Teledentistry: A Tool to Promote Continuing Education Actions on Oral Medicine for Primary Healthcare Professionals.

BACKGROUND: Difficulties in diagnosis of oral mucosal lesions are a significant cause of delayed oral cancer diagnosis, and this difficulty may be due to gaps in knowledge. This study evaluated the diagnostic skills of primary healthcare professionals regarding oral cancer and presented them with an e-learning course. MATERIALS AND METHODS: Forty-seven primary healthcare professionals (32 dentists and 15 nondentists) enrolled in a 24-h course on oral medicine delivered through an e-learning platform. A test, based on 33 clinical images of oral lesions, was used to evaluate the diagnostic skills of participants. The participants were requested to classify each lesion as benign, potentially malignant, or malignant as well as to inform their clinical impression. Three specialists also took the test as the gold standard. RESULTS: Twenty-seven participants completed the test. Nondentists and dentists showed a comparable sensitivity of 68.8 ± 11.1 and 63.7 ± 15.8, respectively. Specialists performed somewhat better; however, the difference was not statistically significant (81.0% ± 4.1%, p = 0.16). Dentists and specialists (70.0% ± 16.6% and 95.5% ± 3.1%, respectively) showed higher specificity than nondentists (39.3 ± 20.6, p < 0.01). Nondentists had a higher number of unanswered questions (p < 0.01) for classification and clinical impression (50.0% ±45.1% and 72.0% ± 25.0%, respectively) than dentists (5.7% ±11.9% and 19.8% ± 20%, respectively). Both dentists and nondentists had low attendance in the course (44.57% ± 37.38% and 26.53% ± 26.53%, respectively, p = 0.26). To the best of our knowledge, this is the first study to assess the diagnostic skills of public health workers belonging to different professional categories. CONCLUSION: Both dentists and nondentists have a fairly good capacity for discriminating the nature of oral lesions. Early squamous cell carcinoma is the most challenging situation and remains an issue to be addressed.

Intrastriatal injection of hypoxanthine alters striatal ectonucleotidase activities: a time-dependent effect.

The aim of this study was to investigate the effects of intrastriatal injection of hypoxanthine on ectonucleotidase (E-NTPDases and ecto-5'-nucleotidase) activities and expressions in the striatum of rats. The effect of pre-treatment with vitamins E and C on the effects elicited by this oxypurine on enzymatic activities and on thiobarbituric reactive substances (TBARS) was also investigated. The effect of pre-incubation with hypoxanthine on nucleotide hydrolysis in striatum homogenate was also determined. Adult Wistar rats were divided into (1) control and (2) hypoxanthine-injected groups. For ectonucleotidase activity determination, the animals were sacrificed at 30 min, 24 h and 7 days after drug infusion. For the evaluation of the expression of NTPDase 1-3 and also ecto-5'-nucleotidase, TBARS assay and the influence of the pre-treatment with vitamins on ectonucleotidase activities, the animals were sacrificed 24 h after hypoxanthine infusion. Results show that hypoxanthine infusion significantly inhibited ectonucleotidase activities and increased TBARS only 24 h after administration. Pre-treatment with vitamins was able to prevent these effects. Moreover, ecto-5'-nucleotidase expression was increased (80%) at 24 h after hypoxanthine infusion. We suggest that these hypoxanthine-induced biochemical modifications could, at least in part, participate in the pathophysiology of Lesch Nyhan disease.

Protective effect of nitric oxide synthase inhibition or antioxidants on brain oxidative damage caused by intracerebroventricular arginine administration.

We have previously demonstrated that acute arginine administration induces oxidative stress and compromises energy metabolism in rat hippocampus. In the present study, we initially investigated the effect of intracerebroventricular infusion of arginine (0.1, 0.5 and 1.5 mM solution) on Na(+),K(+)-ATPase activity and on some parameters of oxidative stress, namely thiobarbituric acid-reactive substances (TBA-RS) and total radical-trapping antioxidant parameter (TRAP) in the hippocampus of rats. Results showed that 1.5 mM arginine solution significantly increases TBA-RS and reduces Na(+),K(+)-ATPase activity and TRAP in the rat hippocampus. We also evaluated the influence of the nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME), and antioxidants, namely alpha-tocopherol plus ascorbic acid, on the effects elicited by arginine on Na(+),K(+)-ATPase activity, TBA-RS and TRAP. Results showed that treatment with alpha-tocopherol plus ascorbic acid per se did not alter these parameters but prevented these effects. Furthermore, intracerebroventricular infusion of L-NAME prevented the inhibition caused by arginine on Na(+),K(+)-ATPase activity, as well as the increased of TBA-RS. Our findings indicate that intracerebroventricular infusion of arginine induces oxidative stress in rat hippocampus and that the inhibition of Na(+),K(+)-ATPase activity caused by this amino acid was probably mediated by NO and/or its derivatives ONOO(-) and/or other free radicals. Finally, we suggest that the administration of antioxidants should be considered as an adjuvant therapy to specific diets in hyperargininemia.

