Background and Aim. Chemokine/receptor axis is a predominant actor of clinical disorders. They are key factors of pathogenesis of almost all clinical situations including asthma. Correspondingly, CXCL12 is involved in the immune responses. Therefore, this study was designed to explore the association between gene polymorphism at position +801 of CXCL12, known as SDF-1α3′A, and susceptibility to asthma in Iranian patients. Material and Methods. In this experimental study, samples were taken from 162 asthma patients and 189 healthy controls on EDTA. DNA was extracted and analyzed for CXCL12 polymorphisms using PCR-RLFP. The demographic information was also collected in parallel with the experimental part of the study by a questionnaire which was designed specifically for this study. Findings. Our results indicated a significant difference (P < 0.0001) between the A/A, A/G, and G/G genotypes and A and G alleles of polymorphisms at position +801 of CXCL12. We also showed an elevated level of CXCL12 circulating level in Iranian asthma patients. Conclusion. Our findings suggest that SDF-1α3′A (CXCL12) polymorphism plays a role in pathogenesis of asthma. It can also be concluded that circulatory level of CXCL12 presumably can be used as one of the pivotal biological markers in diagnosis of asthma. Asthma is characterized as the airways chronic inflammatory disease with mostly airway obstruction, which is accompanied by increased bronchial high responsiveness to various external and internal stimuli. Asthmatic patients suffer from varying symptoms from breathlessness, wheezing, and cough to chest tightness. It is sometimes coupled with allergy, an acquired potency for progression of unwanted reactions that are controlled via immunological mediators . ApproxiAlthough several lines of evidence have indicated the central role of chemokines in regulating multiple aspects of the asthmatic response, the full picture of chemokines' involvement in allergic airway inflammation, the exact role of specific chemokines at various stages in the evolution of allergic lung inflammation and their regulatory pathways in vivo are only partially identified. In the process of allergic airway disease, at least one part of diagnosis is the infiltration of blood leukocytes, including eosinophils, neutrophils, lymphocytes, and macrophages in the lung . The CD4CXCL12 as a homeostatic CXC chemokine is widely expressed with a broad range of functions (from recruitment of mature B and T cells to migration of haematopoietic progenitor cells from the bone marrow). It is well documented that CXCL12 plays a wide variety of roles in two unrelated set of diseases such as human immunodeficiency virus (HIV) and cancer, because its known receptor (CXCR4) is a coreceptor used by HIV T-tropic strains, and it is also the most widely expressed chemokine receptor in many different types of cancers and asthma , 8. “TheThe association between this polymorphism and a variety of diseases from lung and breast cancer, acute myeloid lymphoma, leukemia, and chronic myeloproliferative disease to viral infections such as acute hepatitis is well documented .α 3′A polymorphism and pathogenesis of asthma. Therefore, the authors of this study hypothesized that this polymorphism may play a critical role in development of asthma in Iranian population; thus, this study investigated the predicted role of the +801G/A polymorphism in pathogenesis and progression of asthma in Iranian population.There are no reports describing any association between SDF-1In this study, we recruited 162 unrelated asthmatic patients with a mean age of 51 ± 11 years (ranged between 15 and 79 years) using easy convenience method. Women being pregnant, patients having a history of inflammatory and infectious diseases rather than asthma, and patients with a history of cigarette smoking were excluded from the study. The occurrence of asthma was diagnosed by an expert infectious diseases specialist according to the American Thoracic Society (ATS) criteria. According to the clinical findings and history, patients were classified into 2 groups, that is, allergic and nonallergic groups. We also enrolled 189 genetically unrelated subjects having normal spirometric values and no respiratory symptoms which were matched for sex and similar ethnicity origin with the patients. The study protocol was approved by the ethical committee at our institution , and written informed consent was obtained from all participants, either patients or controls, prior to sample collection. Some characteristics of the subjects are summarized in Asthmatic and control subjects' peripheral blood samples were collected on Ethylenediaminetetraacetic acid (EDTA) precoated tubes and subjected to genomic DNA extraction using a commercial kit . The extracted DNA samples were then stored at −20°C for further use.The CXCL12 gene polymorphism at the position +801 was analyzed by polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method. As previously described by Hassanshahi et al., Azin et al., and Ryabov et al. –12, a PCμL of taq DNA polymerase buffer (10X), 0.5 μL of MgCl2 (stock concentration 1.5 mM), 0.5 μL of each dNTP  stock concentration of 10 mM), 1 μL of each primer (forward: CAGTCAACCTGGGCAAAGCC and reverse: AGCTTTGGTCCTGAGAGTCC), stock concentration of 25 ng/μL, 1 μL of prepared DNA, and sterile double-distilled water to a final volume of 25 μL. The following program was used for fragment amplification: one cycle of 93°C for 2 min, 93°C for 1 min (denaturation), 1 min at 57°C for annealing of CXCL12, 72°C for 40 sec (elongation) followed by 30 cycles of 93°C for 20 sec, 55°C for 20 sec, and 72°C for 40 sec. Within the last 45 sec of the first stage, approximately 0.3 μL of Taq DNA polymerase was added to the mixture. μL which contains 2 μL of primer mix of forward and reverse, 10 μL of Master mix , 3  The amplified PCR product of CXCL12 gene covers +801 regions with a molecular size of 302 bp. The Sac-1  restriction enzyme has only one restriction site on this region, and is capable to digest the fragment and produce two fragments of 202 and 100bp following digestion. In fact, in case of heterozygotic form (A/G), 3 different fragments with 302, 202, and 100 bp are then visible, while in homozygotic form, a 302 bp fragment (without any digestion (A/A)) or two 202 and 100 bp ) was then observed  and studied on ChemiDoc model XRS  subsequent to ethidium bromide staining.2.5 observed . The digThe serum level of CXCL12 was measured by ELISA  in patients and healthy controls immediately after blood collection. Assays were performed as per manufacturer's guidelines. The sensitivity of kits was 2 pg/mL, and inter- and intra-assay assessments of reliability of the kit were conducted.χ2, t-test, and ANOVA using SPSS software version 17 which power of test was %90. P value of less than 0.05 was considered significant.Hardy-Weinberg equilibrium was assessed using genotype data. Allele and genotype frequencies were calculated in patients and healthy controls by direct gene counting. Statistical analysis of the differences between groups was determined by P = 0.85), and the gender variation of patients was 129 (56.58%) female and 99 (43.42%) male and for control group was 101 (61%) female and 88 (38.6%) male (P = 0.9)  . Sac-1 restriction enzyme showed that the frequency of A/A genotype was 18 (4.3%) in patients and 27 (6.5%) in controls . Our results also revealed that the frequency of A/G genotype was 190 (45.6%) and 60 (14.4%) in patients and controls, respectively .Analysis of the polymorphisms in +801 of CXCL12 by G/G genotype in patients was 20 (4.8%) and in controls was 102 (24.5%) (P < 0.0001) . The frequency of A allele was 226 (27.1%) and 114 (13.7%) in patients and controls, respectively. In case of G allele, 230 (27.6%) were observed in patients, while the frequency of this allele was 264 (31.7%) in controls (P < 0.0001). The results of this study also showed that the plasma level of CXCL12 was 312.53 ± 28.75 and 83.57 ± 6.74 pg/mL in asthma patients and healthy controls, respectively (P < 0.0001). Our results indicated that the protein levels of SDF-1 in AG genotype in asthma and control were 448.75 ± 281 and 171.81 ± 54.11, respectively, which the difference was significant (P < 0.0001). Analysis of our data showed that the protein levels of SDF-1 in AA genotype in asthma and control were 22.84 ± 8.41 and 21.62 ± 17.61, respectively, which the difference was not significant (P < 0.822). Analysis of data by Eta test did not show significant difference between genotype and protein level of SDF-1 in patient group ; it also showed that there is no significant difference between FEV1 percent and genotype in patient group  and that there is no significant difference between genotype and FEV1/FVC percent in patient group . Analysis data by Spearman's test (Spearman's correlation coefficient) showed significant difference between protein level of SDF-1 and FEV1 percent and between protein level of SDF-1 and FEV1/FVC percent in patient group (P < 0.0001); this is an inverse relationship between protein level of SDF-1 and FEV1 percent and between protein level of SDF-1 and FEV/FVC percent ( The frequency of the  (24.5%) , where scontrols . Statistectively . Statist percent . P < 0.05) for protein level of SDF-1, FEV1 percent, and FEV1/FVC percent  showed that AA, AG, and GG genotypes have the mean rank of 71.38, 46.81, and 56.79 for the protein level of SDF-1, 32.63, 57.19, and 47.21 for the FEV1 percent and 32.63, 57.19, and 47.21 for the FEV1/FVC percent, respectively, in patient group. For all of them, = 0.021, which showed significant difference ( percent . α (CXCL12) angiogenic alterations associated with asthma which was reported by Hoshino et al., 2003 [The current study was undertaken to investigate the role of CXCL12 + 801 G/A SNP polymorphism in susceptibility to asthma in Iranian asthma patients. All of study groups (patients and controls) had same ethnic background origin and shared a common geographic origin in Southeast part of Iran. We demonstrated a closed association between this polymorphism and asthma in Iranian asthma patients. To our best knowledge, this is the first study to report an association between this polymorphism and asthma. The paramount importance of chemokines and their receptors, as the central actors in the initiation and development of asthma, has been evidenced , 14. Resl., 2003 .Hoshino et al. showed that immunoreactivity of CXCL12 is increased within the airways of asthmatic patients and reported that expression of CXCL12 plays an important role in angiogenesis in the airways of asthmatic patients . The expanded vascularity that has been reported in the asthmatic mucosa might possibly confirm that angiogenesis is a component of the chronic inflammatory response in asthmatic patients, and increased CXCL12 circulatory level in asthmatics may confirm that angiogenesis is a specific characteristic of asthma, which is probably at least partially mediated by CXCL12 –18. α 3′A polymorphism that presents in autoimmune disorders. However, angiogenesis function of chemokines is currently the focus of intense investigations; little information is known on the roles of chemokines in pathological remodeling of the asthmatic airway. Mice lacking the CXCL12 gene have defective vascular development, suggesting that CXCL12 plays an important role in organ vascularisation . Recent Nevertheless, our study was affected by some limitations. It should be considered that all of our study population (both control and asthma patients) were within Iranian population, and thus, the possibility of ethnicity as a confining factor could be excluded. However, it should be noted that different geographical and ethnic backgrounds of the individuals can influence the consequences of the association studies. Therefore, findings of the current study need to be examined and confirmed in other populations and ethnic groups. Moreover, further replication studies using a wider sample size and different population would be essential for further evaluation of the relationship between the CXCL12 genetic variations and the risk of asthma.


