Safety and efficacy of candesartan versus valsartan combined with amlodipine on peripheral and central blood pressure / Seguridad y eficacia de candesartán frente a valsartán combinado con amlodipino sobre la presión arterial periférica y central

Introduction: “Amlodipine/valsartan” or “amlodipine/candesartan” combinations represent two effective antihypertensive agents with complementary mechanisms of action. Nevertheless, a study has yet to be done to evaluate the effect of amlodipine/candesartan on central blood pressure and compare it with amlodipine/valsartan combination. To see how “amlodipine plus candesartan combination” reduces peripheral and central blood pressure compared to the most studied combination, “amlodipine plus valsartan”. Material and methods: Eighty-six patients were randomized in an open-label, prospective study by 1:1 ratio to two groups. Group I (n=42) received the amlodipine and valsartan combination, and group II (n=44) received the amlodipine and candesartan combination. Peripheral and central blood pressure (CBP) was measured at baseline, at 6 and 12 weeks of follow-up. Discussion: Both treatment groups reduced peripheral systolic, diastolic, and mean blood pressure. There was no significant difference between and within both groups. The amlodipine/candesartan combination showed more reduction in peripheral systolic blood pressure (PSBP) after 12 weeks of treatment (p=<0.001). Both groups decreased CBP without significant differences between groups. The amlodipine/candesartan combination showed additional efficacy in decreasing CSBP after 12 weeks (p=<0.001). The two treatment groups did not exert significant efficacy in lowering heart rate (HR) and augmentation index% (AIx%). Conclusion: To conclude, the amlodipine 10mg/candesartan 16mg combination was non-inferior to the amlodipine 10mg/valsartan 160mg combination in terms of reducing peripheral and CBP over time.(AU)

Introducción: «Las combinaciones de amlodipino/valsartán» o «amlodipino/candesartán» representan 2 agentes antihipertensivos efectivos con mecanismos de acción complementarios. Sin embargo, aún no se ha realizado un estudio para evaluar el efecto del amlodipino/candesartán en la presión arterial central y compararlo con la combinación amlodipino/valsartán. En este estudio, se comparó la reducción de la presión arterial periférica y central entre estas 2 combinaciones. Materiales y métodos: Ochenta y seis pacientes fueron asignados aleatoriamente a 2 grupos: el Grupo I (n=42) recibió amlodipino y valsartán, y el Grupo II (n=44) recibió amlodipino y candesartán. Se midió la presión arterial periférica y central al inicio, a las 6 y 12 semanas de seguimiento. Discusión: Ambos grupos redujeron la presión arterial periférica de manera similar, pero la combinación amlodipino/candesartán mostró una mayor reducción en la presión arterial sistólica periférica después de 12 semanas de tratamiento. Ambas combinaciones también disminuyeron la presión arterial central, pero nuevamente, la combinación amlodipino/candesartán tuvo una mayor eficacia en la reducción de la presión arterial sistólica central después de 12 semanas. No se observaron diferencias significativas en la frecuencia cardíaca ni en el índice de aumento entre los grupos. Conclusión: En conclusión, la combinación de amlodipino 10mg/candesartán 16mg demostró ser tan efectiva como la combinación de amlodipino 10mg/valsartán 160mg en la reducción tanto de la presión arterial periférica como central a lo largo del tiempo.(AU)

Safety and efficacy of candesartan versus valsartan combined with amlodipine on peripheral and central blood pressure.

