Systemic Inflammatory Reaction Research Articles

Analysis of fatty acid-derived lipids in critically ill patients after cardiac surgery yields novel pathophysiologically relevant mediators with possible relevance for systemic inflammatory reactions

IntroductionCritically ill patients suffer from a wide variety of clinical events, most of them leading to pro-inflammatory states such as sepsis or simply as consequence of major surgery. Many of these patients develop forms of acute kidney injury, heart or acute liver failure during intensive care. Lipid signaling is critically involved in triggering systemic inflammation processes, pain and vascular tone. We therefore hypothesized that fatty-acid-derived lipid mediators might be regulated during inflammatory stages and other clinical events in critically ill patients and might serve as potential biomarker candidates.Methods and study designUsing liquid chromatography-tandem mass spectrometry (LC-MS/MS), we determined the levels of 53 lipid mediators in plasma from nine patients. These patients were hospitalized at Frankfurt University Hospital’s intensive care unit (ICU) after cardiac surgery. Inflammatory stages were illustrated over time using clinically established biomarkers such as interleukin-6 (IL-6) and leukocyte count. Normal range values of the lipids were obtained from healthy volunteers.ResultsPlasma levels clearly outside the normal range were observed for 22 of 53 lipid mediators, of which 13 were increased (including ceramides Cer (d18:0/18:0), Cer (d18:1/16:0), Cer (d18:1/18:1), glucosyl-ceramide GluCer (d18:1/24:1), lactosylceramide LacCer (d18:1/18:0), and LacCer (d18:1/24:1), 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha), 11,12- and 14,15-DHET and 1- and 2-arachidonoyl glycerol (1-AG and 2-AG), Sphingosine SPH (d18:1) and 20-HETE. Furthermore, nine lipids were decreased (Cer (d18:1/24:0), LacCer (d18:1/16:0), LacCer (d18:1/24:0), sphingosine-1-phosphate S1P (d18:1), S1P (d18:0), the lysophosphatidic acids LPA (16:0), LPA (18:0), LPA (18:1) and 9-HODE. Among increased lipids, the remarkable changes in 1-AG, 2-AG, and to a lower extent of 6-keto-PGF1-alpha plasma levels showed a certain agreement with inflammatory phases. Furthermore, 6-keto-PGF1alpha had its peak shortly before initiation of continuous veno-venous hemodialysis (at least in 5 of the observed patients), 2-AG was elevated in all our nine patients during (right) heart failure in the context of either re-opening patient’s chest, implementation of veno-arterial ECMO or at least while significantly increasing the amount of catecholamines.DiscussionIn this pilot trial we identified several evaluated lipids in critically ill patients representing either potentially (patho-) physiologically relevant mediators of the pro-inflammatory processes and during heart failure or possible markers preceding veno-venous hemodialysis.

Preliminary study on the preparation of lyophilized acellular nerve scaffold complexes from rabbit sciatic nerves with human umbilical cord mesenchymal stem cells.

The gold standard of care for patients with severe peripheral nerve injury is autologous nerve grafting; however, autologous nerve grafts are usually limited for patients because of the limited number of autologous nerve sources and the loss of neurosensory sensation in the donor area, whereas allogeneic or xenografts are even more limited by immune rejection. Tissue-engineered peripheral nerve scaffolds, with the morphology and structure of natural nerves and complex biological signals, hold the most promise as ideal peripheral nerve "replacements". To prepare allogenic peripheral nerve scaffolds using a low-toxicity decellularization method, and use human umbilical cord mesenchymal stem cells (hUC-MSCs) as seed cells to cultivate scaffold-cell complexes for the repair of injured peripheral nerves. After obtaining sciatic nerves from New Zealand rabbits, an optimal acellular scaffold preparation scheme was established by mechanical separation, varying lyophilization cycles, and trypsin and DNase digestion at different times. The scaffolds were evaluated by hematoxylin and eosin (HE) and luxol fast blue (LFB) staining. The maximum load, durability, and elastic modulus of the acellular scaffolds were assessed using a universal material testing machine. The acellular scaffolds were implanted into the dorsal erector spinae muscle of SD rats and the scaffold degradation and systemic inflammatory reactions were observed at 3 days, 1 week, 3 weeks, and 6 weeks following surgery to determine the histocompatibility between xenografts. The effect of acellular scaffold extracts on fibroblast proliferation was assessed using an MTT assay to measure the cytotoxicity of the scaffold residual reagents. In addition, the umbilical cord from cesarean section fetuses was collected, and the Wharton's jelly (WJ) was separated into culture cells and confirm the osteogenic and adipogenic differentiation of mesenchymal stem cells (MSCs) and hUC-MSCs. The cultured cells were induced to differentiate into Schwann cells by the antioxidant-growth factor induction method, and the differentiated cells and the myelinogenic properties were identified. The experiments effectively decellularized the sciatic nerve of the New Zealand rabbits. After comparing the completed acellular scaffolds among the groups, the optimal decellularization preparation steps were established as follows: Mechanical separation of the epineurium, two cycles of lyophilization-rewarming, trypsin digestion for 5 hours, and DNase digestion for 10 hours. After HE staining, no residual nuclear components were evident on the scaffold, whereas the extracellular matrix remained intact. LFB staining showed a significant decrease in myelin sheath composition of the scaffold compared with that before preparation. Biomechanical testing revealed that the maximum tensile strength, elastic modulus, and durability of the acellular scaffold were reduced compared with normal peripheral nerves. Based on the histocompatibility test, the immune response of the recipient SD rats to the scaffold New Zealand rabbits began to decline3 weeks following surgery, and there was no significant rejection after 6 weeks. The MTT assay revealed that the acellular reagent extract had no obvious effects on cell proliferation. The cells were successfully isolated, cultured, and passaged from human umbilical cord WJ by MSC medium, and their ability to differentiate into Schwann-like cells was demonstrated by morphological and immunohistochemical identification. The differentiated cells could also myelinate in vitro. The acellular peripheral nerve scaffold with complete cell removal and intact matrix may be prepared by combining lyophilization and enzyme digestion. The resulting scaffold exhibited good histocompatibility and low cytotoxicity. In addition, hUC-MSCs have the potential to differentiate into Schwann-like cells with myelinogenic ability following in vitro induction.

