Unstimulated Rate Research Articles

Salivary Flow Rate Before, During and After Insertion of Complete Denture in Different Age Group: a Comparative study

Background: Increase in salivary flow accompanying complete denture insertion is a well-known phenomenon. Complete dentures act as a mechanical stimulant, increasing salivary flow after insertion. This study compares unstimulated and stimulated salivary flow rates before, during, and after insertion of a complete denture in different age groups.Method: Forty subjects meeting inclusion criteria were selected. Unstimulated saliva was collected first. Stimulation was produced by chewing paraffin wax and collected of saliva in a graduated container. This procedure was carried out before insertion, during insertion, 24 hours after, and 7 days after insertion of complete dentures. The study data were entered, edited, and coded in Microsoft Excel. The unstimulated and stimulated salivary flow rate was compared at different time intervals.Result: Unstimulated salivary rate was lowest before denture insertion with mean values of 0.27ml/minute. The highest flow rate was recorded during denture insertion (mean values of 0.60 ml/minute). The mean unstimulated salivary flow rate 24 hours after denture insertion was higher than 7 days after denture insertion. The mean stimulated salivary rate was lowest before denture insertion (mean values of 0.59 ml/minute). The highest flow rate was recorded during denture insertion (mean values of 0.93 ml/minute). There was a declining trend in salivary flow rate after denture insertion i.e. 0.86 ml/minute to 0.75ml/min after 24 hours and 7 days of denture insertion. A gradual decrease in salivary flow rate with an increase in age was noted but the association of age with salivary flow rates was not statistically significant.Conclusion: There was an increase in both unstimulated and stimulated salivary flow rate after insertion of complete denture and age does not affect salivary flow rates before, during (immediately after), 24 hours after, and 7 days after denture insertion.

Identification of immunologic and clinical characteristics that predict inflammatory response to C. Novyi-NT bacteriolytic immunotherapy

BackgroundClostridium novyi-NT (CNV-NT), has shown promise as a bacterolytic therapy for solid tumors in mouse models and in dogs with naturally developing neoplasia. Factors that impact the immunologic response to therapy are largely unknown. The goal of this pilot study was to determine if plasma immune biomarkers, immune cell function, peripheral blood cytological composition and tumor characteristics including evaluation of a PET imaging surrogate of tumor tissue hypoxia could predict which dogs with naturally developing naïve neoplasia would develop an inflammatory response to CNV-NT.ResultsDogs that developed an inflammatory response to CNV-NT had a higher heart rate, larger gross tumor volume, greater tumor [64Cu]ATSM SUVMax, increased constitutive leukocyte IL-10 production, more robust NK cell-like function and greater peripheral blood lymphocyte counts compared to dogs that did not develop an inflammatory response to CNV-NT. Of these, unstimulated leukocyte IL-10 production, heart rate, and gross tumor volume appeared to be the best predictors of which dogs will develop an inflammatory response to CNV-NT.ConclusionsDevelopment of inflammation in response to CNV-NT is best predicted by pretreatment unstimulated leukocyte IL-10 production, heart rate, and gross tumor volume.

Lubiprostone stimulates small intestinal mucin release

BackgroundLubiprostone is a synthetic bicyclic fatty acid derivative of prostaglandin E1 (PGE1) used for chronic constipation. The best known action of lubiprostone is simulation of Cl- dependent fluid secretion. In a mouse model of the genetic disease cystic fibrosis, we previously showed that in vivo administration of lubiprostone resulted in greater mucus accumulation in the small intestine. The aim of this study was to directly test whether lubiprostone stimulates intestinal mucin release.MethodsMucin release was measured by mounting segments (4-5 cm) of mouse proximal-mid small intestine in an organ bath, allowing access to the perfusate (luminal) and the bath (serosal) solutions. Nifedipine (10-6 M) and indomethacin (10-5 M) were included in all solutions to inhibit smooth muscle activity and endogenous prostaglandin production, respectively. The tissue was equilibrated under flow for 30 min, using the perfusate collected during the final 10 min of the equilibration period to measure unstimulated release rate. Stimulus was then added to either the perfusate or the bath and the perfusate was collected for another 30 min to measure the stimulated mucin release rate. Mucin in perfusates was quantified by periodic acid-Schiff's base dot-blot assay, using purified pig gastric mucin as a standard.ResultsWhen applied luminally at 1 μM lubiprostone was ineffective at stimulating mucin release. When added to the serosal solution, 1 μM lubiprostone stimulated mucin release to ~300% of the unstimulated rate. As a positive control, serosal 1 μM prostaglandin E2 increased mucin release to ~400% of the unstimulated rate.ConclusionsThese results support the idea that lubiprostone has prostaglandin-like actions on the intestine, which includes stimulation of mucin release. Stimulation of mucin release by lubiprostone may be protective in gastrointestinal conditions where loss of mucus is believed to contribute to pathogenesis. Thus, in addition to chronic constipation, there is greater potential for the therapeutic applications of lubiprostone.