Sulfite increases lipoperoxidation and decreases the activity of catalase in brain of rats.

The main objective of this study was to investigate the in vitro effects of sulfite, a metabolite accumulated in isolated sulfite oxidase deficiency, on Na (+), K (+)-ATPase activity and on some parameters of oxidative stress, namely thiobarbituric acid-reactive substances (TBARS) and catalase activity (antioxidant enzyme) in cerebral cortex, striatum and hippocampus from 10- and 60-day-old rats. Results showed that 500 microM sulfite significantly increased TBARS and reduced catalase activity in the cerebral structures studied from neonates and adults rats; in contrast, sulfite did not alter Na(+), K(+)-ATPase activity. Our present findings show that sulfite increases lipid peroxidation and decreases antioxidant enzyme defenses in rat brain, suggesting an induction of oxidative stress. We presumed that oxidative stress might be, at least in part, associated with the neuronal dysfunction of patients affected by isolated sulfite oxidase deficiency.

alpha-Tocopherol and ascorbic acid prevent memory deficits provoked by chronic hyperprolinemia in rats.

In the present study we investigated the action of alpha-tocopherol and ascorbic acid on the effects elicited by chronic hyperprolinemia on rat performance in the Morris water maze. Rats received subcutaneous injections of proline (experimental group) twice a day, with 10 h-interval, from the 6 to 28th days of age or an equivalent volume of 0.9% saline solution (controls). Half of the proline-treated group also received intraperitoneal administration of alpha-tocopherol (40 mg/kg) and of ascorbic acid (100 mg/kg) from the 6 to 28th days of life. On the 60th day of life, rats were subjected to testing in the water maze. Results show that chronic proline administration provokes impairment on spatial learning in reference memory task, as revealed by the increase of latency in acquisition, in the probe trial and in crossing over the platform location, as well as by the number of crossings, when compared to saline-treated animals. Proline-treated rats also demonstrated a reduced efficiency to find the platform position in the working memory task. Rats chronically treated with proline plus alpha-tocopherol and ascorbic acid had above effects prevented, suggesting the participation of oxidative stress in such effects. Our findings lend support to a novel therapeutic strategy, based on these vitamins, to the cognitive dysfunction associated with hyperprolinemia type II.

Vitamins E and C pretreatment prevents ovariectomy-induced memory deficits in water maze.

We investigated whether the pretreatment with vitamins E (alpha-tocopherol) and C (ascorbic acid) would act on ovariectomy-induced memory deficits in Morris water maze tasks. Adult female Wistar rats were divided into three groups: (1) naive (control), (2) sham (submitted to surgery without removal of ovaries) and (3) ovariectomized. Thirty days after surgery, they were trained in the Morris water maze in order to verify ovariectomy effects both on reference and working memory tasks. Results show that ovariectomized rats presented impairment in spatial navigation in the acquisition phase, as well as in the time spent in target quadrant and in the latency to cross over the location of the platform in test session, when compared to naive and sham groups (controls), in the reference memory task. Ovariectomy did not affect performance in the working memory task. Confirming our hypothesis, ovariectomized rats pretreated for 30 days with vitamins E and C had those impairments prevented. We conclude that ovariectomy significantly impairs spatial reference learning/memory and that pretreatment with vitamins E and C prevents such effect. Assuming this experimental memory impairment might mimic, at least in part, the cognitive deficit sometimes present in the human condition of lack of reproductive hormones, our findings lend support to a novel therapeutic strategy, based on vitamins E and C, to cognitive impairments in post-menopausal women.

Protective effect of antioxidants on brain oxidative damage caused by proline administration.

We have previously demonstrated that acute and chronic hyperprolinemia induce oxidative stress in cerebral cortex of rats. In the present study, we investigated the action of Vitamins E and C on the oxidative damage elicited by acute and chronic administration of proline (Pro) in rat cerebral cortex. Results showed that treatment with Vitamins E and C prevented the alterations caused by acute and chronic administration of proline on chemiluminescence, total radical-trapping antioxidant potential (TRAP) and on the activities of catalase and glutathione peroxidase. If these effects also occur in the human condition, it is possible that antioxidant administration might serve as a potential adjuvant therapy to avoid the progression of the neuropsychiatric dysfunction observed in hyperprolinemic patients.