The use of human saphenous vein grafts (HSVGs) as a bypass conduit is a standard procedure in the treatment of coronary artery disease while their early occlusion remains a major problem.ex vivo perfusion system, which uses standardized and strictly controlled hemodynamic parameters for the pulsatile and non-static perfusion of HSVGs to guarantee a reliable analysis of molecular parameters under different pressure conditions. Cell viability of HSVGs (n = 12) was determined by the metabolic conversion of 3--2,5-diphenyl-tetrazolium bromide (MTT) into a purple formazan dye.We have developed an Under physiological flow rates (10 mmHg) HSVGs remained viable for two weeks. Their exposure to arterial conditions (100 mmHg) was possible for one week without important reduction in viability. Baseline expression of matrix metalloproteinase-2 (MMP-2) after venous perfusion  was strongly up-regulated after exposure to arterial conditions for three days (19.8 ± 4.3) or five days . Zymographic analyses confirmed this increase on the protein level. Our results suggest that expression and activity of MMP-2 are strongly increased after exposure of HSVGs to arterial hemodynamic conditions compared to physiological conditions.Therefore, our system might be helpful to more precisely understand the molecular mechanisms leading to an early failure of HSVGs. Gusic and colleagues investigated the role of the mechanical environment in vein remodeling in a higher developed ex vivo perfusion system with a main focus on medial and intimal growth in the perfused veins. They ran their perfusions system with five different ex vivo hemodynamic environments and showed that pressure and shear stress act independently to regulate vein remodeling [ex vivo perfusion system which can be used to perfuse HSVGs with tightly controlled, steady and standardized perfusion profiles. We have defined the viability time course of perfused HSVGs exposed to arterial and venous perfusion profiles. In addition, we provide evidence that our system is suitable to detect alterations of molecular markers such as MMP-2 as a consequence of preparative injury or increased arterial perfusion pressure.Coronary artery bypass grafting (CABG) using venous grafts is a standard procedure in the treatment of advanced coronary artery disease. However, vein graft occlusion implanted in an arterial pressure environment is still a major problem . Approxi2 . After gac cycle ,10. Beyoac cycle . Formatiac cycle -14 whereac cycle . It is gac cycle . Berceliac cycle . The groac cycle . Comparemodeling . Yet, thNonvaricose HSVGs were obtained from 35 patients  undergoing CABG surgery in the German Heart Center Munich. The endoscopically harvested vein grafts were kept in autologous blood at room temperature until implant. One part of the graft was immediately stored in Ringer solution on ice and transferred from the operating room to the laboratory. One small piece was directly snap-frozen in liquid nitrogen and stored at -80°C until further use as unperfused control tissue. This piece served as a reference to determine relative gene expression. The other part of the vein was mounted into the perfusion device as described. The procedure was acknowledged by the local ethical committee .The circuit of the perfusion system is driven by a roller pump ISMATEC S2  producing a pulsatile and non-static flow. All silicon tubings and the vessel chamber are sterilized prior to use. The vessel mounting procedure is carried out under a biological safety cabinet . Constant pressure conditions are maintained using a syringe pump . The entire system is placed into a styrofoam-isolated chamber to maintain a constant temperature of 37°C. Disposable pressure sensors  are placed on both sides of the vessel chamber to permanently monitor and facilitate the control of pressure conditions of the circuit. All functions and settings are controlled by a PC with a program written in java. Pressure is controlled by a PID-algorithm, data are logged continuously.in vivo conditions. Total time from operating room to perfusion was less than one hour. The perfusion medium was DMEM/Ham's F-12  supplemented with 10% FCS, 2 mM glutamine and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin). Veins were perfused with venous conditions  or with arterial conditions  for various time periods. At the end of each experiment vein ends were discarded. The other part of the vein was snap-frozen in liquid nitrogen and stored at -80°C until further use. In long-term experiments the medium was replaced every two days. The pH of the medium remained stable within this period.HSVGs were fixed in the perfusion device by suture ligation  and adjusted to a length matching the To verify tissue viability, a staining with MTT  was performed. In the presence of metabolically active viable cells the yellow MTT is converted into a water-insoluble purple formazan product due to reduction by mitochondrial dehydrogenases and other cellular enzymes ,19. MTT Taq polymerase for 15 min at 95°C specific products were amplified during 40 cycles using the following conditions: 15 sec at 94°C (denaturation), 20 sec at 60°C  and 20 sec at 72°C (elongation). The relative expression levels of MMP-2 in individual samples were calculated in relation to the expression of the β-actin housekeeping gene. To compare independent samples the ratios of MMP-2/β-actin were calculated.Frozen tissue pieces were minced using a Precellys24 lysis and homogenization system  and total RNA was extracted using Trifast according to the manufacturer's recommendation (Peqlab). All RNA preparations were digested with DNase I prior to cDNA synthesis using Omniscript RT kit . One μl of cDNA was amplified on a LightCycler 1.5 thermo cycler  using the QuantiTect SYBR Green Kit (Qiagen) and BSA (0.5 mg/ml) in a final volume of 20 μl. All primers were used in a final concentration of 0.5 μM. The following primers were used: β-actin forward 5' CCA ACC GCG AGA AGA TGA 3', β-actin reverse 5' CCA GAG GCG TAC AGG GAT AG 3', MMP-2 forward 5' TGC TGG AGA CAA ATT CTG GA 3', MMP-2 reverse 5' GAT GGC ATT CCA GGC ATC 3'. They amplify fragments of 96 and 90 bp, respectively. After an initial activation of 2, 1.5 mM NaN3, 20 mM CaCl2, 0.01% Triton X-100, 10 mM cacodylic acid, pH 5.0). Samples were centrifuged at 4°C for 10 min at 20.000 × g. The supernatant containing proteins was removed and stored at -80°C until further use. Ten μg of extracted protein were mixed with zymogram loading buffer  and separated in 15% SDS-PAGE gels containing 1 mg/ml type A gelatine from porcine skin . To renature proteins, gels were washed two times in 2.5% Triton X-100 for 15 min at room temperature and subsequently incubated in developing buffer, pH 7.5  overnight at 37°C. Gels were stained with 0.5% Coomassie Blue R250 in 40% methanol/10% acetic acid for 15 min and destained in 40% methanol/10% acetic acid until clear bands of lytic activity appeared. The reaction was stopped by transfer of gels in aqua bidest. Gelatinolytic activity was quantified using ImageJ software . The pixel intensities of bands within each gel were normalized against the respective control of unperfused venous tissue.MMP-2 protein activities were evaluated by a standard gelatine zymography. Briefly, 100 mg of frozen HSVG tissue were homogenized in ice cold zymogram buffer . Thereafter, the viability dropped Figure . We thenTo further explore potential pathological changes in HSVGs upon perfusion, we investigated tissue sections from veins after perfusion with venous or arterial pressure profiles at different time points by a hematoxylin/eosin staining. As a reference, we used an unperfused section of the same vein. Exposure to venous pressure for three days did not change the histology and even after five days a minor thickening of the intimal layer was evident . After arterial perfusion for one day also no major changes could be noticed. However, after three days the intimal layer started to visibly thicken and after five days extensive hyperproliferative areas were seen .ex vivo system. Perfusion of HSVGs with 100 mmHg for one day yielded an MMP-2 gene expression ratio which was similar to the reference   Figure . Exposur) Figure . However) Figure . Quantif) Figure . Gelatinex vivo perfusion system proved its ability to monitor alterations in the expression of genes which are expected to increase their activity due to elevated pressure conditions on the RNA and protein level.Thus, our novel ex vivo perfusion system developed to mimic the arterialization of HSVGs. The culture system provides the ability to reproduce the initial events taking place when the grafted vein is exposed to arterial hemodynamic conditions. Therefore, our system may represent a valuable and reasonable approach to identify molecular mechanism underlying the early stages of bypass grafting. Several in vitro and in vivo studies have demonstrated changes in graft morphology, viability, cellular density or gene expression under arterial conditions [ex vivo vein support system to perfuse HSVGs with arterial conditions regarding shear stress, flow rate and pressure during a period of 7 and 14 days. They found significant IH and a marked increase in plasminogen activator inhibitor 1 (PAI-1) expression in the human veins after 7 and 14 days of perfusion [A major problem with HSVGs remains their occlusion after a certain time. Transposition of a vein segment and exposure to the arterial hemodynamic environment leads to an acute increase in flow rates and intraluminal pressure and is thought to be a potential trigger for the pathological remodeling of HSVGs ,23. Genenditions ,22,27,28erfusion . A matheerfusion . Previouerfusion ,21.de novo synthesis of MMP-2 in HSVGs perfused with artertial conditions [ex vivo perfusion system. Thus, the results of our perfusion system perfectly reflect the in vivo situation suggesting that genes which are involved in vascular remodeling are activated by arterial pressure. Therefore, our system can be used to analyze molecular parameters involved in such events in detail under standardized, tightly controlled and reproducible conditions.Within three days of perfusion under arterial pressure conditions in our perfusion system the expression of MMP-2 increased more than nine-fold and reached an even higher value after five days, similar to the activation of PAI-1 . Our datnditions  or in annditions ,16. Bercnditions , as we fex vivo system with autologous blood lacking blood cells, platelets, plasma, blood-surface interaction and the multitude of inflammatory and coagulation mediators playing an important role in the pathophysiology of IH development. However, because of technical reasons, we were not able to perfuse veins with blood by using a roller pump for perfusion to achieve a pulsatile flow. Platelets would be inevitably activated and blood cells destroyed during passage through the pump. Hemolysis could be avoided or highly reduced by using a centrifugal pump instead, which in turn produces a nonpulsatile flow. Inclusion of blood would provide exposure of the vein to a more physiological state, but may also confound the