INTRODUCTION: "Amlodipine/valsartan" or "amlodipine/candesartan" combinations represent two effective antihypertensive agents with complementary mechanisms of action. Nevertheless, a study has yet to be done to evaluate the effect of amlodipine/candesartan on central blood pressure and compare it with amlodipine/valsartan combination. To see how "amlodipine plus candesartan combination" reduces peripheral and central blood pressure compared to the most studied combination, "amlodipine plus valsartan". MATERIAL AND METHODS: Eighty-six patients were randomized in an open-label, prospective study by 1:1 ratio to two groups. Group I (n=42) received the amlodipine and valsartan combination, and group II (n=44) received the amlodipine and candesartan combination. Peripheral and central blood pressure (CBP) was measured at baseline, at 6 and 12 weeks of follow-up. DISCUSSION: Both treatment groups reduced peripheral systolic, diastolic, and mean blood pressure. There was no significant difference between and within both groups. The amlodipine/candesartan combination showed more reduction in peripheral systolic blood pressure (PSBP) after 12 weeks of treatment (p=<0.001). Both groups decreased CBP without significant differences between groups. The amlodipine/candesartan combination showed additional efficacy in decreasing CSBP after 12 weeks (p=<0.001). The two treatment groups did not exert significant efficacy in lowering heart rate (HR) and augmentation index% (AIx%). CONCLUSION: To conclude, the amlodipine 10mg/candesartan 16mg combination was non-inferior to the amlodipine 10mg/valsartan 160mg combination in terms of reducing peripheral and CBP over time.

Breed-typical front limb angular deformity is associated with clinical findings in three chondrodysplastic dog breeds.

Angular front limb deformity (ALD) refers to an excessively curved limb conformation, which is seen in some chondrodysplastic dog breeds. Common characteristics of ALD include carpal valgus (VALG), front limb rotation (ROT), elbow incongruity, and lateral radial head subluxation. These may cause lameness and discomfort in affected dogs. The clinical impact and breed-specific characteristics of front limb conformation in chondrodysplastic breeds are unknown. This prospective and cross-sectional study aimed to investigate differences in front limb conformation between three chondrodysplastic breeds. We further evaluate whether front limb conformation is associated with clinical findings and limb function. We propose novel methods to classify findings in the interosseous space and to quantify lateral radial head subluxation. Data from a total of 224 front limbs from 112 dogs of three chondrodysplastic dog breeds (30 Standard Dachshunds, 29 Skye terriers, and 53 Glen of Imaal terriers) were included in the study. Front limb VALG and ROT were measured with a goniometer. From the radiographs, the elbow joint was graded for incongruity (INC), and the humeroradial angle (HRA) was measured to assess lateral radial subluxation. The association of front limb conformation with clinical signs and limb function was investigated using orthopedic examination, goniometric and kinetic measurements, and radiography. The breeds differed significantly in their front limb conformation. The Dachshund had the least ROT and the least radial head subluxation. The Skye terrier had the most VALG, the most radial head subluxation, and the largest prevalence of moderate and severe INC. The Glen of Imaal terrier had the most ROT. In addition, INC, ROT, VALG, and HRA were found to be independent of each other and were associated with several measurable clinical abnormalities and limb function such as pain, lameness, limited range of motion, and elbow joint osteoarthritis. This implies that VALG, ROT, and HRA could be used in addition to INC grading when choosing musculoskeletal characteristics of dogs suitable for breeding.

The intra- and intertester repeatability of radiographic elbow incongruity grading is high in chondrodystrophic dog breeds.

Elbow incongruity is a form of elbow dysplasia that causes osteoarthritis, pain, and lameness, and it is common in chondrodystrophic dog breeds. The objective of this retrospective secondary analysis study was to evaluate the intra- and interobserver repeatability of a novel radiographic incongruity grading system for assessing elbow incongruity in three chondrodystrophic dog breeds-the dachshund, Skye Terrier, and Glen of Imaal Terrier. We conducted an observer agreement study that included 220 mediolateral antebrachial radiographs from 110 dogs with the elbow in 90° flexion. The radiographs were independently assessed by three observers at three time points, using a four-stepped grading scale. The proportion of agreement and Kappa coefficient were calculated. Both the intra- and interobserver proportions of agreement were substantial when three grades were required to be identical (.705-.777 and .609, respectively), and almost perfect for two identical grades (.991-1.000 and .991, respectively). Some differences in repeatability between breeds were noted; specifically, the intraobserver repeatability was higher in the dachshund, and the interobserver repeatability was lower in the Glen of Imaal Terrier. Our study showed that the radiographic imaging protocol and incongruity grading system have high repeatability when assessing elbow incongruity in chondrodystrophic dog breeds.

Targeted deletion of the CD59 gene causes spontaneous intravascular hemolysis and hemoglobinuria.