The function of miRNAs in the immune system's inflammatory reaction to heart failure.

Heart failure is the end stage of cardiovascular disease, with high morbidity and mortality rates worldwide. Heart failure is associated with long-term and insufficient inhibition of inflammatory response. miRNA is a class of endogenous, non-coding, single-stranded small RNA molecules, that can regulate gene expression through translational inhibition or degradation of targeted mRNA, widely regulate myocardial remodeling, inflammatory response, and other pathological processes, and play an important regulatory role in the occurrence and development of cardiovascular diseases. This article reviews the role of miRNA in the inflammatory response in heart failure.

Bacteriophage therapy and prevention of infectious and inflammatory complications after endoscopic urologic surgery

Relevance. Given the growing antibiotic resistance of pathogens of urologic infection to antimicrobial drugs, one of the urgent problems is the search for alternative methods of antimicrobial prevention of infectious and inflammatory complications after surgical interventions, as well as an adequate therapy regimen during the postoperative period. Bacteriophage preparation is an alternative method for the treatment and prevention of infectious and inflammatory urological diseases.Objective. To determine an effective method for preoperative antimicrobial prevention of infectious and inflammatory complications in patients with kidney stones subjected to percutaneous nephrolithotripsy (PNLT).Methods. The study included 90 patients with coralloid or multiple large kidney stones who underwent PNLT. Before PNLT, all patients underwent bacteriological urinalysis in order to determine sensitivity not only to antibiotics and bacteriophage preparations. Urine sampling was performed for further microbiological cultural examination during renal pelvic puncture on the 3rd and 7th days after PNLT. Depending on the technique of perioperative prophylaxis, three groups of 30 were formed. Group 1 patients were intravenously injected with 1000 mg of ciprofloxacin throughout the operation and then intravenously dipped with 1000 mg once a day for 3–5 days. Group 2 received intramuscular cefotaxime + sulbactam (1.0+0.5) twice a day 2 h before surgery intramuscularly once. In group 3, patients received oral mycobacteriophage 40 ml polyvalent purified orally 1 h before surgery and 40 ml orally 3 times a day for 3–5 days after surgery.Results. In all three groups, the development of infectious complications in patients was assessed: acute pyelonephritis, systemic inflammatory reaction syndrome (SERS) or urosepsis. No serious infectious or inflammatory complications were observed in any group during the early postoperative period. The development of CVD after PNLT was noted on the 1st–3rd postoperative day (in 26.6 %, 20 and 20 % of patients of the 1st, 2nd and 3rd groups, respectively). However, on the 4th–7th day after PNLT, normal blood parameters (leukocytes, rod-shaped neutrophils), temperature, and general well-being were noted.Conclusion. The same effectiveness of different antimicrobial regimens for preventing infectious and inflammatory complications after PNLT was similar. Bacteriophage preparations are effective and can prevent infectious and inflammatory complications following PNLT. The development of CVD after PNLT on the 1st–3rd day after surgery cannot be correlated only with the antimicrobial drugs used and the method of their administration (intravenously, intramuscularly and orally). Most likely, the development of CVD is associated with operational injury.

Changes in the macrophage polarization in the kidneys of rats with acute respiratory distress syndrome after transplantation of human umbilical cord-derived mesenchymal stem cell

Acute respiratory distress syndrome (ARDS) is a critical state of the body characterized by severe hypoxemia and multiple organ failure. One of the organs that is most often additionally affected due to the systemic inflammatory reaction that occurs in the lungs during ARDS is the kidney. It is known that mesenchymal stem cells (MSCs) can exhibit nephroprotective effects due to their anti-inflammatory and immunomodulatory abilities, but the mechanisms of such therapeutic effects require further elucidation. The purpose is to analyze macrophage polarization in the damaged kidneys of rats with modeled ARDS following the transplantation of human umbilical cord-derived MSCs. Materials and methods. Seventy-two sexually mature male Wistar rats were randomly divided into nine groups: intact animals, 3 days, 7 days and 28 days after ARDS modeling, control-MSC and four correction groups: 24 h ARDS + 2 days MSCs, 4 days of ARDS + 3 days of MSCs, 14 days of ARDS + 14 days of MSCs, 21 days of ARDS + 7 days of MSCs. ARDS was modeled by intranasal administration of lipopolysaccharide (LPS) at a dose of 5 mg/kg. A suspension of MSCs was obtained from the umbilical cord of a healthy donor after a normal delivery by an enzymatic method and administered intraperitoneally at a dose of 1 million cells/kg of body weight. For immunohistochemical determination of the total number of macrophages, pro-inflammatory M1 and anti-inflammatory M2 cells, monoclonal antibodies to CD68, CD86 and CD163 were used, respectively, and the percentage of visual fields occupied by the immunoprecipitate was calculated. Results. Comparative analysis of the percentage of areas occupied by the immunoprecipitate with the corresponding antibodies showed a statistically significant increase in the total number of macrophages (3.8-fold on day 3 of ARDS, 2.8-fold on day 7 of ARDS and 2.4-fold on day 28 of ARDS, respectively), as well as a sharp increase in pro-inflammatory M1 (8.8-fold on day 3 of ARDS, 4.9-fold on day 7 of ARDS, and 4.8-fold on day 28 of ARDS, respectively) and a moderate increase in the percentage of anti-inflammatory M2 cells (1.9-fold on day 3 of ARDS, 2-fold on day 7 of ARDS, and 1.6-fold on day 28 of ARDS, respectively) in modeled ARDS compared to intact animals. This indicates the activation of the inflammatory cascade in the kidneys caused by the development of ARDS. On the other hand, in groups of animals that received MSC correction, a significant decrease, compared to untreated animals, in the total number of macrophages was detected (at 24 h of ARDS + 2 days of MSCs, it was 3.2 times lower than on day 3 of ARDS and ≥ 2 times at later times of the experiment), among which anti-inflammatory cells of the M2 phenotype predominated. Conclusion. Transplanted human umbilical cord-derived MSCs have a pronounced immunomodulatory effect in the affected kidneys of rats with acute respiratory distress syndrome, which is manifested in their ability to change the polarization of macrophages in the anti-inflammatory direction.