In vivo and in vitro effects of homocysteine on Na+, K+-ATPase activity in parietal, prefrontal and cingulate cortex of young rats.

In the present study we determined the effect of chronic administration of homocysteine on Na+,K+-ATPase activity in synaptic membranes from parietal, prefrontal and cingulate cortex of young rats. We also studied the in vitro effect of homocysteine on this enzyme activity and on some oxidative stress parameters, namely thiobarbituric acid-reactive substances (TBA-RS) and total radical-trapping antioxidant potential (TRAP) in the same cerebral structures. For the in vivo studies, we induced elevated levels of homocysteine in blood (500 microM), comparable to those of human homocystinuria, and in brain (60 nmol/g wet tissue) of young rats by injecting subcutaneously homocysteine (0.3-0.6 micromol/g of body weight) twice a day at 8 h intervals from the 6th to the 28th postpartum day. Controls received saline in the same volumes. Rats were killed 12 h after the last injection. Chronic administration of homocysteine significantly decreased (50%) Na+,K+-ATPase activity in parietal, increased (36%) in prefrontal and did not alter in cingulate cortex of young rats. In vitro homocysteine decreased Na+,K+-ATPase activity and TRAP and increased TBA-RS in all cerebral structures studied. It is proposed that the alteration of Na+,K+-ATPase and induction of oxidative stress by homocysteine in cerebral cortex may be one of the mechanisms related to the neuronal dysfunction observed in human homocystinuria.

Evaluation of the mechanism underlying the inhibitory effect of guanidinoacetate on brain Na+, K+-ATPase activity.

Guanidinoacetate methyltransferase deficiency (GAMT-deficiency) is an inherited neurometabolic disorder clinically characterized by epilepsy and mental retardation and biochemically by accumulation of guanidinoacetate (GAA) and depletion of creatine. Although the neurological symptoms are predominant, the pathogenesis of the brain dysfunction in this disorder is not yet established. In the present study we investigated the in vitro effect of GAA on Na+, K+-ATPase and Mg2+-ATPase activities in synaptic plasma membrane from hippocampus of young rats. Results showed that GAA significantly inhibited Na+, K+-ATPase activity without affecting Mg2+-ATPase activity. We also evaluated the effect of glutathione (GSH), trolox, Nomega-nitro-L-arginine methyl ester (L-NAME) and taurine (Tau) on the inhibition elicited by GAA on Na+, K+-ATPase activity. GSH, trolox, L-NAME and Tau per se did not alter Na+, K+-ATPase activity. However, L-NAME and taurine prevented the inhibitory effect of GAA on this enzyme activity. Our findings suggest that the inhibition of Na+, K+-ATPase activity caused by GAA is possibly mediated by nitric oxide (NO) formation and/or synaptic membrane alteration. The present data may contribute to the understanding of the neurological dysfunction characteristic of GAMT-deficient patients.

Inhibition of Na+, K+-ATPase activity in rat striatum by the metabolites accumulated in Lesch-Nyhan disease.

In the present study, we investigated the in vitro effect of hypoxanthine, xanthine and uric acid, metabolites accumulating in tissue of patients with Lesch-Nyhan disease, on Na(+), K(+)-ATPase activity in striatum of neonate rats. Results showed that all compounds significantly inhibited Na(+), K(+)-ATPase activity. We also studied the kinetics of the inhibition of Na(+), K(+)-ATPase activity caused by hypoxanthine. The apparent K(m) and V(max) of Na(+), K(+)-ATPase activity for ATP as the substrate and hypoxanthine as the inhibitor were 0.97 mM and 0.69 nmol inorganic phosphate (Pi) released per min per mg of protein, respectively. K(i)-value was 1.9 microM, and the inhibition was of the non-competitive type. We also observed that the inhibitory effects of hypoxanthine, xanthine and uric acid probably occur through the same mechanism, suggesting a common binding site for these oxypurines on Na(+), K(+)-ATPase. Therefore, it is conceivable that inhibition of brain Na(+), K(+)-ATPase activity may be involved at least in part in the neuronal dysfunction characteristic of patients with Lesch-Nyhan disease.

Training in inhibitory avoidance causes a reduction of Na+,K+-ATPase activity in rat hippocampus.