The glycolipid-anchored glycoprotein CD59 inhibits assembly of the lytic membrane attack complex of complement by incorporation into the forming complex. Absence of CD59 and other glycolipid-anchored molecules on circulating cells in the human hemolytic disorder paroxysmal nocturnal hemoglobinuria is associated with intravascular hemolysis and thrombosis. To examine the role of CD59 in protecting host tissues in health and disease, CD59-deficient (CD59(-/-)) mice were produced by gene targeting in embryonic stem cells. Absence of CD59 was confirmed by staining cells and tissues with specific antibody. Despite the complete absence of CD59, mice were healthy and fertile. Erythrocytes in vitro displayed increased susceptibility to complement and were positive in an acidified serum lysis test. Despite this, CD59(-/-) mice were not anemic but had elevated reticulocyte counts, indicating accelerated erythrocyte turnover. Fresh plasma and urine from CD59(-/-) mice contained increased amounts of hemoglobin when compared with littermate controls, providing further evidence for spontaneous intravascular hemolysis. Intravascular hemolysis was increased following administration of cobra venom factor to trigger complement activation. CD59(-/-) mice will provide a tool for characterizing the importance of CD59 in protection of self tissues from membrane attack complex damage in health and during diseases in which complement is activated.

Role and regulation of pig CD59 and membrane cofactor protein/CD46 expressed on pig aortic endothelial cells.

BACKGROUND: Hyperacute rejection in xenotransplantation is caused by activation of complement (C) on endothelium. We have previously shown that purified C-regulators of the pig (CD59 and membrane cofactor protein [MCP]) are efficient regulators of human C (HuC). The aim of this study was to clarify the role of endogenously expressed C-regulatory molecules on pig endothelium in the protection against hyperacute rejection. METHODS: Porcine aortic endothelial cells (PAEC) were harvested and cultured for various passages. PAEC were examined for the expression of endogenous pig CD59 and MCP by flow cytometry. PAEC were assessed for their susceptibility to lysis by HuC. The effect of phorbol 12-myristate 13-acetate and various cytokines on the expression of MCP and CD59 and C-susceptibility was assessed. RESULTS: Primary PAEC showed an initial high level of expression of pig CD59, however, upon culturing, CD59 levels decreased dramatically to about 20% after five passages. In contrast, levels of MCP doubled upon culturing of PAEC to confluency and remained stable during at least five passages. Primary cells and cells in the early passages were more resistant to HuC than cells that were cultured for longer. Blocking the function of CD59 but not of MCP using monoclonal antibody increased the susceptibility to HuC. Purified human CD59 incorporated to a level of expression similar to that of pig CD59 reversed the increased C-susceptibility, suggesting that pig and human CD59 are similarly protective against HuC. Increase of C-resistance and of expression of pig MCP, but not of CD59, was achieved upon incubation with phorbol 12-myristate 13-acetate. Tumor necrosis factor-alpha, interleukin-1beta, interleukin-4, or interferon-gamma had no effect on C-regulator expression or C-susceptibility. CONCLUSIONS: These data demonstrate the importance of using primary PAEC or cells in the first passages of culturing in in vitro models of xenotransplantation and show that pig MCP and, in particular, pig CD59 play an important role in protection of PAEC from HuC.

Distribution of membrane cofactor protein (MCP/CD46) on pig tissues. Relevance To xenotransplantation.

Membrane cofactor protein (MCP; CD46) is a 50-60 000 MW glycoprotein, expressed on a wide variety of cells and tissues in man, which plays an important role in regulating complement activation. Human MCP has also been shown to be the receptor for measles virus. We have recently identified the pig analogue of MCP and demonstrated that pig MCP has cofactor activity for factor I-mediated cleavage of C3b when these components are derived either from pig or human. As a consequence, pig MCP is an efficient regulator of the classic and alternative pathways of human and pig complement. In order to define the potential importance of MCP in protecting against complement activation in the pig, we have conducted a comprehensive survey of its distribution in pig cells and organs. As in humans, MCP in the pig is broadly and abundantly distributed. Pig MCP is highly expressed on all circulating cells, including erythrocytes, in contrast to its absence on human erythrocytes. Multiple isoforms of MCP are found on cells and in tissues, probably representing products of alternative splicing analogous to those found in man. MCP is abundantly expressed throughout all tissues examined with particularly strong staining on the vascular endothelium. Connective tissue elements within liver and testis are also strongly stained by anti-pig MCP antibodies. Pig MCP is expressed only weakly on skeletal muscle cells and expression is absent from smooth muscle cells in the lung and vessel walls, sites at which human MCP is expressed. Of particular note, MCP is not expressed in B-cell areas of the germinal centres of lymph nodes.

