Fernando Neves Nogueira

In vitro approach to evaluate potential harmful effects of beer on teeth

OBJECTIVE The purpose of the present work was to examine some properties of different brands of beer manufactured in Brazil that may be important to oral health. METHODS Samples from seven different beer brands were analyzed for pH, titratable acidity, calcium and phosphate concentrations. Demineralization experiments were carried out by incubating samples with crown tooth particles (40-80 mesh) at 37 degrees C under agitation (100 strokes/min). RESULTS The pH was lower than 4.0 for three of the seven samples and higher than 4.0 for the others. The amount of titratable acidity, expressed as the volume of 0.1N NaOH solution consumed to raise the initial pH to 7.0, and the concentrations of calcium and phosphate varied. Calcium concentration ranged from 0.21 to 1.59 micromol/ml, while phosphate concentration varied from 0.048 to 0.094 micromol/ml. Calcium released to the incubation medium was proportional to the time of incubation up to 5min. Maltose, a disaccharide, was detected in all samples studied. CONCLUSION Differences in the properties examined indicated that some brands of beer studied may have potential dental effects.

Antioxidant enzymatic defense in salivary glands of streptozotocin-induced diabetic rats: a temporal study.

Hyperglycemia induces overproduction of superoxide and it is related to diabetic complications. In this study, we analyzed the antioxidant enzymatic defense and the lipid peroxidation of rat salivary glands in six different periods of diabetic condition. Ninety‐six rats were divided into 12 groups: C7/14/21/28/45/60 (non‐diabetic animals) and D7/14/21/28/45/60 (diabetic animals). Diabetes was induced by streptozotocin and the rats were euthanized after 7, 14, 21, 28, 45, or 60 days. Their parotid (PA) and submandibular (SM) glands were removed soon after the sacrifice and the total protein and malondialdehyde (MDA) concentrations, as well as, the superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities were determined. Twenty‐one days after the diabetes induction, the SM glands showed an increase in SOD, CAT, and GPx activities, as well as, MDA concentration. Concerning the PA glands, an increase in the CAT activity and MDA content was observed throughout the observation period. The results suggest that diabetes can cause alterations on the salivary glands and that PA and SM glands react differently when exposed to diabetes condition. However, no impairment of antioxidant system was observed in the group whose diabetic condition had been induced 60 days earlier, herein named 60‐day group. Copyright

Activity, distribution and regulation of phosphofructokinase in salivary gland of rats with streptozotocin-induced diabetes

Although the influence of diabetes on salivary glands is well studied, it still presents conflicting results. In this work, the regulation of the phosphofructokinase-1 enzyme (PFK-1) was studied utilizing the salivary glands of rats. Diabetes was induced by a single intraperitoneal injection of streptozotocin (60 mg/Kg of body weight) in rats (180-200 g). The animals were killed 30 days after the induction of diabetes and the submandibular and parotid salivary glands were used. Hyperglycemia was evaluated by blood sugar determination. The distribution of PFK-1 between the soluble and cytoskeleton fractions, the phosphate content of PFK-1, the content of fructose-2,6-bisphosphate and the activity of the PFK-2 enzyme were determined. The calculated relative glandular weight showed a higher value for the parotid gland in comparison with the control, but not for the submandibular gland. The activity of PFK-1 expressed per gland showed no variation between diabetic and control animals. However, considering the specific activity, the soluble enzyme presented a value 50% higher than that of the control and the cytoskeleton bound form increased by 84% compared to the control. For the parotid gland, no difference in the specific activity between diabetic and control animals was observed. On the other hand, the activity per gland of the soluble enzyme increased in the diabetic animals. The phosphate content of PFK-1 increased in the submandibular and parotid glands of diabetic rats. Both the content of fructose-2,6-bisphosphate and the active form of PFK-2 were reduced in the diabetic glands. In conclusion, the increase in the activity of PFK-1 observed in the salivary glands of rats with streptozotocin-induced diabetes does not seem to be due to its modulator fructose-2,6-bisphosphate.

Influence of streptozotocin-induced diabetes on hexokinase activity of rat salivary glands