results with numerous other variables. Another limitation is the time-restricted viability of grafts in the ex vivo perfusion system which would not be prolonged by blood perfusion due to the accumulated metabolic waste products and inflammatory reactions. Despite these limitations, the findings of the current study highlight important potential in our understanding of the healing and adaptation of veins transplanted to the arterial environment. From the beginning of the development we tried to keep the total volume of the circuit relatively small. At present it comprises approximately 20 ml which is substantially lower compared to other systems which use volumes up to 500 ml [in vitro [An important advantage of our system is the possibility to mount vessels of variable length and diameter. The sliding unit allows a very flexible adjustment to guarantee that the vessel maintains its natural length and tension throughout the experiment. Our main focus was to setup an experimental system, which is suitable to reliably analyze molecular parameters as a function of altered pressure and flow conditions. Therefore, the most important point was to control the pressure conditions very stringently and also to keep them very stable. In pilot experiments we experienced a continuously decreasing pressure in the circuit, despite any leakage. Knowing that pressure affects gene expression such a behavior would be fatal for a desired molecular readout. With regard to this a unique feature of our perfusion system is the regulation of the mean pressure in the circuit by a computer controlled syringe pump. Decreasing pressure due to diffusion processes through out the silicone tubing  or relaxo 500 ml ,34. If no 500 ml ,36. Anotin vitro . Our sysex vivo perfusion system which maintains human veins viable for up to two weeks under a low pressure profile. The setup guarantees a tightly controlled and stable perfusion rate and the system proved to be suitable to record alterations in gene and protein expression induced by different perfusion profiles. Further advantages of our system are a total flexibility concerning the size of potential vessels and almost infinite possibilities in various research areas by the addition of defined amounts of exogenous substances into the circuit. Our ex vivo perfusion system and its applications may, therefore, help to improve the long-term patency of human bypass grafts.In summary, we have developed a novel The authors declare that they have no competing interests.- SD prepared the human saphenous vein grafts and mounted them into the perfusion system, tested the viability of the vein grafts, performed quantitative RT-PCR-analysis and wrote the major part of the manuscript- SE constructed the set-up of the perfusion system and helped with the development of the software- CT helped to prepare and mount the HSVGs, participated in the quantitative RT-PCR-analysis and performed the zymographic studies- US participated in the design and helped to build the perfusion device- BV harvested the HSVGs for the perfusion system and viability experiments during CABG surgery and helped to draft the manuscript- MAD participated in harvesting the HSVGs for the perfusion system and viability experiments during CABG surgery and helped to draft the manuscript- HH participated in the design and coordination of the study and helped to draft the manuscript- HL participated in the design of the study, performed statistical analyses and participated in writing of the manuscript- RL harvested the HSVGs for the perfusion system, participated in the design of the study and its coordination and helped to draft and improve the manuscript- MK initiated and designed major parts of the study, participated in coordination and writing of the manuscriptAll authors read and approved the final manuscript.Figure S1. Histological analysis of a representative formalin fixed and paraffin-embedded HSVG after perfusion with different pressure profiles and hematoxylin/eosin staining. The control represents the unperfused vessel. The other parts of the vein were perfused with physiological venous (10 mmHg) or arterial pressure (100 mmHg) for the time indicated.Click here for file


The 10-year risk of general CV disease, as predicted with the Framingham Risk Score, was similar in patients and controls (p=0.18). IMT of both common carotid and common femoral artery were increased in EOA , but the frequency of atherosclerotic plaques was not increased. There was no difference in FMD and NMD between the two groups, but the difference between FMD and NMD was increased in EOA. In conclusion, this small controlled study showed an association between EOA and subclinical atherosclerosis that cannot be fully attributed to traditional CV risk factors, as assessed by the Framingham score. These results suggest that chronic, low-grade inflammation is implicated in atherosclerosis in EOA.Chronic inflammatory disorders have been associated with accelerated atherosclerosis and increased cardiovascular (CV) risk. Recent evidence suggests that erosive hand osteoarthritis (EOA) has considerable inflammation; therefore, we examined the presence of subclinical atherosclerosis and endothelial dysfunction in EOA. Twenty-four patients with EOA and 24 age- and sex-matched healthy individuals without clinical OA were included in the study. No subject had a history of CV disease. Intima-media thickness (IMT) and atheromatous plaques in the common carotid and common femoral arteries were measured by Doppler ultrasonography. The endothelium-dependent, flow-mediated dilatation (FMD) and endothelium-independent, sublingual glyceryl trinitrate (NTG)-induced dilatation (NMD) of the brachial artery were assessed. The EOA patients had significantly elevated systolic and diastolic blood pressure ( Erosive hand osteoarthritis (EOA) is a subset of hand osteoarthritis (HOA) which is defined by erosions in interphalangeal (IP) joints on plain radiographs and occurs primarily in postmenopausal women . TraditiA growing body of evidence supports that endothelial dysfunction and atherosclerosis are significantly more prevalent in patients with chronic inflammatory disorders, compared to age-matched controls . This hast carpometacarpal joint squaring. In addition, they had no history or clinical signs and symptoms of osteoarthritis in other joints. All participants, in both groups, had no history of CV disease, such as an acute coronary syndrome, stroke or peripheral arterial disease. History of diabetes mellitus (DM), hypertension (HT), hyperlipidemia (HL) and smoking, and any relevant medication were recorded.Patients participating in the study were followed at the Rheumatology outpatient clinic of the University General Hospital of Larissa. All fulfilled the 1990 American College of Rheumatology criteria for the classification of HOA .  In addThe study protocol was approved by the Institutional Scientific and Ethical Committee, all participants gave a written informed consent, and the study was conducted according to good clinical practice.Ultrasonographic (US) examination was performed by a single trained and senior vascular surgeon (GA). Common carotid and common femoral artery intima-media thickness (IMT), were measured using high resolution B-mode ultrasonography, using a 12 L, 5 Hz linear transducer (GE Logic equipment), according to recently published recommendations . MeasureThe presence and type of atheromatous plaques was recorded for both sites. A plaque was defined as a focal structure encroaching into the lumen, of at least 0.5 mm, or 50% of the surrounding IMT. Alternatively, a plaque should demonstrate thickness >1.5 mm, measured from the media-adventitia to the media-lumen interface . CarotidTwo definitions were used for subclinical atherosclerosis: plaque formation and a common carotid artery IMT >1 mm. The cut-off of 1 mm was used because this level is associated with coronary artery disease and increased CV risk . As commAn US “biopsy score” (USBS) has been assessed in all subjects . This isEndothelial function was assessed by measuring flow-mediated endothelium-dependent dilatation (FMD). The end-diastolic diameter of both brachial arteries was measured at rest and within 15 sec after release of a sphygmomanometer cuff inflated at a pressure of at least 50 mmHg above the subject’s systolic blood pressure (BP) for 3 minutes . SimilarAs the vasodilator potential is not the same across individuals, NMD can be used as a means of measuring the maximum obtainable vascular wall reaction to a vasodilatory stimulus . TherefoAll assessments were conducted in a quiet, dark room and subjects were examined in the supine position. Patients remained at the supine position for at least 10 minutes before the examination.Serum total cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL) and triglycerides were measured by standard methods. The cut-off values for total cholesterol, LDL and HDL are in accordance with the Third Report of the National Cholesterol Education Program (NCEP) .The Framingam 10-year risk of CVD was estimated according to published method . This alQualitative variables were compared with the Fischer’s exact test and the chi-square test, as appropriate. Comparisons between means were carried out with the Student’s t-test and the Mann-Whitney U, as appropriate. Statistical analysis has been performed using the SPSS, version 13, software .For each group the risk of preclinical atherosclerosis was separately assessed with regard to age, sex, history of hypertension, hyperlipidaemia, diabetes and smoking, total cholesterol (whether >240 or not), LDL (whether >160 or not), HDL (whether <50 or not), systolic blood pressure  and diastolic blood pressure (whether > or <90 mmHg), and regular use of vasoactive and hypolipidaemic agents , with the Fisher’s exact test and the Mann-Whitney U test.The two groups were comparable for all demographic variables. No significant difference was found in the frequency of diabetes mellitus, hypertension, hyperlipidaemia and smoking status between EOA patients and controls. Similarly, no difference was found in the rate of prescription of vasoactive and hypolipidemic medications between the two groups Table . Fourteep<0.001 for both)  , compared to the control group  (Table p=0.012). Similarly, common femoral artery IMT was elevated in the EOA compared to the control group (p<0.01) (Table p=0.77). For each type of plaque, the difference between the two groups was not statistically significant (data not shown). One patient in the EOA group had occluded internal carotid artery. All other plaques were not hemodynamically significant. EOA was not associated with an increased risk of plaque, compared to controls.EOA was associated with a marginally significant increase in the risk of IMT>1mm    and hypotension . No difference was found between the two groups in both flow-mediated, endothelium-dependent vasodilatation and nitroglycerine-induced, endothelium-independent vasodilatation. However, the difference between FMD and NMD was significantly greater in EOA patients compared to healthy controls  in patients with EOA that could not be attributed to traditional CV risk factors, as assessed by the 10-year Framingham risk score. Moreover, patients with EOA exhibited evidence of endothelial dysfunction, as estimated by the difference between FMD and NMD, that could not be attributed to differences in systolic or diastolic blood pressure.The link between OA and atherosclerosis could be explained by shared pathogenetic mechanisms. Inflammation appears to play a role in OA and EOA in particular , 5, 8-10A limitation of our study is the small sample size, and therefore, our results should be confirmed in a larger study. However, US examinations were performed by a single, experienced assessor, ruling out inter-observer variations. In addition, 14 out of 24 EOA patients (58.3%) were receiving HCQ for a mean period of 23.5 months. HCQ might have improved vascular function in our study population, since it has been shown to exert a mild protective effect against atherosclerosis in patients with systemic lupus erythematosus . This meIn conclusion, in this small, cross-sectional, controlled study, we found evidence of subclinical atherosclerosis and endothelial dysfunction in patients with EOA.