Salivary reservoir denture – A novel approach to battle xerostomia

Xerostomia is the subjective feeling of dryness in the mouth, often referred to as reduced salivary flow. Individuals usually detect they have a “dry mouth” when the flow of saliva decreases to approximately half the normal unstimulated rate of around 0.3 ml/min.1 Causes of xerostomia are anxiety, Sjogren's syndrome, salivary gland diseases, drug induced side effects, sequelae to head and neck radiation and general medical conditions such as diabetes mellitus. Patients suffering from xerostomia may complain of not only a dry mouth, but also of difficulty in normal oral and oropharyngeal functions including eating, speaking and swallowing. Extreme discomfort in wearing dentures is a common complaint.2,3 Depending upon the cause, a variety of treatment options are available. In cases where all treatment modalities have proven unsuccessful, the incorporation of reservoirs containing salivary substitutes into dentures has been proposed. This paper presents a case report where a reservoir denture was used successfully in rehabilitating a xerostomic complete edentulous patient.

A SLC4-like anion exchanger from renal tubules of the mosquito (Aedes aegypti): evidence for a novel role of stellate cells in diuretic fluid secretion

Transepithelial fluid secretion across the renal (Malpighian) tubule epithelium of the mosquito (Aedes aegypti) is energized by the vacuolar-type (V-type) H(+)-ATPase and not the Na(+)-K(+)-ATPase. Located at the apical membrane of principal cells, the V-type H(+)-ATPase translocates protons from the cytoplasm to the tubule lumen. Secreted protons are likely to derive from metabolic H(2)CO(3), which raises questions about the handling of HCO(3)(-) by principal cells. Accordingly, we tested the hypothesis that a Cl/HCO(3) anion exchanger (AE) related to the solute-linked carrier 4 (SLC4) superfamily mediates the extrusion of HCO(3)(-) across the basal membrane of principal cells. We began by cloning from Aedes Malpighian tubules a full-length cDNA encoding an SLC4-like AE, termed AeAE. When expressed heterologously in Xenopus oocytes, AeAE is both N- and O-glycosylated and mediates Na(+)-independent intracellular pH changes that are sensitive to extracellular Cl(-) concentration and to DIDS. In Aedes Malpighian tubules, AeAE is expressed as two distinct forms: one is O-glycosylated, and the other is N-glycosylated. Significantly, AeAE immunoreactivity localizes to the basal regions of stellate cells but not principal cells. Concentrations of DIDS that inhibit AeAE activity in Xenopus oocytes have no effects on the unstimulated rates of fluid secretion mediated by Malpighian tubules as measured by the Ramsay assay. However, in Malpighian tubules stimulated with kinin or calcitonin-like diuretic peptides, DIDS reduces the diuretic rates of fluid secretion to basal levels. In conclusion, Aedes Malpighian tubules express AeAE in the basal region of stellate cells, where this transporter may participate in producing diuretic rates of transepithelial fluid secretion.

Preganglionic parasympathectomy decreases salivary SIgA secretion rates from the rat submandibular gland

Immunoglobulin A (IgA) is transported into saliva by salivary cells expressing the polymeric immunoglobulin receptor (pIgR). In rat salivary glands, autonomic nerves stimulate this process. To examine how nerves affect pIgR-mediated IgA secretion, the chorda-lingual nerve was sectioned. One week after preganglionic parasympathectomy, both the stimulated and unstimulated rates of salivary IgA secretion were reduced, despite similar glandular amounts of IgA. Biochemical analysis of cells from parasympathectomised and control glands indicated reduced membrane expression of pIgR. It appears the removal of long-term parasympathetic input has affected the routing of pIgR within salivary cells and reduced the SIgA transport into saliva.

