Evaluation of a digital learning object (DLO) to support the learning process in radiographic dental diagnosis.

AIMS: Studies have shown that inappropriate therapeutic strategies may be adopted if crown and root changes are misdiagnosed, potentially leading to undesirable consequences. Therefore, the aim of this study was to evaluate a digital learning object, developed to improve skills in diagnosing radiographic dental changes. MATERIALS AND METHODS: The object was developed using the Visual Basic Application (VBA) software and evaluated by 62 undergraduate students (male: 24 and female: 38) taking an imaging diagnosis course. Participants were divided in two groups: test group, which used the object and control group, which attended conventional classes. After 3 weeks, students answered a 10-question test and took a practice test to diagnose 20 changes in periapical radiographs. RESULTS: The results show that test group performed better that control group in both tests, with statistically significant difference (P = 0.004 and 0.003, respectively). In overall, female students were better than male students. Specific aspects of object usability were assessed using a structured questionnaire based on the System Usability Scale (SUS), with a score of 90.5 and 81.6 by male and female students, respectively. CONCLUSIONS: The results obtained in this study suggest that students who used the DLO performed better than those who used conventional methods. This suggests that the DLO may be a useful teaching tool for dentistry undergraduates, on distance learning courses and as a complementary tool in face-to-face teaching.

CBCT-based volume of simulated root resorption - influence of FOV and voxel size.

AIM: To examine the influence of the field of view (FOV) and voxel size on the measurement of the volume of simulated internal root resorption (IRR) lesions through cone-beam computed tomography (CBCT). METHODOLOGY: Eleven single-rooted teeth with IRR simulated by acid demineralization were studied. CBCT images were acquired using large FOV (voxel sizes of 0.200, 0.250 and 0.300 mm) and limited FOV (voxel sizes of 0.076, 0.100 and 0.200 mm). The IRR volumes were calculated using the Dolphin(®) software. Volumetric measurements were validated using IRR silicone putty casts. The analysis of variance (anova) for randomized block design complemented with the Tukey's test was employed. RESULTS: IRR volumes obtained using voxel sizes of 0.200 and 0.250 mm were similar (P > 0.05). However, both these values were significantly different from that obtained using the 0.300-mm voxel (P < 0.05). There was no significant difference between IRR volumes measured through voxel sizes of 0.076 and 0.100 mm (P > 0.05), but both differed significantly from that obtained through the 0.200-mm voxel (P < 0.05). There was no significant difference between the volumetric measurements of the 0.200-mm voxel images of the restricted and large FOV protocols. The mean volumes of the silicone casts were smaller than those calculated using a 0.200-mm voxel, but were similar to those obtained using voxel sizes of 0.076 and 0.300 mm. CONCLUSIONS: Despite the FOV protocol, voxel size can influence measurement of simulated IRR volumes. The importance of standardization of CBCT image acquisition protocols is emphasized, especially during follow-up of an IRR lesion, to prevent misinterpretation of its extent, which can create a bias in clinical decisions.

CBCT for the assessment of second mesiobuccal (MB2) canals in maxillary molar teeth: effect of voxel size and presence of root filling.

AIM: To compare detection of the second mesiobuccal (MB2) canal in maxillary molar teeth using cone-beam computed tomography (CBCT) with different voxel sizes against conventional radiographic examination when the MB1 was unprepared, prepared and filled. METHODOLOGY: Radiographic examination and 0.2-, 0.25- and 0.3-mm CBCT (n = 89) were performed in 3 stages: S1, no first mesiobuccal (MB1) canal preparation or filling; S2, after MB1 preparation and filling; and S3, after MB1 root canal filling removal and canal repreparation. Images were analysed using the i-Cat software. After RE and CBCT acquisition in S3, all the samples were clarified to directly visualize the presence of the MB2 canal. All images were analysed by a blinded, previously calibrated examiner. Accuracy, specificity and sensitivity were compared using analysis of variance (P < 0.05 level of significance). RESULTS: MB2 root canals were detected in 67% of the samples. Overall, radiographic examination was associated with lower mean accuracy values for detecting MB2 than CBCT regardless of the MB1 condition. The MB1 root canal condition did not influence MB2 detection in 0.2-mm voxel images. The presence of root fillings in the MB1 canals reduced the detection of MB2 canals, especially in the 0.3- and 0.25-mm voxel-size images (P < 0.05). CONCLUSION: CBCT was associated with higher mean values of specificity and sensibility than radiographic examination for the detection of MB2 canals. When endodontic retreatment is necessary removal of the root filling prior to the CBCT examination eliminates artefacts, thereby permitting the use of the 0.3-mm voxel protocol that has good diagnostic performance and lower radiation dose.

