizartrosis (55,3%) y la erosiva (18,4%). El 21,4% y el 43,1% tienen sanas la rodilla y la cadera, respectivamente. Se observa un grado 1 en el 26% y 37%; un grado 2 en el 26,7% y 11,5%; un grado 3 en el 14,9% y 4%; y un grado 4 en el 9,4% y 3,7%, respectivamente. El 44,1% de la población tiene una articulación afectada, el 39,9% tiene 2 y el 13,4% tiene las 3 articulaciones afectadas. La edad (OR = 1,11; p < 0,001), el IMC (OR = 1,11; p = 0,002) y el WOMAC total (OR = 1,03; p = 0,005) son los únicos factores de riesgo si comparamos la afectación de una sola ubicación frente a 3. También se detectó una discrepancia entre el dolor y el daño radiográfico articular en pacientes con grado KL ≤ 2, de modo que un porcentaje significativamente mayor de pacientes con osteoartritis de rodilla experimentaron dolor (66,1%) en comparación con los pacientes de cadera (21,1%) (p < 0,001). La cohorte PROCOAC es un instrumento que permite realizar estudios de incidencia y progresión en la osteoartritis de mano, rodilla y cadera, así como conocer factores que se asocian con los diferentes fenotipos de osteoartritis. The use of well characterized osteoarthritis cohorts is mandatory for the study and knowledge of this disease. Currently, there is no prospective cohort in this pathology in Spain. The objective of this work is to describe the first osteoarthritis cohort in Spain, PROCOAC (Cohort PROspectiva de A Coruña). The Unit of Rheumatology of the University Hospital of A Coruña started a prospective follow-up study in 2006. The patient inclusion criteria were: a) patients older than 55 years who underwent an abdominal x-ray to study both hips; b) patients diagnosed with radiographic hand osteoarthritis according to ACR criteria; c) patients diagnosed with radiographic knee or hip osteoarthritis according to ACR criteria. Follow-up was performed every 2 years collecting clinical, analytical, genetic and radiographic information. The cohort consists of 937 patients, 873 have radiographic knee osteoarthritis, 783 hip osteoarthritis and 679 hand osteoarthritis. The mean age of the population is 63.9 ± 8.9 years and the average BMI is 29.6 ± 5.1. More than half of the population has high blood pressure and 17% diabetes. The predominant osteoarthritis in the hand is nodular (78.1%), followed by trapeziometacarpal (55.3%) and erosive (18.4%). Of them, 21.4% and 43.1% are healthy at knee and hip level respectively; observing a grade 1 in 26% and 37%; a grade 2 in 26.7% and 11.5%; a grade 3 in 14.9% and 4%; and a grade 4 in 9.4% and 3.7%, respectively. Of the population, 44.1% has only one joint affected, 39.9% has 2 and 13.4% has 3 joints affected. Age (OR = 1.11; P <.001), BMI (OR = 1.11; P =.002) and total WOMAC (OR = 1.03; P =.005) are the only risk factors if we compare the involvement of a single location versus 3. A discrepancy between pain and radiographic damage at the joint level was also detected in patients with KL ≤ 2 grade, and therefore a significantly higher percentage of patients with knee osteoarthritis experienced pain (66.1%) compared to patients with osteoarthritis hip (21.1%) (P <.001). The PROCOAC cohort is an instrument that allows studies of incidence and progression in hand, knee and hip osteoarthritis; as well as determining factors that are associated with the different osteoarthritis phenotypes. [ABSTRACT FROM AUTHOR]

Objective. To conduct a replication study and meta-analysis involving the study of mtDNA variants in the radiographic progression of OA in different cohorts worldwide, including Cohort Hip and Cohort Knee (CHECK), the OA Initiative and a cohort from Spain. Methods. The influence of the haplogroups in the rate of radiographic progression at 96 months in 431 subjects from CHECK was assessed in terms of Kellgren and Lawrence (KL) grade. Progression was defined as a change from KL ≥ 1 at baseline to any higher grade during the follow-up. Extended Cox proportional hazard models were used to analyse the influence of mtDNA variants in the rate of radio-graphic knee OA progression. A subsequent meta-analysis of 1603 subjects following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines was conducted to combine the data of individual studies. A sensitivity analysis was performed to validate the stability of the results. Results. CHECK subjects carrying the haplogroup T showed the lowest rate of radiographic knee OA progression [hazard ratio (HR) 0.645 (95% CI 0.419, 0.978); P < 0.05]. When pooled, subjects within the superhaplogroup JT showed