Evaluation of effect of cyclosporine A on the bone tissue with induced periodontal disease to ligature in rats

The administration of cyclosporine A (CsA) has been associated with significant bone loss and increased bone remodeling. The present investigation was designed to evaluate the effects of CsA on alveolar bone of rats subjected to experimental periodontitis, using histomorphometric and histological analysis. Twenty-four rats were divided into groups with 6 animals each: 1, control; 2, rats with ligature around the lower first molars; 3, rats with ligature around the lower first molars and that were treated with 10 mg CsA/kg of body weight/d; and 4, rats treated with 10 mg CsA/kg of body weight/d. At the end of 30 days, rats were humanely killed and subjected to a histological processing, with analysis of the distance cemento-enamel junction and alveolar bone crest, bone area, eroded bone area, and cemento surface. All of them were assessed at the mesial region of the alveolar bone. The CsA therapy combined with ligature placement decreased bone area and increased the eroded bone area around the tooth surface. The results at the histological analysis showed the same combination and changes. Therefore, in spite of the lack of a direct effect on the alveolar bone height, the CsA therapy intensified the imbalance of the alveolar bone homeostasia in a rat model of experimental periodontitis. © 2013 Elsevier Inc.

Cyclosporine improves remyelination in diabetic rats submitted to a gliotoxic demyelinating model in the brainstem

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Clinicopathologic features and outcome of mycophenolate-induced colitis in renal transplant recipients

Reports on the clinical course of mycophenolic acid (MPA)-related colitis in kidney transplant recipients are scarce. This study aimed at assessing MPA-related colitis incidence, risk factors, and progression after kidney transplantation. All kidney transplant patients taking MPA who had colonic biopsies for persistent chronic diarrhea, between 2000 and 2012, at the Kidney Transplantation Unit of Botucatu Medical School Hospital, Brazil, were included. Cytomegalovirus (CMV) immunohistochemistry was performed in all biopsy specimens. Data on presenting symptoms, medications, immunosuppressive drugs, colonoscopic findings, and follow-up were obtained. Of 580 kidney transplant patients on MPA, 34 underwent colonoscopy. Colonoscopic findings were associated with MPA usage in 16 patients. The most frequent histologic patterns were non-specific colitis (31.3%), inflammatory bowel disease (IBD)-like colitis (25%), normal/near normal (18.8%), graft-versus-host disease-like (18.8%), and ischemia-like colitis (12.5%). All patients had persistent acute diarrhea and weight loss. Six of the 16 MPA-related diarrhea patients (37.5%) showed acute dehydration requiring hospitalization. Diarrhea resolved when MPA was switched to sirolimus (50%), discontinued (18.75%), switched to azathioprine (12.5%), or reduced by 50% (18.75%). No graft loss occurred. Four patients died during the study period. Late-onset MPA was more frequent, and no correlation with MPA dose or formulation was found.

Predictive Usefulness of Urinary Biomarkers for the Identification of Cyclosporine A-Induced Nephrotoxicity in a Rat Model

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of protein kinase A signaling pathway in cyclosporine nephrotoxicity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cytotoxic and genotoxic effects of high concentrations of the immunosuppressive drugs cyclosporine and tacrolimus in MRC-5 cells

Immunosuppressive drugs are used to suppress immune system activity in transplant patients and reduce the risk of organ rejection. The present study evaluated the potential cytotoxic, genotoxic and mutagenic of the immunosuppressive drugs cyclosporine (CsA) and tacrolimus (FK-506) on normal human fibroblasts (MRC-5 cells). Based on plasma concentrations of the immunosuppressive drugs, which were obtained from the records of kidney transplant patients at the Kidney Institute of Londrina, Brazil, 11 concentrations of each immunosuppressive were chosen to evaluate cell viability using the MIT assay. From these results, CsA and FK-506 concentrations of 135, 300, 675, and 1520 ng/ml and 8, 16, 24, and 32 ng/ml, respectively, were evaluated using (i) the comet assay, (ii) the nuclear division index (NDI), (iii) the micronucleus test (CBMN) and (iv) cell proliferation curves generated by quantifying cell numbers and protein levels. In this study, 1520 to 3420 ng/ml CsA decreased cell viability after 48 h of exposure. Genotoxic effects were observed only with a concentration of 1520 ng/ml after 3 h of exposure and with concentrations of 675 and 1520 ng/ml after 24 h of exposure. Mutagenic effects were observed only for the concentration of 1520 ng/ml. FK-506 decreased cell viability after 72 h of exposure for concentrations up to 20 ng/ml; genotoxic effects were observed with concentrations up to 8 ng/ml for both treatment times (3 and 24 h) and mutagenic effects were observed with concentrations of 24 and 32 ng/ml after 24 h of treatment. The cell proliferation curves demonstrated the absence of cytostatic effects of these drugs, and these data were confirmed by the NDI analysis. Our results suggest that concentrations lower than 300 ng/ml of CsA and 16 ng/ml of FK-506 are safe for use, as they did not induce genotoxic and mutagenic damage or affect MRC-5 cell viability and proliferation. (C) 2014 Elsevier GmbH. All rights reserved.