Histopathological and Immunohistochemical Investigation on Effects of Boric Acid Used in Treatment of Rats with Knee Osteoartritis on Kidney Tissues

Osteoarthritis (OA) is a disease that often occurs in the knee joints and is characterized by disruption of cartilage homeostasis. Due to the systemic inflammation it creates, it affects not only the joint area, but also many tissues and organs. In this study, the damage caused by systemic inflammatory reactions due to OA in kidney tissue and the protective effect of boric acid were investigated. Wistar albino rats were used in the study. An experimental knee OA model was created by intraarticular injection of monosodium iodoacetate (MIA) in rats. It was formed from 4 groups as Control, OA, OA + 4 mg Boric Acid, OA + 10 mg Boric Acid to work. At the end of the study, kidney tissues were taken from the rats and TNF-α, IL-1β, NOS2 and MMP13 analyzes were performed by histopathological and immunohistochemical methods. Histopathological examinations revealed severe degenerative and necrotic changes in tubular epithelial cells in the OA groups, and these changes decreased in the boric acid-administered group depending on the dose. In immunohistochemical analyzes, it was determined that systemic inflammatory reactions occurring in OA application decreased in a dose-dependent manner with boric acid application. In conclusion; It was determined that kidney tissues were damaged due to systemic inflammatory reactions in rats with OA and boric acid had a protective effect against this damage.

Assessment of systemic reaction to inflammation induced by photodynamic therapy in cervical intraepithelial neoplasia

Photodynamic therapy is a curative treatment of human papillomavirus-associated diseases that provides a selective effect leading to the destruction of pathological cells containing the virus. The retrospective study aimed to determine the role of systemic inflammatory reaction induced by photodynamic stimulation in low- and high-squamous intraepithelial lesion treatment. Methods143 patients with confirmed human papillomavirus-associated cervical intraepithelial neoplasia underwent photodynamic therapy with Photolon and activation in the range of 662 nm. All patients underwent colposcopy, histologic study, HPV DNA analysis, CBC, and immunogram. The chi-square criterion was used to evaluate differences before, 5 days post, and 3 months after PDT; a P value <0.05 was considered significant. ResultsA complete regression in patients with low-squamous intraepithelial lesion (n=117) was achieved 3 months after PDT in 89.7% of cases (105/117) while it persisted in 12 patients. Complete regression in patients with high-squamous intraepithelial lesion (n=26) was achieved in 92.3% (24/26), one patient retained with the lesion, and another one had partial regression in the form of lower grade lesion. On the 5th day post-PDT, the formation of dense fibrin in photodynamic effect was noted on colposcopy, and changes in CBC, accompanied by statistically significant neutrocytosis, increased ESR and other indicators of reactive inflammation were recorded. Patients noted increased body temperature up to 37.8±0.5°C in 88% of cases. ConclusionOne of the mechanisms of PDT`s antitumor and antiviral action is the development of acute inflammation in response to cytotoxic action on cells and vascular response in the form of pathological area devascularization. Such reactive inflammation activates all parts of the immune system. Changes on day 5 post-PDT in inflammatory parameters in CBC and IL-1, IL-6 and TNF-α markers correlate with the clinical picture of the post-photodynamic effect.

Long-term monocyte activation after coronary artery bypass grafting: An exploratory prospective observational study

Major surgery such as coronary artery bypass grafting (CABG) is associated with an increased post-operative risk of atherosclerotic cardiovascular events. Cells of the innate immune system can adopt a long-lasting pro-inflammatory and atherogenic phenotype after brief exposure to exogenous or endogenous inflammatory stimuli, a process called “trained immunity”. We hypothesized that the surgery-induced inflammation leads to sustained alterations in monocyte function, which promote the subsequent occurrence of cardiovascular events. Blood from 13 patients undergoing elective CABG was obtained before, 3–7 days (median 4) after, and 6–8 weeks (median 6) weeks after surgery. At 3–7 days postoperatively, circulating C-reactive protein (CRP) concentration, leukocyte counts and ex vivo Peripheral Blood Mononuclear Cell (PBMC) IL-6, TNFα and IL-1Ra production after stimulation (with various inflammatory stimuli) were significantly increased. Simultaneously, there was a reduction in monocyte HLA-DR expression. 6–8 weeks after CABG there was an ongoing systemic pro-inflammatory state with higher CRP concentrations, increased stimulated ex vivo PBMC IL-6 production, changes in monocytes subsets, and a higher expression of CCR2 on monocytes compared to baseline. In conclusion, CABG induces a persistent systemic inflammatory reaction with a sustained activated monocyte phenotype. This might contribute to the increased atherosclerotic cardiovascular event risk observed in cardiac surgery patients.

Alpha-Pinene-encapsulated lipid nanoparticles diminished inflammatory responses in THP-1 cells and imiquimod-induced psoriasis-like skin injury and splenomegaly in mice.