In Vitro Therapy with Dobutamine, Isoprenaline and Sodium Nitroprusside Protects Vascular Smooth Muscle Metabolism from Subarachnoid Haemorrhage Induced Cerebral Vasospasm

The cerebrospinal fluid (CSF) from subarachnoid haemorrhage (SAH) patients with cerebral vasospasm stimulates vasoconstriction and oxygen consumption in the porcine carotid artery in vitro. Stimulation of oxygen consumption has been used as an in vitro model of vasospasm to assess the relative benefits of nimodipine, isoprenaline, dobutamine, and sodium nitroprusside (SNP). Samples of human CSF were obtained from SAH patients and applied to de-endothelialised porcine carotid artery. Stimulation of oxygen consumption (as an in vitro marker for a stimulation of the vessels) was monitored and the effects of SNP, isoprenaline, dobutamine or nimodipine were measured. The CSF from SAH patients with evidence of vasospasm stimulated oxygen consumption to 0.91 +/- 0.17 (microM O2/min/g dry wt, +/- SD p < or = 0.01) and CSF from SAH patients without vasospasm did not significantly stimulate oxygen consumption 0.27 +/- 0.02, with 0.23 +/- 0.03 (microM O2/min/g dry wt) being an unstimulated rate of respiration for the porcine carotid artery. SNP, isoprenaline or dobutamine significantly (p < or = 0.01) decreased the stimulation of oxygen consumption of the porcine carotid artery whereas nimodipine did not. In a cohort of 41 SAH patients who received nimodipine alone or nimodipine and dobutamine, the in hospital mortality rate of the patients who received only nimodipine was 42% as compared to an in hospital mortality rate of 17% in the nimodipine plus dobutamine group P < or = 0.076). The in vivo data on the 41 patients is not statistically significant, so further studies are required to determine if the differences are important. SNP, isoprenaline and dobutamine significantly decreased oxygen consumption of the porcine carotid arteries exposed to CSF from SAH patients who had vasospasm whereas nimodipine did not. Our in vitro results suggest that these compounds require further study in patients with SAH who are at risk for vasospasm because they may have a direct benefit for the vasospastic arteries.

Analysis of residual saliva and minor salivary gland secretions

Residual saliva and minor salivary gland secretions are important for the maintenance of oral mucosal wetness. Salivary proteins and glycoproteins are the major components of the oral mucosal film, which functions as a moisture retainer and a protective barrier. Here, the correlations between the amounts of residual saliva and minor salivary gland secretions and their protein concentrations were investigated in 30 normal healthy individuals. The thickness of the mucosal film was measured at six mucosal surfaces and minor salivary gland secretion rate was measured at two mucosal surfaces. The thickness of residual saliva was determined by placing filter-paper strips against the mucosa at each site for 5 s and then measuring the volume electronically with a Periotron 8000 ® micro-moisture meter. The unstimulated rate of minor salivary gland secretion was measured for 30 s by the same method. Unstimulated whole salivary flow rate was measured with the spitting method. The total protein concentration of all salivary samples was measured by bicinchoninic acid assay. Before the experiment, the intra-/inter-examiner reliability of the method using the Periotron and the filter-paper strips was investigated. With a range of 0.4256–0.8846, the intraclass correlation coefficient, measured within and between examiners, was indicative of good reliability. The oral mucosal site with the thinnest coat of residual saliva was the anterior hard palate. Mucosal wetness on the hard palate and buccal mucosa showed significant positive correlations with the unstimulated whole salivary flow rate and significant negative correlations with the total protein concentration of residual saliva. Mucosal wetness on the upper and lower labial mucosa also showed significant negative correlations with the total protein concentration of residual saliva. Mucosal wetness on the soft palate was correlated with the minor salivary gland secretion rate ( r=0.477, P<0.01). Among the minor salivary glands, the secretion rate of soft palate glands in females showed a significant correlation with the unstimulated whole salivary flow rate ( r=0.563, P<0.05) and a significant negative correlation with its total protein concentration ( r=−0.525, P<0.05). These data suggested that oral mucosal wetness and minor salivary gland secretions could be influenced by various factors differently according to mucosal sites.

