Methotrexate associated to lipid core nanoparticles improves cardiac allograft vasculopathy and the inflammatory profile in a rabbit heart graft model.

Coronary allograft vasculopathy is an inflammatory-proliferative process that compromises the long-term success of heart transplantation and has no effective treatment. A lipid nanoemulsion (LDE) can carry chemotherapeutic agents in the circulation and concentrates them in the heart graft. The aim of the study was to investigate the effects of methotrexate (MTX) associated to LDE. Rabbits fed a 0.5% cholesterol diet and submitted to heterotopic heart transplantation were treated with cyclosporine A (10 mg·kg-1·day-1 orally) and allocated to treatment with intravenous LDE-MTX (4 mg/kg, weekly, n=10) or with weekly intravenous saline solution (control group, n=10), beginning on the day of surgery. Animals were euthanized 6 weeks later. Compared to controls, grafts of LDE-MTX treated rabbits showed 20% reduction of coronary stenosis, with a four-fold increase in vessel lumen and 80% reduction of macrophage staining in grafts. Necrosis was attenuated by LDE-MTX. Native hearts of both LDE-MTX and Control groups were apparently normal. Gene expression of lipoprotein receptors was significantly greater in grafts compared to native hearts. In LDE-MTX group, gene expression of the pro-inflammatory factors tumor necrosis factor-α, monocyte chemoattractant protein-1, interleukin-18, vascular cell adhesion molecule-1, and matrix metalloproteinase-12 was strongly diminished whereas expression of anti-inflammatory interleukin-10 increased. LDE-MTX promoted improvement of the cardiac allograft vasculopathy and diminished inflammation in heart grafts.

Specific activation of CD4- CD8- double-negative T cells by Trypanosoma cruzi-derived glycolipids induces a proinflammatory profile associated with cardiomyopathy in Chagas patients.

Cardiomyopathy is the most severe outcome of Chagas disease, causing more than 12 000 deaths/year. Immune cells participate in cardiomyopathy development either by direct tissue destruction, or by driving inflammation. We have shown that CD4- CD8- [double-negative (DN)] T cells are major sources of inflammatory and anti-inflammatory cytokines, associated with the cardiac (CARD) and indeterminate (IND) forms of Chagas disease, respectively. Here, we sought to identify Trypanosoma cruzi-derived components that lead to activation of DN T cells in Chagas patients. Glycolipid (GCL), lipid (LIP) and protein-enriched (PRO) fractions derived from trypomastigote forms of T. cruzi were utilized to stimulate cells from IND and CARD patients to determine DN T cell activation by evaluating CD69 and cytokine expression. We observed that GCL, but not LIP or PRO fractions, induced higher activation of DN T cells, especially T cell receptor (TCR)-γδ DN T, from IND and CARD. GCL led to an increase in tumour necrosis factor (TNF) and interleukin (IL)-10 expression by TCR-γδ DN T cells from IND, while inducing IFN-γ expression by TCR-γδ DN T cells from CARD. This led to an increase in the ratio IFN-γ/IL-10 in TCR-γδ DN T cells from CARD, favouring an inflammatory profile. These results identify GCL as the major T. cruzi component responsible for activation of DN T cells in chronic Chagas disease, associated predominantly with an inflammatory profile in CARD, but not IND. These findings may have implications for designing new strategies of control or prevention of Chagas disease cardiomyopathy by modulating the response to GCL.

The monoclonal antibody against the major diagnostic antigen of Paracoccidioides brasiliensis mediates immune protection in infected BALB/c mice challenged intratracheally with the fungus.

The protective role of specific antibodies against Paracoccidioides brasiliensis is controversial. In the present study, we analyzed the effects of monoclonal antibodies on the major diagnostic antigen (gp43) using in vitro and in vivo P. brasiliensis infection models. The passive administration of some monoclonal antibodies (MAbs) before and after intratracheal or intravenous infections led to a reduced fungal burden and decreased pulmonary inflammation. The protection mediated by MAb 3E, the most efficient MAb in the reduction of fungal burden, was associated with the enhanced phagocytosis of P. brasiliensis yeast cells by J774.16, MH-S, or primary macrophages. The ingestion of opsonized yeast cells led to an increase in NO production by macrophages. Passive immunization with MAb 3E induced enhanced levels of gamma interferon in the lungs of infected mice. The reactivity of MAb 3E against a panel of gp43-derived peptides suggested that the sequence NHVRIPIGWAV contains the binding epitope. The present work shows that some but not all MAbs against gp43 can reduce the fungal burden and identifies a new peptide candidate for vaccine development.

Experimental paracoccidioidomycosis: alternative therapy with ajoene, compound from Allium sativum, associated with sulfamethoxazole/trimethoprim.

Ajoene has been described as an antithrombotic, anti-tumour, antifungal, antiparasitic and antibacterial agent. This study deals with the efficacy of ajoene to treat mice intratracheally infected with Paracoccidioides brasiliensis. The results indicate that ajoene therapy is effective in association with antifungal drugs (sulfametoxazol/trimethoprim), showing a positive additive effect. Ajoene-treated mice developed Th1-type cytokine responses producing higher levels of IFN-gamma and IL-12 when compared to the infected but untreated members of the control group. Antifungal activity of ajoene involves a direct effect on fungi and a protective pro-inflammatory immune response. Reduction of fungal load is additive to chemotherapy and therefore the combined treatment is mostly effective against experimental paracoccidioidomycosis.

