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Stimulation of insulin release by thiols.

The effects of thiol compounds on insulin release were studied in microdissected pancreatic islets of non-inbred ob/ob micemin control experiments the reactivity of thiols against 6,6'-dithiodinicotinic acid and the degradation of mouse insulin were measured. At a concentration of 0.1 mM, 1-thio-D-glucose or reduced glutathione potentiated the insulin-releasing action of 10 mM D-glucose without affecting glucose oxidation. When tested at a concentration equivalent to about 0.1 mM reactive thiol, dextran-linked L-cysteine also potentiated the glucose-induced insulin secretion. In microperifusion experiments the insulin-releasing action of 1-thio-D-glucose was found to exhibit a rapid onset followed by a decline of the secretory rate to values lower than those observed with 10 mMD-glucose alone. No thiol stimulated insulin release in the absence of glucose. It is suggested that thiol compounds stimulate insulin release by splitting membrane disulphides in the beta-cells.

Animals↗

Stressful life events and affective disorders inhibit pulsatile LH secretion in hypothalamic amenorrhea.

The aim of this study was to evaluate relationships between emotional state and hypothalamic activity in patients with hypothalamic secondary amenorrhea. Sixty-seven normal weight patients with hypothalamic amenorrhea were submitted to concomitant psychological and LH pulsatility evaluation. Structured clinical interview for anxiety and depressive disorders (DSM III-R) as well as life events investigations (Paykel test) were performed. LH pulses (blood sampling every 10 min for 4 hr) were analyzed through DETECT program and Instantaneous Secretory Rate were computed. Twenty-one patients reporting life events associated to the onset of amenorrhea had LH pulse frequency (2.28 +/- 1.10 pulses/4 hr) lower than those without life events (3.40 +/- 1.46 p = .007). LH pulses amplitude was lower in patients meeting a DSM III-R (21 cases: 1.22 +/- 0.96 mIU/ml) diagnosis than in those without (1.99 +/- 1.20 p = .04) diagnosis. Plasma estradiol and FSH levels as well as duration of amenorrhea and Body Mass Index were similar among groups. It is concluded that psychogenic factors (namely the presence of life events related to the onset of menstrual disorder) are associated with significant and specific changes of hypothalamic activity which could be involved in determining hypogonadism.

Adult↗

Gastric emptying and secretion in monkeys during unavoidable shock.

Gastric fluid and acid secretory rates as well as concurrent fractional emptying rates were determined in monkeys subjected to repeated sessions of unavoidable shock. None of these variables changed from control levels during the first session but, by the last session, acid output was significantly reduced.

Animals↗

Effect of acetaminophen on expression and activity of rat liver multidrug resistance-associated protein 2 and P-glycoprotein.

We evaluated the effect of acetaminophen (APAP), given as a single, 1g/kg body weight dose, on expression and activity of rat liver multidrug resistance-associated protein 2 (Mrp2) and P-glycoprotein (P-gp), two major canalicular drug transporters. The studies were performed 24h after administration of the drug. APAP induced an increase in plasma membrane content of Mrp2 detected by western blotting, consistent with increased detection of the protein at the canalicular level by immunoflourescence microscopy. In vivo biliary excretion of dinitrophenyl-S-glutathione, a well known Mrp2 substrate, was slightly but significantly increased by APAP, agreeing well with upregulation of the transporter. Basal biliary excretion of oxidized glutathione, an endogenous Mrp2 substrate, was also increased by APAP, likely indicating increased hepatic synthesis as a result of APAP-induced oxidative stress followed by accelerated canalicular secretion mediated by Mrp2. APAP also increased the expression of P-gp detected by western blotting and immunofluorescence microscopy as well as the in vivo biliary secretory rate of digoxin, a model P-gp substrate. Because specific APAP-conjugated metabolites are Mrp2 substrates, we postulate that induction of Mrp2 by APAP may represent an adaptive mechanism to accelerate liver disposition of the drug. In addition, increased Mrp2-mediated elimination of oxidized glutathione may be essential in maintaining the redox equilibrium in the hepatocyte under conditions of APAP-induced oxidative stress.

ATP Binding Cassette Transporter, Subfamily B↗

Hepatic overexpression of caveolins increases bile salt secretion in mice.