Dopamine DRD1 receptor regulates renal function and vascular resistance. It plays a role in the pathogenesis of hypertension in animal models. In humans, the DRD1 gene presents a A-48G polymorphism associated to hypertension in a Japanese population. To explore the role of this polymorphism on blood pressure and renal function in Caucasian hypertensive patients (H), we evaluated the allele frequencies in a populations of 697 H and 100 blood volunteers, and found no difference in the distribution of the alleles between the two groups . In H, we found a significant difference between AA and GG in serum creatinine . Treatment restored serum creatinine at levels comparable between genotypes . To replicate the finding, in a case control study of 8 AA and 7 GG hypertensive patients matched for age, sex and body mass index, in pharmacological wash out for 30 days, we evaluated serum  and urinary  biochemistry and renal hemodynamic assessed by ultrasound. Once again, the AA group showed higher serum creatinine, Na, Uric acid and urea, reduced creatinine clearance and a higher level of urinary protein excretion. These changes occurred while no differences were observed in diuresis and renal vascular resistances. In conclusions, the DRD1 A-48G polymorphism identifies a class of H that is prone to hypertension related kidney alterations. Renal dopamine receptors are important regulator of renal hemodynamic and sodium reuptake, and participate in the pathogenesis of hypertension1-like receptor agonists increase renal blood flow and glomerular filtration rate, as well as urinary excretion of sodium and water1-like receptor agonist involves changes in kidney hemodynamic and direct tubular action1-like receptor agonistsThe dopamine receptors are G-protein coupled receptors subdivided into two families: D1-like (includes D1 and D5) and D2-like  grounded on the stimulation or inhibition of adenylyl cyclase, respectivelyIn particular, renal dopamine receptor number and expression have been studied in animal models of hypertension. Watanabe et al. reported that renal cortical dopamine receptor density is diminished in spontaneously hypertensive rats compared with normotensive Wistar-Kyoto rats at 18 weeks of age, but it was similar in both rats at 3 and 7 weeks of ageThe human dopamine receptor 1 (DRD1) gene has been cloned In order to perform an observation of the distribution of the DRD1 variants among hypertensive patients, we selected a study population of 253 essential hypertensive patients referring to the Hypertension Diagnosis and Care Outpatient Clinic of the “Federico II” University of Naples. Anamnesis, clinical data and biochemical features of the patients enrolled in this ambulatory are routinely stored in a computerized database that comprehends over 25000 patients with an average follow-up of 6 yearsAt enrolment in the study, blood and urine samples were collected for determination of serum concentration of creatinine, uric, acid, urea, Na, K and analysis of the concentration of creatinine, proteins, Na, K, in the urine collected in 24 hours were measured by standard methods. Based on results, clearance of creatinine was calculated according to the Cockcroft-Gault formula: [(140-Age)*weight in kilograms/]/ serum creatinine*72*(sex correction factor)*BSA/1.73, where sex correction facto is equal 0.85 for women and 1 for men.2 for females and males, respectivelyUltrasounds were performed using a SONOS 5500 (Philips) by two independent trained experienced physicians . Intraoperator and interassay variability were 5% and 6%, respectively. Echocardiography was performed using standard technique, with a dedicated ultrasound machineCarotid artery B-mode ultrasonography was performed with patients in the supine position with the neck extended in mild rotation. The scanning protocol was performed with an ultrasound device  equipped with a 7.5-MHz high-resolution transducer with an axial resolution of 0.1 nm and examinations were recorded on S-VHS videotapes. The maximum arterial intima-media thickness (IMT) in up to 12 arterial walls, including the right and the left, near and far distal common (1 cm) bifurcation, and proximal internal carotid artery was estimated offline with an image processing workstation. Subjects with a maximum IMT of >1.5 mm were considered to have plaque according to previous studiesFrom each patient we collected 15 ml of venous blood sample from an antecubital vein for DNA extraction. Genomic DNA was extracted from whole blood samples using a commercially available column extraction kit . A restriction fragment length polymorphism assay was developed to detect a G allele at nucleotide -48 using the following forward (5-GGC TTT CTG GTG CCC AAG ACA GTG-3) and reverse (5-AGC ACA GAC CAG CGT GTT CCC CA-3) primers2 analysis. Association between genotype and hypertension was evaluated by multiple logistic linear regression analysis. Differences in the clinical data between the subjects with and without hypertension and between subjects with A/A or G/G were assessed by ANOVA followed by Bonferroni’s test. A p value < 0.05 was considered significant.Data are presented as mean ± SD. Allele frequencies were calculated from the genotypes of all subjects. Significant differences between the total number of alleles on all chromosomes for the hypertensive and normotensive groups were assessed by χOur population is largely composed of young male patients and in 75% of cases had a family history of hypertension . We compWe then stratified our population based on the DRD1 A-48G polymorphism. There was no effect of the A-48G polymorphisms on anthropometric, hemodynamic and cardiovascular ultrasound parameters . InteresTo further explore whether DRD1 variant can differently interfere with the effects of high blood pressure on kidney function, we selected 15 patients carrying 2 copies of either the A-48 or the G-48 DRD1 alleles. Patients were chosen to match for age, sex, BMI, and blood pressure, and did not present kidney failure or left ventricle hypertrophy. They had similar history of high blood pressure levels and did not present diabetes or dyslipidemia . After 3The major finding of our work is the identification of the -48A/A variant of the DRD1 gene as a potential risk factor for kidney damage in hypertension. This conclusion relies on the evidences gathered in two different cohorts of patients, using two different experimental approaches. In the first population, using an observational design, in a relatively large cohort of hypertensive patients we report that this polymorphism has same frequency in hypertensive patients and a control population of normotensive blood donors, thus suggesting that this polymorphism, at least in white caucasic does not associate with high blood pressure. More importantly, though, hypertensive patients bearing two copies of the A-48 variant presented a significant increase in serum creatinine levels, which is an indicator of somewhat impaired kidney function. So far, hypertension related organ damage has been considered uniquely a consequence of the exposure to elevated blood pressure levels. Nevertheless, the observation that hypertensive patients with similar anthropometric parameters and blood pressure levels are more prone than others to develop hypertension related target organ damage as well the large familiarity for cerebrovascular events related to high blood pressure levels have provided the necessary background for the investigation of genetic determinants of target organ damage. We have also produced recent evidence that the cardiac damage related to hypertension is indeed determined by a naturally occurring polymorphism of the β2 adrenergic receptorTo further detail the nature and characteristics of these kidney alterations, we decided to circumscribe the observation to a limited number of selected patients in which a deeper exploration of kidney function was performed. This part of the study was conducted in selected hypertensive patients that were chosen among the homozygous for the A or the G-48 variant of the DRD1 gene, since no differences were observed between heterozygous and G/G homozygous patients. This case control study, produced evidence that the A/A population is more prone to kidney damage, as suggested by larger microalbuminuria and reduced creatinine clearance. Given the distribution on both vascular smooth muscle cells and tubular epithelium of the DRD1 receptor within the kidney, the altered kidney function can be explained by differences in the kidney hemodynamic or the tubular epithelium sodium reabsorption. Our data support the notion that the A-48ADRD1 mutation leads to an impairment in the kidney filtration function since no differences were observed in the renal artery and parenchymal artery hemodynamic.Although our data are suggestive that the A-48 mutation leads to an impairment in the function of the DRD1 receptor, the mechanism underlying this alterations are not known. It is possible to speculate that the mutation falls within a regulatory region of the promoter of the gene, but so far there aren’t study available in the literature proving this hypothesis. In this sense, our observation is the first to report an impairment of a physiological function associated with this receptor, therefore supporting the hypothesis that this mutation causes a reduced expression of the DRD1 receptor at the tubular level. Previous studies have explored the impact of this polymorphism on behavior and dependences, since the large abundance of this receptor in the central nervous system and its role in dopamine and serotonine metabolismIn conclusion, our data support the role of the DRD1 as a genetic marker for hypertension related kidney damage and provide molecular bases for the notion that the hypertension related target organ damage is modified by genotypes. This evidence will lead to development of new therapeutic strategies tailored on the genetic profile of hypertensive patients.