Peptide immunization as an adjuvant to chemotherapy in mice challenged intratracheally with virulent yeast cells of Paracoccidioides brasiliensis.

Immunization with peptide P10, derived from gp43, and chemotherapy were used together in an attempt to improve treatment of paracoccidioidomycosis and prevent relapses. The combined treatment showed an additive protective effect when administered at 48 h or 30 days after intratracheal challenge. Its use is recommended to improve regular chemotherapy and reduce the duration of treatment.

Immediate reconstruction of the central segment of the mandible using the masseter osteomuscular flap.

We describe a new technique for reconstruction of the central segment of the mandible using a masseter osteomuscular flap. The advantages of this method are that bone viability is preserved, both form and function of the mandible are maintained, it is a local flap with low donor-site morbidity, and the operative time is shortened.

Total reconstruction of the alar cartilage en bloc using the ear cartilage: a study in cadavers.

To determine whether the lamina tragi, isthmus, and cavum conchae are a donor area for reconstruction of the alar cartilage with all its elements (medial crus, junction of the medial and lateral crura, and lateral crus), with the same dimension and en bloc, 40 alar cartilages and 40 lower parts of ear cartilages of 20 cadavers were dissected. Several measurements were taken in the alar cartilages, such as distance, thickness, and angle. Then they were compared with the measurements performed in the ear cartilages and segments removed from the lamina tragi, isthmus, and cavum conchae. This study, done with cadavers, shows that from the lamina tragi, isthmus, and cavum conchae, en bloc resection is possible with characteristics of form and dimension similar to those of the homolateral alar cartilage. The segment removed en bloc from the intermediate part of the lamina tragi, isthmus, and cavum conchae replaces, respectively, the medial crus, junction of the medial and lateral crura, and lateral crus.

Total en bloc reconstruction of the alar cartilage using autogenous ear cartilage.

A new technique is described for increasing the nasal tip by using an en bloc autogenous ear cartilage graft in such a manner as to create, in essence, a replacement of the entire ipsilateral alar cartilage.

Capsular hematoma as a late complication in breast reconstruction with silicone gel prostheses.

This paper presents a rare complication of breast reconstruction with silicone gel prostheses in which formation of intracapsular hematomas in the mammary region occurred 3 years after surgery. Treatment by bilateral capsular excision was successful. The rarity of this occurrence and the progress of the case are the major reasons for this publication.

Carcinoma of the vulva. Clinicopathologic factors involved in inguinal and pelvic lymph node metastasis.

Eighty-five women with vulvar squamous cell carcinoma were subjected to radical vulvectomy with bilateral inguinal and femoral node dissection or to radical vulvectomy with bilateral inguinofemoral and deep pelvic node dissection. The association between lymph node status (metastatic or not) and several parameters was analyzed: tumor location, size and clinical stage; tumor thickness, histologic grade and mitotic index; blood vessel, lymphatic and perineural infiltration; and lymphocytic and plasma cell infiltrates. There were no metastases to the pelvic lymph nodes without previous inguinal lymph node involvement. Unilateral vulvar carcinomas did not have contralateral metastatic nodes when there was no ipsilateral nodal involvement. Lymphatic vessel infiltration showed a statistically significant correlation with inguinal node metastases (P less than .05). No correlation was found between lymph node metastasis and tumor size, clitoral invasion, tumor thickness, histologic grade, blood vessel and perineural infiltration, lymphocytic and plasma cell infiltrates, and mitotic index.

Great omentum free flap in the repair of extensive loss of scalp / Uso de retalho livre do grande epíploo na reparaçäo de extensas perdas de couro cabeludo

Os autores apresentam um caso de grande exposiçäo de calota craniana. Como soluçäo terapêutica foi empregado o retalho livre do grande epíploo. A anastomose microcirúrgica foi feita apenas com vasos temporais superficiais direitos, devido à ausência congênita dos vasos gastrepiplóicos esquerdos. A cobertura do retalho foi obtida com enxerto de pele parcial. A paciente evoluiu para a cura, sem complicaçöes

Parada cardiaca anoxica hipotermica (20 graus) e alteracoes enzimaticas mitocondriais. / Anoxic hypothermic (20 degrees) heart arrest and mitochondrial enzymatic changes

Foram estudados 24 caes distribuidos nos seguintes grupos: Grupo I - Oito animais, cujos coracoes foram estudados imediatamente apos anestesia e toratocomia (controle).Grupo II Oito animais submetidos a 60 minutos de parada cardiaca anoxica normotermica. Grupo III Oito animais submetidos a 60 minutos de parada cardiaca anoxica hipotermica (20 graus).Os resultados obtidos possibilitaram as seguintes conclusoes: A) O esfriamento seletivo do miocardio impediu a diminuicao das concentracoes de ATP. B) As alteracoes nos sistemas enzimaticos estudados (alfa-ceco-glutario-desidrogenase: succino-desidrogenase; beta-hidroxi-butirico-desidrogenase; glutamico-desidrogenase e malico-desidrogenase), determinadas pela parada anoxica normotermica, excetuando-se o controle da beta-hidroxi-butiricodesidrogenase, nao foram prevenidas pelo esfriamento do miocardio a 20 graus C