In vitro study on the cell adhesion ability of immobilized lactobacilli on natural supports

The aim of the present study was to investigate the effect of probiotic immobilization onto wheat grains, both wet and freeze dried, on the adhesion properties of the probiotic cells and make comparisons with wet and freeze dried free cells. Lactobacillus casei ATCC 393 and Lactobacillus plantarum NCIMB 8826 were used as model probiotic strains. The results showed satisfactory adhesion ability of free cells to a monolayer of Caco-2 cells (> 1000 CFU/100 Caco-2 cells for wet cells). Cell immobilization resulted in a significant decrease in adhesion, for both wet and freeze dried formulations, most likely because immobilized cells did not have direct access to the Caco-2 cells, but it still remained in adequate levels (> 100 CFU/100 Caco-2 cells for wet cells). No clear correlation could be observed between cell adhesion and the hydrophobicity of the bacterial cells, measured by the hexadecane adhesion assay. Most notably, immobilization enhanced the monolayer integrity of Caco-2 cells, demonstrated by a more than 2-fold increase in transepithelial electrical resistance (TEER) compared to free cells. SEM micrographs ascertained the adhesion of both immobilized and free cells to the brush border microvilli. Finally, the impact of the food matrix on the adhesion properties of probiotic bacteria and on the design of novel functional products is discussed.

In vitro and clinical evaluation of specific dentifrices for complete denture hygiene

To study the physical properties of two experimental dentifrices for complete denture hygiene, their effect on denture biofilm removal and antimicrobial properties by means of a clinical trial. The experimental dentifrices comprised two compositions. One was based on the addition of 1% chloramine T (D1) and the other on the presence of 0.01% fluorosurfactant (D2). Measurements of density, pH, consistency, rheological features and abrasiveness were conducted. Sixty complete denture wearers were randomly assigned to three groups and were instructed to brush their dentures with a specific toothbrush: (1) Water (control); (2) D1; or (3) D2. Each method was used for 21 days. Denture biofilm was disclosed by a 1% neutral red solution and quantified by means of digital photos taken from the internal surface. Microbiological assessment was conducted to quantify Candida sp. and mutans streptococci. Data were evaluated by one-way anova and Tukey HSD, or Kruskal-Wallis (alpha = 0.05). Both dentifrices decreased biofilm coverage when compared with the control group. D1 was the most efficacious treatment to reduce mutans streptococci, whereas D2 showed an intermediate outcome (anova, p < 0.040). No treatment influenced Candida albicans or non-albicans species (Kruskal-Wallis, p = 0.163 and 0.746, respectively). It can be concluded that brushing complete dentures with the experimental dentifrices tested could be effective for the removal of denture biofilm.

Yerba Mate (Ilex paraguariensis) Aqueous Extract Decreases Intestinal SGLT1 Gene Expression but Does Not Affect Other Biochemical Parameters in Alloxan-Diabetic Wistar Rats

Yerba mate (Ilex paraguariensis) is rich in polyphenols, especially chlorogenic acids. Evidence suggests that dietary polyphenols could play a role in glucose absorption and metabolism. The aim of this study was to evaluate the antidiabetic properties of yerba mate extract in alloxan-induced diabetic Wistar rats. Animals (n = 41) were divided in four groups: nondiabetic control (NDC, n = 10), nondiabetic yerba mate (NDY, n = 10), diabetic control (DC, n = 11), and diabetic yerba mate (NDY, n = 10). The intervention consisted in the administration of yerba mate extract in a 1 g extract/kg body weight dose for 28 days; controls received saline solution only. There were no significant differences in serum glucose, insulin, and hepatic glucose-6-phosphatase activity between the groups that ingested yerba mate extract (NDY and DY) and the controls (NDC and DC). However, the intestinal SGLT1 gene expression was significantly lower in animals that received yerba mate both in upper (p = 0.007) and middle (p < 0.001) small intestine. These results indicate that bioactive compounds present in yerba mate might be capable of interfering in glucose absorption, by decreasing SGLT1 expression.