the same trend [HR 0.707 (95% CI 0.501, 0.965); P < 0.05]. BMI [HR 1.046 (95% CI 1.018, 1.073); P < 0.05] and bilateral OA [HR 2.266 (95% CI 1.733, 2.954); P < 0.05] at baseline are risk factors for radiographic knee OA progression as well. In the meta-analysis there was a reduced rate of radiographic progression in subjects with haplogroup T [HR 0.612 (95% CI 0.454, 0.824); P = 0.001] or in the superhaplogroup JT [HR 0.765 (95% CI 0.624, 0.938); P = 0.009]. Sensitivity analysis revealed that the results were robust. Conclusion. The mtDNA variants in the superhaplogroup JT associate with a reduced rate of radiographic OA progression. The mtDNA polymorphisms in the superhaplogroup JT emerge as potential complementary genetic biomarkers for disease progression. [ABSTRACT FROM AUTHOR]

Not all patients with osteoarthritis (OA) show the same disease progression, as some of them remain relatively stable over time, while others progress to severe structural deterioration of the joint. In this sense, the main goal of both genetic and protein biomarkers in OA is to predict not only the risk of OA at an earlier stage of the disease but also which OA patients are more likely to progress to severe disease. Taking into account the incidence of the mitochondria and the mtDNA haplogroups in the pathogenesis of OA, the main objective of this work was to evaluate the incidence of the mtDNA haplogroups in the radiographic progression of the OA disease in a well-characterized follow-up cohort of Spanish patients. DNA from 281 OA patients from Hospital Universitario A Coruña was isolated to determine the European mtDNA haplogroups. Knee or hip radiographs from all affected joints were obtained at two time points with at least 36 months apart. Radiographs were evaluated using the Kellgren/Lawrence (K/L) scale; radiographic OA progression was defined as any radiographic worsening of the K/L joint score. Statistical analyses included Kaplan-Meier survival curves and Cox regression models. Patients belonging to the cluster TJ showed a slower radiographic OA progression than patients in the cluster KU ( p = 0.036). Moreover, patients carrying the most common mtDNA haplogroup H are more apt to require total joint replacement surgery than non-H patients ( p = 0.049). The inherited mitochondrial variants influence the radiographic progression of OA and could be considered among the genetic variants taken into account when the radiographic progression of OA is analyzed. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the influence of the mtDNA haplogroups on knee osteoarthritis progression in Osteoarthritis Initiative (OAI) participants through longitudinal data from radiographs and magnetic resonance imaging (MRI). Methods: Four-year knee osteoarthritis progression was analyzed as increase in Kellgren and Lawrence (KL) grade, in addition to increase in OARSI atlas grade for joint space narrowing (JSN), osteophytes and subchondral sclerosis in the tibia medial compartment of 891 Caucasian individuals from the progression subcohort. The influence of the haplogroups on the rate of structural progression was also assessed as the four-year change in minimum joint space width (mJSW in millimetres) in both knees of (n = 216) patients with baseline unilateral medial-tibiofemoral JSN. Quantitative cartilage measures from longitudinal MRI data were those related to cartilage thickness and volume with a 24 month follow-up period (n = 381). Results: During the four-year follow-up period, knee OA patients with the haplogroup T showed the lowest increase in KL grade (Hazard Risk [HR] = 0.499; 95% Confidence Interval [CI]: 0.261–0.819; p<0.05) as well as the lowest cumulative probability of progression for JSN (HR = 0.547; 95% CI: 0.280–0.900; p<0.05), osteophytes (HR = 0.573; 95% CI: 0.304–0.893; p<0.05) and subchondral sclerosis (HR = 0.549; 95% CI: 0.295–0.884; p<0.05). They also showed the lowest decline in mJSW (standardized response means (SRM) = −0.39; p = 0.037) in those knees without baseline medial JSN (no-JSN knees). Normalized cartilage volume loss was significantly lower in patients carrying the haplogroup T at medial tibia femoral (SRM = −0.33; p = 0.023) and central medial femoral (SRM = −0.27; p = 0.031) compartments. Cartilage thickness loss was significantly lower in carriers of haplogroup T at central medial tibia-femoral (SRM = −0.42; p = 0.011), medial tibia femoral (SRM = −0.32; p = 0.018), medial tibia anterior (SRM = +0.31; p = 0.013) and central medial femoral (SRM = −0.19; p = 0.013) compartments. Conclusions: Mitochondrial genome seems to play a role in the progression of knee osteoarthritis. mtDNA variation could improve identification of patients predisposed to faster or severe progression of the disease. