Role of pituitary adenylyl cyclase-activating polypeptide in intracellular calcium dynamics of neurons and satellite cells in rat superior cervical ganglia.

Pituitary adenylyl cyclase-activating polypeptide (PACAP) is a bioactive peptide with diverse effects in the nervous system. The present study investigated whether stimulation of PACAP receptors (PACAPRs) induces responses in neurons and satellite cells of the superior cervical ganglia (SCG), with special reference to intracellular Ca2+ ([Ca2+]i) changes. The expression of PACAPRs in SCG was detected by reverse transcription-PCR. PACAP type 1 receptor (PAC1R), vasoactive intestinal peptide receptor type (VPAC)1R, and VPAC2R transcripts were expressed in SCG, with PAC1R showing the highest levels. Confocal microscopy analysis revealed that PACAP38 and PACAP27 induced an increase in [Ca2+]i in SCG, first in satellite cells and subsequently in neurons. Neither extracellular Ca2+ removal nor Ca2+ channel blockade affected the PACAP38-induced increase in [Ca2+]i in satellite cells; however, this was partly inhibited in neurons. U73122 or xestospongin C treatment completely and partly abrogated [Ca2+]i changes in satellite cells and in neurons, respectively, whereas VPAC1R and VPAC2R agonists increased [Ca2+]i in satellite cells only. This is the first report demonstrating the expression of PACAPRs specifically, VPAC1 and VPAC2 in SCG and providing evidence for PACAP38-induced [Ca2+]i changes in both satellite cells and neurons via Ca2+ mobilization.

Gut Microbiota Promotes Obesity-Associated Liver Cancer through PGE2-Mediated Suppression of Antitumor Immunity.

Obesity increases the risk of cancers, including hepatocellular carcinomas (HCC). However, the precise molecular mechanisms through which obesity promotes HCC development are still unclear. Recent studies have shown that gut microbiota may influence liver diseases by transferring its metabolites and components. Here, we show that the hepatic translocation of obesity-induced lipoteichoic acid (LTA), a Gram-positive gut microbial component, promotes HCC development by creating a tumor-promoting microenvironment. LTA enhances the senescence-associated secretory phenotype (SASP) of hepatic stellate cells (HSC) collaboratively with an obesity-induced gut microbial metabolite, deoxycholic acid, to upregulate the expression of SASP factors and COX2 through Toll-like receptor 2. Interestingly, COX2-mediated prostaglandin E2 (PGE2) production suppresses the antitumor immunity through a PTGER4 receptor, thereby contributing to HCC progression. Moreover, COX2 overexpression and excess PGE2 production were detected in HSCs in human HCCs with noncirrhotic, nonalcoholic steatohepatitis (NASH), indicating that a similar mechanism could function in humans.Significance: We showed the importance of the gut-liver axis in obesity-associated HCC. The gut microbiota-driven COX2 pathway produced the lipid mediator PGE2 in senescent HSCs in the tumor microenvironment, which plays a pivotal role in suppressing antitumor immunity, suggesting that PGE2 and its receptor may be novel therapeutic targets for noncirrhotic NASH-associated HCC. Cancer Discov; 7(5); 522-38. ©2017 AACR.This article is highlighted in the In This Issue feature, p. 443.

P2Y purinoceptors mediate ATP-induced changes in intracellular calcium and amylase release in acinar cells of mouse parotid glands.

Adenosine 5'-triphosphate (ATP) can act as an extracellular signal that regulates various cellular functions. The present study aimed to determine which purinoceptors play a role in ATP-induced changes in intracellular Ca(2+) ([Ca(2+)]i) and amylase secretion in mouse parotid glands. ATP induced a steep increase in [Ca(2+)]i in acinar cells. The removal of extracellular Ca(2+) or the use of Ca(2+) channel blockers slightly inhibited this increase. Inhibition of PLCγ by U73122 and of IP3 by xestospongin C did not completely block this increase. The purinoceptor antagonists suramin and reactive blue-2 strongly inhibited the ATP-induced changes in [Ca(2+)]i. 2-MeSATP induced a strong increase in [Ca(2+)]i, while Bz-ATP induced a small [Ca(2+)]i increase, and UTP and α,ß-MeATP had no effect. The potency order of ATP analogs (2-MeSATP > ATP >> UTP) suggested that P2Y1 and P2Y12 play a significant role in the cellular response to ATP. RT-PCR revealed that P2X2,4,7 and P2Y1,2,10,12,14 were expressed in acinar cells. Ca(2+)-dependent exocytotic secretion of amylase was detected in parotid glands. These findings indicated that ATP activates P2Y receptors more than P2X receptors at low concentrations. Thus, P2Y receptors were found to be the main receptors involved in Ca(2+)-related cell homeostasis and amylase secretion in mouse parotid glands.

Evaluation of transcutaneous and end-tidal carbon dioxide levels during inhalation sedation in volunteers.

