Structure of human uropepsin at 2.45 angstrom resolution

The molecular structure of human uropepsin, an aspartic proteinase from the urine produced in the form of pepsinogen A in the gastric mucosa, has been determined by molecular replacement using human pepsin as the search model. Crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 50.99, b = 75.56, c = 89.90 Angstrom. Crystallographic refinement led to an R factor of 0.161 at 2.45 Angstrom resolution. The positions of 2437 non-H protein atoms in 326 residues have been determined and the model contains 143 water molecules. The structure is bilobal, consisting of two predominantly beta -sheet lobes which, as observed in other aspartic proteinases, are related by a pseudo-twofold axis. A model of the uropepsin-pepstatin complex has been constructed based on the high-resolution crystal structure of pepsin complexed with pepstatin.

Response of Corriedale and Crioula Lanada sheep to artificial primary infection with Haemonchus contortus

Clinical, parasitological and biochemical parameters were evaluated in Corriedale and Crioula Lanada sheep after a single experimental infection with Haemonchus contortus. Ten 4-month-old worm-free lambs, of each breed, were infected with 200 L-3 H. contortus per kg live weight,and four uninfected animals of each breed were used as controls. Every week, the animals were weighed and blood and faecal samples were collected for measurement of packed cell volume (PCV), total serum protein (TSP) and albumin (ALB), and the number of eggs per gram of faeces (EPG), respectively. Twelve weeks after infection, the animals were slaughtered. The worm burden was determined and samples of the abomasal mucosa were processed for determination of the number of eosinophils, mast cells and globule leukocytes. No significant differences in PCV, TSP, ALB, parasite burden or the cell populations of the abomasal mucosa were observed between breeds, but Crioula lambs had a lower EPG count. The comparison of the infected groups with their respective controls revealed significant alterations in PCV, TSP and ALB in the Corriedale lambs and in PCV, TSP, ALB and the density of eosinophils and mast cells in the Crioula lambs.

Bacteriocin-like substances production by Streptococcus-salivarius against oral bacteria

Samples of tongue and bucal mucosa surfaces were obtained from six healthy subjects with the purpose of isolating S. salivarius. It was verified that 47 out of 48 S. salivarius strains produced bacteriocin-like substances against at least one of the indicator species: Actinomyces viscosus, Rothia dentocariosa, Streptococcus pyogenes, Staphylococcus aureus, Streptococcus mutans and Streptococcus sanguis. The method employed to test for bacteriocin production was that of deferred antagonism. The results showed that there was a high antagonism against R. dentocariosa, S. pyogenes and A. viscosus; extremely low against S. mutans and S. sanguis and no inhibition for S. aureus.

Preliminary studies of Mammea americana L. (Guttiferae) bark/latex extract point to an effective antiulcer effect on gastric ulcer models in mice

Plant extracts are some of the most attractive sources of new drugs and have shown promising results for the treatment of gastric ulcers. Several folk medicinal plants and herbs have been used to treat gastrointestinal disorders, including gastric ulcers. Mammea americana L. (Guttiferae) fruit is very common in the diet of the population of northern South America. Our research interest in this plant arose because of its potential medicinal value as a tonic and against stomachache, as used in folk medicine. In this paper we evaluated three different extracts (ethanolic/EtOH, methanolic/MeOH and dichloromethane/DCM) obtained from M. americana L., for their ability to protect the gastric mucosa against injuries caused by necrotizing agents (0.3 M HCI/60% EtOH), hypothermic restraint stress, nonsteroidal anti-inflammatory drugs (NSAID, indomethacin) and pylorus ligation. In the HCI/EtOH-induced gastriculcer model, EtOH and DCM extracts demonstrated significant inhibition of the ulcerative lesion index by 54% (12.0 +/- 2.6 mm) and 86% (3.7 +/- 1.8 mm), respectively, in relation to the control value (26.0 +/- 1.4 mm) (p < 0.000 1). In the NSAID/cholinomimetic-induced lesion model, both EtOH and DCM extracts showed antiulcerogenic effects with significant reduction in the damage to these gastric lesions of 36% (8.3 +/- 2.0 mm) and 42% (7.5 +/- 1.4 mm), respectively, as compared to the control group (13.0 +/- 0.9 mm) (p < 0.0001). In the gastric ulcer induced by hypothermic-restraint stress, both extracts also showed significant activity, and inhibited the gastric lesion index by 58% and 75%, respectively. The EtOH and DCM extracts also changed gastric juice parameters as well as those of cimetidine, decreased gastric acid secretion significantly (p < 0.0001), increased pH values and promoted reduced acid output (p < 0.0001). In all gastric-ulcer-induced models, MeOH extract did not show any significant antiulcerogenic activity, nor did it change gastric-juice parameters (p > 0.05). The results suggest that EtOH and DCM extracts obtained from M. americana possess excellent antisecretory and/or gastrotective effect in all gastric ulcer models. These results suggest that the antiulcerogenic compound(s) present in M. americana may be clustered in the apolar fraction, which will be investigated by our group for the probable mechanisms of action. (c) 2004 Elsevier GmbH. All rights reserved.

