Effects of the probiotic Bacillus cereus GM on experimental schistosomiasis mansoni.

Probiotics contribute to the integrity of the intestinal mucosa and preventing dysbiosis caused by opportunistic pathogens, such as intestinal helminths. Bacillus cereus GM obtained from Biovicerin® was cultured to obtain spores for in vivo evaluation on experimental schistosomiasis. The assay was performed for 90 days, where all animals were infected with 50 cercariae of Schistosoma mansoni on the 15th day. Three experimental groups were formed, as follows: G1-saline solution from the 1st until the 90th day; G2-B. cereus GM (105 spores in 300 µL of sterile saline) from the 1st until the 90th day; and G3-B. cereus GM 35th day (onset of oviposition) until the 90th day. G2 showed a significant reduction of 43.4% of total worms, 48.8% of female worms and 42.5% of eggs in the liver tissue. In G3, the reduction was 25.2%, 29.1%, and 44% of the total number of worms, female worms, and eggs in the liver tissue, respectively. G2 and G3 showed a 25% (p < 0.001) and 22% (p < 0.001) reduction in AST levels, respectively, but ALT levels did not change. ALP levels were reduced by 23% (p < 0.001) in the G2 group, but not in the G3. The average volume of granulomas reduced (p < 0.0001) 65.2% and 46.3% in the liver tissue and 83.0% and 53.2% in the intestine, respectively, in groups G2 and G3. Th1 profile cytokine (IFN-γ, TNF-α, and IL-6) and IL-17 were significantly increased (p < 0.001) stimulated with B. cereus GM in groups G2 and G3. IL-4 showed significant values when the stimulus was mediated by ConA. By modulating the immune response, B. cereus GM reduced the burden of worms, improved some markers of liver function, and reduced the granulomatous inflammatory reaction in mice infected with S. mansoni, especially when administered before infection.

Intraurethral injection with LPS: an effective experimental model of prostatic inflammation.

OBJECTIVE: Chronic inflammation has been recognized as having a prominent role pathogenesis of benign prostatic hyperplasia (BPH) and cancer. It is believed that chronic inflammation induces prostatic fibromuscular growth. This correlation has been clearly illustrated by both in vivo and in vitro studies; however, current experimental models of BPH require complex surgery or hormonal treatment. Therefore, the aim of the present study was to propose a new murine model of BPH/prostatitis induced by intraurethral injection of LPS. METHODS: Male Swiss and C57Bl/6 mice were then sacrificed 3, 7, 10, and 14 days after intraurethral injection of LPS. The prostates were quickly dissected and fixed for morphological and immunohistochemical analyses. RESULTS: The results showed that LPS played an important role in the cell proliferation of the prostate. Histological and ultrastructural analysis showed epithelial hyperplasia, clear stromal cells, little inflammatory infiltration, and heavy bleeding. Treatment with LPS also promoted the increase of growth factor (FGF-7 and TGF-ß), α-actin, and proinflammatory cytokines (IL-1, IL-6, IL-17), both in the stroma and epithelium. CONCLUSION: According to the present findings, it can be concluded that the intraurethral administration of LPS promotes tissue remodeling, as well as stimulating the pattern of pro-inflammatory cytokines, and therefore, constitutes an effective experimental model of BPH/inflammation.

Chemiluminescent detection of glycocode alterations in hepatic granulomatous lesions of experimental schistosomiasis.