Cardiovascular disease is the leading cause of death in patients with end stage renal disease (ESRD). The vasodilator mechanisms in small resistance arteries are in earlier studies shown to be reduced in patients with end stage renal disease. We studied whether endothelium dependent vasodilatation were diminished in ESRD patients and the interaction between the macro- and microcirculation.Eleven patients with ESRD had prior to renal transplant or insertion of peritoneal dialysis catheter measured pulse wave velocity. During surgery, a subcutaneous fat biopsy was extracted. Resistance arteries were then dissected and mounted on a wire myograph for measurements of dilator response to increasing concentrations of acetylcholine after preconstriction with noradrenaline. Twelve healthy kidney donors served as controls.Systolic blood pressure was elevated in patients compared to the healthy controls; no difference in the concentration of asymmetric dimethyl arginine was seen. No significant difference in the endothelium dependent vasodilatation between patients and controls was found. Correlation of small artery properties showed an inverse relationship between diastolic blood pressure and nitric oxide dependent vasodilatation in controls. Pulse pressure was positively correlated to the total endothelial vasodilatation in patients. A negative association between S-phosphate and endothelial derived hyperpolarisation-like vasodilatation was seen in resistance arteries from controls.This study finds similar vasodilator properties in kidney patients and controls. However, correlations of pulse pressure and diastolic blood pressure with resistance artery function indicate compensating measures in the microcirculation during end stage renal disease. Cardiovascular disease (CVD) is overrepresented in patients with reduced kidney function, and the risk increases with declining kidney function, why CVD is the major cause of death in patients with end stage renal disease (ESRD) Arterial stiffness, defined as decreased ability and degree of the conductive arteries to absorb the pulse pressure is related to mortality in dialysis and non-dialysis patients The function of the large arteries' viscoelasticity reduces the pulsative pressure and flow that results from the intermittent ventricular ejection, securing a stabile pressure and flow at the level of small arteries. When stiffness increases, an augmentation of the pulse wave is present because of changes in the timing and size of reflection. This might induce damage and change the functionality of the vascular microcirculation as it is exposed to a higher pulsative pressure and flow.Endothelium-dependent vasodilatation occurs via three main pathways; cyclooxygenase (COX) products, nitric oxide (NO) and endothelium-derived hyperpolarisation (EDH) Investigations of the micro circulation (the resistance arteries) of ESRD patients show no difference from similar arteries from healthy controls when comparing morphology and sensitivity to vasoconstrictors In this study we wanted to evaluate differences in vascular function in arteries from patients with ESRD and normal controls and to study the interaction between the macro- and micro-circulation. Especially we wanted to test whether endothelium dependent vasodilatation was diminished in ESRD patients with well controlled blood pressure.Eleven patients with ESRD were enrolled in the study. Nine had living related donor renal transplant and two insertion of peritoneal dialysis catheter. Patients included were above 18 years of age. Exclusion criteria were persisting cardiac arrhythmias, severe congestive heart failure, reduced pulmonary function, severe psychiatric disease, acute infection and leg-amputation. Five patients were treated with peritoneal dialysis (vintage 252 days (101-568) median (range)); four with hemodialysis (vintage 769 days (14-1201)) and two were not on dialysis. Patient's kidney diseases were glomerulonephritis (n = 5), adult polycystic kidney disease (n = 2), obstructive nephropathy (n = 2), type 1 diabetes mellitus (n = 1) and vasculitis (n = 1). No patient had diabetes mellitus as comorbidity. Nine out of eleven were treated with antihypertensive medication. Two were treated with angiotensin converting enzyme inhibitors, four with angiotensin receptor blockade, six with calcium antagonists, four with beta-blockers, six with diuretics, ten with erythropoietin analogs and one with statins.The majority of patients were cardiovascular assessed as part of the pre-transplantation assessment or ESRD regular control with echocardiography, coronary angiogram and CT scans of the iliac arteries. All investigation was performed in the year before the study and none had any cardiovascular event between the cardiovascular assessment and transplantation. All patients had an echocardiography; ten had normal ejection fraction (EF) while one patient had an EF of 40%. One of the patients had indication of a light aortastenosis and five patients had light left ventricular hypertrophy. Nine patients had a coronary angiogram; seven patients had normal coronary arteries, one had earlier coronary by-pass and one had some calcification but no stenosis. Seven had a CT scan of the iliac arteries; five had no calcification, 2 had mild to moderate calcification. Cholesterol was measured in six patients, with a mean value of 4.9 mmol/l. Inflammatory profile were assessed with c-reactive protein in ten of eleven patients prior to surgery; all values were less than 10 mg/l.Twelve kidney donors with no preexisting medical conditions or drug intake served as healthy controls. Before surgery the kidney recipients received 2 days of immunosuppressive therapy consisting of 0.2 mg/kg tacrolimus, 1.5 g mycophenolat mofetil acid and 20 mg prednisolone. The study was performed in accordance with the Declaration of Helsinki. Protocol and consent forms were approved by the local research Ethics Committee , and all participants gave written informed consent before inclusion.Participant's body weight, height and blood pressure were measured and hemodynamic data were calculated the day before surgery. Patient's type of dialysis, vintage, underlying condition and drug intake were obtained from medical records. Blood samples were taken on the day of surgery. During surgery a 2×3 cm biopsy containing skin and subcutaneous fat was removed from the abdominal wall with scalpel without the use of diathermy.2and 21% O2 in a nitrogen based gas to keep pH at 7.4 at all times. Upon mounting, the arteries were left 20 min to equilibrate before stepwise stretching, characterizing the elastic properties as described by Mulvany and Halpern 100 . Viability of the arteries was tested using 10 μM noradrenaline (NA) twice before beginning the experiments. The arteries were preconstricted with 3 μM NA and subsequent relaxation using increasing concentrations of the endothelium-dependent vasodilator ACh. The experiment were repeated after incubation (20 min) with first the COX-inhibitor indomethacin (3 μM) and then the combination of the NOS inhibitor L-NAME (100 μM) and indomethacin.Subcutaneous fat biopsies were immediately after extraction placed in 5°C physiological salt solution (PSS), transported to the lab where 2 mm long segments of arteries were isolated and mounted on two stainless steel wires (40 μm diameter) in the organ baths of a 4-channel wire myograph , Aarhus, Denmark) or in a double channel myograph  for isometric force measurement. The myograph contained PSS at 37°C, continuously bubbled with a gas mixture containing 5% COTo ensure that the endothelium was viable no artery was submitted to more than 3 endothelium-dependent relaxation curves and were left resting for 20 minutes between curves during incubation.2PO4 1.18, MgSO4 1.17, NaHCO3 25, CaCl2 1.6, EDTA 0.026, and glucose 5.5. The chemicals were obtained from Sigma . Indomethacin was dissolved in ethanol, the other chemicals in distilled water.The composition of PSS was (mM): NaCl 119, KCl 4.7, KH−1 and a volume of 1.8 l after a rest of at least 10 min Cardiac output (CO) was measured by rebreathing technique in a closed system containing a gas mixture of sulfahexafloride and nitrous oxide in a mixture of oxygen and nitrogen . Rebreathing was performed in the sitting position during 15 s with a breathing rate of 14–16 minPWV and AI were measured in the supine position after 10 min of rest. Carotid –femoral PWV was measured with SphygmoCor, AtCor Medical, Texas, US, using the integral software.Augmentation pressure was calculated as the difference between the second and first systolic peaks, and AI was calculated as the augmentation pressure expressed as percentage of pulse pressure. AI was measured for aorta. All of the measurements were made in duplicate by one trained study nurse, and the mean values were used in the subsequent analysis.Asymmetric dimethyl arginine (ADMA) was measured using ELISA . All other biochemistry was analysed at the Department of Clinical Biochemistry, Aarhus University Hospital. EGFR was calculated from the 4 point MDRD formula.50 values are calculated by non-linear regression for variable slope for each vessel and presented as the negative logarithmic value in mol/L (pEC50). NO dependent response was calculated as the difference between EC50 values of indomethacin and indomethacin/L-NAME curves respectively (ΔpEC50). Mean pEC50, ΔpEC50 and baseline characteristics on patients were compared using unpaired student's t-test. Associations between variables were calculated using Pearson's correlation. P<0.05 were considered significant. All statistical analyses were performed with GraphPad Prism Data in figures are presented as mean ± SEM, in tables as mean ± SEM or median (range). Concentration-response curves to ACh are given as percentage