Prospective evaluation of aminopeptidase activities in plasma and peripheral organs of streptozotocin-induced diabetic rats.

The cleavage of peptides by aminopeptidase enzyme types could be among the mechanisms related to certain disruptions on mediator and modulatory functions in diabetes mellitus. In order to examine this hypothesis, we measured representative aminopeptidase activities in tissues of peripheral organs of control and streptozotocin-diabetic rats. None of the examined aminopeptidase activities differed between diabetics and controls in plasma, ileum, stomach or lung. Soluble and membrane-associated alanyl, and membrane-associated cystyl aminopeptidase activities were higher in the kidney of diabetics. Decreased activity was observed in soluble and membrane-associated aspartyl and soluble dipeptidyl-peptidase IV, while increased activity was observed in soluble alanyl, arginyl, and cystyl aminopeptidases in the pancreas of diabetics. In the jejunum, soluble cystyl aminopeptidase increased in diabetics. Soluble arginyl and type-1-pyroglutamyl aminopeptidase and membrane-associated dipeptidyl-peptidase IV activities increased in the liver of diabetics. Membrane-associated dipeptidyl-peptidase IV and alanyl aminopeptidase activities in the spleen were higher in diabetics than in controls. Membrane-associated alanyl aminopeptidase activity also increased in the heart of diabetics. All these changes in streptozotocin-treated rats were avoided by the administration of insulin. Our comparative analysis of a diverse array of aminopeptidase activities supported the proposal that the regulation of peptide cleavage by these enzyme types is associated with the effects of streptozotocin-diabetes mellitus on peripheral organs.

Methotrexate and cyclosporine treatments modify the activities of dipeptidyl peptidase IV and prolyl oligopeptidase in murine macrophages.

Analysis of the effects of cyclosporine A (25-28 mgkg(-1)) and/or methotrexate (0.1 mgkg(-1)) treatments on dipeptidyl peptidase IV (DPPIV) and prolyl oligopeptidase (POP) activities and on algesic response in two distinct status of murine macrophages (Mphis) was undertaken. In resident Mphis, DPPIV and POP were affected by neither individual nor combined treatments. In thioglycolate-elicited Mphis, methotrexate increased DPPIV (99-110%) and POP (60%), while cyclosporine inhibited POP (21%). Combined treatment with both drugs promoted a rise (51-84%) of both enzyme activities. Only cyclosporine decreased (42%) the tolerance to algesic stimulus. Methotrexate was revealed to exert prevalent action over that of cyclosporine on proinflammatory Mphi POP. The opposite effects of methotrexate and cyclosporine on POP activity might influence the availability of the nociceptive mediators bradykinin and substance P in proinflammatory Mphis. The exacerbated response to thermally induced algesia observed in cyclosporine-treated animals could be related to upregulation of those mediators.

Galanin-like immunoreactivity in the brain of the snake Bothrops jararaca.

The distribution of galanin-like immunoreactive perikarya and nerve fibers in the brain of the snake Bothrops jararaca was studied by means of immunohistochemistry using an antiserum against porcine galanin. Immunoreactive neurons were only detected in the infundibular recess nucleus. Immunoreactive fibers were found in the telencephalic, diencephalic and mesencephalic areas such as the dorsal cortex, nucleus accumbens, lamina terminalis, preoptic area, mediodorsal region of the supraoptic nucleus, subfornical organ, nucleus of the paraventricular organ, subcommisural organ and periventricular grey region. The habenula, paraventricular nucleus, infundibular recess nucleus and hypothalamo-hypophyseal tract presented denser innervations. The outer layer of the median eminence displayed numerous fibers located close to the portal system, while scarce fibers were seen in the inner median eminence and neural lobe of the hypophysis. The distribution of labelled neurons in the brain of this snake was more restricted than that described in a turtle. The wide hypothalamic and extrahypothalamic distribution of labelled fibers suggests that galanin peptides may have hypophysiotropic, neuromodulator and neurotransmitter roles in the snake B. jararaca.

Distribution of somatostatin immunoreactivity in the brain of the snake Bothrops jararaca.