Sympathetic decentralization abolishes increased secretion of immunoglobulin A evoked by parasympathetic stimulation of rat submandibular glands

Salivary secretion of immunoglobulin A (IgA) in response to electrical stimulation of the parasympathetic nerve supply was assessed bilaterally in the submandibular glands of anaesthetized rats 1 week following unilateral pre-ganglionic sympathectomy (decentralization). Nerve-mediated stimulation on the non-denervated side increased IgA secretion several fold above an unstimulated rate of secretion whereas sympathetic decentralization reduced the parasympathetically stimulated secretion of IgA without affecting the basal rate. Glandular levels of IgA were increased following decentralization compared to the control glands. Salivary levels of free secretory component (FSC), the cleaved polymeric immunoglobulin receptor (plgR), were increased by parasympathetic stimulation and reduced by sympathectomy, though not as much as IgA. The decreased secretion of FSC suggests a reduced production of plgR and may account in part, for reduced IgA secretion following long-term removal of sympathetic nerve impulses.

Stimulation of fluid secretion of malpighian tubules of drosophila melanogaster meig. by cyclic nucleotides of inosine, cytidine, thymidine and uridine

External application of the 3',5'-cyclic monophosphates of inosine, cytidine, uridine and thymidine stimulated the fluid secretion rate (FSR) of Malpighian tubules isolated from Drosophila melanogaster. The evidence suggested that the cyclic nucleotides acted intracellularly in some capacity. Receptors of the 'purinergic' type appeared not to be major contributors to fluid secretion; of three purinergic agonists tried, adenosine, adenosine 5'-monophosphate (AMP) and adenosine 5'-triphosphate (ATP), only adenosine had an effect, but this was not observed consistently. None of the purinergic agonists interfered with the stimulation of the FSR by adenosine 3',5'-cyclic monophosphate (cAMP). The maximum stimulation of the fluid-secretion rate by any cyclic nucleotide was approximately double the unstimulated (control) rate. Tubules stimulated to less than maximal FSR by one cyclic nucleotide could be stimulated maximally by an appropriate concentration of another cyclic nucleotide. Malpighian tubules bathed in solutions that contained either [3H]cAMP or [3H]cGMP accumulated radioactivity to a level many times that in the medium. Accumulation of radioactivity by tubules bathed in 430 nmol l-1 [3H]cAMP was suppressed by 1 mmol l-1 non-radioactive cyclic nucleotides in the order cAMP>>cGMP>cIMP>cCMP; neither cTMP nor cUMP suppressed the accumulation of [3H]cAMP. Approximately 35 % of the [3H]cAMP and 80 % of the [3H]cGMP that entered the Malpighian tubule cells was metabolised to compounds that were not identified. It was concluded that cyclic nucleotides enter the Malpighian tubule cells by at least one transport mechanism which is particularly sensitive to purine-based nucleotides.

The influence of nerves on the secretion of immunoglobulin A into submandibular saliva in rats.

1. The influence of sympathetic and parasympathetic nerve stimulations on salivary secretion of immunoglobulin A (IgA) was studied in the submandibular glands of anaesthetized rats by stimulating the nerve supplies with bipolar electrodes. 2. Although the flow of saliva from sympathetically stimulated glands was only 23% of that from parasympathetically stimulated glands the output of IgA was over 2-fold greater. This difference was attributable to influences of the nerves on IgA secretion through the epithelial cell polymeric immunoglobulin receptor-mediated pathway, as Western blotting with specific antibodies to IgA and secretory component revealed that secretory IgA (SIgA) dominated in all saliva samples. 3. Study of saliva secreted in sequential periods of nerve stimulation or following rest pauses suggested that SIgA secretion occurred in the absence of stimulation but this was upregulated 2.6- and 6-fold by parasympathetic and sympathetic nerve stimulations, respectively, compared with the calculated unstimulated rate. 4. The IgA content of extensively stimulated glands was 77% of levels in unstimulated contralateral control glands despite a secretion into saliva equivalent to almost 90% of the glandular IgA content. The IgA may be synthesized and secreted by glandular plasma cells at a rate which exceeds demand and/or such synthesis may be upregulated by nerve impulses. 5. The results indicate that salivary secretion of SIgA is upregulated by nerve impulses and that sympathetic nerves induce a greater effect than parasympathetic nerves.