relaxation of the pre-constriction to NA. ECKidney function was significant lower in ESRD patient, as expected. Age, weight and BMI were similar, whereas systolic, but not diastolic, blood pressure was significantly elevated in ESRD patients , despiteRelaxation of subcutaneous resistance arteries with ACh was not significantly different between ESRD patients and controls . Both maThe EDH-like relaxation was recorded after blockade of eNOS and COX . The max50 for concentration response curves incubated with indomethacin and indomethacin + L-NAME respectively .The NO-dependent vasodilatation was calculated as ΔpECectively . No signTo assess whether three concentration response curves to acetylcholine can be obtained, time control experiments were performed. Three concentration response curves, with 20 minutes of rest in between, were conducted; without use of any blocking agent . As seenPWV and SVRi were not significantly different in the two groups, while AI was lower in the ESRD group , which wDBP was inversely related to the NO-dependent vasorelaxation in controls  . The invPulse pressure was positively correlated to ACh induced vasodilatation in ESRD , this was not the case in control subjects  .Correlation of ADMA and NO-response showed a trend in controls , althougAs expected S-phosphate levels were different in the two groups. A negative association was found between S-phosphate and EDH-like response in controls, but not in ESRD  .Cardiac output and vasodilatation were inversely related in controls , higher In the present study, we evaluated micro- and macrovascular differences in ESRD patients and healthy controls and for the first time correlated these findings. The ESRD patients had little comorbidity and well controlled blood pressure. In the microvascular function no significant different response to endothelium dependent vasodilatation was seen between ESRD and healthy controls. In the macrovascular arteries a significant higher SBP was present in the ESRD patients. Micro- and macrovascular function was correlated, DBP and the NO-dependent vasorelaxation was inversely related in controls. Pulse pressure was positively correlated to endothelial vasodilatation in ESRD and interestingly a negative correlation between S-phosphate and EDH-like response was found in controls.The microvascular endothelial dependent vasodilator response to ACh has earlier been shown to be diminished in ESRD, while attempts to determine the affected pathway, has yielded diverging results Our patients have substantial lower blood pressure than the patients in the aforementioned papers. Since diastolic hypertension is known to cause an attenuated ACh induced vasorelaxation in the resistance arteries Interestingly we found a positive correlation between pulse pressure and micro-vascular vasodilatation in ESRD patients, which was not present in control subjects. This indicates that higher pulse pressure, which could translate into arterial stiffness, generates a process in the microcirculation that increases the vasodilator properties. Knowing that our patients are well treated in terms of blood pressure and with exposure to uremic toxins, we can speculate, that with longer exposure to renal deficiency, this compensation reaches its maximum and instead diminishing of the vasodilatation starts, as shown in other studies As expected the pulse wave velocity was increased in the ESRD population Interestingly, we showed that the EDH-like response in healthy controls is dependent on the S-phosphate level, with a decreased vasodilator response with higher S-phosphate, even within the normal range of S-phosphate. We believe that further studies in this field are required to finally establish this correlation. Interestingly is has recently been shown that incubating resistance vessels from both healthy controls and patients with chronic kidney disease with a higher concentration of phosphate decrease the endothelium dependent vasodilatation When assessing cardiac output and sensitivity of resistance arteries to ACh, opposing associations were seen in controls and ESRD . This coThe sample size of study groups was relatively small which possess a risk of committing type II errors and overinterpretation of the correlations. The majority of patients was receiving kidney transplants, and was therefore with less comorbidity than most patients in dialysis. This could explain why we see little difference between healthy controls and kidney patients in some of our assays.In conclusion this study finds similar vasodilator properties in small resistance arteries of ESRD and control subjects. ESRD patients were well controlled in terms of blood pressure and uremic exposure. However correlations of pulse pressure, diastolic blood pressure and cardiac output with micro-vascular functions, indicate compensating measures in the microcirculation of ESRD patients.


The overall objective of this pilot study was to study the direct effects of tactile massage (TM) on blood pressure, heart rate and blood glucose in a sample of women suffering from primary insomnia.The study had an experimental prospective design, with a total of 10 women . The participants underwent TM twice a week for six weeks resulting in a total of 120 treatments. For short-term effects, systolic and diastolic blood pressure, heart rate and blood glucoses were assessed by the therapist before and after each treatment. Long-term assessments were made at baseline, week 7, and week 13.As a short term result after the treatment with TM, the participants reached a statistically significant reduction of systolic blood pressure , diastolic blood pressure , heart rate  and blood glucose . No long-term effects with respect to the studied variables can be observed.In summary, we have shown in a normotensive but highly stressed sample of women, that TM has beneficiary effects on parameters of stress and cardiovascular function. In total, 120 TM treatments were analyzed with respect to the objective of the study, but in order to more understand the practical effects, and to more deeply evaluate TM’s place in the modalities of stress reduction, we recommend further studies with larger samples.


Measurement of blood hemoglobin (Hb) concentration is a routine procedure. Using a non-invasive point-of-care device reduces pain and discomfort for the patient and allows time saving in patient care. The aims of the present study were to assess the concordance of Hb levels obtained non-invasively with the Pronto-7 monitor  or with the NBM-200MP monitor  and the values obtained from the usual colorimetric method using blood samples and to determine the source of discordance.−1  with an upper agreement limit at 2.94 g.L−1 , a lower agreement limit at -1.84 g.L−1  and an intra-class correlation coefficient at 0.80 . The corresponding values for the NBM-200MP™ were 0.21 , 3.42 , -3.01  and 0.69 . Multivariate analysis showed that age and laboratory values of hemoglobin were independently associated with the bias when using Pronto-7™, while perfusion index and laboratory value of hemoglobin were independently associated with the bias when using NBM-200MP™.We conducted two consecutive prospective open trials enrolling patients presenting in the emergency department of a university hospital. The first was designed to assess Pronto-7™ and the second NBM-200MP™. In each study, the main outcome measure was the agreement between both methods. Independent factors associated with the bias were determined using multiple linear regression. Three hundred patients were prospectively enrolled in each study. For Pronto-7™, the absolute mean difference was 0.56 g.LDespite a relatively limited bias in both cases, the large limits of agreement found in both cases render the clinical usefulness of such devices debatable. For both devices, the bias is independently and inversely associated with the true value of hemoglobin.NCT01321580NCT01321593ClinicalTrials.gov  After the introduction of pulse oximetry Indeed, blood hemoglobin is routinely assessed mainly for two purposes, to diagnose anemia, and then to pursue more invasive testing, and to assess the need for blood transfusion. The cut-off values leading to the diagnosis of anemia are widely accepted To date, four peer-reviewed publications Accordingly, we report the results of two subsequent studies using the same design, assessing two devices, the monitor Pronto-7™  and the monitor NBM-200MP™. The two studies are hereafter referred to as the “Masimo-Study” and the “Orsense-Study”. The aims of the studies were i) to assess the concordance of hemoglobin levels obtained non-invasively with the values obtained from the usual colorimetric method in the hospital laboratory and using blood samples and ii) to determine the source of errors in the measurements.th 2010 to February 15th 2011 for the first and from February 15th 2011 to March 31st for the second).Design and data analysis were strictly similar for the “Pronto-Study”  and the “Orsense-Study”  which were conducted consecutively (from December 15These studies were conducted in accordance with the STARD guidelines (STAndards for the Reporting of Diagnostic accuracy studies) These prospective open studies, performed in the emergency department of a university hospital, were approved by the Ethics Committee (CPP Ile-de-France VIII). Consecutive patients examined by the same senior emergency nurse and requiring a hemoglobin measurement were enrolled after they gave their informed written consent. Only the laboratory results were used for subsequent patient care.