Psoriasis, a persistent skin condition caused by the disorder of the immune system, impacts approximately 1.25 million individuals globally. Nevertheless, the presence of adverse effects in conventional clinical drugs necessitates further exploration of novel medications or combination therapies to mitigate these reactions and enhance their effectiveness. Hence, our intention here in this paper is to utilize the lipid nanoparticle delivery system for overcoming the volatility and hydrophobic properties of α-pinene, a naturally occurring compound renowned for its anti-inflammatory and antiviral effects, and further explore its potential pharmacological applications both in vitro and in vivo. The production of α-pinene lipid nanoparticles (APLNs) was achieved through the utilization of high pressure homogenization methods. APLNs was successfully fabricated with enhanced stability and water solubility. Meanwhile, the application of APLNs could drastically reduce the expression of lipopolysaccharide (LPS)-induced inflammation-related factors in THP-1 cells. Administration of APLNs to a mouse model of auricular swelling could effectively reduce redness and swelling in the auricles of mice as well. Furthermore, APLNs were also found to alleviate skin damage in mice with Imiquimod (IMQ)-induced psoriasis model, as well as decrease the levels of psoriasis-related protein nuclear factor kappa-B (NF-κB) and interleukin-17 (IL-17), interleukin-23 (IL-23), and other inflammation-related cytokines. More importantly, utilization of APLNs successfully mitigated the systemic inflammatory reactions in mice, resulting in the reduction of spleen-to-body ratio (wt%) and of inflammatory cytokines' expression in the serum. Overall, our results suggest that with the help of lipid nanoparticle encapsulation, APLNs possess a better pharmacological effect in anti-inflammation and could potentially serve as an anti-psoriasis drug.

Wire injury induced moderate aortic valve stenosis in mice is accompanied by a chronic systemic inflammatory reaction

Abstract Background Patients with aortic valve stenosis (AS) have elevated serum levels of inflammation markers, and blood cytokine levels correlate with ventricular recovery after transcatheter aortic valve replacement. While presence of inflammatory processes in stenotic aortic valves is acknowledged, no systematic characterization of the systemic immune reaction upon AS has been performed, yet. Purpose Hypothesis of this study was that AS induces a systemic inflammatory reaction linked with local processes in the heart. Methods AS was induced by wire injury (WI) or sham surgery was performed in 3-4 months old male C57Bl/6J mice. AS severity, left ventricular (LV) function and hypertrophy indices were assessed with echocardiography (echo). After four weeks, explanted organs were weighted, and flow cytometry identified total leukocyte numbers in blood, heart, and peripheral immune organs (spleen, liver, lymph nodes) and uptake of Cy5-labelled perfluorocarbon nanoemulsions by whole blood leukocytes. Costimulatory molecules (flow cytometry, RT-qPCR) and cytokines (RT-qPCR, Multiplex Immunoassay) were analysed in heart, peripheral immune organs and blood. Results R-values were positive for spleen weight correlation with AS severity, cardiac function and mass suggesting cardiac hypertrophy to preserve heart function. Myocardial myeloid and T cell r-values were positive or negative, respectively, for correlation with spleen and liver weight, hinting at T cell recruitment from peripheral stores. Immune cell numbers in peripheral lymph nodes and spleen showed negative r-values for correlation with heart hypertrophy indices; number of liver myeloid cells correlated negatively with LV wall thickness (monocytes=-0.8571, p=0.0238) suggesting immune cell recruitment to hypertrophic hearts. Cytokine mRNA levels trended mainly towards increase in heart and regional lymph nodes and reduction in spleen and liver after WI. Correlation with echo was more homogeneous after WI, with r-values mainly positive, except of T cell activation markers, for aortic valves, but negative for myocardium, hinting at mechanisms preserving heart function. Correlating unchanged cytokine protein levels in myocardium and plasma with echo, r-values were mostly positive for myocardium, and were more positive than in sham for plasma. Echo and costimulatory molecule correlation showed a more homogeneous pattern in WI, with mostly positive r-values in myocardium and periphery, while most r-values were negative in sham. Together with reduced PFC-uptake by lymphatic cells, this hints at systemic immune cell activation in AS. Conclusion The results suggest that number and activation state of leukocytes in peripheral organs are directly linked to cardiac processes in AS. Considering the pathological value of inflammation, it is crucial to in future studies find out if modulation of the systemic inflammatory reaction relieves severity of AS and prevents development of heart failure.

Role of Preoperative Inflammatory Blood Cell Indexes as a Postoperative Risk Predictor Among Patients Undergoing On-Pump Cardiac Surgery.

Estimating patient risk before heart surgery (HS) is crucial. Perioperative inflammation is associated with several complications and mortality. This study investigated blood cell count inflammatory indices (BCCII) to predict risks, including neutrophil-to-lymphocyte ratio (NLR), derivate NLR (DNLR), neutrophil-to-platelet-lymphocyte ratio (NLPR), lymphocyte-to-monocyte ratio, platelet-to-lymphocyte ratio (PLR), Systemic Inflammatory Index (SII), Systemic Inflammatory Reaction Index (SIRI), and Aggregate Index of Systemic Inflammation (AISI). Data from a cohort of patients undergoing on-pump HS at a single center in Brazil were retrospectively analyzed. Data were obtained from medical records and a laboratory analyzer, and SPSS version 20.0 was used for index calculations and statistical analyses. In total, 444 surgeries were performed, and 40 in-hospital deaths occurred. Except for PLR, all other indexes were independent predictors of death after multivariate adjustment (all p < 0.05). Discrimination performance was absent for PLR and AISI, and NLR, NLPR, and DNLR demonstrated the best area under the receiver operating characteristic curve (AUC > 0.7; all p < 0.0001). For survivors (n = 404), all indexes exhibited a correlation with the length of hospital stay (all p < 0.03), and NLR, NLPR, and DNLR were predictors (p < 0.026) of poor operative outcomes (acute myocardial infarction, cerebrovascular attack, cardiac arrest, low cardiac output, prolonged mechanical ventilation, renal failure, and sepsis). All BCCII scores were associated with length of hospital stay. Apart from PLR, all indexes were independent predictors of in-hospital mortality. Accuracy was highest for NLR, NLPR, and DNLR; for survivors, these three factors were good predictors of poor operative outcomes.