In vitro study of cyclosporine A 0.05 % on primary and recurrent pterygium fibroblasts

To compare the cyclosporine 0.05 % exposure effect on fibroblasts from primary and recurrent pterygium. Primary culture of fibroblasts from primary and recurrent pterygium was performed until the third passage, which was exposed to cyclosporine 0.05 % in a group and the other remaining unexposed (control group), in triplicates. After 3, 6, 12, and 17 days of exposure the viable cell counting was performed by hemocytometer. The results were statistically analyzed using the technique of analysis of non-parametric variance model for repeated measures with three factors. There was a significant reduction in both fibroblast proliferation, in primary as in the recurrent pterygium cultures exposed to cyclosporine when compared not exposed cultures, with statistical significance (P < 0.05). Comparing primary and recurrent pterygium that received the drug, there was no significant difference in cell proliferation in relation to primary or recurrent pterygium. Cyclosporine 0.05 % is effective in inhibiting fibroblast proliferation in culture, both in primary and as in recurrent pterygium.

Subcutaneous phaeohyphomycosis caused by Phaeoacremonium parasiticum in a renal transplant patient

A 49-year-old renally transplanted man, under a five-year course of immunosuppressive therapy with prednisone and cyclosporine A, experienced a subcutaneous phaeohyphomycosis caused by Phaeoacremonium parasiticum. The clinical presentation consisted of impressive, large, inflammatory and draining cystic tumors on the left foot that had been present for one year. A significant improvement was obtained with itraconazole plus intralesional injection with amphotericin B. Drug interaction was observed between itraconazole and cyclosporine A causing a severe hypertensive crisis and requiring a temporary sharp reduction in cyclosporine administration. Subcutaneous phaeohyphomycosis caused by P. parasiticum is uncommon among major organ transplant patients but several cases have previously been published and some patterns are emerging, e. g., limbs are generally involved but no known traumatic event has preceded lesion development. The identification of the case isolate was confirmed using a recently published online system based in part on beta-tubulin sequence comparison.

Histoplasmosis presenting as cellulitis 18 years after renal transplantation

A 49-year-old renal transplant patient, under an 18-year course of immunosuppressive therapy with prednisone and azathioprine and, more recently, prednisone plus mycophenolate sodium, developed a cutaneous-subcutaneous infection caused by Histoplasma capsulatum. The clinical presentation consisted of a slowly enlarging, erythematous and infiltrative 25 cm plaque in the major axis on the arm. There was no involvement of the lungs or any other organ. Cure was obtained with itraconazole treatment after 12 months. Histoplasmosis is an uncommon opportunistic infection among solid organ transplanted patients with incidence of 0% to 2.1% observed in a large number of cases. This report describes an atypical cutaneous clinical presentation of a potentially fatal disease in immunosuppressed patients.

Role of Peritubular Capillaries and Vascular Endothelial Growth Factor in Chronic Allograft Nephropathy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A Brazilian registry of juvenile dermatomyositis: onset features and classification of 189 cases

