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.

Development of animal model for studying deep second-degree thermal burns.

Thermal lesions were produced in 12 male Wistar rats, positioning a massive aluminum bar 10 mm in diameter (51 g), preheated to 99°C ± 2°C/10 min. on the back of each animal for 15 sec. After 7, 14, 21, and 28 days, animals were euthanized. The edema intensity was mild, with no bubble and formation of a thick and dry crust from the 3rd day. The percentage of tissue shrinkage at 28 days was 66.67 ± 1.66%. There was no sign of infection, bleeding, or secretion. Within 28 days reepithelialization was incomplete, with fibroblastic proliferation and moderate fibrosis and presence of modeled dense collagen fibers. It is concluded that the model established is applicable in obtaining deep second-degree thermal burns in order to evaluate the healing action of therapeutic agents of topical use.

Topical application effect of the isolectin hydrogel (Cramoll 1,4) on second-degree burns: experimental model.

This study aimed at evaluating the use of hydrogel isolectin in the treatment of second-degree burns. Twenty male rats were randomly divided into two groups (G1 = treatment with hydrogel containing 100 µg/mL Cramoll 1,4 and G2 = Control, hydrogel). After 7, 14, 21, 28, and 35 days, animals were euthanized. On the 7th day, G1 showed intense exudates, necrosis and edema. On the 14th day, G1 showed tissue reepithelialization and moderate autolysis. On the 21st day, G1 showed intense fibroblastic proliferation, presence of dense collagen, and moderate fibrosis. On the 28th day, G1 showed complete tissue epithelialization. On the 35th day, G1 showed modeled dense collagen. The significant wound contraction was initiated from day, 14 in the G1. There were no significant differences in biochemical and hematological parameters analyzed. These results extend the potential of therapeutic applications for Cramoll 1,4 in the treatment of thermal burns.

Immunohistopathological changes in murine Schistosomiasis mansoni under the influence of N-acetyl-L-cysteine.

The main pathology associated with Schistosomiasis mansoni is granulomatous inflammation that may develop into hepatosplenic disease with fibrosis and hepatoesplenomegaly. It is known that N-acetyl-L-cysteine (NAC) reduces tissue damage in chronic liver diseases owing to its anti-inflammatory, antioxidant, and detoxifying properties. In this study, we investigated the imunohistopathological changes in murine schistosomiasis mansoni under the influence of NAC, in combination with Praziquantel (PZQ) or not. Three groups of mice were formed to evaluate the effects of NAC during infection in the acute, intermediate, and chronic phases. Each group was further subdivided into four subgroups: NAC, PZQ, NAC + PZQ and control (without treatment). Oral administration of NAC (200 mg/kg/day) was carried out on the first day after infection for the acute phase and on the 45th for the intermediate and chronic phases for 59 and 45, 75 days, respectively. PZQ (100 mg/kg/day), was given orally by gavage from the 45th to 49th day after infection. Histopathological analysis of liver tissue provided evidence that combined NAC + PZQ treatment reduced the development of granulomas observed in the chronic phase. Animals treated with NAC and/or PZQ showed a reduction in the size of granulomas and all those treated with NAC exhibited a lower degree of fibrosis. In all groups, NAC decreased the synthesis of interferon-γ and nitric oxide, while increasing the levels of interleukin-10, but it did not influence the production of interleukin-4. On the whole, NAC treatment induced an immunomodulatory effect and reduced liver damage during the granulomatous inflammation in S. mansoni-infected mice.

[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.

Evaluating the effectiveness of gel formulation of irradiated seed lectin Cratylia mollis during bone repair in rats.

CONTEXT: Regeneration corresponds to the replacement of damaged cells with ones that have the same morphology and function. For experimental evaluation of materials that may favor the process of bone healing, defects are created with dimensions that prevent spontaneous regeneration. For the development and use of new drugs, it is necessary to study its effects in vitro, which depends on the formulation, concentration, and rate of irradiation in vivo and the route and frequency of administration; thus, it is possible to characterize the physiological and molecular mechanisms involved in the response and cellular effects. OBJECTIVE: The objective of this study was to assess the effect of Cramoll-1,4 on the process of bone repair. MATERIALS AND METHODS: A formulation of biopharmaceutical lectin Cramoll-1,4 at a concentration of 300 mg/100 mL was applied in a single application via gamma radiation and its effect on the process of bone repair in rats was assessed. RESULTS: Histologically, it was observed that the bone defect is coated by loose connective tissue rich in fibroblasts, providing a range similar to the thick bone original and competing with site of new bone formation. This prevented direct contact between the formulation and experimental bone tissue, as, despite its proven effectiveness in experiments on the repair of skin lesions, the formulation used did not promote bone stimulation that would have promoted the tissue repair process. CONCLUSION: Because of the direct interference of loose tissue repair that prevented direct contact of the implant with the bone interface, the formulation did not promote bone stimulation.

The incorporation of Brazilian propolis into collagen-based dressing films improves dermal burn healing.

ETHNOPHARMACOLOGICAL RELEVANCE: Hydroalcoholic solutions of propolis, a resinous product produced by bees, have been currently employed in improving the cicatricial repair. Biological activity of propolis might be related to its antimicrobial, anti-inflammatory and immunomudalatory properties. AIM OF THIS STUDY: Investigate the suitability of the collagen-based films containing hydroalcoholic extracts of two different varieties of Brazilian propolis (green and red ones) on the dermal burn healing in rodent model. MATERIALS AND METHODS: The hydroalcoholic extracts of red propolis (RP) or Green propolis (GP) were incorporated into collagen-based dressing films (COL). Burn wounds were performed in the dorsum of Wistar rats and dressing with COL, COL+GPa (0.5%), COL+GPb (1,0%) or COL+RP (0.5%). A control group (CTR) was performed keeping the wound undressed. The histological analyses were carried out after 3, 7, 14, 21 and 30 days for histological assessment of the inflammatory response, epithelization rates (ER), myofibroblastic count (MC) and collagenization pattern. RESULTS: GPa, GPb and RP provided significant decrease of the inflammatory severity, improved the ER in GPa in 7 (p=0.000), 14 (p=0.000), 21 (p=0.005) and 30 days (p=0.015), and induced earlier replacement of type-III for type-I collagen (p<0.05) than COL and CTR. In all the groups, the MC increased progressively from 3 to 14 days, and then started to decrease slowly until 21 days. Although no significant difference was observed among the groups in 3, 7 and 30 days, the MC was significantly increased in RP in 14 (p=0.0001) and 21 days (p=0.04), as well as grosser interlacement of the collagen bundles compared with the other groups. CONCLUSION: The incorporation of hydroalcoholic extracts of Brazilian propolis improved the biological events associated to burn healing without toxic effects, but the red variety provided the best results. Therefore, these collagen-based containing natural apicultural products films may be considered a promising new dressing for wound occlusion and tissue repairing.

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.