This work describes the glycophenotype evaluation of mice liver granulomatous lesion caused by infection of Schistosoma mansoni by using lectins labeled to acridinium ester (AE). The employed lectins were Concanavalin A (Con A), wheat germ agglutinin (WGA) and Sambucus nigra agglutinin (SNA) that specifically recognize α-D-glucose/mannose, N-acetyl-D-glucosamine and α-NeuNAc-[2→6]-Gal/GalNAc. The chemiluminescence expressed in relative light unit (RLU) obtained from the hepatic granuloma tissues (0.25 cm2) treated with the lectins-AE was compared with control tissues. Con A-AE infected tissues showed higher statistically significant values (1,501,182 ± 163,450 RLU) compared with the control tissue (575,280 ± 97,216 RLU). WGA-AE results also showed higher values (189,654 ± 20,686 RLU) than that found for the controls (82,878 ± 24,411). SNA-AE results did not present statistical difference between granulomatous tissues (198,990 ± 15,131) and controls (167,290 ± 25,194). There is a significant increase in glucose/mannose residues and N-acetyl-D-glucosamine in hepatic granuloma caused by S. mansoni, while the sialic acid remains virtually unchanged. The understanding of schistosome glycophenotype is relevant for the development of new diagnostic methods for schistosomiasis, design of new drug targets and preparation of glycan-based vaccines.

Histopathological and Digital Morphometrical Evaluation of Uterine Leiomyoma in Brazilian Women.

The current study aims to evaluate histopathological and digital morphometrical aspects associated with uterine leiomyomas in one hundred and fifty (150) patients diagnosed with leiomyoma. Uterine tissues were subjected to the histopathological and digital morphometric analyses of the interstitial collagen distribution. The analysis of medical records indicates that most of the women diagnosed with uterine leiomyomas (68.7%) are between 37 and 48 years old. As for the anatomic location of the tumors, approximately 61.4% of the patients had intramural and subserosal lesions. In 50% of the studied cases, the patients developed uterine leiomyomatosis (with more than eight tumors). As for the morphometric study, the average size of the interstitial collagen distribution held approximately 28.53% of the capture area, whereas it was of 7.43% in the normal tissue adjacent to the tumor. Another important aspect observed in the current study was the high rate of young women subjected to total hysterectomy, a fact that resulted in early and definitive sterility.

Immunocytochemical study of TOP2A and Ki-67 in cervical smears from women under routine gynecological care.

BACKGROUND: Cervical cancer is one of the most common female cancers and is caused by human papillomavirus (HPV). Viral infection leads to cell cycle deregulation by inactivating p53 and retinoblastoma protein by viral oncoproteins E6 and E7, respectively. Then, nuclear proteins such as DNA topoisomerase type IIa (TOP2A) and Ki-67 show increased expression because of increased cell division. These molecules are used as biomarkers for immunohistochemistry analysis of cervical tissue. METHODS: In this cross-sectional study, we recruited 110 women receiving regular gynecological surveillance at public health centers in Olinda - PE, Brazil. Cervicovaginal cells were collected to determine the presence of cytological abnormalities and HPV infection. Pap smear slides were used to evaluate the expression of TOP2A and Ki-67 using immunocytochemistry techniques. RESULTS: Of the 110 women, 75.4 % showed HPV-DNA(+) infection (83/110) and 29.1 % showed cellular abnormalities (32/110). Two atypical cells of undetermined significance, one low-grade squamous intraepithelial lesion, and one high-grade squamous intraepithelial lesion samples showed no HPV-DNA. TOP2A was positive in 71.9 % of samples, while Ki-67 was positive in 81.2 %. Immunocytochemistry results were positive in 4 of 5 atypical cells of undetermined significance samples. In HPV-DNA(+) samples with cytological abnormalities, immunocytochemistry results were positive 96.4 % of samples (p < 0.0001; odds ratio = 28.0). Among the samples infected with HR-HPV, TOP2A(+) was effective in 71 % samples, while and Ki-67(+) was 77.4 %. Ki-67 and TOP2A were positive for all samples infected with HPV6, HPV11, and HPV18. Ki-67 was also positive for all HPV16 samples, except for one negative sample in cytopathology analysis. CONCLUSIONS: TOP2A and Ki-67 antibodies may be used in combination for cervical cancer screening in immunocytochemistry assays.

Biocompatibility and cutaneous reactivity of cellulosic polysaccharide film in induced skin wounds in rats.

