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The endogenous cannabinoid receptor agonist anandamide is present in central and peripheral tissues. As the kidney contains both the amidase that degrades anandamide and transcripts for anandamide receptors, we characterized the molecular components of the anandamide signaling system and the vascular effects of exogenous anandamide in the kidney. We show that anandamide is present in kidney homogenates, cultured renal endothelial cells (

Self financingThe first outbreak of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) occurred in Wuhan, Hubei Province, China, in late 2019. The subsequent COVID-19 pandemic rapidly affected the health and economy of the world. The global approach to the pandemic was to isolate populations to reduce the spread of this deadly virus while vaccines began to be developed. In March 2020, the first phase I clinical trial of a novel lipid nanoparticle (LNP)-encapsulated mRNA-based vaccine, mRNA-1273, which encodes the spike protein (S protein) of SARS-CoV-2, began in the United States (US). The production of mRNA-based vaccines is a promising recent development in the production of vaccines. However, there remain significant challenges in the development and testing of vaccines as rapidly as possible to control COVID-19, which requires international collaboration. This review aims to describe the background to the rationale for the development of mRNA-based SARS-CoV-2 vaccines and the current status of the mRNA-1273 vaccine.

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It is now well accepted that endogenous morphine is present in animals, both in invertebrates and vertebrates. It is a key signaling molecule that plays an important role in downregulating physiological responses, such as those in the immune system, including immune elements in the CNS. It has been demonstrated that a specific mu-opiate-receptor subtype, mu3, mediates these downregulatory effects through release of NO. This article examines morphine as an endogenous signaling molecule, in terms of its role in neural and immune regulation.

Studies from our laboratory have revealed a novel μ opiate receptor, μ3, which is expressed in both vascular tissues and leukocytes. The μ3 receptor is selective for opiate alkaloids and is insensitive to opioid peptides. We now identify the μ3 receptor at the molecular level using a 441-bp conserved region of the μ1 receptor. Sequence analysis of the isolated cDNA suggests that it is a novel, alternatively spliced variant of the μ opiate receptor gene. To determine whether protein expressed from this cDNA exhibits the biochemical characteristics expected of the μ3 receptor, the cDNA clone was expressed in a heterologous system. At the functional level, COS-1 cells transfected with the μ3 receptor cDNA exhibited dose-dependent release of NO following treatment with morphine, but not opioid peptides (i.e., Met-enkephalin). Naloxone was able to block the effect of morphine on COS-1 transfected cells. Nontransfected COS-1 cells did not produce NO in the presence of morphine or the opioid peptides at similar concentrations. Receptor binding analysis with [3H]dihydromorphine further supports the opiate alkaloid selectivity and opioid peptide insensitivity of this receptor. These data suggest that this new μ opiate receptor cDNA encodes the μ3 opiate receptor, since it exhibits biochemical characteristics known to be unique to this receptor (opiate alkaloid selective and opioid peptide insensitive). Furthermore, using Northern blot, RT-PCR, and sequence analysis, we have demonstrated the expression of this new μ variant in human vascular tissue, mononuclear cells, polymorphonuclear cells, and human neuroblastoma cells.

The presence of morphine-like and codeinelike substances was demonstrated in the pedal ganglia, hemolymph, and mantle tissues of the mollusc Mytilus edulis. The pharmacological activities of the endogenous morphine-like material resemble those of authentic morphine. Both substances were found to counteract, in a dose-dependent manner, the stimulatory effect of tumor necrosis factor ax or interleukin la on human monocytes and Mytilus immunocytes, when added simultaneously to the incubation medium. The immunosuppressive effect of this opiate material expresses itself in a lowering of chemotactic activity, cellular velocity, and adherence. Codeine mimics the activity of authentic morphine, but only at much higher concentrations. Specific high-affinity receptor sites (jx3) for morphine have been identified on human monocytes and Mytilus immunocytes. In Mytilus recovering from experimentally induced stress, the return of "alerted" immunocytes to a more inactive state appears to be due to a significant rise in the content of morphine-like material in the pedal ganglia and hemolymph at this time. Thus, morphine may have a role in calming or terminating the state of immune alertness.The occurrence throughout the animal kingdom of messenger molecules identical with, or closely related to, those known in mammals has been amply documented by biochemical identification, functional analysis, and the demonstration of specific receptor sites (1-6). This applies in particular to classical neurotransmitters, neuropeptides, and cytokines. The demonstration by Spector and coworkers (9-11) and others (7, 8) added to the sample in a 6:10 (vol/vol) ratio (HCl/sample) to a final strength of 10%]. The samples were then heated at 100°C for 30 min, cooled, and centrifuged at 10,000 x g for 20 min. The resulting precipitate was discarded and the supernatant was extracted with 5 vol of 10% (vol/vol) 1-butanol in chloroform. The supernatant was adjusted to pH 8.8-9.0. The organic phase was back-extracted into 2 vol of 0.01 M HCl. Morphine recovery was 50-70%. The aqueous phase was evaporated to dryness and dissolved in 10 ml of phosphate-buffered saline (PBS, pH 7.4) and then the pH was adjusted to 8.5-9.0. Samples were passed through a Sep-Pak C18 cartridge. Morphine and codeine were eluted from the cartridge with 7 ml of 0.1 M pyridine in acetic acid (pH 6.2) containing 2.5% (vol/vol) 1-propanol. The eluate was evaporated to dryness and dissolved in 250 ,ul of 1 mM HCl. An aliquot of 200 ,ul was injected onto a C18 reverse-phase HPLC column (LiChrosorb, 0.4 x 25 cm, Merck). The samples were eluted with 0.1 M pyridine in acetic acid (pH 6.2) followed by a linear gradient of 0-25% 1-propanol/0.1 M pyridine in acetic acid at a flow rate of 1.5 ml/min. Four 1-min fractions were collected during morphine and codeine elution as determined by known standards. These fractions were then evaporated, dissolved, and assayed for morphine and codeine by a sensitive radioimmunoassay (RIA) described by . Polyclonal antibodies generated against 3...