SIR-EN-New Biomarker for Identifying Patients at Risk of Endometrial Carcinoma in Abnormal Uterine Bleeding at Menopause.

This study aimed to evaluate the efficacy of a new biomarker, termed SIR-En, in identifying patients at risk of endometrial carcinoma among those presenting with abnormal uterine bleeding during menopause. A retrospective case-control analysis was conducted on 242 women with menopausal abnormal uterine bleeding and endometrial thickness ≥ 4 mm. Peripheral blood samples were collected within 7 days before histological diagnosis. systemic inflammatory reaction (SIR) indices were calculated, including NLR, MLR, PLR, and SII. SIR-En was derived by multiplying SII and endometrial thickness. Statistical analyses, including multivariate linear regression and ROC curve analysis, were performed to assess the diagnostic capability of SIR-En. Patients were categorized into endometrial hyperplasia (50 patients) and endometrial cancer (192 patients) groups. The SIR-En index was significantly higher in the carcinoma group (8710 vs. 6420; p = 0.003). The ROC curve for SIR-En had an AUC of 0.6351 (95% CI: 0.5579-0.7121). Using Youden's method, the optimal SIR-En cutoff was 13,806, showing a specificity of 0.940 and a positive predictive value of 0.957. Combining systemic inflammatory indices with endometrial thickness, the SIR-En index can effectively distinguish between endometrial hyperplasia and carcinoma in menopausal women with abnormal uterine bleeding. Despite the retrospective design, the identified cutoff's high specificity and positive predictive value support its potential utility in clinical practice. Further prospective studies are required to validate these findings and optimize clinical application.

Correlation between gut microbiota and musculoskeletal diseases

Dysbiosis of the microbiota is linked to an increase in intestinal permeability and a simultaneous reduction in antioxidant functions, contributing to the creation of a chronic inflammatory state that results in weakening and fragility of the musculoskeletal system. Dysbiosis can, in fact, negatively influence the immune system, leading to an increase in the production of pro-inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α), which can damage joint tissue. In this regard, the European Society for Clinical and Economic Aspects of Osteoporosis, Osteoarthritis and Musculoskeletal Diseases (ESCEO) recently stated that several risk factors for osteoarthritis (OA) can interact with the intestinal microbiota, which appears to be a critical determinant of metabolism and bioavailability of OA drugs. Recent scientific evidence shows that changes in the composition of the microbiota, altering intestinal permeability and thus allowing leakage of bacteria and their components into the bloodstream, can trigger a systemic inflammatory reaction capable of contributing to the progression of joint degeneration. It is known that physical exercise can play a key role not only in improving the functioning of patients suffering from OA, but also in terms of modulating the composition of the microbiota, strengthening the immunity of the intestinal mucosa; furthermore, obesity, which is an important risk factor for the development of OA, has been linked to both dysbiosis of the gut microbiota and chronic inflammation, suggesting that complex connections exist between microbiota, obesity, and musculoskeletal pathologies. Intestinal dysbiosis can have a negative impact on bone health by influencing the RANK/RANKL/OPG pathway; in this context, it has been hypothesized that food supplements, prebiotics and probiotics can support an adequate balance of the intestinal microbiota and consequently bone health. In conclusion, the correlation between gut dysbiosis and musculoskeletal diseases, such as OA, sarcopenia, and osteoporosis, is still the subject of research and further studies are necessary to fully understand the underlying mechanisms. KEY WORDS: Microbiota, musculoskeletal diseases, gut, osteosarcopenia.

Biomarkers as Predictors of Severity Multiple Organ Dysfunctions After Cardiac Surgery (Case Series Study)

Introduction. The question of the possibility of predicting the severity of multiple organ failure that developed in the postoperative period in cardiac surgery patients is not sufficiently covered in the literature.The aim of the study is determine the prospects of studying the relationship between the level of biomarkers and the severity of multiple organ failure in patients undergoing cardiac surgery.Materials and methods. A series of seven observations was carried out on the clinical course of the immediate postoperative period in patients who underwent elective cardiac surgery. In the preoperative period, the risk of in-hospital mortality was assessed using the EuroSCORE II scale and the presence of multiple organ failure using the SOFA scale. Before surgery and at the beginning of the first postoperative day, the plasma level of presepsin was studied. Also, at the beginning of the first postoperative day, plasma levels of transferases and troponia T were studied, and the severity of multiple organ failure was analyzed using the SOFA scale. The length of stay of patients in the ICU was assessed. The results of the study are presented in the form of tables and graphs and subjected to visual analysis.Results. The obtained data do not allow us to accurately link the level of tissue damage markers (aspartate aminotransferase, alanine aminotransferase, troponin T) and the marker of the systemic inflammatory reaction presepsin with the severity of multiple organ failure in patients after cardiac surgery. At the same time, they do not exclude the existence of such a connection.

Complete Blood Collection-based Systemic Inflammation Biomarkers as a Severity Biomarker in Alopecia Areata: A Cross-sectional Study.