The influence of diabetes on the enzyme hexokinase (HK) was examined in the salivary glands of rats. Diabetes was induced by an intraperitoneal injection of streptozotocin (60 mg/Kg body weight) in overnight fasted rats (180–200 g). The animals were killed 48 hours and 30 days after the induction of diabetes and the submandibular and parotid salivary glands extracted for use. Hyperglycemia was evaluated by determining the blood sugar. The area occupied by each intralobular component, acini, ducts, total parenchyma and stroma was measured, and no differences were observed compared with control. In the soluble fraction of the submandibular gland, no difference in the specific activity of HK was observed, between the diabetic and control animals, however, the activity per gland and per g of tissue showed lower values than control. The specific activity of the bound form was reduced in the diabetic gland. The results obtained for the parotid gland were different from the submandibular. The specific activity of both the soluble and bound forms were increased in the diabetic animals. The DEAE-cellulose column chromatography of the soluble and bound forms of the enzyme from both glands showed a first peak appearing during the washing of the column and two other peaks were eluted by the gradient. Thus, three isoenzymes in the submandibular and parotid salivary glands for the control and diabetic rats have been found.ResumenSe estudia la influencia de la diabetes sobre la actividad hexoquinasa (HK) en glándulas salivales de rata. La diabetes se induce con una inyección intraperitoneal de estreptozotocina (60 mg/Kg de peso corporal) en animales mantenidos en ayunas durante una noche (180–200 g). Los animales se sacrificaron 48 h y 30 d tras inducción de la diabetes y se obtuvieron las glándulas submandibulares y las parótidas. La hiperglucemia se valoró determinando el azúcar sanguíneo. El área ocupada por cada componente intralobular, acinos, conductos, parénquima total y estroma no varió entre los animales tratados y control. La actividad específica de la HK (mU/g proteína) en la fracción soluble de la glándula submandibular no varió entre los animales del grupo control y los diabéticos. Sin embargo, la actividad HK referida a g de tejido era menor en diabéticos que en el grupo control. Respecto de la forma ligada, se observó disminución también en la actividad específica en las diabéticas. Los resultados en las glándulas parótidas fueron diferentes que en las submandibulares, ya que se observó incremento de la actividad HK específica, tanto de la forma soluble como de la ligada, en los animales diabéticos. La columna cromatográfica DEAE-celulosa de las formas soluble y ligada de las enzimas de ambas glándulas presentaron la aparición del primer pico durante el lavado de la columna. Otros dos picos fueron eluídos por un gradiente. Por tanto, se han encontrado tres isoenzimas en las glándulas submandibulares y parótidas tanto en los animales control como en los diabéticos.

Sialic acid reduction in the saliva of streptozotocin induced diabetic rats

OBJECTIVE Diabetes causes changes in the salivary glands and in the composition of saliva, as well as symptoms such as dry mouth and hyposalivation. Therefore, this study aimed at investigating changes in salivary secretion and composition, in response to parasympathetic stimuli, in diabetic rats induced with streptozotocin. DESIGN Diabetes was induced by a single intraperitoneal injection of streptozotocin. Thirty days after diabetes induction, the animals were anaesthetized and salivation was stimulated by an intraperitoneal injection of Pilocarpine (0.6mg/kg body weight) dissolved in distilled water. Saliva was collected for 40min and immediately stored at -80°C until analysis. The salivary flow rate, amount of total protein, amylase and peroxidase activities, and free and total sialic acid contents were measured. RESULTS Salivary flow rate was reduced in the diabetic group (p<0.05). Moreover, increases in total protein amount and in amylase and peroxidase activities were observed in diabetic animals. No difference was observed for free sialic acid content between groups. On the other hand, a significantly decrease in the total sialic acid content was observed in the diabetic group (p<0.05). CONCLUSIONS Our findings suggest that a decrease in sialic acid in the saliva of diabetic animals can be related to xerostomia reported by diabetic patients. However, further clinical trials are needed to verify if the decrease in sialic acid also occurs in human saliva.

Effects of Single Exposure of Sodium Fluoride on Lipid Peroxidation and Antioxidant Enzymes in Salivary Glands of Rats

Many studies suggest that fluoride exposure can inhibit the activity of various enzymes and can generate free radicals, which interfere with antioxidant defence mechanisms in living systems. To further the understanding of this issue, this present study examined the effects of low-dose fluoride treatment on the activity of enzymatic antioxidant superoxide dismutase (SOD) and catalase (CAT), as well as the levels of lipid peroxidation (LPO) in the parotid (PA) and submandibular (SM) salivary glands of rats. Rats were injected with a single dose of sodium fluoride (NaF) (15 mg F−/kg b.w.) then euthanized at various time intervals up to 24 hours (h) following exposure. NaF exposure did not cause significant differences in SOD or CAT activity or LPO levels in PA glands compared to control. Conversely, SM glands presented increased SOD activity after 3 h and decreased SOD activity after 1, 12, and 24 h, while LPO was increased after 6, 12, and 24 h of the NaF injection. There were no significant differences in the CAT activity in the groups studied. Our results demonstrated that NaF intoxication caused oxidative stress in salivary glands few hours after administration. These changes were more pronounced in SM than in PA gland.