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this study was to identify those proteins relatively more abundant in the synovial fluid (SF) of patients suffering from rheumatoid arthritis (RA) and osteoarthritis (OA) using high performance liquid chromatography coupled to mass spectrometry. 20 individual SF samples from each disease were pooled into two groups (RA and OA) to reduce the contribution of extreme individual values. Prior to the proteomic analysis, samples were immunodepleted from the top 20 most abundant plasma proteins, to enrich the lower-abundance protein fractions. Then, they were subjected to protein size fractioning and in-gel digestion, followed by reversed-phase peptide separation in a nano-LC system and subsequent peptide identification by MALDI-TOF/TOF. This strategy led to the identification of 136 different proteins in SF, which is the largest number of SF proteins described up to date by proteomics. A relative quantification of the proteins between RA and OA was carried out by spectral counting analysis. In RA, our results show a greater relative abundance of proteins related to complement activation, inflammation and the immune response, such as the major matrix metalloproteinases and several neutrophil-related proteins. In OA, we detected an increase in proteins involved in the formation and remodeling of the extracellular matrix (ECM), such as fibronectin, kininogen-1, cartilage acidic protein 1 and cartilage oligomeric matrix protein. The results obtained for MMP-1, BGH3, fibronectin and gelsolin were verified by immunoblotting analyses. Some of the novel proteins identified in this work might be relevant not only for increasing knowledge on the etiopathogenesis of RA and OA processes, but also as putative disease biomarkers, as their presence in SF is a prior step to their dilution in serum. This article is part of a Special Issue entitled: Proteomics: The clinical link. [Copyright &y& Elsevier]

Background: Osteoarthritis (OA) is a multifactorial disease characterized by destruction of the articular cartilage due to environmental, mechanical and genetic components. The genetics of OA is complex and is not completely understood. Recent works have demonstrated the importance of microRNAs (miRNAs) in cartilage function. MiRNAs are a class of small noncoding RNAs that regulate gene expression and are involved in different cellular process: apoptosis, proliferation, development, glucose and lipid metabolism. The aim of this study was to identify and characterize the expression profile of miRNAs in normal and OA chondrocytes and to determine their role in the OA. Methods: Chondrocytes were moved to aggregate culture and evaluated using histological and qPCR techniques. miRNAs were isolated and analyzed using the Agilent Human miRNA Microarray. Results: Of the 723 miRNAs analyzed, 7 miRNAs showed a statistically significant differential expression. Amongst these 7 human miRNAs, 1 was up-regulated in OA chondrocytes (hsa-miR-483-5p) and 6 were up-regulated in normal chondrocytes (hsa-miR-149*, hsa-miR-582-3p, hsa-miR-1227, hsa-miR-634, hsa-miR-576-5p and hsa-miR-641). These profiling results were validated by the detection of some selected miRNAs by qPCR. In silico analyses predicted that key molecular pathways potentially altered by the miRNAs differentially expressed in normal and OA chondrocytes include TGF-beta, Wnt, Erb and mTOR signalling; all of them implicated in the development, maintenance and destruction of articular cartilage. Conclusions: We have identified 7 miRNAs differentially expressed in OA and normal chondrocytes. Our potential miRNA target predictions and the signalling cascades altered by the differentially expressed miRNAs supports the potential involvement of the detected miRNAs in OA pathology. Due to the importance of miRNA in mediating the translation of target mRNA into protein, the identification of these miRNAs differentially expressed in normal and OA chondrocyte micropellets could have important diagnostic and therapeutic potential. Further studies are needed to know the function of these miRNAs, including the search of their target mRNA genes, which could lead to the development of novel therapeutic strategies for the OA treatment. [ABSTRACT FROM AUTHOR]