® 4D DC sensor (revision B)  or by the monitor NBM-200MP™ , with the probe placed on the patient's finger, while a nurse collected a venous blood sample in a EDTA (EthyleneDiamineTetraacetic Acid) tube, which was sent immediately to the hematology laboratory for hemoglobin measurement by the ADVIA® 2120 . Three consecutive values of SpHb were collected; the first two minutes after the probe had been placed on the patient's finger and two others after two minute intervals, all the measurements were obtained over a 6 minute period. Only one simultaneous blood sampling was performed for each patient. The measure of hemoglobin level by the ADVIA® 2120 method was considered to be the gold-standard measure.Investigators recorded the hemoglobin level determined non-invasively by the Monitor Pronto-7™ (version 2.1.9) with the RainbowAt the same time, body temperature, systolic and diastolic blood pressure, peripheral oxygen saturation, perfusion index (displayed on the Pronto-7™ monitor or on the NBM-200MP™ monitor) and heart rate were collected. Of note, the Perfusion Index (PI) of the Pronto-7 monitor provides a numeric indication of the pulse strength at the measurement site. It is a calculated percentage between the pulsatile signal and non-pulsatile signal of arterial blood moving through the site. A similar index exists for the NBM-200MP. However, concerning the Orsense device, unlike standard sensors, the NBM sensor is located on the root of the finger, where perfusion is better than in the fingertip and as the measurement is based on occlusion spectroscopy technology it is thought to rely less on the pulse signal.During the procedure, the anxiety and the pain of the patients were assessed using in both cases a visual analog scale (VAS). The VAS was graphically represented as a horizontal line, 100 millimeter (mm) in length, with word descriptors at each end . The patient marked on the line the point that they felt represented their perception of their current state. The VAS score was determined by measuring in mm from the left hand end of the line to the point that the patient marked. The evaluation was performed 2 min after the placement of the probe on the patient's finger and 2 min after the blood sampling.Results are expressed as median and first and third quartiles [Q1 to Q3] or counts and percentages. Comparison of values obtained by the two methods was performed with a paired Student t test. The individual values of SpHb and perfusion index (PI) were defined as the mean of the three measurements. For the concordance analysis, in both studies, we used the mean of the three consecutive non-invasive measurements to define the value of SpHb.For agreement between the two hemoglobin determination methods (invasive and non-invasive), a Bland–Altman analysis was applied calculating bias as the mean difference between both methods and limits of agreement as the range in which 95% of the differences between the two methods are expected to lie Multivariate linear regression was used to determine a set of variables independently associated with bias. Bias was defined as the absolute difference between the two measurements and was expressed in g/dL. For continuous covariates, the linearity assumption was checked using additive regression models with splines. Model goodness-of-fit was assessed by examination of residuals .The percentage of variation over the three non-invasive measurements was determined. First, the variation was calculated for each of the three measurements, then the highest value was used to define the individual coefficient of variation of the non-invasive measurement.The within-subject coefficient of variation of each noninvasive and invasive technique was calculated by analyzing 15 different samples in duplicate. The coefficient of variation is defined as the ratio of the standard deviation of the absolute difference between the duplicates divided by the mean of the average value of the duplicate According to the methods developed for reliability study Comparisons of pain and anxiety VAS scores were also performed using non-linear mixed effect models, in order to take into account the intra-subject correlation.A two-sided p-value of 0.05 was considered significant. All analyses were performed using R 2.10.1 statistical software .Three hundred patients were prospectively included in each study. The flow chart of the studies is shown in −1  with an upper agreement limit at 2.94 g.L−1 (95% CI 2.70 to 3.19), a lower agreement limit at -1.84 g.L−1 (95% CI -2.08 to -1.58) and an intra-class correlation coefficient at 0.80 (95% CI 0.74 to 0.84). The corresponding values for the NBM-200MP™ monitor were 0.21 , 3.42 , -3.01  and 0.69 .The Bland-Altman graphical representation of the concordance is reported in In univariate analysis, age, diastolic blood pressure and the hemoglobin value when assessed in the laboratory were significantly associated with the bias associated with Pronto-7™. Similarly, in univariate analysis, gender, heart rate, body temperature, perfusion index and the hemoglobin value when assessed in the laboratory were significantly associated with the bias associated with Orsense™ Table 3.Similarly, The within-subject coefficients of variation were 1.3% for the ADVIA 2120, 3.5% for the Pronto 7™ and 5.9% for the NBM-200MP™. The pain and anxiety VAS scores were significantly lower with the non-invasive measurement in both studies. The differences for pain on the VAS scale were 1.62  (p<0.0001) and 1.62  (p<0.0001) in the Pronto-Study and the Orsense-Study respectively. Similarly, the differences for anxiety were 1.00  (p<0.0001) and 0.8  (p<0.0001).Compared to previous reports Compared to previous reports This first study of the monitor NBM-200MP™ shows a small bias but limits of agreement higher than 3 g/dl and an ICC coefficient lower than 0.8. Moreover, the measure seems to be unstable, with a mean coefficient of variation at 5.9% and an absolute difference between two consecutive measurements lower than 2.5% in less than 50% of the cases. For this monitor also, the bias is independently affected by the true level of hemoglobin and the index of perfusion is also independently associated.The two studied monitors showed slight differences in performance. These differences could be explained by the fact that the technology used is not the same. Indeed, Pronto-7™ and NBM-200MP™ are both non-invasive solution for on-line, continuous and spot hemoglobin measurements combined with oximetry measurements, but the first uses more than seven wavelengths of light to acquire blood constituent data based on light absorption through a finger probe. The second involves a ring-shaped sensor fitted on the patient's finger that temporarily gently squeezes the finger to over-systolic pressure, similar to blood pressure measurements. Moreover, the signal processing algorithms and filters used are not the same in both cases. In contrast with these differences in performance, both devices were associated with a reduction in pain and anxiety scales with the non-invasive measure, regardless of the monitor used. Even if this result is to be expected, it is interesting for the improvement in patient care associated with the use of such point-of-care methods.A noninvasive and accurate estimation of hemoglobin could have many roles in patient care, for example as an early warning system for bleeding, as a way of monitoring high risk patients for bleeding , and as a screening tool for people who have difficult venous access. It also has many theoretical advantages, such as its ability to perform repeated sampling without causing iatrogenic blood loss linked to anemia, particularly in the intensive care unit and also its ability to monitor hemoglobin concentrations in a pediatric population.®  provides a quick and acceptable estimation of hemoglobin compared to laboratory measurements but requires taking a blood sample®.Hemoglobin can also be measured through the use of other transportable, portable, and handheld instruments at or near the site of patient care (point-of-care testing). HemoCueEven if the two populations involved in the two studies reported in the present article seem comparable, the two devices were not compared in a single trial. Therefore, no direct comparison of their performance is possible. A comparative trial was not conducted for technical reasons, as the two devices were not available at the same time in our center. However, because of the identical design, the large sample size and the comparability of the two samples, results can be compared. The study was performed in a single center; this could limit generalization of the results. However, the large sample size led to include patients with various medical conditions. Even if the rate of anemic patients was around 30% in both studies, few patients presented extreme values of hemoglobin and the results of the studies may not be generalizable to them. Thus, further studies are needed to address this issue. Moreover, it would be interesting to test theses devices in the setting of significant fluid shifts, blood loss or active bleeding, as they are common situations occurring in the operating room, or after major trauma. The performance of the studied devices needs to be assessed in these situations.Finally, although we studied the latest version of the device produced by Masimo, we assessed the first one produced by Orsense and another version, specifically designed for spot-check measurement, namely NBM-200™, exists but was not assessed in the present study. This could explain the slight difference between the two devices and so further investigations are needed to evaluate this alternative version.In short, two devices dedicated to non-invasive measurement of hemoglobin were assessed in the present study. Bias was found to be small but independently and inversely associated with the true value of hemoglobin. Of more importance is that limits of agreement are large in both cases making the clinical usefulness of such devices debatable.