The distribution of perikarya and fibers containing somatostatin was studied in the brain of the snake Bothrops jararaca by means of immunohistochemistry using an antiserum against synthetic somatostatin. Immunoreactive perikarya and fibers were localized in telencephalic, diencephalic and mesencephalic areas. In the telencephalon, numerous immunoreactive perikarya were found in the medial, dorsomedial, dorsal and lateral cortex, mainly in the deep plexiform layer, less so in the cellular layer, but not in the superficial plexiform layer. Immunoreactive perikarya were also observed in the dorsal ventricular ridge, the nucleus of the diagonal band of Broca, amygdaloid complex, septum and lamina terminalis. In the diencephalon, labelled cells were observed in the paraventricular, periventricular hypothalamic and in the recessus infundibular nuclei. In the mesencephalon, immunoreactive perikarya were seen in the mesencephalic reticular formation, reticular nucleus of the isthmus and torus semicircularis. Labelled fibers ran along the diencephalic floor and the inner zone of the median eminence, and ended in the neural lobe of the hypophysis. Other fibers were observed in the outer zone of the median eminence close to the portal vessels and in the septum, lamina terminalis, retrochiasmatic nucleus, deep layers of the tectum, periventricular gray and granular layer of the cerebellum. Our data suggest that somatostatin may function as a mediator of adenohypophysial secretion as well as neurotransmitter and/or neuromodulator which can regulate the neurohypophysial peptides in the snake B. jararaca.

Galanin-like immunoreactivity in the brain of the snake Bothrops jararaca

The distribution of galanin-like immunoreactive perikarya and nerve fibers in the brain of the snake Bothrops jararaca was studied by means of immunohistochemistry using an antiserum against porcine galanin. Immunoreactive neurons were only detected in the infundibular recess nucleus. Immunoreactive fibers were found in the telencephalic, diencephalic and mesencephalic areas such as the dorsal cortex, nucleus accumbens, lamina terminalis, preoptic area, mediodorsal region of the supraoptic nucleus, subfornical organ, nucleus of the paraventricular organ, subcommisural organ and periventricular grey region. The habenula, paraventricular nucleus, infundibular recess nucleus and hypothalamo-hypophyseal tract presented denser innervations. The outer layer of the median eminence displayed numerous fibers located close to the portal system, while scarce fibers were seen in the inner median eminence and neural lobe of the hypophysis. The distribution of labelled neurons in the brain of this snake was more restricted than that described in a turtle. The wide hypothalamic and extrahypothalamic distribution of labelled fibers suggests that galanin peptides may have hypophysiotropic, neuromodulator and neurotransmitter roles in the snake B. jararaca.

Reproductive cycle of the Neotropical Crotalus durissus terrificus: I. Seasonal levels and interplay between steroid hormones and vasotocinase.

Crotaline snakes present delayed fertilization and sperm storage because secondary vitellogenesis is not completed by the time of mating. The release of vitellogenesis and synchrony between ovulation and fertilization suggest a steroidal modulation. We investigated changes of sexual steroid levels during reproduction in the Neotropical rattlesnake Crotalus durissus terrificus, analyzing macroscopical variations of reproductive condition (vitellogenesis, pregnancy, and post-partum) and plasma levels of estradiol, progesterone, and vasotocinase cystine aminopeptidase (CAP) activity over 2 years. Data showed 44.4% non-reproductive snakes (40.1% primary vitellogenesis and 4.3% post-partum) and 55.6% reproductive (36.8% secondary vitellogenesis and 18.8% pregnant). Estradiol was low in spring and summer, increasing in autumn till it peaked in winter. Estradiol in secondary vitellogenesis was significantly higher than in primary vitellogenesis, or in pregnant and post-partum females, Progesterone dropped significantly in autumn compared to summer, winter, and spring. Pregnant females showed the highest levels of progesterone compared to primary or secondary vitellogenesis, or post-partum females. CAP activity showed lowest values in reproductive females in autumn and greatest levels in post-partum females. A significant negative linear relationship was obtained between CAP activity and estradiol. The combination of morphological observations, levels of steroids and CAP activity allowed us to suggest a similar morphological reproductive pattern between temperate and tropical rattlesnakes, and to infer the role of estradiol, progesterone and CAP activity on vitellogenesis, gestation and sperm storage, respectively.

Reproductive cycle of the Neotropical Crotalus durissus terrificus: II. Establishment and maintenance of the uterine muscular twisting, a strategy for long-term sperm storage.