The development of a skin substitute suitable for immediately performing the function of the lost dermis and epidermis could result in a positive impact on the treatment of patients with extensive skin lesions. A biopolymer film was applied to skin wounds to investigate the biocompatibility and cutaneous reaction and to test its activity as a mechanical barrier and conductor in the healing process. Forty Wistar rats of both sexes were used in the present study. Two excisions were performed in the dorsal part of the skin flaps. The polysaccharide film was applied over one of the incisions and other incision was washed with saline. The time spent for complete healing of both lesions was virtually the same in both groups, during 21 days of observation. The film remained attached to the bed of the exposed wound for an average period of 6 days. There were no statistically significant differences with regard to lesion measurement area at assessment times of 2nd, 7th and 14th postoperative days. At day 21, the scar area showed a significant difference (0.0229). After 40 days, all wounds were completely healed. No statistically significant differences were found between the histological parameters assessed in the experimental and control groups. The cellulosic polysaccharide film integrated well with the tissue showing high biocompatibility and low skin reactivity.

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.

Comparative analysis of extracellular matrix and cellular carbohydrate expression in the sporotrichosis and chromoblastomycosis.

This work was based on the analysis of digital images of histochemical profile from subcutaneous lesions in sporotrichosis (ST) and chromoblastomycosis (CM) patients. An additional aim was the detection of carbohydrate expression using lectin histochemical analysis of the different carbohydrates in the fungal cell wall from four different species (Sporothrix schenckii, Fonsecaea pedrosoi, Phialophora verrucosa, and Cladophialophora carrionii) associated with diseases mentioned earlier. Slides from tissue biopsies from ST and CM positive patients (n=10, each) were stained according to routine techniques. Slides were incubated with 25 µg/ml of Con A lectins and WGA conjugated to peroxidase. Digital image analysis was carried out in a workstation using OPTIMAS™ software system. Routine histochemistry results indicated that there is significantly higher collagen deposition and elastic fibers in ST characteristic lesions compared with that found in CM cases. The ST interstitial fibrosis area was larger than in CM lesions. Comparative lectin binding showed a positive and intense lectin staining pattern in the cell wall of S. schenckii, suggesting a higher expression of glucose/mannose and N-acetyl glucosamine in their cell surface as evidenced by Con A and WGA, respectively. However, these lectins were not effective to recognize some carbohydrates moieties in the F. pedrosoi, P. verrucosa, and C. carrionii. Such findings contribute to additional information about specific recognition processes between fungal parasites and their host cell targets may be mediated by the interaction of carbohydrate-binding proteins, such as lectins, on the surface of one type of cell that combine with complementary sugars on the surface of another cells into fibro-connective tissues associated with lesions.

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 Masson's 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.

Immobilization of anti-galectin-3 onto polysiloxane-polyvinyl alcohol disks for tumor prostatic diseases diagnosis.

This work aimed to immobilize the antibody anti-galectin-3 onto polysiloxane-polyvinyl alcohol (POS-PVA) support, to evaluate its capacity to capture the serum antigen galectin-3 and to quantify by ELISA the antigen levels in sera from patients with prostatic adenocarcinoma (PA) and benign prostatic hyperplasia (BPH) and healthy individuals. Also, for comparative effect, the galectin-3 expression in the prostate tissue through immunohistochemistry was evaluated. The optical density (galectin-3 level) values established for the sera from PA and BPH patients were lower compared with those found for the healthy individuals. Galectin-3 immunohistochemically showed a significant increase and reduction of the cytoplasmatic protein expression in BPH and PA, respectively, compared with the normal prostate. These results showed that POS-PVA disks could be used as solid phase to immobilize serum galectins and in immunoassays procedures for the correspondent IgG anti-galectins detection in human sera.

[Hepatitis C serum prevalence in hemodialyzed patients]. / Soroprevalência da hepatite C em pacientes hemodialisados.

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.

[Portal hypertension in mansonic schistosomiasis: repercussions of surgical treatment on the histomorphometric profile of the gastric mucosa]. / 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.