Invasive monitoring in critically ill patients allows a continuous measurement of arterial pressure, cardiac output, and the derivation of dynamic predictors of fluid responsiveness. However, the pressure signal may be altered by the dynamic characteristics of the fluid-filled tubing. The aim of the present study was to evaluate the reliability of radial artery blood pressure measurement and derived indexes during the early period after cardiac surgery.t test was used for statistical comparison, P < 0.05 was considered statistically significant.After IRB approval, 30 patients admitted to the ICU after elective cardiac surgery  with a radial artery catheter were included. In the ICU, an independent continuous recording of arterial pressure during at least 18 hours was started via a double-head pressure transducer  for a retrospective analysis and three fast flushes were performed. First, the whole record was examined for episodes of overdamping (Ov) or attenuation (At). Ov was defined as a decrease in systolic (sAP), an increase diastolic (dAP), and an unchanged mean pressure (mAP). At was defined as a decrease in sAP, dAP and mAP. Second, three periods of 10 minutes during the first hour were analysed assuming that the dynamic characteristics remained constant. This allowed the correction of the distorted raw signals and the study of the consequences of an underdamped signal on sAP, pulse pressure variation (PPV) and dP/dt as an estimate of left ventricular contractility. A paired P < 0.0001) (range: 0.3 to 5.9) or by 4.3 ± 0.9% (range: 0.4 to 15.9%), raw PPV was 9.5 ± 7.3 versus 10.0 ± 7.8 for the corrected PPV (range from -2.6 to 4.3); raw dP/dt was overestimated by 134 ± 47 mmHg/second (P < 0.0001) (range: -13 to 353) or by 24 ± 6%.Mean age was 69 ± 13 years, 14 patients received noradrenaline, eight patients dobutamine, and nine patients volume expansion. During the whole record, the number of episodes of Ov or At ranged from 0 to 15 with a duration of 0 to 6 hours: 17 patients had at least one episode of Ov and/or At tracing, 10 patients had at least two episodes, eight patients had at least five episodes. Seven episodes lasted more than 20 minutes and three more than 1 hour. During the first hour, sAP was overestimated by 5.0 ± 1.4 mmHg (These results showed that frequent artefacts and distortions induced by the fluid-filled tubing could modify the arterial waveform and could lead to inaccurate therapy . More at


Introduction: Hypertension is a chronic disease, which represents one of the most common public health problems in the world and afflicts about 28% of the North Americans, 44% of the Europeans and 26% of the East Mediterranean people aged 35 to 64 years. In Iran, about 11.5% of the people who are over 15 years old are afflicted. Hypertension is the most prevalent cardiovascular disease; it is the leading cause of stroke, heart attack, kidney disease and aortic aneurysm.Materials and Methods: This is a cross–sectional study, carried out on 200 patients who were referred to the Health centers of Yazd city in center of Iran. We adapted tools from previous studies. All patients were asked to express their personal goals with respect to their hypertension values. The reliability and validity of the questionnaire were determined and their Alfa cronbach was (a=0.83).Results: The mean number of years with hypertension was 8.7[+/–]7.6. Men and women showed moderately high means for attitudes, subjective norms, and positive anticipated emotions toward trying to reduce or maintain their blood pressure. The data indicated there is significantly different between the personnel and housewives, with p<0.001. In addition, the personnel's mean grade scores  for trying the self–regulation of their blood pressure was (15.9[+/–]4.08) and, in the housewives was (12.49[+/–]4.33).Conclusion: The results of this study showed that the Model of Goal Behavior can explain more than 52% of the self–regulation of hypertension, so, this Model can be a basic Model for intervention in education, in order to decrease and control hypertension in patients. Hypertension is a chronic cardiovascular disease, which is the leading cause of morbidities in human population, becoming a major public health problem . About 2Being a primary contributor to disability and mortality, hypertension cannot be cured but must be managed. Medication is the leading way to treating hypertension. Pressure control can substantially reduce the risk of stroke and heart attack. More than 100 different drugs are available for its treatment; all have proven efficacy, most have few side effects and many are formulated for once–daily dosing. Then, why do the reported rates of blood pressure control are so disappointing, and apparently not getting any better? Indeed, the success rate for the control of hypertension in the United States is only 27%, being even lower in England, France, and Germany . Despite major financial and time investment, the treatment compliance of hypertension has remained low. The majority of hypertensive patients do not reach the current target Blood Pressure (BP) levels set by the international guidelines (self–monitoring).One promising method to improve BP control in hypertensive patients is self–measurement of BP .Medical Clearly, the management of hypertension requires self-regulation, which is defined as the mental and physical process that a person manages in order to achieve a goal. Unfortunately, until now, studies have had limited inquiry to compliance with medication regimens and, have focused on correlations such as the health status. Little research exists about decision- making processes and social psychological aspect of the self-regulation of hypertension. Most of the discussions in the literature have been conceptual rather than empirical in nature . The purTo better explain goal striving, Bagozzi, Baumgartner, and Pieter  introducThe processes behind the functioning of anticipated emotions are counterfactual thinking but might be termed as prefectural to stress the expected, forward–looking aspects of the thought processes. Individuals may think about imaginary alternatives to events in terms of the implications of these events for the future. People's behaviors may well be determined by what they imply for the future . Thus, aThe MGB also includes past behavior as a predictor of desires, intentions, and trying. This yields two benefits. First, past behavior serves as a control for unmeasured determinants of behavior that may have been stable over time in their effects. Second, past behavior may reflect habitual processes. A recent meta-analysis for the role of past behavior supports these claims, as do a number of recent studies . This was a cross–sectional study implemented on 200 patients who were referred to Health centers of Yazd city in center of Iran. We used some tools from the previous studies –17. PatiFor data collection, we used a questionnaire that was previously used by Taylor et al  but we cTo ensure the clarity of the questionnaire, pilot testing of the questionnaire was also performed by using the coherence and consistency of 30 participants who were not included in the survey.Afterwards, the questionnaire was modified based on their feedback. Content validity was established by 5 experts who were academic staff and health educators. The reliability and validity of the questionnaire was determined and its Alfa cronbach was (a=0.83).All the data were transferred directly into SPSS . The level of confidence interval was of 95%We obtained an informed consent from all the participants and, in addition, the managers of Health centers assured the participants that their responses were confidential.Of 200 participants, 45.5% were men and 54.5% were women. The mean age was (59.8[+/–]11.9) 30–86 years old. A total of 8.5% had diploma or completed some university education, 9.5% studied 6–12 years of school (did not graduate with diploma), 28.5% studied in primary school (less than six years of school) and 53.5% were illiterate. The mean number of years with hypertension was (8.7[+/–]7.6). 86.5% reported adherence to their anti–hypertensive medications. With respect to stated goals, 61.5% of them indicated that their goal was to reduce their blood pressure and 35.5% had the goal to maintain their current blood pressure. The participants in this study were approximately 52% housewives, 10% government personnel, 19.5% free workers and 18.5% unemployedMen and women showed moderately high means for attitudes, subjective norms, and positive anticipated emotions toward trying to reduce or maintain their blood pressure. Men showed moderate means for perceived behavioral control, past behavior, desire, trying and intention to reduce or maintain their blood pressure. Women showed lower moderate mean in perceived behavioral control, past behavior, desire, trying and intention to reduce or maintain their blood pressure. The mean of negative anticipated emotion was lower than the mean in men and women. There was a significant difference between all the constructs of MGB, gender and self–regulation of their hypertension, unless the negative anticipated emotion. With respect to education, the data showed there is a significant difference between the trying intention for self–regulation of blood pressure, and the level of education of participants with p<0.001, it is significant for a perceived behavior control which is (p<0.008) and for positive anticipated emotion which is (p<0.039).The mean grade scores for trying, of the subjects in guidance and high school education, in self–regulation for decreasing or maintaining their blood pressure, was more than (17.05[+/–]3.82) others and in illiterate participants it was lower than others (13.12[+/–]4.23).. The data showed there is a significant difference between the trying of the government personnel and that of the housewives' with p<0.001, so the mean grade scores of trying for the self-regulation of their blood pressure in government personnel was (15.9[+/–]4.08) and in housewives was (12.49[+/–]4.33)Significant effects on desire were found for attitude (p<0.01), for PAE (p<0.01), for subjective norms (p<0.01), for PBC (P<0.01), for past behavior (p<0.01) and for trying (p<0.01). There was no significant difference between desire and NAE. The data showed there was a significant difference between all the constructs of MGB and trying and the intention for self–regulation of blood pressure (P<0.01) unless NAE. . ResultsIn this study, we understood how decision–making processes occur in the self–regulation of blood pressure. In this study, the mean number of years with hypertension was (8.7[+/–]7.6). 86.5% reported adherence to their anti–hypertensive medications. With respect to the stated goals, 61.5% indicated that their goal was to reduce their blood pressure and 35.5% had the goal to maintain their current blood pressure. The results of Stephanie  revealedWe saw a significant difference between all the constructs of MGB and the gender of the participants, unless a negative anticipated emotion occurred. Men and women showed moderately high means for attitudes, subjective norms, and positive anticipated emotions toward trying to reduce or maintain their blood pressure. Perceived behavioral control, past behavior, desire, trying and intention to reduce or maintain blood pressure mean scores in women were lower than in men. The mean of negative anticipated emotion was lower in men and women. However, Stephanie et al  describeIn a study, carried out by Jackson , it was Our results showed that the past behavior, attitude, subjective norms, perceived behavior control and desire to self–regulation have an influence on the intention of reducing or maintaining their blood pressure. These results are consistent with the results of Hagger et al , they reIn the present study, negative anticipated emotions had not influence on the intention to control the blood pressure; these are the similar results to the ones of Chapman , who shoResults of our study indicate that the MGB parameter estimates and explains variance. The explained variances in perceived behavior control , past behavior  and intention  in trying to decrease and maintain blood pressure was of 52%. The explained variances in perceived behavior control , past behavior  and desire  for the intention of decreasing and maintaining blood pressure was of 50% and explained variances in perceived behavior control , past behavior  NAE , PAE , subjective norms  and attitude  for the desire of decreasing and maintaining blood pressure was of 29%.In a study carried out by Bagozzi et al , the expWe can indicate that the behavior of hypertension control, could improve about 52% of the blood pressure, but we must pay attention to the hidden aspect of this behavior and improve them. This way, if we manipulate the Attitude, Subjective Norm and Emotion of patients and they improve their Desire, we cannot only control the hypertension 30% more than before, but we can also make a better substructure to change the  patients' behavior