Caveolins are cholesterol-binding proteins involved in the regulation of several intracellular processes, including cholesterol transport. Because hepatocytes express caveolin-1 and caveolin-2, these proteins might modulate hepatic lipid metabolism and biliary lipid secretion. Our aim was to investigate the potential physiologic role of caveolins in hepatic cholesterol and bile salt (BS) metabolism and transport using adenoviral gene transfer. C57BL/6 mice were infected with recombinant human caveolin-1 and caveolin-2 adenoviruses. Mice infected with adenovirus lacking the transgene were used as controls. Hepatic caveolin expression was evaluated by immunochemical methods. Reverse-transcription polymerase chain reaction (RT-PCR) and immunoblotting were used to assess messenger RNA (mRNA) levels and protein mass of BS transporters (sodium taurocholate cotransporting polypeptide [Ntcp] and bile salt export pump [Bsep]). Serum, liver, biliary, and fecal biochemical determinations and BS maximal secretory rate (SRm) were performed by standard methods. Ad.Cav-1- and Ad.Cav-2-infected mice exhibited a 10- and 7-fold increase in hepatic caveolin-1 and caveolin-2 protein expression, respectively. Caveolin-1-overexpressing mice had a significant increase in plasma high-density lipoprotein (HDL) cholesterol and hepatic free cholesterol content, whereas total plasma cholesterol and triglyceride levels remained unchanged. Hepatic caveolin-1 and/or caveolin-2 overexpression significantly increased bile flow and secretion of all biliary lipids. Caveolin-1-overexpressing mice showed a 2.5-fold increase in taurocholate (TC) SRm, indicating increased canalicular BS transport capacity. BS pool size and fecal BS excretion remained within the normal range in mice with Cav-1 overexpression. No changes were seen in the protein mass of BS transporters Ntcp and Bsep. In conclusion, our findings indicate that caveolins may play an important role in regulating hepatic BS and cholesterol metabolism.

Adenoviridae↗

The phorbol ester induced atrial natriuretic peptide secretion is stimulated by forskolin and Bay K8644 and inhibited by 8-bromo-cyclic GMP.

The role of intracellular signals in the regulation of atrial natriuretic peptide (ANP) release was studied using the isolated perfused rat heart. The phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), known to activate the protein kinase C pathway, produced a dose-dependent increase in perfusate ANP immunoreactivity. Bay k8644, a putative calcium channel activator, and forskolin, which stimulates adenylate cyclase, induced a sustained increase in ANP secretory rate. TPA in combination with either Bay k8644 or forskolin induced higher ANP secretion than the calculated additive value for each agent. 8-bromo-cyclic GMP and sodium nitroprusside, when given alone, had no effect on ANP secretion, but delayed the TPA-stimulated increase in perfusate ANP. ANP secretion appears therefore to be mediated both by the phosphoinositide and the cAMP system, whereas the cGMP pathway may be inhibitory.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Atrial natriuretic peptide gene expression and its secretion by pneumocytes derived from neonatal rat lungs.

Using a primary culture of pneumocytes derived from neonatal rat lungs, we investigated the synthesis and secretion at transcriptional and peptide levels of pulmonary rat(r) atrial natriuretic peptide (ANP). Total RNA extracted from pneumocytes contained a hybridizing RNA band of the same size as atrial rANP mRNA. Immunoreactive (IR)-rANP content in pneumocytes was 0.5% of that in atrial myocytes, and 8.6% of that in ventricular myocytes, while the secretory rate from pneumocytes was about 7% of atrial and ventricular myocytes. Triiodothyronine (T3, 5 x 10(-10) to 5 x 10(-8) M), dexamethasone and testosterone (5 x 10(-9) to 5 x 10(-8) M) significantly stimulated the synthesis of IR-rANP by pneumocytes in a dose-dependent manner. However, the stimulatory effect exerted by T3 on rANP synthesis, unlike in the case of cardiocytes, was much more potent than that of dexamethasone, as evidenced by the significant difference in potency at both transcriptional and peptide levels. The present study suggests that ANP secreted from lungs may at least in part contribute to circulating ANP pool, and that the tissue-dependent difference of sensitivity to thyroid hormone may play an important role in the regulation of developmental ANP gene expression in mammalian lungs.

Animals↗

Brain natriuretic peptide is a novel cardiac hormone.