Use of comet assay to assess DNA damage in patients infected by Helicobacter pylori - Comparisons between visual and image analyses

Studies of DNA damage in gastric epithelial cells of Helicobacter pylori (H. pylori)-infected patients are conflicting, possibly due to different methods used for scoring DNA damage by Comet assay. Therefore, we compared the sensitivity of visual microscopic analysis (arbitrary units-scores and comets%) and image analysis system (tail moment), in the gastric epithelial cells from the antrum and corpus of 122 H. pylori-infected and 32 non-infected patients. The feasibility of cryopreserved peripheral blood lymphocytes and whole-blood cells for DNA damage biomonitoring was also investigated. In the antrum, the levels of DNA damage were significantly higher in H. pylori-infected patients with gastritis than in non-infected patients with normal mucosa, when evaluated by image analysis system, arbitrary units and comets%. In the corpus, the comets% was not sufficiently sensitive to detect the difference between H. pylori-infected patients with gastritis and non-infected patients with normal mucosa. The image analysis system was sensitive enough to detect differences between non-infected patients and H. pylori-infected patients with mild gastritis and between infected patients with moderate and severe gastritis, in both antrum, and corpus, while arbitrary units and comets% were unable to detect these differences. In cryopreserved peripheral blood lymphocytes, the levels of DNA damage (tail moment) were significantly higher in H. pylori-infected patients with moderate and severe gastritis than in non-infected patients. Overall, our results indicate that the image analysis system is more sensitive and adequate to measure the levels of DNA damage in gastric epithelial cells than the other methods assayed. (c) 2005 Elsevier B.V. All rights reserved.

Picrosirius-polarization staining method as an efficient histopathological tool for collagenolysis detectin in vesical prolapse lesions

The Picrosirius-polarization method has been indicated as a selective histochemical stain for collagen detection in tissue sections. This method can also be of value for studying collagen degradation given that, under polarized light, collagen displays birefringence due to its molecular order. The aim of this study is to highlight this staining method as an additional instrument for a rapid and excellent confirmatory diagnosis of the presence of collagenolysis in connective tissue in the vaginal wall with vesical prolapse lesion, in tissue sections. Dramatic changes in collagen morphology were found in vaginal mucosa in vesical prolapse disorder: they were weakly stained by Sirius red and under polarized light appeared as thin, pale (weakly birefringent), greenish, and with fibers more scattered, while the histoarchitecture of the organ showed a disrupted appearance. Thus, in the present study, we showed in vaginal mucosa in the vesicle prolapse that corroded collagenous framework appears as fragmentary and irregularly separated collagenous structures, that are weakly birefringent, corresponding to a molecular disorganization of these fibers caused by collagenolysis. (C) 2006 Elsevier Ltd. All rights reserved.

Effect of Mouriri pusa extracts on experimentally induced gastric lesions in rodents: Role of endogenous sulfhydryls compounds and nitric oxide in gastroprotection

Several plants are used in folk medicine to treat gastrointestinal disorders. Mouriri pusa Gardn. (Melastomataceae) is a medicinal plant commonly used in the central region of Brazil against gastric ulcer. Two organic extracts methanolic (MeOH) and dichloromethane (DCM) obtained by sequential extraction from the leaves of Mouriri pusa were evaluated for their ability to protect the gastric mucosa against injuries caused by necrotizing agents (0.3 M HCl/60% EtOH, absolute ethanol, non-steroidal anti-inflammatory drug, stress and pylorus ligature) in mice and rats. The best results were obtained after pretreatment with MeOH extract whereas the DCM extract did not show the same significant antiulcerogenic activity. No acute toxicity was observed in animals treated with 5 g/kg, p.o. of MeOH extract. The mechanism involving the antiulcerogenic action of MeOH extract seemed to be related to NO generation and also suggested the effective participation of endogenous sulfhydryl group in the gastroprotective action. Phytochemical investigation of the MeOH extract of Mouriri pusa yielded tannins, flavonoids and (-)-epicatechin. The presence of these phenolic compounds probably would explain the antiulcerogenic effect of the polar extract of Mouriri pusa leaves. (c) 2006 Elsevier B.V.. All rights reserved.