Tissue distribution of the rat analogue of decay-accelerating factor.

In humans, decay-accelerating factor (DAF) is a widely distributed, cell-bound inhibitor of the complement activation enzymes and plays a key role in regulating complement activation, preventing the generation of anaphylotoxins and opsonins, and protecting against complement-mediated lysis. Rodent analogues of DAF have recently been identified, providing a new avenue for the analysis of function. Rat DAF was cloned in our laboratory. Here we describe the generation of monoclonal antibodies (mAbs) against rat DAF, using transfected cells as immunogen, and their use in the analysis of the distribution of DAF in the rat by flow cytometry, Western blot analysis and immunohistochemistry. One of the mAbs was found to block the complement inhibitory function of rat DAF, offering the prospect of neutralization of DAF function in vivo. The antibodies have also been used for purification of DAF from rat erythrocytes by affinity chromatography. Rat DAF purified in this manner was similar in molecular mass to human DAF. The purified protein incorporated into lipid membranes, confirming the presence of a glycolipid anchor, and incorporated protein strongly inhibited the rat C3 convertase. Rat DAF was strongly expressed on endothelia throughout the animal and was also present in most tissues and organs. DAF expression was weak or absent in the brain and on circulating and spleen-resident T cells. Strong DAF expression observed in the kidney was restricted to the glomerulus and Bowman's capsule. DAF expression in the testis was found only in association with the later stages of spermatogenesis.

Characterization in vitro and in vivo of the pig analogue of human CD59 using new monoclonal antibodies.

CD59 is the sole characterized regulator of the complement membrane attack complex in humans. It is very widely and abundantly distributed, being present on all circulating cells, endothelia and epithelia, and in most tissues. CD59 analogues in rodents are distributed similarly. Interest in complement regulation in the pig has developed out of the current enthusiasm to exploit this species as a donor in xenotransplantation of organs to humans. We have recently isolated and cloned the pig analogue of human CD59. We here report the development and characterization of monoclonal antibodies against pig CD59. We have used these antibodies to develop efficient methods for the purification of pig CD59 to homogeneity from erythrocyte membranes and have obtained new information on the structure and function of the purified protein. The antibodies were found to function well in immunohistochemistry and have been used to perform a comprehensive survey of the expression and distribution of pig CD59 on cells and in organs of normal pigs. Pig CD59, like human CD59, is broadly expressed but there are some striking differences in tissue distribution, notably the apparent lack of pig CD59 on circulating platelets and on a subset of leucocytes in blood and lymphoid organs. The reported findings have important implications for the current approaches to avoiding complement-mediated hyperacute rejection in pig-to-human xenografts.

Detection of human TNF-alpha mRNA by NASBA.

A method to amplify and detect TNF-alpha mRNA from primed Mono Mac 6 cells is described. A silica-based extraction system was utilised for preparation of cell extracts and specific oligonucleotide primers were designed for amplification of TNF-alpha mRNA by the NASBA process. Amplification products were detected using either a liquid hybridisation assay, with analysis by polyacrylamide gel electrophoresis, or a plate hybridisation system. The method has many potential applications for the study of inflammatory cytokines and cellular mRNAs in cell culture and clinical samples.

Molecular cloning and functional characterization of the pig analogue of CD59: relevance to xenotransplantation.

