Anandamide metabolism by human liver and kidney microsomal cytochrome p450 enzymes to form hydroxyeicosatetraenoic and epoxyeicosatrienoic acid ethanolamides

ABSTRACT The endocannabinoid anandamide is an arachidonic acid derivative that is found in most tissues where it acts as an important signaling mediator in neurological, immune, cardiovascular, and other functions. Cytochromes P450 (P450s) are known to oxidize arachidonic acid to the physiologically active molecules hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs), which play important roles in blood pressure regulation and inflammation. To determine whether anandamide can also be oxidized by P450s, its metabolism by human liver and kidney microsomes was investigated. The kidney microsomes metabolized anandamide to a single monooxygenated product, which was identified as 20-HETE-ethanolamide (EA). Human liver microsomal incubations with anandamide also produced 20-HETE-EA in addition to 5,6-, 8,9-, 11-12, and 14,15-EET-EA. The EET-EAs produced by the liver microsomal P450s were converted to their corresponding dihydroxy derivatives by microsomal epoxide hydrolase. P450 4F2 was identified as the isoform that is most probably responsible for the formation of 20-HETE-EA in both human kidney and human liver, with an apparent K m of 0.7 M. The apparent K m values of the human liver microsomes for the formation of the EET-EAs were between 4 and 5 M, and P450 3A4 was identified as the primary P450 in the liver responsible for epoxidation of anandamide. The in vivo formation and biological relevance of the P450-derived HETE and EET ethanolamides remains to be determined

Follow-Up After Hip and Knee Arthroplasty: a Review of the Literature and a Report on a Pilot Project at the Vreden National Medical Research Center of Traumatology and Orthopedics

Background. Today in our country, the follow-up of patients after arthroplasty is carried out in accordance with clinical guidelines, the wording of which is based on monographs from 2006, 2008, and 2014, in addition, clinical guidelines for follow-up do not take into account the results of treatment assessed by the patient himself. The purpose of this study was to examine existing systems and develop a proprietary follow-up system for patients after hip and knee arthroplasty. Results. A review of the literature revealed that follow-up of patients after arthroplasty is an unsolved problem, within which there is low coverage, reluctance or forgetfulness of the asymptomatic patient, the problem of accessibility of medical examinations, and an excessive financial burden on the health care system. Since 2022, fixed recommendations for follow-up after arthroplasty have been used in the clinical practice of our center in discharge epicrisis. Recommendations for the frequency of follow-up were formulated by experts based on a comprehensive review of the literature and their own experience. In the first three months, 221 hip and 235 knee evaluation questionnaires were collected through the proposed mechanism, with a progressive increase in the number of questionnaires based on weekly monitoring data. Conclusion. Unfortunately, the outpatient clinic system is not always able to provide qualitative monitoring of patients after arthroplasty due to various reasons, therefore, in our opinion, the implementation of the mechanism of remote monitoring of patients will allow detecting various complications at the stage of early diagnosis, which will contribute to prompt solution of these problems. The remote monitoring system is also an important source of scientific data

Revising the structure of a new eicosanoid from human platelets to 8,9-11,12-diepoxy-13-hydroxy-eicosadienoic acid

Eicosanoids are critical mediators of fever, pain, and inflammation generated by immune and tissue cells. We recently described a new bioactive eicosanoid generated by cyclooxygenase-1 (COX-1) turnover during platelet activation that can stimulate human neutrophil integrin expression. On the basis of mass spectrometry (MS/MS and MS3), stable isotope labeling, and GC-MS analysis, we previously proposed a structure of 8-hydroxy-9,11-dioxolane eicosatetraenoic acid (DXA3). Here, we achieved enzymatic synthesis and 1H NMR characterization of this compound with results in conflict with the previously proposed structural assignment. Accordingly, by using LC-MS, we screened autoxidation reactions of 11-hydroperoxy-eicosatetraenoic acid (11-HpETE) and thereby identified a candidate sharing the precise reverse-phase chromatographic and MS characteristics of the platelet product. We optimized these methods to increase yield, allowing full structural analysis by 1H NMR. The revised assignment is presented here as 8,9–11,12-diepoxy-13-hydroxyeicosadienoic acid, abbreviated to 8,9–11,12-DiEp-13-HEDE or DiEpHEDE, substituted for the previous name DXA3. We found that in platelets, the lipid likely forms via dioxolane ring opening with rearrangement to the diepoxy moieties followed by oxygen insertion at C13. We present its enzymatic biosynthetic pathway and MS/MS fragmentation pattern and, using the synthetic compound, demonstrate that it has bioactivity. For the platelet lipid, we estimate 16 isomers based on our current knowledge (and four isomers for the synthetic lipid). Determining the exact isomeric structure of the platelet lipid remains to be undertaken