(65)Zn(2+) transport by isolated gill epithelial cells of the American lobster, Homarus americanus

Gills are the first site of impact by metal ions in contaminated waters. Work on whole gill cells and metal uptake has not been reported before in crustaceans. In this study, gill filaments of the American lobster, Homarus americanus, were dissociated in physiological saline and separated into several cell types on a 30, 40, 50, and 80% sucrose gradient. Cells from each sucrose solution were separately resuspended in physiological saline and incubated in (65)Zn(2+) in order to assess the nature of metal uptake by each cell type. Characteristics of zinc accumulation by each kind of cell were investigated in the presence and absence of 10 mM calcium, variable NaCl concentrations and pH values, and 100 mu M verapamil, nifedipine, and the calcium ionophore A23187. (65)Zn(2+) influxes were hyperbolic functions of zinc concentration (1-1,000 mu M) and followed Michaelis-Menten kinetics. Calcium reduced both apparent zinc binding affinity (K (m)) and maximal transport velocity (J (max)) for 30% sucrose cells, but doubled the apparent maximal transport velocity for 80% sucrose cells. Results suggest that calcium, sodium, and protons enter gill epithelial cells by an endogenous broad-specificity cation channel and trans-stimulate metal uptake by a plasma membrane carrier system. Differences in zinc transport observed between gill epithelial cell types appear related to apparent affinity differences of the transporters in each kind of cell. Low affinity cells from 30% sucrose were inhibited by calcium, while high affinity cells from 80% sucrose were stimulated. (65)Zn(2+) transport was also studied by isolated, intact, gill filament tips. These intact gill fragments generally displayed the same transport properties as did cells from 80% sucrose and provided support for metal uptake processes being an apical phenomenon. A working model for zinc transport by lobster gill cells is presented.

Histological and functional renal alterations caused by Bothrops alternatus snake venom: Expression and activity of Na(+)/K(+)-ATPase

Background: Acute renal failure is a serious complication of human envenoming by Bothrops snakes. The ion pump Na(+)/K(+)-ATPase has an important role in renal tubule function, where it modulates sodium reabsorption and homeostasis of the extracellular compartment. Here, we investigated the morphological and functional renal alterations and changes in Na(+)/K(+)-ATPase expression and activity in rats injected with Bothrops alternatus snake venom. Methods: Male Wistar rats were injected with venom (0.8 mg/kg, iv.) and renal function was assessed 6.24, 48 and 72 h and 7 days post-venom. The rats were then killed and renal Na(+)/K(+)-ATPase activity was assayed based on phosphate release from ATP; gene and protein expressions were assessed by real time PCR and immunofluorescence microscopy, respectively. Results: Venom caused lobulation of the capillary tufts, dilation of Bowman`s capsular space. F-actin disruption in Bowman`s capsule and renal tubule brush border, and deposition of collagen around glomeruli and proximal tubules that persisted seven days after envenoming. Enhanced sodium and potassium excretion, reduced proximal sodium reabsorption, and proteinuria were observed 6 h post-venom, followed by a transient decrease in the glomerular filtration rate. Gene and protein expressions of the Na(+)/K(+)-ATPase alpha(1) subunit were increased 6 h post-venom, whereas Na(+)/K(+)-ATPase activity increased 6 h and 24 h post-venom. Conclusions: Bothrops alternatus venom caused marked morphological and functional renal alterations with enhanced Na(+)/K(+)-ATPase expression and activity in the early phase of renal damage. General significance: Enhanced Na(+)/K(+)-ATPase activity in the early hours after envenoming may attenuate the renal dysfunction associated with venom-induced damage. (C) 2011 Elsevier B.V. All rights reserved.

Dipeptidyl peptidase IV inhibition downregulates Na(+)-H(+) exchanger NHE3 in rat renal proximal tubule