Gender Difference in the Basal Intracellular Ca2+Concentration in Rat Valvular Endothelial Cells

Intracellular Ca2+concentration ([Ca2+]i) was measured by Fura 2/AM fluorescence imaging microscopy in freshly isolated valvular endothelial cells taken from female and male rats. The basal level of [Ca2+]iwas significantly elevated in female valvular endothelial cells when compared to males (P< 0.05). Inhibition of the sarco-endoplasmic reticulum Ca2+-ATPase with cyclopiazonic acid (CPA, 10 μM) caused a greater increase in the [Ca2+]iin female than male endothelial cells. Removal of extracellular Ca2+returned the [Ca2+]ito the basal level. The rate of [Ca2+]idecline was significantly slower in female endothelial cells compared to males. There were no differences in the unstimulated rate of Mn2+quenching between two groups. These results demonstrate that estrogen affects NOS at least in part, by an alteration in Ca2+homeostasis in endothelial cells.

Destabilization of Peptide Binding and Interdomain Communication by an E543K Mutation in the Bovine 70-kDa Heat Shock Cognate Protein, a Molecular Chaperone

We have compared 70-kDa heat shock cognate protein (Hsc70) isolated from bovine brain with recombinant wild type protein and mutant E543K protein (previously studied as wild type in our laboratory). Wild type bovine and recombinant protein differ by posttranslational modification of lysine 561 but interact similarly with a short peptide (fluorescein-labeled FYQLALT) and with denatured staphylococcal nuclease-(Delta135-149). Mutation E543K results in 4. 5-fold faster release of peptide and lower stability of complexes with staphylococcal nuclease-(Delta135-149). ATP hydrolysis rates of the wild type proteins are enhanced 6-10-fold by the addition of peptide. The E543K mutant has a peptide-stimulated hydrolytic rate similar to that of wild type protein but a higher unstimulated rate, yielding a mere 2-fold enhancement. All three versions of Hsc70 possess similar ATP-dependent conformational shifts, and all show potassium ion dependence. These data support the following model: (i) in the presence of K+, Mg2+, and ATP, the peptide binding domain inhibits the ATPase; (ii) binding of peptide relieves this inhibition; and (iii) the E543K mutation significantly attenuates the inhibition by the peptide binding domain and destabilizes Hsc70-peptide complexes.

Impairment by interleukin 1 beta and tumour necrosis factor alpha of the glucagon-induced increase in phosphoenolpyruvate carboxykinase gene expression and gluconeogenesis in cultured rat hepatocytes.

The influence of the inflammatory mediators interleukin 1 beta (IL1 beta) and tumour necrosis factor alpha (TNF alpha) on the glucagon-induced expression of phosphoenolpyruvate carboxykinase (PCK) and on glucose formation via gluconeogenesis was investigated in cultured rat hepatocytes. Gene expression was monitored by determination of mRNA levels and of enzyme activity. Glucose formation was estimated with newly synthesized radioactive glucose derived from a radiolabelled lactate precursor. Glucagon (0.1 or 1 nM) induced PCK mRNA transiently to a maximum 2 h after its application. In the presence of recombinant human (rh) IL1 beta or rhTNF alpha the increase in PCK mRNA levels was totally inhibited at 0.1 nM glucagon, whereas at 1 nM glucagon the maximal increase was inhibited by only 25%. Glucagon (0.1 or 1 nM) induced PCK activity to a maximum after 4 h (4-fold and 6-fold over prestimulatory activity respectively). In the presence of rhIL1 beta or rhTNF alpha the maximal increase was inhibited by approx. 50%. Addition of rhIL1 beta or rhTNF alpha 2 h after glucagon, at the maximal glucagon-induced PCK mRNA levels, accelerated the decay of PCK mRNA. Glucagon (1 or 10 nM) [corrected] increased glucose formation from lactate by 1.3-fold and 1.7-fold respectively over unstimulated rates. In the presence of rhIL1 beta or rhTNF alpha this increase in glucose formation was inhibited by 60-90%. At 0.1 nM, glucagon doubled the intracellular cAMP concentration. This increase was prevented by rhIL1 beta or rhTNF alpha. At 1 nM, glucagon increased cAMP concentrations by 10-fold. In the presence of rhIL1 beta or rhTNF alpha this increase was inhibited by 70%. From the results it is suggested that rhIL1 beta and rhTNF alpha prevented glucagon-stimulated PCK gene expression and gluconeogenesis at least in part by inhibition of the glucagon-stimulated increase in cAMP concentrations.