Crotaline snakes store sperm by means of a uterine musculature twisting (UMT). We investigated the influence of plasma levels of estradiol and progesterone and vasotocinase cystine aminopeptidase (CAP) activity on UMT formation and maintenance, and the in vitro uterine reactivity for AVT in Crotalus durissus terrificus in primary or secondary vitellogenesis with or without UMT. Frequency of females in secondary vitellogenesis with UMT is significantly higher than in primary one. Estradiol levels did not vary in all conditions studied, however, significantly low levels of progesterone were found in snakes in secondary vitellogenesis with UMT compared to those without it. UMT is always observed when high levels of estradiol and low levels of progesterone are detected. CAP activity did not change in the presence of UMT. AVT produced concentration-response contractions of the isolated uterus of snakes in all stages analysed and the pD2 value and maximum contractile response were significantly higher in primary vitellogenesis without UMT than in other reproductive conditions, indicating that uterus of those snakes presents a higher contractile capacity which may favour UMT establishment. In conclusion, we show a relationship of UMT and estradiol/progesterone balance and a possible participation of AVT in UMT formation and maintenance in the Neotropical rattlesnake.

Peptide metabolism and the control of body fluid homeostasis.

In this paper, we review our current understanding of the medicinal chemistry of the major peptide systems, which influence body fluid homeostasis. Electrolytes play pivotal roles in intra- and intercellular communication, acid-base equilibrium and, when bound to several macromolecules, they regulate a myriad of enzymatic proteins, receptors and transcription factors. Cell turgor influences the plasma membrane, which activates mechanically-gated ion channels or mechanoreceptors, and the expression of a number of genes which underlie long-term metabolic responses to hormones, substrates and reactive oxygen intermediates. The altered kinetics and enzymatic cleavage of peptides during water-electrolyte imbalance can contribute to cardiac and renal damage associated with elevated blood pressure. Identification of the enzymes which are responsible for cleavage, together with emerging information about the mechanisms of action and structures of regulatory and effector peptides, has laid a foundation for the discovery of novel drugs, some of which are in use or are now undergoing evaluation in experimental trials. The development of models of hydrosaline challenge with relative efficiency to induce selective water-electrolyte imbalance has permitted the identification of kallikrein-kinin, renin-angiotensin-aldosterone, vasopressin-oxytocin, thyrotropin-releasing hormone and luteinizing hormone-releasing hormone as susceptible substrates. At present, the angiotensin-I converting enzyme inhibitors are well-known efficacious, orally active, blood pressure-lowering agents which have been used in hypertensive patients. In addition to several new analogues of this class of drug, some selective dual inhibitors of angiotensin-I converting enzyme and neutral endopeptidase and inhibitors of aminopeptidases are now also being rationally assayed and their beneficial effects on hypertension and hydromineral balance indicate that this type of drug may have powerful therapeutic effects for disorders of body fluid homeostasis.

Effects of changes in hydromineral balance on rat brain aspartyl, arginyl, and alanyl aminopeptidase activities.

In order to examine the possible relationship between the processing and inactivation roles of aminopeptidases and the disruption of water-electrolyte balance, we measured the activities of aspartyl aminopeptidase (Asp-Ap), arginyl aminopeptidase (Arg-Ap) and alanyl aminopeptidase (Ala-Ap) in certain brain areas (hypothalamus, hippocampus, thalamus and brain cortices) and in the pituitary gland in several models of hydrosaline change. The activity of hypothalamic membrane-bound Asp-Ap significantly decreased (more than 50%) following treatments which induced a hypovolemic state. Aminopeptidase M activity (membrane-bound Ala-Ap activity with low sensitivity to puromycin) was also significantly decreased by 53 % in the thalamus of rats under conditions of hypovolemia plus hyperosmolality in comparison to the control group. These results indicate that aminopeptidases in the central nervous system may be involved in the physiological regulation of hydromineral balance.

Effect of the disruption of body fluid balance on pyroglutamyl aminopeptidase (Type-1) in rat brain structures.