In the microvillar microdomain of the kidney brush border, sodium hydrogen exchanger type 3 (NHE3) exists in physical complexes with the serine protease dipeptidyl peptidase IV (DPPIV). The purpose of this study was to explore the functional relationship between NHE3 and DPPIV in the intact proximal tubule in vivo. To this end, male Wistar rats were treated with an injection of the reversible DPPIV inhibitor Lys [Z(NO(2))]-pyrrolidide (I40; 60 mg center dot kg(-1)center dot day(-1) ip) for 7 days. Rats injected with equal amounts of the noninhibitory compound Lys[ Z(NO(2))]-OH served as controls. Na(+) -H(+) exchange activity in isolated microvillar membrane vesicles was 45 +/- 5% decreased in rats treated with I40. Membrane fractionation studies using isopycnic centrifugation revealed that I40 provoked redistribution of NHE3 along with a small fraction of DPPIV from the apical enriched microvillar membranes to the intermicrovillar microdomain of the brush border. I40 significantly increased urine output ( 67 +/- 9%; P < 0.01), fractional sodium excretion ( 63 +/- 7%; P < 0.01), as well as lithium clearance ( 81 +/- 9%; P < 0.01), an index of end-proximal tubule delivery. Although not significant, a tendency toward decreased blood pressure and plasma pH/HCO(3)(-) was noted in I40-treated rats. These findings indicate that inhibition of DPPIV catalytic activity is associated with inhibition of NHE3-mediated NaHCO(3) reabsorption in rat renal proximal tubule. Inhibition of apical Na(+) -H(+) exchange is due to reduced abundance of NHE3 protein in the microvillar microdomain of the kidney brush border. Moreover, this study demonstrates a physiologically significant interaction between NHE3 and DPPIV in the intact proximal tubule in vivo.

Genotoxic effects of X-rays on keratinized mucosa cells during panoramic dental radiography

Objectives: The aim of this study was to evaluate the genotoxic effects of X-rays on epithelial gingival cells during panoramic dental radiography using a differentiated protocol for the micronucleus test. Methods: 40 healthy individuals who underwent this procedure for diagnostic purposes on request from their dentists agreed to participate in this study. All of them answered a questionnaire before the examination. Epithelial gingival cells were obtained from the keratinized mucosa of the upper dental arcade by gentle scraping with a cervical brush immediately before exposure and 10 days later. Cytological preparations were stained according to the Feulgen-Rossenbeck reaction, counterstained with fast green 1% for 1 min and analysed under a light microscope. Micronuclei, nuclear projections (broken eggs) and degenerative nuclear alterations (pyknosis, karyolysis, karyorrhexis and condensed chromatin) were scored. Results: The frequency of micronuclei was significantly higher after exposure (P < 0.05), as were frequencies of nuclear alterations indicate of apoptosis (P < 0.001). Conclusions: These results indicate that X-ray radiation emitted during panoramic dental radiography induces a genotoxic effect on epithelial gingival cells that increases the frequency of chromosomal damage and nuclear alterations indicative of apoptosis.

HOST-MEDIATED INDUCTION OF alpha-AMYLASES BY LARVAE OF THE MEXICAN BEAN WEEVIL Zabrotes subfasciatus (COLEOPTERA: CHRYSOMELIDAE: BRUCHINAE) IS IRREVERSIBLE AND OBSERVED FROM THE INITIATION OF THE FEEDING PERIOD

Larvae of Zabrotes subfasciatus secrete alpha-amylases that are insensitive to the alpha-amylase inhibitor found in seeds of Phaseolus vulgaris. By analyzing amylase activities during larval development on P. vulgaris, we detected activity of the constitutive amylase and the two inducible amylase isoforms at all stages. When larvae were transferred from the non alpha-amylase inhibitor containing seeds of Vigna unguiculata to P. vulgaris, the inducible alpha-amylases were expressed at the same level as in control larvae fed on P. vulgaris. Interestingly, when larvae were transferred from seeds of P. vulgaris to those of V. unguiculata, inducible alpha-amylases continued to be expressed at a level similar to that found in control larvae fed P. vulgaris continuously. When 10-day-old larvae were removed from seeds of V. unguiculata and transferred into capsules containing flour of P. vulgaris cotyledons, and thus maintained until completing 17 days ( age when the larvae stopped feeding), we could detect higher activity of the inducible alpha-amylases. However, when larvae of the same age were transferred from P. vulgaris into capsules containing flour of V. unguiculata, the inducible alpha-amylases remained up-regulated. These results suggest that the larvae of Z. subfasciatus have the ability to induce insensitive amylases early in their development. A short period of feeding on P. vulgaris cotyledon flour was sufficient to irreversibly induce the inducible alpha-amylase isoforms. Incubations of brush border membrane vesicles with the alpha-amylase inhibitor 1 from P. vulgaris suggest that the inhibitor is recognized by putative receptors found in the midgut microvillar membranes. (C) 2010 Wiley Periodicals, Inc.