Measurement of end-tidal carbon dioxide (PETCO2) is useful because of its noninvasiveness, continuity, and response time when sudden changes in ventilation occur during inhalation sedation. We compared the accuracy of PETCO2 using a nasal mask and nasal cannula with the accuracy of transcutaneous carbon dioxide (TC-CO2) and determined which method is more useful during inhalation sedation in volunteers. We used a modified nasal mask (MNM) and modified nasal cannula (MNC) for measurement of PETCO2. The capnometer measured PETCO2 in the gas expired from the nasal cavity by means of two devices. The volunteers received supplemental O2 by means of each device at a flow rate of 6 L/min. After the volunteers lay quietly for 5 min with a supply of 100 % O2, they received supplemental N2O by means of each device at concentrations of 10, 20, and 25 % for 5 min and 30 % for 25 min. The correlation coefficient was poorer in the MNM than in the MNC, and the mean difference between TC-CO2 and PETCO2 in the MNM was greater than that in the MNC. The difference between the TC-CO2 and PETCO2 ranged from 3 to 6 mmHg in the MNM and from 2 to 5 mmHg in the MNC. The difference between two variables against the TC-CO2 and the CO2 waveforms obtained by means of the two devices were within the clinically acceptable range. Our two devices can provide continuous monitoring of PETCO2 with a supply of N2O/O2 in patients undergoing inhalation sedation.

The effect of mepivacaine on swine lingual, pulmonary and coronary arteries.

BACKGROUND: Although mepivacaine has a known biphasic action on the aortic and coronary artery in several animal species, its effects on the lingual and pulmonary artery are not well understood and it is not yet known whether mepivacaine produces vasoconstriction in these vessels. The present study aims to investigate the direct effects of mepivacaine on swine lingual, pulmonary and coronary arterial endothelium-denuded rings. METHODS: Artery rings were perfused with isotonic 40 mM KCl until a stable constricted plateau was reached. The rings were then perfused with isotonic 40 mM KCl plus a particular concentration of mepivacaine (0.4 µM, 4.0 µM, 40 µM, 0.4 mM and 4.0 mM). The isometric tension strengths in each experiment were normalized to the strength of the isometric tension immediately before mepivacaine perfusion and expressed as a percentage. RESULTS: Mepivacaine at 0.4 to 40 µM did not significantly alter 40 mM KCl-induced contraction in the lingual, pulmonary and coronary arterial rings. In contrast, mepivacaine at 4 mM produced attenuated vasoconstriction in the lingual, pulmonary and coronary arterial compared with isotonic 40 mM KCl. CONCLUSIONS: Mepivacaine produced vasoconstriction at lower concentrations, followed by attenuated vasoconstriction at higher concentrations on swine lingual, pulmonary and coronary arterial endothelium-denuded rings. Mepivacaine (4 µM) appeared to increase isotonic 40 mM KCl-induced contraction, followed by attenuated vasoconstriction at 4 mM. Dentists using 3 % mepivacaine should take into consideration that the risk of complications may be increased if more than six mepivacaine cartridges are used in dental treatment or minor surgery, or if over 15 ml of mepivacaine is administered to a patient with cardiovascular complications during general anesthesia for oral maxillofacial surgery.

Severe Bradycardia Possibly due to a Local Anesthetic Oral Mucosal Injection during General Anesthesia.

Local anesthesia may induce systemic complications leading to parasympathetic activity leading to bradycardia and hypotension. We report a case of a 50-year-old man undergoing dental surgery under general anesthesia who experienced severe bradycardia and hypotension after local anesthesia infiltration. Concerns regarding the utilization of a relatively large lumen injection needle for local anesthesia during general anesthesia are discussed.

Effect of lidocaine on swine lingual and pulmonary arteries.

PURPOSE: Lidocaine has a biphasic action on smooth muscle of peripheral blood vessels, with vasoconstriction at low concentrations and vasodilation at higher concentrations. Many in vivo studies have demonstrated the effects of lidocaine on aortic or coronary arteries in several animals, but there are few reports about the effect on peripheral vessels. This study was designed to investigate the direct effects of lidocaine on peripheral vessels, namely swine lingual and pulmonary arterial rings. METHODS: Swine lingual artery and pulmonary artery segments, about 2-3 mm in diameter, were cut into 3-mm-long rings, and the lumen surface was gently rubbed to remove the endothelium. Isometric tension was measured using a displacement transducer and recorded. After a stable constriction was developed with 5 µM noradrenaline, 5 µM noradrenaline containing lidocaine (0.5, 1.0, 10, 20, 50 or 100 µg/ml) was perfused for 5 min, and then all drug perfusion was stopped. The strength of any isometric tension during an experiment was normalized to the strength of the isometric tension immediately before lidocaine perfusion, and expressed as a percentage. RESULTS: Lidocaine elicited a concentration-dependent biphasic response of lingual and pulmonary arterial rings. The lidocaine concentration at 1 µg/ml caused mild contraction. Dilation occurred at 10 µg/ml and increased with increasing dose. CONCLUSIONS: Lidocaine-induced vasoconstriction of swine lingual and pulmonary smooth muscle may occur at low concentration when lidocaine is infiltrated into the oral submucosa or administered intravenously for the treatment of ventricular arrhythmia.