The role of allergic rhinitis in nasal responses to sudden temperature changes

Background: Air conditioning-induced rhinitis in allergic individuals is a common epidemiologic finding, but its physiopathology,is still controversial. The aim of this study was to describe and compare the effects of experimental air conditioning temperature changes on the nasal mucosa of individuals with persistent allergic rhinitis compared with a control group.Methods: A case-control challenge study was performed in a laboratory of thermal comfort with experimental twin challenge chambers set at a 12 C difference in temperature. A group of 32 patients with persistent allergic rhinitis and a group of 16 control subjects were exposed for 30 minutes, 3 times alternately in each chamber. Nasal symptom scores were recorded and nasal samples collected before, immediately after, and 24 and 48 hours after the challenge.Results: the rhinitis group showed a higher symptom score, epithelial shedding, percentage of eosinophils, total inflammatory cells, leukotriene C-4, eosinophil cationic protein, albumin, and tryptase levels compared with controls. There was also a significant increase in symptom score, total cells recovered, percentage of eosinophils, epithelial shedding, albumin, myeloperoxidase, and soluble intercellular adhesion molecule 1 in both groups compared with baseline levels.Conclusion: Sudden temperature changes led to a more pronounced inflammatory nasal response in the rhinitis group with the recruitment and activation of eosinophils.Clinical implications: Persistent allergic rhinitis is a risk factor for developing sudden temperature change-related rhinitis even in the absence of allergen exposure.

Localized lymphatic sporotrichosis after fish-induced injury (Tilapia sp.)

Localized lymphatic sporotrichosis generally develops after the fungus Sporothrix schenckii is traumatically introduced into skin or mucosa by contaminated plant material. An 18-year-old male fisherman was injured by spines of the dorsal fin of a fish on the left third finger. The lesion became ulcerated, edematous and suppurative and did not respond to tetracycline and cephalexin. Fifteen days after the accident, a nodular lymphangitic pattern of swelling was observed. Histopathological findings and an intradermal test were suggestive of sporotrichosis and mycological cultures confirmed the diagnosis. The lesions resolved after oral treatment with potassium iodide. Sporotrichosis is a common subcutaneous mycosis in Brazil, and there is a previous report in the literature of this disease being acquired via trauma involving fish spines.

Paradental cyst: Report of two cases

Background: the paradentat cyst is an odontogenic lesion of inflammatory origin that has few clinical signs and symptoms apart from recurring acute episodes. A well-defined radiolucency associated with the roots or distal to the crown may be seen radiographically. The purpose of this article is to report on different aspects of two cases involving paradental cysts. In the first case, the patient complained about recurring pericoronitis. A semilunar-shaped radiolucency on the distal aspect of the mandibular third molar was noted on the periapical radiograph. In the second case, the patient's main complaint was chronic trauma of the overlying mucosa. Radiographs revealed an enlarged pericoronal space.Methods: In both cases, the mandibular third molar was extracted due to a lack of space. Lesional samples were sent for histopathologic analysis.Results: In the first case, the drainage of cystic fluid and a regular concavity were found during tooth removal. In the second case, a nodular lesion was found adhering to the disto-buccal surface of the tooth arising from the distal wall of a periodontal pocket. The histopathologic analysis revealed a hyperplastic stratified squamous epithelium with arcading lining a fibrous capsule with inflammatory infiltrate, resulting in a final diagnosis of a paradental cyst.Conclusions: the presence of a paradental cyst should be considered when recurrent inflammatory periodontal processes are associated with partially erupted vital teeth, even when characteristic radiographic findings are absent. Definitive diagnosis requires a clinicopathologic correlation incorporating surgical, radiographic, and histologic findings.

Oral cysticercosis: case report

Frequent in developing countries, cysticercosis is a parasitic infection that rarely involves the mouth. This study reports a case of oral cysticercosis in a 13-year-old female patient who had an asymptomatic nodule in the right labial mucosa. An excisional biopsy was carried out and the histopathologic examination revealed a cystic space containing a Taenia solium larva.