Noninvasive Evaluation of Oral Lesions Using Depth-sensitive Optical Spectroscopy

BACKGROUND: Optical spectroscopy is a noninvasive technique with potential applications for diagnosis of oral dysplasia and early cancer. In this study, we evaluated the diagnostic performance of a depth-sensitive optical spectroscopy (DSOS) system for distinguishing dysplasia and carcinoma from non-neoplastic oral mucosa. METHODS: Patients with oral lesions and volunteers without any oral abnormalities were recruited to participate. Autofluorescence and diffuse reflectance spectra of selected oral sites were measured using the DSOS system. A total of 424 oral sites in 124 subjects were measured and analyzed, including 154 sites in 60 patients with oral lesions and 270 sites in 64 normal volunteers. Measured optical spectra were used to develop computer-based algorithms to identify the presence of dysplasia or cancer. Sensitivity and specificity were calculated using a gold standard of histopathology for patient sites and clinical impression for normal volunteer sites. RESULTS: Differences in oral spectra were observed in: (1) neoplastic versus nonneoplastic sites, (2) keratinized versus nonkeratinized tissue, and (3) shallow versus deep depths within oral tissue. Algorithms based on spectra from 310 nonkeratinized anatomic sites (buccal, tongue, floor of mouth, and lip) yielded an area under the receiver operating characteristic curve of 0.96 in the training set and 0.93 in the validation set. CONCLUSIONS: The ability to selectively target epithelial and shallow stromal depth regions appeared to be diagnostically useful. For nonkeratinized oral sites, the sensitivity and specificity of this objective diagnostic technique were comparable to that of clinical diagnosis by expert observers. Thus, DSOS has potential to augment oral cancer screening efforts in community settings. Cancer 2009;115:1669-79. (C) 2009 American Cancer Society.

Estudo morfométrico e estereológico digital da mucosa do intestino delgado de crianças eutróficas e desnutridas com diarréia

Objetivos: Testar a hipótese de que a mucosa do intestino delgado proximal de crianças com diarréia persistente apresenta alterações morfométricas e estereológicas proporcionais ao estado nutricional. Métodos: estudo transversal, incluindo 65 pacientes pediátricos internados no período de maio de 1989 a novembro de 1991, com idade entre 4 meses e 5 anos , com diarréia de mais de 14 dias de duração, que necessitaram realizar biópsia de intestino delgado como parte do protocolo de investigação. A avaliação nutricional foi realizada pelos métodos de Gomez, Waterlow e pelos escores z para peso/ idade (P/I), peso/estatura (P/E) e estatura/idade (E/I), divididos em: eutróficos = z ≥ 2 DP e desnutridos z < -2dp; eutróficos = z ≥ 2 DP, risco nutricional = z < -1DP e desnutridos = z < -2DP; e de maneira contínua em ordem decrescente, utilizando-se as tabelas do NCHS. A captura e análise das imagens por programa de computador foi efetuada com o auxílio do patologista. Nos fragmentos de mucosa do intestino delgado, foram medidas a altura dos vilos, a profundidade das criptas, a espessura da mucosa, a espessura total da mucosa e a relação vilo/cripta, com aumento de 100 vezes. Com aumento de 500 vezes, foram medidas a altura do enterócito, a altura do núcleo e do bordo em escova. O programa computadorizado utilizado foi o Scion Image. A análise estereológica, foi feita com o uso de arcos ciclóides. Resultados: Para os escores z P/I, P/E e E/I, divididos em duas categorias de estado nutricional, não houve diferença estatisticamente significante quanto às medidas da altura dos vilos, profundidade das criptas, espessura da mucosa, espessura total da mucosa e relação vilo/cripta. A altura do enterócito foi a característica que apresentou maior diferença entre os grupos eutrófico e desnutrido, para os índices P/I e P/E, em 500 aumentos, sem atingir significância estatística. Quando os escores z foram divididos em 3 categorias de estado nutricional, a análise morfométrica digitalizada mostrou diferença estatisticamente significante para a relação vilo/cripta entre eutróficos e desnutridos leves e entre eutróficos e desnutridos moderados e graves (p=0,048). A relação vilo/cripta foi maior nos eutróficos. A avaliação nutricional pelos critérios de Waterlow e a análise estereológica não mostraram associação com o estado nutricional. Pelo método de Gomez, houve diferença estatisticamente significante para a altura do enterócito entre eutróficos e desnutridos de Grau III: quanto maior o grau de desnutrição, menor a altura do enterócito (r= -.3330; p = 0,005). As variáveis altura do enterócito, altura do núcleo do enterócito e do bordo em escova apresentaram uma clara associação com os índices P/I (r=0,25;p=0,038), P/E (r=0,029;p=0,019) e com o critério de avaliação nutricional de Gomez (r=-0,33;p=0,007), quando foram avaliadas pelo coeficiente de correlação de Pearson. A altura do núcleo mostrou associação com o índice P/I (r=0,24;p=0,054). A altura do bordo em escova mostrou associação com o índice P/I (r=0,26;p=0,032) e a avaliação nutricional de Gomez (r=-0,28;p=0,020). Conclusões: As associações encontradas entre o estado nutricional - avaliado de acordo com Gomez e os índices P/I e P/E - e as variáveis da mucosa do intestino delgado mostraram relação com o peso dos pacientes. Embora estas associações tenham sido de magnitude fraca a moderada, há uma tendência à diminuição do tamanho do enterócito, seu núcleo e seu bordo em escova à medida que aumenta o grau de desnutrição.