Previous studies have suggested that alopecia areata (AA) is an organ-specific disease characterized by loss of immune privilege of hair follicles. However, an increasing body of research indicates that it not only affects the skin but may also be accompanied by systemic inflammatory reactions. Therefore, searching for simple and easily available biomarkers to describe the underlying systemic inflammation in AA patients is of great clinical significance. Complete blood collection-based systemic inflammation biomarkers have been shown to be associated with the severity and prognosis of various skin and autoimmune diseases. They involve multiple cell lineages and can reveal different pathways of immune-inflammatory responses. The aim of this study was to investigate the level of complete blood collection-based systemic inflammation biomarkers in patients with AA, and to analyse their relationship with the disease severity. A total of 302 AA patients and 296 healthy controls were included in this study and the neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), systemic immune inflammation index (SII), and white blood cell/lymphocyte ratio (WLR) were calculated. The differences in these indicators between the 2 groups were compared, and the relationship between NLR, PLR, SII, WLR, and the risk of severe AA were analysed. AA patients had higher NLR, SII, and WLR compared with healthy controls (p = 0.004, 0.002, and 0.002 respectively). PLR and SII were higher in the severe AA group compared with the mild-to-moderate AA patients (p = 0.005 and 0.011 respectively). The risk of severe AA increased with the increasing of PLR, SII, NLR, and WLR (p for trend was 0.001, 0.006, 0.022, and 0.021, respectively). The levels of systemic inflammation biomark-ers in AA patients are higher than in healthy people. NLR, PLR, SII, and WLR are risk factors for severe AA, suggesting a close association between systemic inflammation and disease occurrence in AA patients.

Role of oxidative stress in the occurrence of periodontitis in the background of metabolically associated diseases

Oxidative stress is an important component of slow pathological inflammation in metabolic diseases and chronic periodontitis. Periodontal diseases are interrelated with somatic pathology. This results in both local and systemic inflammatory reactions, which increases the risk of the onset and progression of somatic diseases. Psychological stress associated with oxidative stress with increased production of oxidants and oxidative damage is an important factor in the etiology of both periodontitis and metabolic disorders. Objective — to analyze the possible bond between chronic periodontitis, 8‑hydroxydeoxyguanosine (8‑OHdG), as a biomarker of oxidative stress, and cortisol in persons with metabolic‑associated diseases. Materials and methods. 68 people were surveyed, of which 47 had metabolically ‑associated diseases that were diagnosed according to domestic and international criteria. The control group consisted of systematically healthy people, representative by age and article that did not receive dental therapeutic and preventive measures for the last 6 months. The diagnosis of «chronic periodontitis» was carried out in accordance with existing recommendations provided at World Workshop on the Classification of Periodontal and Peri­Implant Diseases and Conditions (2017). The concentration of 8‑OHdG in plasma and the level of saliva cortisol were measured by the immunoenzyme method according to the manufacturer’s instructions. The arithmetic mean, standard error, and Student’s t‑test were calculated to compare the characteristics. Pearson’s correlation test was used to identify the relationship between the indicators. Results. Each of the groups had patients who had a healthy periodontium and those who had chronic periodontitis. The 8‑OHdG level in the main group was much higher (p&lt;0.001) than in the control group, and the difference in the concentration of 8‑OHdG was also observed between those patients who had periodontal disease and those who had a healthy periodontium. The level of salivary cortisol in patients with chronic periodontitis was much higher compared to patients with clinically healthy periodontium (p=0.00). A correlation was found between cortisol levels and the presence of periodontal disease (r=0.619; p=0.00), between plasma 8OHdG levels and the epithelial attachment loss index (r=0.537; p=0.00), the hygiene index (r=0.599; p=0.00), and between 8OHdG levels and salivary cortisol concentrations (r=0.292; p=0.036). Conclusions. Oxidative stress plays an important role in delayed pathological inflammation in both metabolic diseases and chronic periodontitis. 8‑OHdG is a key and universal biomarker for the evaluation of DNA oxidative damage. There is a positive relationship between the oxidative marker, periodontitis and cortisol.

9224 Evaluation of Safety and Efficacy of Administration of Estradiol by Subcutaneous Injection in Transgender Women

Abstract Disclosure: H. Perkins: None. H. Moreau: None. J. LaBudde: None. W. Craig: None. D.I. Spratt: None. Background: Off-label use of subcutaneous (SC) estradiol (E2) by transgender women for gender-affirming hormone therapy is becoming increasingly widespread, despite a lack of published data regarding its efficacy and safety. Methods: This retrospective cohort study used manual data extraction from electronic medical records for patients ages 18-79 years, currently or previously receiving oral (PO) 17-beta E2 or SC E2 valerate therapy at Maine Medical Center Reproductive Endocrinology and Infertility and Gender Clinics between 5/30/14-5/30/23, who completed E2 dose adjustments aimed at obtaining a serum E2 of 75-250 pg/mL. SC E2 was administered weekly and PO E2 daily. Serum testosterone (T), estrone (E1), and sex hormone binding globulin (SHBG) levels were recorded if drawn within 90 days of E2 measurement on an optimized dose. In a subset of patients receiving SC E2, serum E2 levels were drawn 1-2 days post- and pre-injection. All patient-reported local or systemic reactions were recorded. Data are expressed as median (full range) or mean ± standard deviation (full range). Results: 45 patients were included with no significant differences in demographics between PO (n=20) and SC (n=25) groups. Both groups achieved therapeutic E2 levels after dose adjustment without statistical difference in mean serum E2 (pg/mL) between SC and PO groups: 148.7 ± 41.0 (78-226) vs. 151.6 ± 31.7 (97-208); p=0.79 by t-test. In patients without orchiectomy, both SC and PO groups had T (ng/dL) suppressed to normal adult female range (10-55) in 11/12 SC patients and in 17/17 PO patients; median T was 14.0 (7.7-86) for SC vs. 20.6 (3.4-42) for PO; p=0.98 by Mann-Whitney U test. Median E2 dose (mg/week) in the SC group was 3.0 (1.4-6.0) and in the PO group was 42.0 (28-84). Median serum E1 (pg/mL) was lower in the SC group than the PO group: 73.5 (29-115), n=12 vs. 907 (596-1601), n=5, p&amp;lt;0.001 by Mann-Whitney U test. E1/E2 ratio was lower in the SC group than the PO group: 0.4 ± 0.1 vs. 8.0 ± 2.9; p&amp;lt;0.001 by t-test. There was no significant difference in median serum SHBG (nmol/L) between SC and PO groups: 71.7 (19.3-146.7), n=15 vs. 73.4 (51.0-187.4), n=14; p=0.20 by Mann-Whitney U test. Mean post- and pre-dose serum E2 levels in 5 patients were 138.0 ± 24.0 and 88.0 ± 57.6, respectively. There was one transient local reaction and no systemic allergic or inflammatory reactions in the SC group with no systemic reactions in the PO group. Conclusion: These results demonstrate that SC E2 at doses of 1.4-6 mg/week can be administered safely to achieve therapeutic serum E2 levels and suppress serum T levels to within the normal female range. Our preliminary data reveal lower serum E1 levels in the SC group than in the PO group suggesting reduced hepatic first-pass metabolism with SC administration; this pattern was not seen with serum SHBG levels. Post- and pre-dose serum E2 levels revealed only a mild decline in E2 levels between injections. Presentation: 6/3/2024