Compelling evidence has indicated the involvement of Na(+),K(+)-ATPase in the mechanisms of synaptic plasticity. In the present study, we investigated the effect of inhibitory avoidance training on Na(+),K(+)-ATPase activity, at different times after training, in the rat hippocampus. Male adult Wistar rats were trained in a step-down inhibitory avoidance task and compared to those submitted to isolated footshock (0.4 mA) or placed directly onto the platform. Na(+),K(+)-ATPase activity decreased, by 60%, in hippocampus of rats sacrificed immediately after the isolated footshock, as well as immediately (0 min) and 6 h after training; this effect was not present 24 h after training. We also verified that enzyme activity was not altered in rats killed after just being on the platform. These findings suggest that Na(+),K(+)-ATPase activity may be involved in the memory consolidation of step-down inhibitory avoidance in the hippocampus.

Brain Na+,K(+)-ATPase inhibition induced by arginine administration is prevented by vitamins E and C.

Hyperargininemia is a metabolic disorder caused by deficiency of arginase activity resulting in tissue accumulation of arginine and neurological dysfunction. We have previously demonstrated that arginine induces oxidative stress and decreases Na+,K(+)-ATPase in rat midbrain. In the present study we investigated the action of vitamins E and C on the inhibition of Na+,K(+)-ATPase provoked by arginine in the midbrain of 60-day-old rats. Animals were pretreated for 1 week with daily IP administration of saline (control) or vitamins E (40 mg/kg) and C (100 mg/kg). Twelve h after the last injection, animals received one injection of arginine (0.8 micromol/g of body weight) or saline. Chemiluminescence was significantly increased, whereas total antioxidant capacity and Na+,K(+)-ATPase activity were significantly decreased. Furthermore, treatment with vitamins E and C prevented these effects. If these effects also occur in the human condition, it is possible that antioxidant administration might slow the progression of neurodegeneration in this disorder.

Proline induces oxidative stress in cerebral cortex of rats.

In the present study we investigated the in vivo and in vitro effects of proline on some parameters of oxidative stress, such as chemiluminescence, total radical-trapping antioxidant potential (TRAP) and the activity of the antioxidant enzymes catalase, glutathione peroxidase and superoxide dismutase in rat cerebral cortex. Ten-day-old rats received one subcutaneous injection of proline (12.8 micromol/g body weight), while control rats received saline in the same volumes. The animals were killed 1h after injection, the cerebral cortex was isolated and the assays immediately carried out. For the in vitro studies, homogenates from cerebral cortex of 10-day-old untreated rats were incubated for 1h at 37 degrees C with various concentrations of proline (3.0 microM-1.0mM). Results showed that proline-treated rats presented a decrease of TRAP (30%) and an increase of chemiluminescence (78%). In contrast, the activities of catalase, glutathione peroxidase and superoxide dismutase were not modified by proline acute treatment. Furthermore, the presence of proline in the medium increased chemiluminescence, decreased TRAP and the activity of superoxide dismutase at proline concentrations similar to those observed in tissues of hyperprolinemic patients (0.5-1.0mM). However, catalase and glutathione peroxidase activities were not affected by the presence of proline in the medium. The results indicate that proline induces oxidative stress in the brain, which may be related, at least in part, to the neurological dysfunction observed in hyperprolinemia.

Proline reduces acetylcholinesterase activity in cerebral cortex of rats.

In the present study we investigated the in vivo and in vitro effect of proline (Pro) on acetylcholinesterase (AChE) activity in rat cerebral cortex. The action of vitamins E and C on the effects produced by Pro was also tested. Twelve-day-old rats received one s.c. injection of Pro (12.8 micromol/g body weight) or an equivalent volume of 0.9% saline solution (control) and were killed 1 h later. In another set of experiments, 5-day-old rats were pretreated for 1 week with daily i.p. administration of saline (control) or vitamins E (40 mg/kg) and C (100 mg/kg). Twelve hours after the last injection the rats received one s.c. injection of Pro (12.8 micromol/g body weight) or saline (control) and were killed 1 h later. For the in vitro studies, cerebral cortex homogenates of 12-day-old untreated rats were incubated for 1 h with various concentrations of Pro (3.0 microM-1.0 mM) or with 1.0 mM Pro, 1.0 mM trolox, or 1.0 mM Pro plus 1.0 mM trolox. Controls did not contain Pro in the incubation medium. Our results showed that the AChE activity significantly decreased (25%) in rat brain subjected to Pro administration and that the pretreatment with vitamins E and C prevented this effect. Furthermore, Pro (0.5 and 1.0 mM) also inhibits AChE activity in vitro and trolox prevented this effect. The data suggest that the inhibitory effect of Pro on AChE activity is associated with oxidative stress. Although it is difficult to extrapolate our findings to the human condition, our results may be relevant to explain, at least in part, the neurologic dysfunction associated with hyperprolinemia type II.