ObjectiveTo describe onset features, classification and treatment of juvenile dermatomyositis (JDM) and juvenile polymyositis (JPM) from a multicentre registry.MethodsInclusion criteria were onset age lower than 18 years and a diagnosis of any idiopathic inflammatory myopathy (IIM) by attending physician. Bohan & Peter (1975) criteria categorisation was established by a scoring algorithm to define JDM and JPM based oil clinical protocol data.ResultsOf the 189 cases included, 178 were classified as JDM, 9 as JPM (19.8: 1) and 2 did not fit the criteria; 6.9% had features of chronic arthritis and connective tissue disease overlap. Diagnosis classification agreement occurred in 66.1%. Medial? onset age was 7 years, median follow-up duration was 3.6 years. Malignancy was described in 2 (1.1%) cases. Muscle weakness occurred in 95.8%; heliotrope rash 83.5%; Gottron plaques 83.1%; 92% had at least one abnormal muscle enzyme result. Muscle biopsy performed in 74.6% was abnormal in 91.5% and electromyogram performed in 39.2% resulted abnormal in 93.2%. Logistic regression analysis was done in 66 cases with all parameters assessed and only aldolase resulted significant, as independent variable for definite JDM (OR=5.4, 95%CI 1.2-24.4, p=0.03). Regarding treatment, 97.9% received steroids; 72% had in addition at least one: methotrexate (75.7%), hydroxychloroquine (64.7%), cyclosporine A (20.6%), IV immunoglobulin (20.6%), azathioprine (10.3%) or cyclophosphamide (9.6%). In this series 24.3% developed calcinosis and mortality rate was 4.2%.ConclusionEvaluation of predefined criteria set for a valid diagnosis indicated aldolase as the most important parameter associated with de, methotrexate combination, was the most indicated treatment.

Análise quantitativa dos tecidos gengivais de ratos tratados com fenitoína e ciclosporina

Os aumentos gengivais podem ser decorrentes de reações teciduais a estímulos idiopáticos, patológicos e farmacológicos. O objetivo desse trabalho foi avaliar morfometricamente e estereologicamente a ação da fenitoína (Fen) e ciclosporina (CsA) sobre os tecidos gengivais de ratos. Dez ratos receberam, por via intraperitonial, Fen na dose inicial de 2 mg/kg de peso corporal/dia, aumentando 2 mg a cada duas semanas, durante 60 dias. em outros 10 ratos, administraram-se, por via subcutânea, 10mg/kg de peso corporal/dia de CsA, durante o mesmo período do grupo anterior. Os valores morfométricos e estereométricos dos tecidos gengivais dos ratos tratados com CsA foram significativamente maiores quando comparado com os valores dos tecidos gengivais do grupo tratado com Fen. Esses resultados sugerem que a CsA na dose utilizada é mais eficaz no desenvolvimento do aumento gengival em ratos, podendo estar atuando na proliferação de fibroblastos e no desequilíbrio fisiológico da síntese de fibras colágenas.

Mechanism for the uncoupling of oxidative phosphorylation by juliprosopine on rat brain mitochondria

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison of clinical features and drug therapies among European and Latin American patients with juvenile dermatomyositis

ObjectiveTo compare the demographic features, presenting manifestations, diagnostic investigations, disease course, and drug therapies of children with juvenile dermatomyositis (JDM) followed in Europe and Latin America.MethodsPatients were inception cohorts seen between 1980 and 2004 in 27 paediatric rheumatology centres. The following information was collected through the review of patient charts: sex; age at disease onset; date of disease onset and diagnosis; onset type; presenting clinical features; diagnostic investigations; course type; and medications received during disease course.ResultsFour hundred and ninety patients (65.5% females, mean onset age 7.0 years, mean disease duration 7.7 years) were included. Disease presentation was acute or insidious in 57.1% and 42.9% of the patients, respectively. The course type was monophasic in 41.3% of patients and chronic polycyclic or continuous in 58.6% of patients. The more common presenting manifestations were muscle weakness (84.9%), Gottron's papules (72.9%), heliotrope rash (62%), and malar rash (56.7%). Overall, the demographic and clinical features of the 2 continental cohorts were comparable. European patients received more frequently high-dose intravenous methylprednisolone, cyclosporine, cyclophosphamide, and azathioprine, while methotrexate and antimalarials medications were used more commonly by Latin American physicians.ConclusionThe demographic and clinical characteristics of JDM are similar in European and Latin American patients. We found, however, several differences in the use of medications between European and Latin American paediatric rheumatologists.

Pulpal lesions in normal and cyclosporin a treated rats

The objective of this study was to examine the development of pulpal lesions in the lower molar of control and cyclosporin A (CyA) treated rats. The pulps of the first lower molars of 20 normal and 20 CyA treated rats were exposed and left open into the oral cavity. Five animals of each group were killed at 7, 14, 21, and 28 days after the pulp exposure. The specimens were sectioned sagittally at a thickness of 7 μm and stained with hematoxylin and eosin. The pulpal lesions were similar for both normal and CyA treated rats in all studied periods and the differences between both groups were not statistically significant by the Student t test at the 5% (0.05) level of significance, indicating that the immunosuppression did not alter the evolution of the inflammatory process. Copyright © 1997 by The American Association of Endodontists.