Selective inhibition of intestinal guanosine 3,5-cyclic monophosphate signaling by small-molecule protein kinase inhibitors

The guanosine 3,5-cyclic monophosphate (cGMP)-dependent protein kinase II (cGKII) serine/threonine kinase relays signaling through guanylyl cyclase C (GCC) to control intestinal fluid homeostasis. Here, we report the discovery of small-molecule inhibitors of cGKII. These inhibitors were imidazole-aminopyrimidines, which blocked recombinant human cGKII at submicromolar concentrations but exhibited comparatively little activity toward the phylogenetically related protein kinases cGKI and cAMP-dependent protein kinase (PKA). Whereas aminopyrimidyl motifs are common in protein kinase inhibitors, molecular modeling of these imidazole-aminopyrimidines in the ATP-binding pocket of cGKII indicated an unconventional binding mode that directs their amine substituent into a narrow pocket delineated by hydrophobic residues of the hinge and the C-helix. Crucially, this set of residues included the Leu-530 gatekeeper, which is not conserved in cGKI and PKA. In intestinal organoids, these compounds blocked cGKII-dependent phosphorylation of the vasodilator-stimulated phosphoprotein (VASP). In mouse small intestinal tissue, cGKII inhibition significantly attenuated the anion secretory response provoked by the GCC-activating bacterial heat-stable toxin (STa), a frequent cause of infectious secretory diarrhea. In contrast, both PKA-dependent VASP phosphorylation and intestinal anion secretion were unaffected by treatment with these compounds, whereas experiments with T84 cells indicated that they weakly inhibit the activity of cAMP-hydrolyzing phosphodiesterases. As these protein kinase inhibitors are the first to display selective inhibition of cGKII, they may expedite research on cGMP signaling and may aid future development of therapeutics for managing diarrheal disease and other pathogenic syndromes that involve cGKII

Systematic review: cardiovascular safety profile of 5‐ HT 4 agonists developed for gastrointestinal disorders

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/90554/1/apt5011.pd

Gastrointestinal Dysfunction in a Parkinson’s Disease Rat Model and the Changes of Dopaminergic, Nitric Oxidergic, and Cholinergic Neurotransmitters in Myenteric Plexus

This study aims to explore the gastrointestinal dysfunction and the changes of dopaminergic, nitric oxidergic, and cholinergic neurons in the myenteric plexus of a Parkinson’s disease (PD) rat model. A PD rat model was induced through unilateral substantia nigra administration of 6-hydroxydopamine. Four weeks later, the feces in 1 h and residual solid food in stomach at 2 h after feeding were measured. Changes in tyrosine hydroxylase (TH) in substantial nigra, TH, choline acetyltransferase (ChAT), and neuronal nitric oxide synthase (nNOS) in gastric antrum and colon tissue were examined by immunohistochemistry. Reverse transcription (RT) polymerase chain reaction (PCR) and Western blot were used to evaluate and compare the levels of messenger RNA (mRNA) and protein expression of TH, ChAT, and nNOS in the GI tract between normal and 6-hydroxydopamine-lesioned rats. Compared with control samples, the number of TH+ cells in the damaged side of substantia nigra of 6-hydroxydopamine-lesioned rats decreased significantly (P < 0.01). The weight and water content of the fecal matter decreased (P < 0.01), and the percentage of residual solid food increased (P < 0.01). The average integrated optical densities of TH-positive areas in the gastric antrum and colon tissue increased significantly (P < 0.01), nNOS decreased significantly (P < 0.01), and there were no significant changes in ChAT (P > 0.05). TH and nNOS mRNA levels in the gastric antrum and proximal colon decreased (P < 0.01), there were no significant changes in ChAT mRNA levels (P > 0.05). The protein levels of TH in the GI tract were significantly increased (P < 0.01), nNOS significantly decreased (P < 0.01), and ChAT had no significant changes (P > 0.05). 6-Hydroxydopamine-lesioned rats had delayed gastric emptying and constipation that might be related to the gastrointestinal TH increase and nNOS decrease. These symptoms were not related to changes in cholinergic transmitters

Chemistry of a polluted cloudy boundary layer

A one-dimensional photochemical model for cloud-topped boundary layers is developed which includes detailed descriptions of gas-phase and aqueous-phase chemistry, and of the radiation field in and below cloud. The model is used to interpret the accumulation of pollutants observed over Bakersfield, California, during a wintertime stagnation episode with low stratus. The main features of the observations are well simulated; in particular, sulfate accumulates progressively over the course of the episode due to sustained aqueous-phase oxidation of SO2 in the stratus cloud. The major source of sulfate is the reaction S(IV) + Fe(III), provided that this reaction proceeds by a non radical mechanism in which Fe(III) is not reduced. A radical mechanism with SO3 − and Fe(II) as immediate products would quench sulfate production because of depletion of Fe(III). The model results suggest that the non radical mechanism is more consistent with observations, although this result follows from the absence of a rapid Fe(II) oxidation pathway in the model. Even with the non-radical mechanism, most of the soluble iron is present as Fe(II) because Fe(III) is rapidly reduced by O2 −. The S(IV) + Fe(III) reaction provides the principal source of H2O2 in the model; photochemical production of H2O2 from HO2 or O2(−I) is slow because HO2 is depleted by high levels of NOx. The aqueous-phase reaction S(IV) + OH initiates a radical-assisted S(IV) oxidation chain but we find that the chain is not propagated due to efficient termination by SO4 − + Cl− followed by Cl + H2O. A major uncertainty attached to that result is that the reactivities of S(IV)-carbonyl adducts with radical oxidants are unknown. The chain could be efficiently propagated, with high sulfate yields, if the S(IV)-carbonyl adducts were involved in chain propagation. A remarkable feature of the observations, which is well reproduced by the model, is the close balance between total atmospheric concentrations of acids and bases. We argue that this balance reflects the control of sulfate production by NH3, which follows from the pH dependence of the S(IV) + Fe(III) reaction. Such a balance should be a general characteristic of polluted environments where aqueous-phase oxidation of SO2 is the main source of acidity. At night, the acidity of the cloud approaches a steady state between NH3 emissions and H2SO4 production by the S(IV) + Fe(III) reaction. A steady state analysis suggests that [H+] at night should be proportional to (ESO 2/ENH 3)1/2 where ESO 2 and ENH 3 are emission rates of SO2 and NH3, respectively. From this analysis it appears that cloud water pH values below 3 are unlikely to occur in the Bakersfield atmosphere during the nighttime hours. Very high acidities could, however, be achieved in the daytime because of photochemical acid production by the gas-phase reactions NO2 + OH and SO2 + OH