Mario Ribeiro de Melo-Júnior

Healing activity induced by Cramoll 1,4 lectin in healthy and immunocompromised mice.

Cramoll 1,4 is a lectin extracted from seeds of Cratylia mollis Mart. Many assays have shown the cytokine release activity and pro-inflammatory profile of this lectin. Here, we used Cramoll 1,4 in the treatment of cutaneous wounds in normal and immunocompromised mice for available your cicatricial power. Surgical wounds were treated daily with a topical administration of Cramoll 1,4 and parameters as edema, hyperemia, scab, granulation and scar tissues as well as contraction of wounds were analyzed. Cramoll 1,4 wounds showed higher edema and arrival of more polimorphonuclear cells at the site of lesions. Granulation tissue and collagen fiber deposition were observed with higher intensity in all Cramoll 1,4 treated wounds and promoted excellent closing and repair of lesions in less time than other groups. Results showed that Cramoll 1,4 lectin was effective in the repair of experimental lesions in mice and can be used as a future cicatricial compound.

Hepatitis C serum prevalence in hemodialyzed patients

The objectives of this study were to estimate the serum prevalence of hepatitis C virus (HCV) infection in a dialysis center in the greater Recife region, and to correlate HCV serum positivity with some risk factors. Analyses were performed on 250 patients of both sexes, with ages ranging from 17 to 92 years old. Epidemiological data on these patients were obtained in order to determine the risk factors for this infection. Anti-HCV antibodies were investigated using fourth-generation ELISA. Statistically significant associations (p<0.05) were observed in relation to the risk factors of hemodialysis duration, number of blood component transfusions and time taken for transfusions. The prevalence was low (8.4%) in relation to other Brazilian studies. However, more studies in other centers are needed in order to estimate the real prevalence of HCV infection among patients undergoing hemodialysis in the State of Pernambuco.

Potential effects of Cramoll 1,4 lectin on murine Schistosomiasis mansoni.

Cratylia mollis is a natural forage plant from the Northeast of Brazil. C. mollis seed lectin (Cramoll) containing molecular forms 1 and 4 (Cramoll 1,4) has shown anti-inflammatory and wound-healing activities. This work analyzed the effect of Cramoll 1,4 on experimental schistosomiasis in mice. Experimental groups (n=15/group) were composed of female albino Swiss mice, which were subcutaneously and caudally infected with Schistosoma mansoni (BH strain, 100 cercariae/mouse) and were treated with an intraperitoneal dose after infection as follows: (1) Cramoll 1,4 (50 mg kg(-1) single dose - after 40 days of infection), (2) Cramoll 1,4 (7 mg kg(-1) daily dose - for 7 days after infection) and control (untreated mice). Mice were sacrificed 8 weeks after infection and adult worms were recovered from the portal-hepatic system. Livers were fixed in 10% (v/v) formaldehyde/0.15M NaCl and tissue sections were processed for haematoxilin and Massons trichrome stainings. Mice infected subcutaneously harboured no or very few worms and hence the effect of Cramoll 1,4 could not be assessed. Results (P≤0.05) were obtained with Cramoll 1,4 using the two treatments, with reduction of: egg excretion (79 and 80%), adult worm recovery (71 and 79%) and liver granulomas (40 and 73.5%) in relation to control. This study showed the potential anti-helminthic activity of Cramoll 1,4 when tested against Schistosomiasis mansoni infection in mice.

Altered lectin-binding sites in normal colon and ulcerative colitis

O presente trabalho objetivou, atraves de histoquimica com lectinas e analise digital de imagens, avaliar a expressao de carboidratos em amostras de colo normal e com colite ulcerativa. A partir de fragmentos de mucosa intestinal foram obtidos cortes histologicos (4mm) que foram incubados com lectinas (Con A, WGA, LTA e PNA), e os resultados das marcacoes foram avaliados atraves de microscopia optica e sistema de analise de imagens. Os resultados obtidos revelaram uma intensa marcacao para as celulas inflamatorias, principalmente neutrofilos infiltrados no tecido de reto e sigmoide, bem como celulas das glândulas intestinais. As lectinas WGA e LTA exibiram padroes distintos de marcacao entre o epitelio normal e os casos de colite ulcerativa. As lectinas PNA e Con A falharam em reconhecer os carboidratos celulares nos casos estudados em ambos os grupos. Os resultados obtidos foram confirmados pela analise de imagem. As observacoes obtidas sugerem que as lectinas WGA e LTA sao marcadores promissores para diferenciar o epitelio normal do padrao inflamatorio da colite ulcerativa, indicando uma expressao distinta de N-acetilglicosamina e L-fucose nos respectivos casos estudados.

A distribuição dos eosinófilos nas diferentes fases de evolução do granuloma hepático em camundongos infectados pelo Schistosoma mansoni

In the present study, the distribution of eosinophils at different stages of the formation of hepatic granuloma in mice infected with Schistosoma mansoni was evaluated. From the results obtained, we suggest a new classification for the evolution of hepatic granuloma in mice, constructed from the phases described by other authors. In each phase, there is a different pattern of eosinophil distribution. In the exudative-necrotic phase, the eosinophils are concentrated in the periphery and center of the granuloma, and are scarce in the necrotic area; in the productive phase, the eosinophils are dispersed throughout the granuloma; and in the cure due to fibrosis phase, the eosinophils are concentrated in the periphery and center of the granuloma. Eosinophils were found in direct contact with the eggs at all stages of evolution of the granuloma. It was concluded that the dynamics of eosinophils have an important role in forming the granulomatous reaction of the host and in resolving the inflammatory process caused by the parasite egg, as well as adding new data regarding hepatic granuloma classification.