6961 Evaluation of Safety and Efficacy of Administration of Estradiol by Subcutaneous Injection in Transgender Women

Abstract Disclosure: H. Perkins: None. H. Moreau: None. J. LaBudde: None. W. Craig: None. D.I. Spratt: None. Background: Off-label use of subcutaneous (SC) estradiol (E2) by transgender women for gender-affirming hormone therapy is becoming increasingly widespread, despite a lack of published data regarding its efficacy and safety. Methods: This retrospective cohort study used manual data extraction from electronic medical records for patients ages 18-79 years, currently or previously receiving oral (PO) 17-beta E2 or SC E2 valerate therapy at Maine Medical Center Reproductive Endocrinology and Infertility and Gender Clinics between 5/30/14-5/30/23, who completed E2 dose adjustments aimed at obtaining a serum E2 of 75-250 pg/mL. SC E2 was administered weekly and PO E2 daily. Serum testosterone (T), estrone (E1), and sex hormone binding globulin (SHBG) levels were recorded if drawn within 90 days of E2 measurement on an optimized dose. In a subset of patients receiving SC E2, serum E2 levels were drawn 1-2 days post- and pre-injection. All patient-reported local or systemic reactions were recorded. Data are expressed as median (full range) or mean ± standard deviation (full range). Results: 45 patients were included with no significant differences in demographics between PO (n=20) and SC (n=25) groups. Both groups achieved therapeutic E2 levels after dose adjustment without statistical difference in mean serum E2 (pg/mL) between SC and PO groups: 148.7 ± 41.0 (78-226) vs. 151.6 ± 31.7 (97-208); p=0.79 by t-test. In patients without orchiectomy, both SC and PO groups had T (ng/dL) suppressed to normal adult female range (10-55) in 11/12 SC patients and in 17/17 PO patients; median T was 14.0 (7.7-86) for SC vs. 20.6 (3.4-42) for PO; p=0.98 by Mann-Whitney U test. Median E2 dose (mg/week) in the SC group was 3.0 (1.4-6.0) and in the PO group was 42.0 (28-84). Median serum E1 (pg/mL) was lower in the SC group than the PO group: 73.5 (29-115), n=12 vs. 907 (596-1601), n=5, p&amp;lt;0.001 by Mann-Whitney U test. E1/E2 ratio was lower in the SC group than the PO group: 0.4 ± 0.1 vs. 8.0 ± 2.9; p&amp;lt;0.001 by t-test. There was no significant difference in median serum SHBG (nmol/L) between SC and PO groups: 71.7 (19.3-146.7), n=15 vs. 73.4 (51.0-187.4), n=14; p=0.20 by Mann-Whitney U test. Mean post- and pre-dose serum E2 levels in 5 patients were 138.0 ± 24.0 and 88.0 ± 57.6, respectively. There was one transient local reaction and no systemic allergic or inflammatory reactions in the SC group with no systemic reactions in the PO group. Conclusion: These results demonstrate that SC E2 at doses of 1.4-6 mg/week can be administered safely to achieve therapeutic serum E2 levels and suppress serum T levels to within the normal female range. Our preliminary data reveal lower serum E1 levels in the SC group than in the PO group suggesting reduced hepatic first-pass metabolism with SC administration; this pattern was not seen with serum SHBG levels. Post- and pre-dose serum E2 levels revealed only a mild decline in E2 levels between injections. Presentation: 6/3/2024