The activity of soluble and membrane-bound pyroglutamyl aminopeptidase Type-1 (PAP I) was evaluated in the hypothalamus, hippocampus, thalamus, brain cortex, and pituitary gland of rats after applying certain hydromineral challenges. Compared to euhydrated rats, decreased enzyme activity was found in the hypophysis of rats deprived of water for 48 h, or rats drinking ad libitum hypertonic sodium chloride solution (2%) for 6 days or distilled water for 6 days and then submitted to acute water overload. PAP I cleaves the pGlu-amino acid bond of neuropeptides such as thyroliberin, luliberin, neurotensin, and bombesin. The decay of particulate PAP I activity may cause an increase of these pyroglutamate peptides in the whole pituitary. Although the deleterious or pro-homeostatic influence of this decay remains to be elucidated, the present data provide evidence for the involvement of this enzyme activity at this anatomical location in the water-electrolyte imbalance.

The distribution of vasotocin and mesotocin immunoreactivity in the brain of the snake, Bothrops jararaca.

Polyclonal antibodies against vasotocin (AVT) and mesotocin (MST) were used to explore the distribution of these peptides in the brain of the snake Bothrops jararaca. Magnocellular AVT- and MST-immunoreactive (ir) perikarya were observed in the supraoptic nucleus (SON), being AVT-ir neurons more numerous. A portion of the SON, in the lateroventral margin of the diencephalon ventrally to optic tract, showed only AVT-ir perikarya and fibers. However, the caudal most portion displayed only mesotocinergic perikarya. Parvocellular and magnocellular AVT- and MST-ir perikarya were present in the paraventricular nucleus (PVN) being AVT-ir fibers more abundant than MST-ir. Vasotocinergic perikarya were also found in a dorsolateral aggregation (DLA) far from the PVN. Mesotocinergic perikarya were also present in the recessus infundibular nucleus and ependyma near to paraventricular organ. Nerve fibers emerging from supraoptic and paraventricular nuclei run along the diencephalic floor, internal zone of the median eminence (ME) to end in the neural lobe. Also a dense network of AVT- and MST-ir fibers was present in the external zone of the ME, close to the vessels of the hypophysial portal system. As a rule, all regions having vasotocinergic and mesotocinergic perikarya also showed immunoreactive fibers. Vasotocinergic and mesotocinergic fibers but not perikarya were found in the lamina terminalis (LT). Moreover AVT-ir fibers were present in the nucleus accumbens and MST-ir fibers in the septum. In mesencephalon and rhombencephalon MST-ir fibers were more numerous than AVT-ir fibers. Vasotocinergic and mesotocinergic fibers in extrahypothalamic areas suggest that these peptides could function as neurotransmitters and/or neuromodulators in the snake B. jararaca.

Effects of hydrosaline treatments on prolyl endopeptidase activity in rat tissues.

Enzymatic cleavage of some peptide hormones, neurotransmitters and neuromodulators could be implicated in the regulation of extra- and intracellular fluid volume and osmolality. Prolyl endopeptidase is known to hydrolyze several peptides, which act on hydromineral balance, such as angiotensins, bradykinin, vasopressin, oxytocin, thyrotropin-releasing hormone, neurotensin and opioids. In this work, we analyzed the effects of certain volume and/or osmotic changes in the activity of the soluble and membrane-bound prolyl endopeptidase in several brain areas, heart, lungs, kidney and adrenal and pituitary glands of the rat. Soluble prolyl endopeptidase activity was higher in the renal cortex of the chronic salt-loaded rats than in the control rats. In the water-deprived and polyethylene glycol-treated rats, heart particulate prolyl endopeptidase was lower than in the control rats. Particulate prolyl endopeptidase was also lower in the adrenal gland of the acute salt-loaded rats and in the brain cortex of the water-loaded rats than in the control rats. Data suggest that tissue-dependent peptide hydrolysis evoked by prolyl endopeptidase activity is involved in the water-electrolyte homeostasis.

Corticotropin-releasing hormone-like immunoreactivity in the brain of the snake Bothrops jararaca.

The distribution of corticotropin-releasing hormone in the brain of the snake Bothrops jararaca was studied immunohistochemically. Immunoreactive neurons were detected in telencephalic, diencephalic and mesencephalic areas such as dorsal cortex, subfornical organ, paraventricular nucleus, recessus infundibular nucleus, nucleus of the oculomotor nerve and nucleus of the trigeminal nerve. Immunoreactive fibres ran along the hypothalamo-hypophysial tract to end in the outer layer of the median eminence and the neural lobe of the hypophysis. In general, immunoreactive fibres occurred in the same places of immunoreactive neurons. In addition, immunoreactive fibres were observed in the septum, amygdala, lamina terminalis, supraoptic nucleus, nucleus of the paraventricular organ, ventromedial hypothalamic nucleus and interpeduncular nucleus. These results indicate that, as for other vertebrates, corticotropin-releasing hormone in B. jararaca brain, besides being a releasing hormone, may also act as a central neurotransmitter and/or neuromodulator.