Avaliação clínica e microbiológica das condições de saúde oral dos receptores de transplante renal do Hospital Universitário Onofre Lopes, Natal RN

Introduction: The chronic immunosuppression promotes the development of changes in the oral cavity of the kidney transplant recipients (KTR), however with the exception of gingival overgrowth, little is known regarding the prevalence of oral lesions in this population. Objective: To evaluate the prevalence of clinical and microbiological conditions of the oral cavity of the recipients of kidney transplantation and the associated factors. Methods: This was a cross-sectional study which examined 96 clinical KTR and experimental where collected saliva stimulated them to identify Candida sp. Data collection consisted of evaluation stomatologic, socio-demographic, clinical aspects of transplantation, condition of oral hygiene and dental caries, yonder to questions about knowledge of oral alteration after transplantation. Results: Of the total, 66.7% of KTR had some type of oral manifestation. The most common was saburral tongue, followed by gingival overgrowth, with both oral manifestations related to gender and concomitant use of cyclosporine and nifedipine (p <0.05). Tacrolimus showed a protective effect for gingival overgrowth (OR = 0.13). The oral hygiene was associated with saburral tongue(p = 0.03) and severity of gingival overgrowth (p = 0.0001). Oral candidiasis was diagnosed in 17.7% of patients and Candida albicans was isolated most frequently in the saliva of RTR with a colonization of 58.3%. The average DMF-T increased with age. The method of oral hygiene was most used brush and toothpaste to 61.5%. Changes in the oral cavity was seen in 54.2% of KTR, citing as the main growth and gingival ulcers. Instructions for oral hygiene after transplantation were neglected for 61.5% of RTR. Dry mouth and halitosis were reported in 30.2% and 36.5% respectively. Conclusions: More the half of the KTR had at least one injury of the mouth, the immunosuppressive drugs and oral hygiene are associated with these alterations. Prospective cohort studies are needed to elucidate the relationship between oral manifestations and levels of drug and risk of oral manifestations occur over time. The kidney transplant recipients showed to be aware of oral alterations occurred after transplantation and uninformed about the oral hygiene instructions. With regard to hygiene, the incidence of caries was considered high, conditions of risk were identified and improvements in primary attention should be encouraged and reflected in the monitoring of renal transplant

Efeito do manejo cultural e químico na incidência e severidade da mancha-preta dos citros

A citricultura apresenta vários problemas fitossanitários, dentre os quais a mancha-preta dos citros (Guignardia citricarpa). O controle desta doença baseia-se no emprego de práticas culturais e no uso de fungicidas. Avaliou-se o efeito do manejo do mato em conjunto com o químico no controle da doença. Os experimentos foram instalados em pomares de laranjeiras-doces, nos municípios de Matão, Rio Claro e Mogi Guaçu, no Estado de São Paulo. No manejo do mato, comparou-se o uso isolado de roçadeira ecológica com o conjugado rastelo mecânico e trincha, aos 35 dias, após 2/3 de pétalas caídas. No controle químico, foram realizadas duas pulverizações com fungicida protetor e de 2 a 5 pulverizações da mistura de produto sistêmico com protetor, aos 45 dias, após 2/3 de pétalas caídas, em intervalos de aplicação de 35 dias. em todas as aplicações, foi adicionado óleo mineral emulsionável (0,25%). Avaliou-se a área abaixo da curva de progresso da incidência e severidade da doença, com os dados de cinco avaliações realizadas quinzenalmente, a partir da maturidade fisiológica dos frutos. em todas as áreas, o uso do controle químico, associado com o manejo do mato, reduziu a intensidade da doença.