9224 Evaluation of Safety and Efficacy of Administration of Estradiol by Subcutaneous Injection in Transgender Women

Abstract Disclosure: H. Perkins: None. H. Moreau: None. J. LaBudde: None. W. Craig: None. D.I. Spratt: None. Background: Off-label use of subcutaneous (SC) estradiol (E2) by transgender women for gender-affirming hormone therapy is becoming increasingly widespread, despite a lack of published data regarding its efficacy and safety. Methods: This retrospective cohort study used manual data extraction from electronic medical records for patients ages 18-79 years, currently or previously receiving oral (PO) 17-beta E2 or SC E2 valerate therapy at Maine Medical Center Reproductive Endocrinology and Infertility and Gender Clinics between 5/30/14-5/30/23, who completed E2 dose adjustments aimed at obtaining a serum E2 of 75-250 pg/mL. SC E2 was administered weekly and PO E2 daily. Serum testosterone (T), estrone (E1), and sex hormone binding globulin (SHBG) levels were recorded if drawn within 90 days of E2 measurement on an optimized dose. In a subset of patients receiving SC E2, serum E2 levels were drawn 1-2 days post- and pre-injection. All patient-reported local or systemic reactions were recorded. Data are expressed as median (full range) or mean ± standard deviation (full range). Results: 45 patients were included with no significant differences in demographics between PO (n=20) and SC (n=25) groups. Both groups achieved therapeutic E2 levels after dose adjustment without statistical difference in mean serum E2 (pg/mL) between SC and PO groups: 148.7 ± 41.0 (78-226) vs. 151.6 ± 31.7 (97-208); p=0.79 by t-test. In patients without orchiectomy, both SC and PO groups had T (ng/dL) suppressed to normal adult female range (10-55) in 11/12 SC patients and in 17/17 PO patients; median T was 14.0 (7.7-86) for SC vs. 20.6 (3.4-42) for PO; p=0.98 by Mann-Whitney U test. Median E2 dose (mg/week) in the SC group was 3.0 (1.4-6.0) and in the PO group was 42.0 (28-84). Median serum E1 (pg/mL) was lower in the SC group than the PO group: 73.5 (29-115), n=12 vs. 907 (596-1601), n=5, p&amp;lt;0.001 by Mann-Whitney U test. E1/E2 ratio was lower in the SC group than the PO group: 0.4 ± 0.1 vs. 8.0 ± 2.9; p&amp;lt;0.001 by t-test. There was no significant difference in median serum SHBG (nmol/L) between SC and PO groups: 71.7 (19.3-146.7), n=15 vs. 73.4 (51.0-187.4), n=14; p=0.20 by Mann-Whitney U test. Mean post- and pre-dose serum E2 levels in 5 patients were 138.0 ± 24.0 and 88.0 ± 57.6, respectively. There was one transient local reaction and no systemic allergic or inflammatory reactions in the SC group with no systemic reactions in the PO group. Conclusion: These results demonstrate that SC E2 at doses of 1.4-6 mg/week can be administered safely to achieve therapeutic serum E2 levels and suppress serum T levels to within the normal female range. Our preliminary data reveal lower serum E1 levels in the SC group than in the PO group suggesting reduced hepatic first-pass metabolism with SC administration; this pattern was not seen with serum SHBG levels. Post- and pre-dose serum E2 levels revealed only a mild decline in E2 levels between injections. Presentation: 6/3/2024

6961 Evaluation of Safety and Efficacy of Administration of Estradiol by Subcutaneous Injection in Transgender Women

Abstract Disclosure: H. Perkins: None. H. Moreau: None. J. LaBudde: None. W. Craig: None. D.I. Spratt: None. Background: Off-label use of subcutaneous (SC) estradiol (E2) by transgender women for gender-affirming hormone therapy is becoming increasingly widespread, despite a lack of published data regarding its efficacy and safety. Methods: This retrospective cohort study used manual data extraction from electronic medical records for patients ages 18-79 years, currently or previously receiving oral (PO) 17-beta E2 or SC E2 valerate therapy at Maine Medical Center Reproductive Endocrinology and Infertility and Gender Clinics between 5/30/14-5/30/23, who completed E2 dose adjustments aimed at obtaining a serum E2 of 75-250 pg/mL. SC E2 was administered weekly and PO E2 daily. Serum testosterone (T), estrone (E1), and sex hormone binding globulin (SHBG) levels were recorded if drawn within 90 days of E2 measurement on an optimized dose. In a subset of patients receiving SC E2, serum E2 levels were drawn 1-2 days post- and pre-injection. All patient-reported local or systemic reactions were recorded. Data are expressed as median (full range) or mean ± standard deviation (full range). Results: 45 patients were included with no significant differences in demographics between PO (n=20) and SC (n=25) groups. Both groups achieved therapeutic E2 levels after dose adjustment without statistical difference in mean serum E2 (pg/mL) between SC and PO groups: 148.7 ± 41.0 (78-226) vs. 151.6 ± 31.7 (97-208); p=0.79 by t-test. In patients without orchiectomy, both SC and PO groups had T (ng/dL) suppressed to normal adult female range (10-55) in 11/12 SC patients and in 17/17 PO patients; median T was 14.0 (7.7-86) for SC vs. 20.6 (3.4-42) for PO; p=0.98 by Mann-Whitney U test. Median E2 dose (mg/week) in the SC group was 3.0 (1.4-6.0) and in the PO group was 42.0 (28-84). Median serum E1 (pg/mL) was lower in the SC group than the PO group: 73.5 (29-115), n=12 vs. 907 (596-1601), n=5, p&amp;lt;0.001 by Mann-Whitney U test. E1/E2 ratio was lower in the SC group than the PO group: 0.4 ± 0.1 vs. 8.0 ± 2.9; p&amp;lt;0.001 by t-test. There was no significant difference in median serum SHBG (nmol/L) between SC and PO groups: 71.7 (19.3-146.7), n=15 vs. 73.4 (51.0-187.4), n=14; p=0.20 by Mann-Whitney U test. Mean post- and pre-dose serum E2 levels in 5 patients were 138.0 ± 24.0 and 88.0 ± 57.6, respectively. There was one transient local reaction and no systemic allergic or inflammatory reactions in the SC group with no systemic reactions in the PO group. Conclusion: These results demonstrate that SC E2 at doses of 1.4-6 mg/week can be administered safely to achieve therapeutic serum E2 levels and suppress serum T levels to within the normal female range. Our preliminary data reveal lower serum E1 levels in the SC group than in the PO group suggesting reduced hepatic first-pass metabolism with SC administration; this pattern was not seen with serum SHBG levels. Post- and pre-dose serum E2 levels revealed only a mild decline in E2 levels between injections. Presentation: 6/3/2024