Chemiluminescent Detection of Carbohydrates in the Tumoral Breast Diseases

Nowadays, there is an increase of investigations into the fibroadenoma, mainly because some studies have shown that the occurrence of fibroadenoma is linked to an increased risk of developing breast carcinoma. Currently, the chemiluminescence biomarkers are applied for validation methods and screening. Here, a lectin chemiluminescence is proposed as new histochemistry method to identify carbohydrates in mammary tumoral tissues. The lectins concanavalin A (Con A) and peanut agglutinin (PNA) conjugated to acridinium ester were used to characterize the glycocode of breast tissues: normal, fibroadenoma, and invasive duct carcinoma (IDC). The lectin chemiluminescence expressed in relative light units (RLU) was higher in fibroadenoma and IDC than in normal tissue for both lectins tested. The relationship RLU emission versus tissue area described a linear and hyperbolic curve for IDC and fibroadenoma, respectively, using Con A whereas hyperbolic curves for both transformed tissues using PNA. RLU was abolished by inhibiting the interaction between tissues and lectins using their specific carbohydrates: methyl-α-d-mannoside (Con A) and galactose (PNA). The intrinsic fluorescence emission did not change with combination of the lectins (Con A/PNA) to the acridinium ester for hydrophobic residues. These results represent the lectin chemiluminescence as an alternative of histochemistry method for tumoral diagnosis in the breast.

Digital image analysis of skin neoplasms evaluated by lectin histochemistry: potential marker to biochemical alterations and tumour differential diagnosis

The present study aims to evaluate, through lectin histochemistry, the alterations in the expression of cell surface carbohydrate between benign and malignant lesions of skin using computer image analysis. Skin fragments were obtained through biopsies and diagnosed as basal cell carcinoma (BCC), epidermoid carcinoma (EpC), trichoepithelioma (TE), keratoacanthoma (KA), seborrheic keratosis (SK) and actinic keratosis (AK). Lectins Con A, WGA, PNA, UEA-I and LTA were used in histochemistry study. Image analysis was carried out in a workstation using OPTIMASTM software system. PNA strong binding pattern to studied tumours evidenced the high expression of D-galactose residues in the epidermal neoplasms when compared to other sugars recognized by the other lectins. Among benign neoplasms, KA presented a high expression of glucose/mannose, a-fucose and D-galactose residues evidenced by the intense staining of Con A (94.7%), LTA (84.2%) and PNA (89.4%), respectively. Malignant tumours showed distinct binding patterns. EpC presented significant binding only by PNA lectin. BCC was differentially stained in comparison to the staining pattern observed in benign lesions such as TE. Qualitative (lectin histochemistry) and quantitative (digital image analysis) data obtained in this study evidenced those lectins are potential markers to biochemical alterations in skin neoplasms.

Intratracheal co-administration of antioxidants and ceftriaxone reduces pulmonary injury and mortality rate in an experimental model of sepsis

Recent studies showed that both sepsis and antibiotic therapy are associated with cell death and linked to reactive oxygen species generation. This study investigated the effects of intratracheal administration of combinations of antioxidants (n‐acetyl cysteine (NAC), vitamins C and E) in the treatment of sepsis‐induced lung injury.

Lectin staining patterns in human gastric mucosae with and without exposure to Helicobacter pylori

The aim of the present study was to evaluate qualitative changes in the glycoconjugate expression in human gastric tissue of positive and negative patients for Helicobacter pylori, through lectins: Wheat Germ Agglutinin (WGA) and Concanavalin A (Con A). The lectins recognized differently the glycoconjugates in the superficial mucous layer at the gastric tissues. The results suggest a significant change in the carbohydrate moieties present on the surface of the gastric cells during infection.

Hipertensão porta na esquistossomose mansônica: repercussões do tratamento cirúrgico no perfil histomorfométrico da mucosa gástrica

The repercussions from surgical treatment for controlling portal hypertension and its effects on the gastric vasculature of young patients with mansonic schistosomiasis were investigated by digital image analysis. The study included five patients at the preoperative stage and 27 patients who had undergone surgical intervention at different times in the past: 0-2 years ago, n=4; 2-6 years ago, n=13, and more than 6 years ago, n=10. Endoscopic biopsies were taken from the mucosa of the gastric antrum and body endoscopic mucosa and the samples underwent routine histological tests after embedding in paraffin blocks. Histological thin sections were used for histomorphometric analysis of the following parameters: mean number of vessels per field, and mean diameter and thickness of the vessel walls. The results showed that, between the patients whose operation was not more than two years ago and those whose operation was more than six years ago, there was a significant decrease in the density and diameter of the vessels. These findings give support to the concept that the surgical treatment administered decreases specific histological alterations like hemorrhage and ectasia, over the long term.: The repercussions from surgical treatment for controlling portal hypertension and its effects on the gastric vasculature of young patients with mansonic schistosomiasis were investigated by digital image analysis. The study included five patients at the preoperative stage and 27 patients who had undergone surgical intervention at different times in the past: 0-2 years ago, n=4; 2-6 years ago, n=13, and more than 6 years ago, n=10. Endoscopic biopsies were taken from the mucosa of the gastric antrum and body endoscopic mucosa and the samples underwent routine histological tests after embedding in paraffin blocks. Histological thin sections were used for histomorphometric analysis of the following parameters: mean number of vessels per field, and mean diameter and thickness of the vessel walls. The results showed that, between the patients whose operation was not more than two years ago and those whose operation was more than six years ago, there was a significant decrease in the density and diameter of the vessels. These findings give support to the concept that the surgical treatment administered decreases specific histological alterations like hemorrhage and ectasia, over the long term.