In this work, we report the cloning of the cDNA for the porcine analogue of human CD59. Degenerate primers, derived from the N-terminal sequence of pig erythrocyte CD59, were used to obtain the corresponding cDNA sequence. From this sequence, gene-specific primers were designed and used to amplify the 3' and 5' ends of the cDNA using the rapid amplification of cDNA ends (RACE) method. The complete 768-bp cDNA so obtained consisted of a 84-bp 5' untranslated region, a 26-amino-acid NH2-signal peptide, a 98-amino-acid coding region, including putative N-glycosylation sites and a glycosylphosphatidylinositol-anchoring signal, and a 312-bp 3' untranslated region. The mature protein sequence was 48% identical to human CD59 at the amino acid level. Northern blot analysis revealed several distinct CD59 transcripts, and a variability in expression levels of the different transcripts in the panel of tissues screened. Stable expression of pig CD59 in a CD59-negative human cell line conferred protection against lysis by complement from pig and several other species. Separate expression of pig and human CD59 at similar levels in the same cell line allowed a direct functional comparison between these two analogues. Pig CD59 and human CD59 showed similar activity in inhibiting lysis by complement from all species tested; in particular, expressed pig CD59 efficiently inhibited lysis by human complement. The relevance of these data to current work in the engineering of pig organs for xenotransplantation is discussed.

Neutralization of cytomegalovirus virions: the role of complement.

Complement provides a key immunologic defense against invading pathogens; thus, a clear understanding of the interactions between cytomegalovirus (CMV) and complement may permit the development of strategies to enhance CMV neutralization. In the presence of specific anti-CMV antibodies, complement enhanced the neutralizing ability of serum by 2- to 3-fold. However, in the absence of specific anti-CMV antibodies, complement was ineffective in neutralizing CMV virions by plaque assay. Although complement alone did not mediate any neutralizing effect, CMV consumed complement activity from seronegative serum, resulting in the deposition of C3 on the virion. However, only in the presence of specific anti-CMV antibody did complement activation continue to the deposition of C9 on the virions. These results strongly suggest complement regulation by CMV virions that is modulated by anti-CMV antibody; this regulation may be attributed to three host complement regulators on the virions: CD55, CD46, and CD59.

Impassable ureteral strictures: management with percutaneous ureteroneocystostomy.

Sixteen of 227 patients referred for percutaneous placement of a ureteral stent had impassable stenoses. Stenoses were benign (n = 8) or attributed to malignant retroperitoneal neoplasm (n = 8). Electrocautery was used to create a neotract between the stenosed ureter and the bladder or ileal loop. A double-J stent was placed after dilation of the tract by use of angioplasty. Neotracts were established and stents were placed in all patients. Complications (digestive tract fistulas) developed in two patients. This technique is safe if the electrode is placed close to the bladder or ileal loop. The procedure can be used as an alternative to surgery or permanent nephrostomy or in initial treatment of benign anastomotic stenosis.

Electro-oculogram changes at the switch in a manic-depressive patient.

The correlations between eye movements on the EOG of a patient with a 48 hour cycle of manic-depressive type are described. They are used to confirm the fact that his switch from one state to another occurred at the same time of night whether he was awake or asleep.

[The enzyme linked immuno-sorbent assay (ELISA) method applied for human toxoplasmosis diagnosis (author's transl)]. / La méthode ELISA dans le diagnostic sérologique de la toxoplasmose humaine.

The enzyme linked immuno-sorbent assay (ELISA) was applied for the diagnosis of toxoplasmosis in man. Examination of different parameters showed that for the same optic density, the concentration of the antigen varies directly with the serum dilution is a linear pattern. Two hundred and thirty eight serum samples were examined by the direct agglutination test, the indirect fluorescent antibody technique and the ELISA method. The ELISA was found to be more sensitive than the other two. Eighty sera were titrated by using the macro method in tubes and the micro method in plates. The latter procedure was more sensitive and showed better discrimination between low and high positives.

[The enzyme-linked-immunosorbent assay and its application to toxoplasmosis immunity (author's transl)]. / Application de la méthode immunoenzymatique à la détection de l'immunité antitoxoplasmique.

The enzyme-linked-immunosorbent assay (ELISA) was used for the serodiagnosis of toxoplasmosis. 238 sera were tested, and the results compared with those of agglutination and immunofluorescent test. ELISA gives a good correlation (p < 0.001).