Using a radioimmunoassay for brain natriuretic peptide (BNP), we have measured levels of BNP-like immunoreactivity (-LI) in extract of the porcine heart, in perfusate from the isolated porcine heart and in porcine plasma. BNP-LI was detected in the extract of the atrium, though no detectable amount of BNP-LI (more than 1 ng/g) was present in the ventricle. The BNP-LI level in the porcine atrium was 148.7 +/- 23.3 ng/g. BNP-LI was also detected in the perfusate from the heart. Basal secretory rate of BNP was 3.18 +/- 0.76 ng/min. Moreover, BNP-LI was detected in porcine plasma at the concentration of 4.2 +/- 1.3 pg/ml. Gel filtration studies showed that BNP is present in the atrium as a large molecule and is secreted into the circulation as a small molecule. The percentage of BNP-LI to atrial natriuretic peptide (ANP)-LI was almost the same among the extract, the perfusate and the plasma (2-3 percent). These results indicate that BNP is synthesized in and is secreted into the circulation from the heat in a similar fashion as ANP.

Animals↗

Effect of silymarin on biliary bile salt secretion in the rat.

The effect of the hepatoprotector silymarin on bile secretion, with particular regard to bile salt secretion, was studied in Wistar rats. Silymarin (25, 50, 100, and 150 mg/kg/day, i.p., for 5 days) induced a dose-dependent increase in bile flow and bile salt secretion, the maximal effect being reached at a dose of 100 mg/kg/day (+17 and +49%, for bile flow and bile salt output, respectively; P < 0.05). Assessment of bile salt composition in bile revealed that stimulation of the bile salt secretion was accounted for mainly by an increase in the biliary secretion of beta-muricholate and, to a lesser extent, of alpha-muricholate, chenodeoxycholate, ursodeoxycholate, and deoxycholate. The maximum secretory rate (T(m)) of bile salts, as assessed by infusing the non-hepatotoxic bile salt tauroursodeoxycholate i.v. at stepwise-increasing rates, was not influenced by silymarin. The flavonolignan also increased the endogenous bile salt pool size (+53%, P < 0.05) and biliary bile acid excretion after bile acid pool depletion (+54%, P < 0.05), a measure of de novo bile salt synthesis. These results suggest that silymarin increases the biliary excretion and the endogenous pool of bile salts by stimulating the synthesis, among others, of hepatoprotective bile salts, such as beta-muricholate and ursodeoxycholate.

Analysis of Variance↗

Kappa opiate agonist RU 51599 inhibits vasopressin gene expression and osmotically-induced vasopressin secretion in vitro.

Kappa (kappa) opioid agonists induce a water diuresis and inhibit vasopressin (AVP) secretion. Hypothalamic and neurohypophysial sites have both been implicated in the response. The present study was designed to ascertain if kappa-agonist inhibition of osmotically-stimulated AVP secretion is associated with parallel changes in AVP gene expression. Experiments were performed using the selective kappa-agonist RU 51599 (RU) in compartmentalized hypothalamo-neurohypophysial explants. When added to either the hypothalamus or the neural lobe, RU dose dependently inhibited osmotically-induced AVP secretion that was reversed by the highly selective kappa-antagonist nor-binaltorphimine (nor-BNI) only at the hypothalamic, not the neurohypophysial level. AVP mRNA content paralleled the changes in AVP secretory rate induced by hypothalamic kappa-agonism. AVP mRNA levels were unaltered when RU was applied to the neural lobe. Neurohypophysial AVP content did not change. These data indicate that hypothalamic kappa-agonism inhibits osmotically induced AVP secretion and that a non-kappa1 opiate receptor mediates posterior pituitary opioid inhibition of AVP release. Neural or receptor inputs to the hypothalamus or magnocellular cell body may downwardly modulate AVP mRNA content by altering AVP gene transcription and/or message stability. Inhibition of AVP release directly at the neurohypophysis can be uncoupled from the cellular mechanisms that generate changes in AVP mRNA content.

Animals↗

Analysis by mass spectrometry of POMC-derived peptides in amphibian melanotrope subpopulations.

We have previously shown that the melanotrope population of the pituitary intermediate lobe of Rana ridibunda is composed of two subpopulations, of low (LD) and high density (HD), that show distinct ultrastructural features and display different synthetic and secretory rates. To investigate whether LD and HD melanotrope cells also differ in proopiomelanocortin (POMC) processing, we have analyzed the POMC-end products in single cells from both subpopulations by means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The mass spectra revealed the presence of 8 POMC-derived peptides in HD and LD melanotrope cells, indicating a similar processing of the precursor in both subpopulations. However, the relative abundance of three POMC-end products (i.e. lys-gamma1-MSH, acetyl-alpha-MSH, and CLIP fragment) was higher in the HD subset. Moreover, two peptides with molecular weights of 1030 and 1818 Da, respectively, were detected that could not be assigned to any product deduced from the frog POMC sequence. The relative amount of the 1030 Da peptide was higher in LD melanotrope cells. Taken together, our results suggest that POMC processing is differentially regulated in the two melanotrope cell subsets.