MASTER Real-Time Multi-Message Observations of High Energy Phenomena

This review considers synchronous and follow-up MASTER Global Robotic Net optical observations of high energy astrophysical phenomena such as fast radio bursts (FRB), gamma-ray bursts (including prompt optical emission polarization discovery), gravitational-wave events, detected by LIGO/VIRGO (including GW170817 and independent Kilonova discovery), high energy neutrino sources (including the detection of IC-170922A progenitor) and others. We report on the first large optical monitoring campaign of the closest at that moment radio burster FRB 180916.J0158+65 simultaneously with a radio burst. We obtained synchronous limits on the optical flux of the FRB 180916.J0158+65 and FRB 200428 (soft gamma repeater SGR 1935+2154) (The CHIME/FRB Collaboration, Nature 2020, 587) at 155093 MASTER images with the total exposure time equal to 2,705,058 s, i.e., 31.3 days. It follows from these synchronous limitations that the ratio of the energies released in the optical and radio ranges does not exceed 4 × 105. Our optical monitoring covered a total of 6 weeks. On 28 April 2020, MASTER automatically following up on a Swift alert began to observe the galactic soft gamma repeater SGR 1935+2154 experienced another flare. On the same day, radio telescopes detected a short radio burst FRB 200428 and MASTER-Tavrida telescope determined the best prompt optical limit of FRB/SGR 1935+2154. Our optical limit shows that X-ray and radio emissions are not explained by a single power-law spectrum. In the course of our observations, using special methods, we found a faint extended afterglow in the FRB 180916.J0158+65 direction associated with the extended emission of the host galaxy

MASTER Real-Time Multi-Message Observations of High Energy Phenomena

This review considers synchronous and follow-up MASTER Global Robotic Net optical observations of high energy astrophysical phenomena such as fast radio bursts (FRB), gamma-ray bursts (including prompt optical emission polarization discovery), gravitational-wave events, detected by LIGO/VIRGO (including GW170817 and independent Kilonova discovery), high energy neutrino sources (including the detection of IC-170922A progenitor) and others. We report on the first large optical monitoring campaign of the closest at that moment radio burster FRB 180916.J0158+65 simultaneously with a radio burst. We obtained synchronous limits on the optical flux of the FRB 180916.J0158+65 and FRB 200428 (soft gamma repeater SGR 1935+2154) (The CHIME/FRB Collaboration, Nature 2020, 587) at 155093 MASTER images with the total exposure time equal to 2,705,058 s, i.e., 31.3 days. It follows from these synchronous limitations that the ratio of the energies released in the optical and radio ranges does not exceed 4 × 105. Our optical monitoring covered a total of 6 weeks. On 28 April 2020, MASTER automatically following up on a Swift alert began to observe the galactic soft gamma repeater SGR 1935+2154 experienced another flare. On the same day, radio telescopes detected a short radio burst FRB 200428 and MASTER-Tavrida telescope determined the best prompt optical limit of FRB/SGR 1935+2154. Our optical limit shows that X-ray and radio emissions are not explained by a single power-law spectrum. In the course of our observations, using special methods, we found a faint extended afterglow in the FRB 180916.J0158+65 direction associated with the extended emission of the host galaxy