Constitutive steroidogenesis does not require the actions of cAMP on cholesteryl ester hydrolysis or internalization of plasma membrane cholesterol.

The R2C Leydig tumor cells demonstrate constitutive steroidogenesis as well as the constitutive production of the steroidogenic acute regulatory peptide (StAR). Since the introduction of StAR into cells does not by itself reproduce the steroidogenic response of cAMP, the present studies were designed to determine whether processes normally stimulated by cAMP in responsive cells were maximally, constitutively expressed in the R2C cells. Measurements of cholesteryl ester hydrolytic rate and plasma membrane cholesterol internalization revealed that both processes are stimulated by dibutyryl-cAMP in R2C cells. The effect of cAMP on these processes was of the same magnitude as that detected in hormonally responsive cells. Thus, in R2C cells the unstimulated basal rate of either process was sufficient to maintain maximal stimulation of steroidogenesis.

IFN-gamma up-regulates expression of the complement components C3 and C4 by stabilization of mRNA.

We investigated the mechanisms underlying the regulation of complement genes C3 and C4 by IFN-gamma. IFN-gamma (500 U/ml, 24 h incubation) increased steady state mRNA levels for both C3 and C4 in three different cell types (Hep G2, U937, and primary fibroblasts). The response to IFN-gamma in Hep G2 cells was time and dose dependent. At all doses of IFN-gamma and at all incubation times, the transcription rate for these two genes, determined by nuclear run-on assays, was reduced (0.3 +/- 0.1; unstimulated rate = 1.00). The t1/2 of mRNA for C3 and C4 in unstimulated cells was 1.8 +/- 0.3 and 2.2 +/- 0.2 h, respectively. After high-dose IFN-gamma stimulation, both C3 and C4 mRNA levels remained at 100% with respect to baseline at 5 h, but after 12 h, levels fell to 13 +/- 2% (C3) and 8 +/- 3% (C4) of baseline values, giving a half-life for these mRNA species of between 5 and 12 h. IFN-gamma stimulation increased C3 and C4 protein synthesis measured at 24 h. We suggest that it is the increase in mRNA stability that is the major effector mechanism by which IFN-gamma regulates C3 and C4 gene expression.

Substrate-dependent alteration in O2 consumption and energy metabolism in vascular smooth muscle.

Energy metabolism and the substrate utilization pattern of intact porcine carotid artery were investigated in the presence or absence of glucose and/or octanoate during the phases of isometric contraction induced by K+ depolarization. During the early phase of contraction, there was a rapid increase in the rate of O2 uptake that was independent of the rate of force generation but dependent on the availability of intracellular pyruvate, the source of which was glucose and not glycogen. Lactate production increased linearly from the onset of contractile stimulation and was not suppressed by octanoate oxidation. There was no alteration from the basal resting state in the concentrations of the metabolites of the tricarboxylic acid cycle in the presence or absence of octanoate. During the phase of steady-state force maintenance, O2 consumption was increased compared with the basal unstimulated rate but was not increased when glucose and octanoate were present, which is consistent with the Crabtree effect. This was associated with increased aerobic lactic acid production and inhibition of the tricarboxylic acid cycle at the citrate synthase step. Alteration of the high-energy phosphate content could not account for the pattern of O2 consumption during contraction under different substrate conditions. In the absence of glucose, the energy from octanoate oxidation could substitute for the energy ordinarily derived from aerobic glycogen and lactic acid production. It is concluded that energy metabolism of vascular smooth muscle is coordinated during contraction by integration of the pathways of aerobic glycolysis and oxidative phosphorylation.