Interactions among challenges of hydromineral balance, angiotensin-converting enzyme, and cystine aminopeptidase.

Enzymatic cleavage of some peptides could be included among the mechanisms of water-electrolyte homeostasis. To test this hypothesis, the angiotensin-converting activity (ACE) of plasma and the L-cystine-di-beta-naphthylamidase activity (CAP) of plasma and of soluble and particulate fractions from different areas of the central nervous system (CNS) were investigated in rats submitted to treatments eliciting hydromineral imbalance. CAP in the CNS was unchanged by hydromineral challenges. The correlations observed between plasma osmolality and CAP, and plasma CAP and ACE suggested a contribution of these activities to the restoration of basal water-electrolyte and blood pressure conditions through the hydrolysis of vasopressin, oxytocin, angiotensin I and bradykinin.

Plasma arginine-vasotocin and hydroosmotic status of the terrestrial pit viper Bothrops jararaca.

Peaks corresponding to arg-vasotocin obtained by HPLC from Sep-Pak C18 column extracts of Bothrops jararaca plasma were identified by radioimmunoassay and amino acid analysis. Plasma vasotocin and protein levels, osmolality, and L-cystine-di-beta-naphthylamidase were also compared in snakes under normal hydration conditions with or without chronic administration of vasotocin or in the presence of chronic hydroosmotic challenges. Sep-Pak C18 and radioimmunoassay were validated for the extraction and determination of this peptide, respectively (about 80% recovery). EDTA presented a protective action on this recovery compared to the use of heparin as anticoagulant for snake blood. A reduction of vasotocin content related to the time of incubation of this peptide added to snake plasma was detected by radioimmunoassay. Snake plasma activity also on L-cystine-di-beta-naphthylamide indicated that this vasotocin-destroying effect was due to hydrolysis by a cystine-aminopeptidase-like activity. Plasma levels of vasotocin revealed an unexpected dispersion and absent correlation with plasma levels of osmolality. Measurable vasotocin in a large number of snakes associated with lower levels of l-cystine-di-beta-naphthylamidase in acute than in chronic salt loading suggested the role of this enzyme activity in long-term regulation of the vasotocin system in this snake.

L-cystine-di-beta-naphthylamidase activity in plasma of the snake Bothrops jararaca after chronic challenges of water balance.

Plasma protein, osmolality and L-cystine-di-beta-naphthylamidase activity were measured in Bothrops jararaca snakes submitted to normal hydration, water deprivation, water loading, salt loading or argvasotocin injections. Protein and osmolality were similar for male and female snakes in all treatments. Protein content was similar in all treatments, indicating the ability of the animals to maintain fluid balance between the vascular and extravascular compartment under osmotic challenges. Osmolality was increased in water-deprived and salt-loaded and was decreased in water-loaded animals compared with normally hydrated or argvasotocin-injected snakes. Enzymatic activity differed between males and females and was higher in salt-loaded females and in arg-vasotocin-injected females and males than in water-loaded snakes. A positive linear regression between osmolality and L-cystine-di-beta-naphthylamidase was obtained for female snakes (correlation coefficient = 0.6162, P < 0.01). The data show a role of arg-vasotocin in the consistent relationship between water balance and this sex-related aminopeptidase activity.

Circulating vasotocin in the snake Bothrops jararaca.

1. There is biochemical and pharmacological evidence to suggest the presence of vasotocin in the blood and plasma of the snake Bothrops jararaca (Bj). 2. XE-64 extracts from Bj blood showed antidiuretic and hypotensive activities in rats and a contractile effect on rat isolated uterus, which was totally dialysable and inhibited by thioglycollate. 3. Extracts from Bj whole plasma presented an antidiuretic activity which was only partially dialysable. 4. The plasma extracts also showed oxytocic properties. 5. When EDTA and Sep-Pak C18 extraction were used, a better recovery and characterization of vasotocin by HPLC was obtained. 6. These results indicate the occurrence of free and bound circulating vasotocin in Bj, in an equilibrium dependent of its enzymatic hydrolysis.