Animals↗

Nitric oxide modulates salivary amylase and fluid, but not epidermal growth factor secretion in conscious rats.

The involvement of the L-arginine/NO pathway in the control of salivary fluid, amylase and epidermal growth factor (EGF) secretion was investigated in conscious rats. For the collection of saliva, an oesophageal cannula was implanted. To obtain steady secretion, submaximal carbachol background infusion was given. Different treatments included NO synthase inhibitor N(G)-nitro-L-arginine (NOLA; with or without phentolamine, propranolol), L-arginine, D-arginine and NO donor 3-morpholinosydnonimine (SIN-1) administration. Volume, amylase activity and EGF output in the secreted fluid were determined in 30 min mixed saliva samples. Carbachol infusion alone produced a modest, sustained salivary fluid and amylase secretion. NOLA (30 mg/kg) further increased both fluid (p<0.001) and amylase outputs (p<0.001). These latter effects were prevented by L-arginine but not by D-arginine or by phentolamine. Propranolol administration decreased both fluid and amylase secretion below the carbachol plateau, and NOLA did not modify this suppressed secretory rate. SIN-1 did not alter either volume or amylase secretion. Interestingly, NOLA given without carbachol did not modify salivation. Neither carbachol nor NOLA changed salivary EGF output. The present results suggest that the L-arginine/NO pathway has a modulatory role in the cholinergic control of salivary amylase secretion, but not in EGF output. The mechanisms of inhibitory action of NO on salivary fluid and amylase secretion remain to be identified.

Amylases↗

Making sense of refractive surgery in 2001: why, when, for whom, and by whom?

Surgical alteration of the focusing or refractive properties of the eye has been performed on millions of patients. An array of procedures to correct myopia, hyperopia, astigmatism, and presbyopia have been introduced over the past 25 years with varying degrees of success. Improved technology has increased patient and physician satisfaction and enthusiasm. Currently available surgical procedures can be categorized as incisional, surface-altering, lamellar, and intraocular. The choice of procedure depends on individual patient indications and contraindications based on results of ocular examinations, eg, corneal pachymetry to measure corneal thickness, keratometry to measure the corneal curvature, basal tear secretory rate, and dark-adapted pupil size. The postoperative uncorrected visual acuity depends, in large part, on the quality of the preoperative evaluation and refraction. Before scheduling a patient for surgery, the ophthalmologist must ensure that the patient understands the potential risks of the procedure and has realistic expectations for the postoperative level and quality of uncorrected visual acuity. Postoperative complications include corneal flap displacement, undercorrection and overcorrection, and epithelial ingrowth under the corneal flap and inflammatory keratitis. Postoperative dry eye, infection, and inflammation are usually treated medically. Ongoing technological innovations to customize the surgical approach to an individual patient's eye continue to improve outcomes.

Astigmatism↗

Hepatic portal vein cannulation for infusion and blood sampling in freely moving rats.

Chronic portal vein cannulation in the rat is an important technique to study secretory rates of hormones from the endocrine pancreas. Moreover, it can be used for studying the effects of enteric hormones and pharmaca on behavioral and physiological processes. This article contains an extensive description of a cannulation technique of the portal vein that has many advantages over those reported so far in the literature, and that was very successful in several behavioral and physiological studies during the last decade.

Animals↗

Effect of kappa opioid agonist RU 51599 on osmotic and non-osmotic stimulated arginine vasopressin release and gene regulation in small cell lung carcinoma cells.