Factors influencing mercury evaporation rate from dental amalgam fillings.

Factors influencing mercury evaporation from dental amalgam fillings were studied in 11 volunteers. Air was drawn from the oral cavity for 1 min and continuously analyzed with a mercury detector. In six volunteers the median unstimulated evaporation rate was 0.1 ng Hg/s, range 0.09-1.3 ng Hg/s. After chewing gum for 5 min the highest evaporation rate was 2.7 ng Hg/s. Chewing paraffin wax gave only a small increase in evaporation rate. Changes in airflow rates between 1.5 and 2.5 1/min during the 1 min sampling did not change the amount of mercury drawn from the oral cavity. Sampling with different mouthpieces and closed mouth was compared to open mouth sampling with a thin plastic tube. It was found that the latter method could result in lower values for some volunteers due to simultaneous mouth breathing. After placing individual plastic teeth covers in the mouth, the intraoral evaporation of mercury decreased immediately by 89-100% of previous levels. This technique could be used to detect mercury evaporation from separate amalgam fillings or to reduce the intraoral mercury vapor concentration. Rinsing the mouth with heated water for 1 min increased the mean evaporation rate by a factor of 1.7 when the water temperature increased from 35 degrees C to 45 degrees C.

Functional and morphological changes associated with the ageing of primary cultures of embryonic adrenal gland cells derived from the Pekin duck.

The morphological and functional changes associated with ageing were studied in adrenal steroidogenic cells derived from duck embryos. Cells grown for not more than three days had structural characteristics similar to their counterparts in vivo; they contained numerous lipid droplets and mitochondria, an abundant smooth endoplasmic reticulum, an even network of microtubules, and microfilaments that formed extensive and elaborate systems of parallel stress fibers. After the 3rd day of growth in culture, many of the cells started to decrease in size and become elongated; the older cells showed less well-defined actin filaments and contained elongated mitochondria, fewer lipid droplets, less smooth endoplasmic reticulum, and swollen cisternae of rough endoplasmic reticulum. The proliferative capacity of the cells was the same when they were cultured in either the presence or the absence of 1-24 ACTH. After the first day of growth in culture, the steroidogenic capacity of the cells declined and the addition of 1-24 ACTH to the growth medium did not prevent changes in their structure and function. The decline in steroidogenic capacity occurred both in terms of the amount of hormone released into the culture medium and in the ability of the cells to respond when incubated in buffer containing 1-24 ACTH. Since the basal unstimulated rates of corticosteroid production also declined as the cells aged, it is probable that the steroidogenic deficiency occurs at a site distal to the corticotropin receptor; this is also consistent with the ultrastructural observations that suggest a relationship between the morphological changes and the decline in steroidogenic capacity as the cells age.

Cellular Aspects of the Stress Response in Precocial Neonates

The extent to which control over adrenal steroidogenic function is established during embryogenesis, and therefore the ability of precocial neonates to withstand stress, is probably an important determinant of breeding success. Studies on the ontogeny of adrenal steroidogenic function in birds, however, have been limited to the chicken Gallus domesticus and the duckling Anas platyrhynchos. Up to the 21st day of embryogenesis, the temporal patterns of adrenal gland development in these species are similar. By the 15th day the adrenal anlage is encapsulated and consists of islands of chromaffin cells distributed throughout a mass of structurally typical steroidogenic cells; at this time the steroidogenic cells are responsive to 1-24 ACTH and they contain small quantities of corticosterone and aldosterone. During the next six days the corticotropic responsiveness increases although the inducible response of cells from the 21 day-old emerging chick embryo is less than that of cells from the 21 day-old duck embryo. In the duckling, corticotropic responsiveness increases further during the next week preceding emergence. Also the steroidogenic cells in the emerging duck embryo, but not those in the emerging chicken embryo, preferentially synthesize aldosterone in response to angiotensin II. Although catecholamines and acetylcholine each suppress basal unstimulated rates of corticosterone and aldosterone release from the steroidogenic tissue, none has an effect on 1-24 ACTHinducible corticosteroid release.