Structural basis for lysophosphatidylserine recognition by GPR34.

GPR34 is a recently identified G-protein coupled receptor, which has an immunomodulatory role and recognizes lysophosphatidylserine (LysoPS) as a putative ligand. Here, we report cryo-electron microscopy structures of human GPR34-Gi complex bound with one of two ligands bound: either the LysoPS analogue S3E-LysoPS, or M1, a derivative of S3E-LysoPS in which oleic acid is substituted with a metabolically stable aromatic fatty acid surrogate. The ligand-binding pocket is laterally open toward the membrane, allowing lateral entry of lipidic agonists into the cavity. The amine and carboxylate groups of the serine moiety are recognized by the charged residue cluster. The acyl chain of S3E-LysoPS is bent and fits into the L-shaped hydrophobic pocket in TM4-5 gap, and the aromatic fatty acid surrogate of M1 fits more appropriately. Molecular dynamics simulations further account for the LysoPS-regioselectivity of GPR34. Thus, using a series of structural and physiological experiments, we provide evidence that chemically unstable 2-acyl LysoPS is the physiological ligand for GPR34. Overall, we anticipate the present structures will pave the way for development of novel anticancer drugs that specifically target GPR34.

Surface Normals and Shape From Water.

In this paper, we introduce a novel method for reconstructing surface normals and depth of dynamic objects in water. Past shape recovery methods have leveraged various visual cues for estimating shape (e.g., depth) or surface normals. Methods that estimate both compute one from the other. We show that these two geometric surface properties can be simultaneously recovered for each pixel when the object is observed underwater. Our key idea is to leverage multi-wavelength near-infrared light absorption along different underwater light paths in conjunction with surface shading. Our method can handle both Lambertian and non-Lambertian surfaces. We derive a principled theory for this surface normals and shape from water method and a practical calibration method for determining its imaging parameters values. By construction, the method can be implemented as a one-shot imaging system. We prototype both an off-line and a video-rate imaging system and demonstrate the effectiveness of the method on a number of real-world static and dynamic objects. The results show that the method can recover intricate surface features that are otherwise inaccessible.

Non-Rigid Shape From Water.

We introduce a novel 3D sensing method for recovering a consistent, dense 3D shape of a dynamic, non-rigid object in water. The method reconstructs a complete (or fuller) 3D surface of the target object in a canonical frame (e.g., rest shape) as it freely deforms and moves between frames by estimating underwater 3D scene flow and using it to integrate per-frame depth estimates recovered from two near-infrared observations. The reconstructed shape is refined in the course of this global non-rigid shape recovery by leveraging both geometric and radiometric constraints. We implement our method with a single camera and a light source without the orthographic assumption on either by deriving a practical calibration method that estimates the point source position with respect to the camera. Our reconstruction method also accounts for scattering by water. We prototype a video-rate imaging system and show 3D shape reconstruction results on a number of real-world static, deformable, and dynamic objects and creatures in real-world water. The results demonstrate the effectiveness of the method in recovering complete shapes of complex, non-rigid objects in water, which opens new avenues of application for underwater 3D sensing in the sub-meter range.

4-(Dimethylamino)pyridine N-Oxide-Catalyzed Macrolactamization Using 2-Methyl-6-nitrobenzoic Anhydride in the Synthesis of the Depsipeptidic Analogue of FE399.

A depsipeptidic analogue of FE399 was efficiently synthesized mainly through macrolactamization using 2-methyl-6-nitrobenzoic anhydride (MNBA), and a detailed investigation of the desired 16-membered macrolactam core of FE399 was performed. It was determined that the combination of MNBA and a catalytic amount of 4-(dimethylamino)pyridine N-oxide exhibits much higher activity than that of conventionally used coupling reagents such as hexafluorophosphate azabenzotriazole tetramethyl uronium and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate.

Effects of tipepidine on MK-801-induced cognitive impairment in mice.