Doxorubicin as an antioxidant: Maintenance of myocardial levels of lycopene under doxorubicin treatment

The mechanism of doxorubicin-induced cardiotoxicity remains controversial. Wistar rats (n=96) were randomly assigned to a control (C), lycopene (L), doxorubicin (D), or doxorubicin+lycopene (DL) group. The L and DL groups received lycopene (5 mg/kg body wt/day by gavage) for 7 weeks. The D and DL groups received doxombicin (4 mg/kg body wt intraperitoneally) at 3, 4, 5, and 6 weeks and were killed at 7 weeks for analyses. Myocardial tissue lycopene levels and total antioxidant performance (TAP) were analyzed by HPLC and fluorometry, respectively. Lycopene metabolism was determined by incubating H-2(10)-lycopene with intestinal mucosa postmitochondrial fraction and lipoxygenase and analyzed with HPLC and APCI mass spectroscopy. Myocardial tissue lycopene levels in DL and L were similar. TAP adjusted for tissue protein were higher in myocardium of D than those of C (P=0.002). Lycopene metabolism study identified a lower oxidative cleavage of lycopene in D as compared to those of C. Our results showed that lycopene was not depleted in myocardium of lycopene-supplemented rats treated with doxorubicin and that higher antioxidant capacity in myocardium and less oxidative cleavage of lycopene in intestinal mucosa of doxorubicin-treated rats suggest an antioxidant role of doxombicin rather than acting as a prooxidant. (c) 2007 Elsevier B.V. All rights reserved.

EFFECTS OF PROTEIN-DEFICIENCY AND NATURAL INTESTINAL INFECTION WITH GIARDIA-LAMBLIA ON JEJUNAL INTRAEPITHELIAL LYMPHOCYTES IN RATS OF DIFFERENT AGES

1. The interaction between experimental protein deprivation and natural intestinal infection by Giardia lamblia was studied in terms of its effects on the intraepithelial lymphocyte (IEL) population and morphology of the jejunal mucosa of rats of different ages.2. Young, adult and old male Wistar rats received a protein-deficient diet (2% casein) or a control diet (20% casein) for 42 days. Mucosal height and the number of lymphocytes located among 500 consecutive epithelial cells (EC) along the villi or crossing the basement membrane were determined in PAS-stained jejunal fragments.3. The number of IEL increased progressively with animal age, from 14 to 25 per 100 epithelial cells, with significant differences between age ranges. However, the number of IEL did not differ between control and protein-deficient rats in any of the age groups. The proportion of lymphocytes crossing the basement membrane was approximately two-fold greater in young (2.8/100 EC) and adult (5.8/100 EC) protein-deficient animals than in their respective controls (1.6 and 2.8/100 EC). The intensity of parasite colonization was moderate, from 3 to 5/100 EC and did not differ between groups. The pattern of morphologic changes of jejunal mucosa in protozoal infection did not differ between control and protein-deficient animals in any of the three age groups.4. We conclude that intestinal infection with Giardia lamblia probably stimulated the local immune response, masking the reduction of the IEL population induced by protein deficiency. The increase in lymphocyte numbers with age may be related to prolonged antigenic stimulation promoted by infection.

Molecular model for the binary complex of uropepsin and pepstatin

The three-dimensional structure of human uropepsin complexed with pepstatin has been modelled using human pepsin as a template. Uropepsin is an aspartic proteinase from the urine, produced in the form of pepsinogen A in the gastric mucosa. The structure is bilobal, consisting of two predominantly beta -sheet lobes which, as observed in other aspartic proteinases, are related by a pseudo twofold axis. A structural comparison between binary complexes of pepsin:pepstatin and uropepsin:pepstatin is discussed. (C) 2001 Academic Press.

Removal of epulis fissuratum associated to vestibuloplasty with carbon dioxide laser

There have been numerous surgical techniques developed for removing the epulis fissuratum lesions in order to improve alveolar ridge contour and improving adaptation of full prostheses. Most of these techniques can cause postoperative complications, such as oedema, pain, difficulty in swallowing and jaw movements, haemorrhage, infections and slow placement of final prostheses. The use of carbon dioxide (CO2) laser for the surgical removal of epulis lesions has resulted in many significant improvements including convenient mucosa removal, no bleeding or need for sutures, and minimal postoperative pain and oedema. This study is of 15 cases of removal of extensive epulis with vestibuloplasty in the maxilla and mandible that was carried out with CO2 laser, with no postoperative complications, rapid healing, and excellent aesthetic and functional outcome, all of which allowed for more rapid placement of final prosthesis.