Caracterização de escovas de grafite e avaliação do seu desempenho em função da microestrutura

The electric can be converted into thermal, luminous, electromagnetic, and also in mechanical energy. In this context the electric engines plays a fundamental role, specially that they work very below of its nominal capacity, with consequent decrease load density. In industrial environment, these characteristics of work of DC engines had also generated an extreme consumption of coal brushs and attack in the commutator reducing the useful life of the engine and increasing maintenance demand and cost. The general objective of the present work is to study the influence of the granulometry of the coal brush used in DC engines with the resistance to the consumption of the same ones, as well as in the performance presented by the commutator of the engine. Additionally, determining the measurable and not measurable profits when the brush used is adjusted to the application. The brushes had been produced by an industry of the sector and tested in industrial environment to evaluate their performance and consumption. Preliminary results evidence a substantial improvement in the performance of these brushes in function of its microstructure and the application in which it is used

Análise de viabilidade técnica de utilização da Fibra de bananeira com resina sintética em Compósitos

This paper aims to present the feasibility of using a composite using discarded material from the cultivation of banana tree (pseudostem), which is fibrillated together with synthetic resin replacing glass fiber to be used in structural elements that do not demand large mechanical stress such as reservoirs, troughs, domes, sewage pipes etc.. For this, there were studies about the mechanical properties of a composite made with polyester resin and fiber of banana tree (Musa sp, musac), in which the splints were removed from the pseudostem, being made fibrillation by hand, with the aid of a brush steel, followed by natural drying. After treatment for cleaning and removal of wax, the fiber was cut into pieces of approximately 60 mm to 100 mm, for, together with synthetic resin, make cards of a features fiber composite with random orientation relative to the weight of the resin. We used three different percentages of fiber (3%, 6% and 9%), in order to make a comparative study between them and what would be the one with the best performance. Were manufactured specimens of each material and then subjected to uniaxial tensile tests, three point bending, moisture absorption and thermal characteristics. The results show that, in general, the use of banana tree fiber is feasible simply by an improvement in the production process (machining of the procedure) and greater care in the manufacture of parts

Controle profissional de biofilme dentário como estratégia para manutenção da saúde periodontal de pacientes com aparelho ortodôntico fixo: análise comparativa entre diferentes períodos

The presence of fixed orthodontics appliances interfere on sanitation, allowing periodontal diseases to appear, despite the fact patients keep on visiting the dentist every month. This research aims to determine a protocol for the mechanical control of the dental biofilm performed by the professional. A protocol that was able to maintain the periodontal health of the patients under orthodontic treatment with fixed appliances, and in order to do so, it used a non-controlled, randomized and blind clinical essay. The sample involved 40 adolescents who were under the installation of fixed orthodontics appliances and it was divided in three groups, as follows: monthly controlled group (group 1) composed of 11 patients, the quarterly controlled group (group 2) with 16 patients and the semestrial controlled group (group 3) with 13 patients. For data collection, an interview and clinical exams with probing depth measurement, quantity of keratinized mucosa, Gingival Index and the Plaque Index were used. On the initial exam all patients received brushing guidelines as well as the professional control of dental biofilm, with periodontal scaler, Robinson s brush and prophylactic paste. However, Group 1 returned every month for control procedures; Group 2 every three months and Group 3 after six months. The intervention had a six-month duration (for all the three groups), when all the exams were repeated by another examiner who did not know which group each patient was inserted in. Finally, the research concluded that despite the fact there is no statistically significant difference among the three groups, clinically the patients from the monthly group presented a better response to professional control, with less accumulation of dental biofilm and less rate of gingival inflammation. Thus, the mechanical control of the dental biofilm performed by the professional could not avoid gingival increase, characterized by the raise of probing depth measurement, neither the quantity of keratinized mucosa