Arginine vasopressin (AVP) is synthesized in the hypothalamus, stored in the posterior pituitary, and osmotic and non-osmotic stimuli release AVP into the circulation for antidiuretic and vascular actions on target tissue. The kappa-opioid agonist, RU 51599, exhibits a potent diuretic activity in both experimental animals and humans. This diuretic activity is characterized by a water diuresis without an associated increase in electrolyte excretion. Studies with cultured rat hypothalamo-neurohypophysial system explant showed that AVP mRNA level changed in parallel to the RU 51599-induced changes in AVP secretory rate. There are, however, no hypothalamic neuronal cell lines to study AVP gene regulation system, and it is not known whether RU 51599, regulates AVP secretion and biosynthesis under osmotic and non-osmotic stimulatory conditions of AVP release. The effect of RU 51599 on AVP release, AVP mRNA, and AVP gene promoter activity in osmotic and non-osmotic conditions was therefore studied using cultured small cell lung carcinoma (SCLC) cell lines. RU 51599 significantly inhibited AVP release by osmotic stimulation (330 mOsm) and non-osmotic stimulators, angiotensin II (AII) and endothelin 3 (ET3). However, RU 51599 did not show any effect on the AVP mRNA and AVP gene promoter activity stimulated by high osmolality and ET3. These results indicate, therefore, that RU 51599 suppresses AVP secretion by inhibition at the step of AVP release during osmotic and non-osmotic stimulation but does not affect the AVP gene transcription level in the SCLC cells.

Angiotensin II↗

Conjunctival cysts of the orbit following enucleation.

Four patients developed orbital conjunctival cysts following enucleation. The cysts were demonstrated by ultrasonography and computerized tomography and confirmed by histopathology. They were lined by a non-keratinizing stratified squamous epithelium without goblet cells and contained fluid with mucinous strands. They were excised at eight, seven, and seven, and 25 years post-enucleation, respectively, the latter being the longest interval yet recorded. A simple classification of conjunctival cysts of the orbit is proposed and the mechanisms of the development of cysts in anophthalmic sockets are discussed. The effects of secretory rate and cyst growth are reviewed and surgical management in prevention and treatment of these lesions is outlined.

Adult↗

Adaptive regulation of hepatic bile salt transport. Effect of prolonged bile salt depletion in the rat.

Exposure of the liver to increased bile salt flux can increase the bile salt maximum secretory rate (SRm), presumably through the induction of new transport sites. The converse, i.e., the down-regulation of SRm upon bile salt deprivation, has not been demonstrated. We examined the effects of bile salt depletion for 24 h and 48 h on taurocholate SRm and bromsulphalein (BSP) SRm, and on [14C]taurocholate binding to isolated liver surface membranes in unrestrained external biliary fistula rats. Taurocholate SRm was significantly decreased by 35% and 51% in 24-h-depleted and 48-h-depleted rats, respectively, compared with control, sham-operated rats. Maximal taurocholate concentration in bile was also significantly lower in bile salt-deprived rats. In contrast, BSP SRm was not significantly different between depleted animals and controls. Bile salt depletion for 24 h and 48 h did not significantly alter liver surface membrane protein recovery and membrane enzyme specific activity, including Na+ + K+-ATPase. Specific [14C]taurocholate binding to liver surface membranes was significantly decreased by 25% in 24-h-depleted rats compared with control rats. In contrast to taurocholate SRm, bile salt depletion for 48 h did not result in further reduction of specific taurocholate binding sites. This study demonstrates that taurocholate SRm progressively decreased in 24-h- and 48-h-bile salt-depleted rats, this being consistent with adaptive down-regulation of hepatic bile salt transport.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of L-arginine/nitric oxide pathway on salivary amylase secretion in conscious rats.

In a recent study we have demonstrated the presence of nitric oxide synthase immunoreactive neurons and also perivascular, periacinar and periductal nerve fibres in feline submandibular salivary gland. The role of nitric oxide (NO) in salivary vasoregulation has been suggested by other data too, but the effect of NO on salivary amylase secretion has not been investigated yet. Under ether anaesthesia a catheter was introduced into the oesophagus for salivary juice collections, and a cannula was inserted into the jugular vein for infusions. After postanaesthesia recovery, submaximal carbachol infusion was given as a background to obtain steady secretion because of the low basal secretory rate. Then different groups of animals received NO synthase inhibitor NOLA (NG-nitro-L-arginine), L-arginine, D-arginine or NO donor SIN-1 (3-morpholinosydnonimine). Volume and amylase activity were determined in mixed saliva samples collected for 30 min. Carbachol background infusion alone induced an elevated, sustained salivary secretion. NOLA (30 mg/kg) increased both volume and amylase output (P < 0.001). This effect was prevented by L-arginine but not by D-arginine. SIN-1 did not change either volume or amylase secretion. The present results suggest that the L-arginine/NO pathway has a modulatory effect on salivary fluid and amylase secretion, which is probably not related to its effect on salivary blood flow. NO may block certain presently unidentified secretagogue mechanisms and/or may relax myoepithelial cells.

Amylases↗