We previously reported that centrally acting non-narcotic antitussives, including tipepidine, inhibit G-protein-coupled inwardly rectifying potassium (GIRK) channel-activated currents of neurons. In addition, when administered at a cough suppressant dose, the drugs ameliorated the symptoms of various models of intractable brain disease in rodents. In the current study, we investigated whether tipepidine causes recovery from schizophrenia-like cognitive dysfunction, which was induced by MK-801 (0.2 mg/kg, i.p.) in mice. We also examined the effect of tipepidine and clozapine co-administration on the dysfunction. Moreover, we studied whether clozapine inhibits GIRK channel activated currents in single brain neurons using the patch-clamp technique. Tipepidine elicited recovery from MK-801-induced cognitive impairment in the novel objective recognition test and Y-maze test. Further, co-administration of tipepidine and clozapine, at subthreshold doses of each drug, improved MK-801-induced cognitive impairment in the novel objective recognition test. Clozapine (3 × 10-5 M) had a minor effect on baclofen-induced currents in dopamine neurons of the ventral tegmental area.

Enantioselective total synthesis of naturally occurring eushearilide and evaluation of its antifungal activity.

The asymmetric total synthesis of a newly proposed structure of (3S,16E,20E,23S)-(+)-eushearilide was achieved primarily through an asymmetric Mukaiyama aldol reaction, Schlosser-modified Wittig reaction and 2-methyl-6-nitrobenzoic anhydride-mediated macrolactonization. Based on detailed spectroscopic analyses, the obtained synthetic compound was found to be identical to natural eushearilide. Therefore, we were able to determine the true structure of eushearilide. Moreover, the synthetic compound was found to exhibit significant in vitro antifungal activity against various fungi and bacteria.

Effect of tipepidine with novel antidepressant-like action on c-fos-like protein expression in rat brain.

We previously reported that tipepidine, a centrally acting non-narcotic antitussive, has an antidepressant-like effect in normal and imipramine treatment-resistant depression model rats. Recently, mapping the induction of c-fos-like immunoreactivity (FLI) in the rat brain showed FLI-positive neurons in several brain areas after acute administration of different classes of antidepressants. Here, the effect of a single injection of an antidepressive dose of tipepidine on FLI was studied in seven areas of the rat brain including the central nucleus of the amygdala (CeA) and the nucleus accumbens (NAc). Desipramine was also used for comparison. Rats were anesthetized and perfused 2h after injection with tipepidine (20 and 40mg/kg, i.p.), desipramine (10mg/kg, i.p.), or saline. Then, immunostaining of FLI-positive neurons in brain slices was performed with conventional methods. A single injection of tipepidine increased FLI-positive neurons in the CeA, similar to preexisting antidepressants, and induced the characteristic pattern of an increase in FLI-positive neurons in six other brain areas including the NAc, an effect that was different from other antidepressants. In addition, a single injection of desipramine (10mg/kg) or tipepidine (20mg/kg) decreased the immobility time in the forced swimming test to a similar extent. The results obtained from the previous behavioral study and the current immunohistochemical study suggest that tipepidine may be a novel antidepressant.

Pharmacological mechanisms of antidepressant-like effect of tipepidine in the forced swimming test.

We previously reported that the centrally acting non-narcotic antitussive, tipepidine, produces a novel antidepressant-like effect in the forced swimming test in rats, but the mechanism of the antidepressant-like effect of tipepidine is not clear. We investigated the pharmacological mechanism of the antidepressant-like effect of tipepidine in the forced swimming test in rats. A catecholamine-depleting agent, alpha-methyl-p-tyrosine (AMPT; 300 mg/kg, s.c.), was given 6h before the first injection and with the last injection of tipepidine (40 mg/kg, i.p.). A serotonin (5-HT)-depleting agent, p-chlorophenylalanine (PCPA; 350 mg/kg, i.p.), was given 72 h and 48 h before the pretest session. The dopamine D(1) receptor antagonist, SCH23390 (0.02 mg/kg, s.c.) was given 15min before each of the three injections of tipepidine. The dopamine D(2) receptor antagonist raclopride (0.2mg/kg, s.c.), the alpha 1 adrenoceptor antagonist prazosin (1mg/kg, i.p.), the alpha 2 adrenoceptor antagonist yohimbine (2mg/kg, i.p.) and the beta adrenoceptor antagonist propranolol (2mg/kg, i.p.) were given 30 min before each of the three injections of tipepidine. AMPT, but not PCPA, significantly inhibited the immobility time-reducing effect of tipepidine in the forced swimming test. Furthermore, the effect of tipepidine was significantly inhibited by SCH23390 and yohimbine. However, raclopride, prazosin, and propranolol failed to block the effect of tipepidine. The results suggest that the antidepressant-like effect of tipepidine in the forced swimming test may be due at least in part to the effects of dopamine and noradrenaline released at the dopamine D(1) receptor and alpha 2 adrenoceptor, respectively.