The antioxidant effects of green tea reduces blood pressure and sympathoexcitation in an experimental model of hypertension

Background: Oxidative stress is a key mediator in the maintenance of sympathoexcitation and hypertension in human and experimental models. Green tea is widely known to be potent antioxidant. Objective: We aimed to evaluate the effects of green tea in a model of hypertension. Methods: Hypertension was induced by the nitric oxide synthase inhibitor [N-nitro-L-arginine-methyl-ester (L-NAME); 20 mg/kg per day, orally, for 2 weeks] in male Wistar rats. After the first week of L-NAME treatment, animals received green tea ad libitum for 1 week. At the end of the treatment period, blood pressure, heart rate, baroreflex sensitivity, renal sympathetic nerve activity, and vascular and systemic oxidative stress were assessed. Results: L-NAME-treated animals exhibited an increase in blood pressure (165 ± 2 mmHg) compared with control rats (103 ± 1 mmHg) and green tea treatment reduced hypertension (119 ± 1 mmHg). Hypertensive animals showed a higher renal sympathetic nerve activity (161 ± 12 spikes/s) than the control group (97 ± 2 spikes/s), and green tea also decreased this parameter in the hypertensive treated group (125 ± 5 spikes/s). Arterial baroreceptor function and vascular and systemic oxidative stress were improved in hypertensive rats after green tea treatment. Conclusions: Taken together, short-term green tea treatment improved cardiovascular function in a hypertension model characterized by sympathoexcitation, which may be because of its antioxidant properties.

Metabolic evaluations of cancer metabolism by NMR-based stable isotope tracer methodologies

Cancer cells are widely recognized for being able to adapt their metabolism towards converting available nutrients into biomass to increase proliferation rates.

Salivary Alterations in Rats with Experimental Chronic Kidney Disease

Objective This study aimed to analyze changes in saliva composition and salivary secretion process of rats with chronic kidney disease induced by 5/6 nephrectomy to set the foundation for salivary studies related to CKD. Methods CKD was induced in Wistar rats via 5/6 nephrectomy. Blood and saliva samples were collected from Control, Sham and CKD groups at 8 and 12 weeks after the surgery. Salivation was stimulated via intraperitoneal injections of pilocarpine (1.0 mg/Kg body weight) or isoproterenol (5.0 mg/Kg body weight). Saliva was collected and immediately stored at -80°C until analysis. The salivary flow rate, total protein, amylase and peroxidase activities, and urea concentrations were measured. The blood urea nitrogen (BUN) and serum creatinine concentrations were also evaluated. Results Increases in BUN and serum creatinine concentrations were observed in the CKD groups. Amylase activity was significantly reduced in response to both stimuli in the CKD groups at 8 weeks and increased in the CKD groups at 12 weeks in response to isoproterenol stimulus. The peroxidase activities of the CKD groups were significantly reduced in response to isoproterenol stimulation and were increased at 12 weeks in response to pilocarpine stimulation. Salivary urea was significantly increased in the CKD groups at 8 weeks in response to the isoproterenol stimuli and at 12 weeks in response to both salivary agonists. Conclusions The pattern of alterations observed in this experimental model is similar to those observed in patients and clearly demonstrates the viability of 5/6 nephrectomy as an experimental model in future studies to understand the alterations in salivary compositions and in salivary glands that are elicited by CKD.

Effect of mouthrinses with different active agents in the prevention of initial dental erosion.

INTRODUCTION Hydrochloric acid (HCl) from the gastric juice is the only source of intrinsic acid, which can reach the oral cavity in cases of gastroesophageal reflux or chronic vomiting, enhancing the risk of dental erosion. AIM Compare the effects of mouthrinses with different active agents in the prevention of initial dental erosion caused by HCl. SUBJECTS AND METHODS Casein (CAS at 0.2%), sodium hexametaphosphate (HMP at 0.02%), titanium tetrafluoride (TiF4 at 0.34%), and stannous fluoride (SnF2 at 0.87%) were individually added to an experimental mouthrinse. The mouthrinse without additives was used as the negative control (C) and a commercially available mouthrinse for erosion (ELM-Elmex®) as the reference product. Enamel specimens were exposed to human saliva and randomly assigned to 6 experimental groups (n = 8). Specimens were submitted to erosion in HCl for 10 s, followed by to the experimental mouthrinses for 30 s, and artificial saliva for 60 min. This cycle was repeated 3 times. The total amounts of calcium and phosphorus released by the specimens in the 2nd and 3rd erosive challenges were evaluated by atomic emission spectrometry. Statistical analysis used Shapiro-Wilks and Hartley tests, followed by one-way ANOVA and Tukey tests. RESULTS When compared with C, ELM and HMP presented significantly less calcium in solution, with no difference between them. All the groups showed similar and significantly less phosphorus than C, except CAS. CONCLUSIONS HMP was the only agent that could match the protection against initial erosion of the commercially available mouthrinse in both analyses.