Plasma LDH Research Articles

Cucurbitacin-E; Curvularia lunata Secondary Metabolite as a BRCA 1 and 2 Regulator in Mice Associated Breast Cancer

Background: Cucurbitacin-induced apoptosis and inhibition of cell growth can render several cancers ineffective. The microbial transformation of cucurbitacin-E-glucoside to cucurbitacin- E was carried out by Curvularia lunata (NRRL 2178). Moreover, in vitro anticancer activity against the MCF-7 cell line as well as in vivo anticancer activity against dimethylbenz (a) anthracene (DMBA)-induced breast cancer in mice using cucurbitacin-E was evaluated. Methods: The cucurbitacin-E-glucoside was biotransformed by Curvularia lunata to cucurbitacin- E, and the isolated compound was tested in vitro against the MCF-7 cell line, and its IC50 was calculated. LD50 of cucurbitacin-E was estimated in mice, and its protective activity against DMBA-induced cancer in mice was studied. Breast cancer induction was done by a single-dose subcutaneous administration of DMBA (50 mg). Plasma ALT, AST, ALP, and LDH, as well as GSH, SOD, GPx, MDA, TNF-α, IL-6, and tumor suppressor P53 assays, were used to assess cucurbitacin- E's capacity to protect the liver and breast against DMBA-induced toxicity. Moreover, by assessing the gene expression of tumor suppressor genes (BRCA 1 and 2) and conducting histopathological analysis, the suppressive effect of cucurbitacin-E was examined. Results: The IC50 value of cucurbitacin-E against MCF-7 cell lines equals 72.15 ± 0.64 μg/ml. LD50 of cucurbitacin-E given orally in adult mice is equal to 1200 mg/kg b.w. The levels of plasma ALT, AST, ALP, and LDH were decreased significantly in DMBA-treated mice when administered with cucurbitacin-E at 1/50 LD50 (24 mg/kg/b.w.) and 1/20 LD50 (60 mg/kg/b.w.). In breast tissue, the levels of GSH, SOD, GPx, and P53 were significantly increased, as were decreased levels of TNF-α, IL-6, P53, and MDA. Conversely, there was a downregulation in the mRNA expression levels of BRCA1 and BRCA2. The histopathological analysis revealed that cucurbitacin-E management improved the tissue architecture of breast tumors. Conclusions: These findings demonstrate the ability of cucurbitacin-E to inhibit cancer cells in the rat breast by controlling oxidative stress and inflammatory biomarkers, as well as downregulating the mRNA expression levels of BRCA1 and BRCA2.

Characterization and Clinical Significance of Hemolysis After Pulsed Field Ablation for Atrial Fibrillation: Results of a Multicenter Analysis.

Pulsed field ablation (PFA) is increasingly used in clinical practice for the treatment of atrial fibrillation. While the susceptibility of erythrocytes to electroporation is well established, the effect of cardiac PFA technologies on hemolysis has remained underreported. The aim of this study was to investigate the incidence, severity, and clinical impact of PFA-induced hemolysis. We included n=145 patients undergoing atrial fibrillation catheter ablation with a pentaspline PFA catheter (biphasic, bipolar pulses of 2 kV) and n=70 patients receiving radiofrequency ablation (40-90 W) at 4 high-volume European centers. The lesion set comprised pulmonary vein isolation for paroxysmal atrial fibrillation and pulmonary vein isolation±additional lesions for persistent atrial fibrillation. Hemolysis and renal function biomarkers were analyzed in blood samples at baseline, at the end of ablation, and 24 hours after the procedure. Baseline characteristics were well balanced between groups (overall mean, 65.7±9.4 years; 69.3% male). The ablation procedures comprised a mean of 61.6±27.4 PFA deliveries and 26.3±15.0 minute RF duration. Hemolysis was detected in 94.3% versus 6.8% of patients after PFA versus radiofrequency ablation (P<0.001): PFA was associated with significantly lower haptoglobin levels (0.5±0.4 versus 1.0±0.4 g/L), while free plasma hemoglobin (592.8±330.6 versus 147.8±183.0 mg/L), bilirubin (21.3±11.3 versus 14.8±8.8 µmol/L), and LDH (lactate dehydrogenase, 352.7±115.7 versus 253.2±56.5 U/L) were significantly higher after PFA versus radiofrequency ablation (all P<0.001). Hemolysis correlated with the number of PFA deliveries (r=0.62 [95% CI, 0.33-0.80]; P<0.001), with the highest severity occurring ≥54 PFA deliveries. After PFA, hemoglobinuria occurred in 36.4%, while creatinine increase was higher in patients with baseline glomerular filtration rate <50 mL/min versus baseline glomerular filtration rate >50 mL/min (Δcrea, 27.0±103.1 versus -0.2±12.1 µmol/L; P=0.010). Intravascular hemolysis is a frequent finding after PFA and increases with the number of PFA deliveries. Until the clinical impact of PFA-associated hemolysis is fully elucidated, a careful titration of PFA deliveries during the ablation procedure is warranted.

Short-Term Atorvastatin Therapy in Healthy Individuals Results in Unaltered Plasma MMP Levels and Disrupted MMP-7 Correlation with Blood Lipids and Blood Count-Derived Inflammatory Markers.

Background: Matrix metalloproteinases (MMPs) play an important role in the pathophysiology of atherosclerosis. Reportedly, statins can decrease MMP activity in patients with atherosclerotic cardiovascular disease, but this effect has not been studied in healthy individuals. Methods: MMPs 2, 7, and 9 and several other parameters were measured before and after a four-week course of moderate-dose atorvastatin (20 mg/day) in 21 healthy individuals. Results: Atorvastatin treatment resulted in lower total cholesterol, LDL-cholesterol, non-HDL-cholesterol, and triglycerides (p < 0.001 for all), but higher levels of plasma enzymes AST, ALT, CK, and LDH (p < 0.05 for all). No effect of atorvastatin on plasma MMP median concentrations was recorded. Before treatment, moderate positive significant correlations were found between MMP-7 and age, blood lipids, and blood count-derived inflammatory markers. Pre-treatment MMP-7 was best predicted by the total cholesterol-to-HDL cholesterol ratio in a remnant cholesterol-weighted least squares regression model. After atorvastatin treatment, MMP-7 no longer correlated with these markers. Conclusions: While the effect of statins on plasma MMPs in atherosclerosis is controversial, short-term moderate-dose atorvastatin treatment does not seem to affect levels of MMPs 2, 7, and 9 in healthy individuals. However, an intriguing correlation between MMP-7 and atherosclerosis-related blood lipids and neutrophil-associated inflammatory biomarkers seems to be disrupted by atorvastatin independently of hsCRP, possibly via pleiotropic effects.

Diallyl trisulfide induces pyroptosis and impairs lung CSC-like properties by activating the ROS/Caspase 1 signaling pathway

Lung cancer stem cells (CSCs) drive continuous cancer growth and metastatic dissemination; thus, there is an urgent requirement to acquire effective therapeutic strategies for targeting lung CSCs. Diallyl trisulfide (DATS), a garlic organosulfide, possesses suppressive potential in lung cancer; however, its underlying mechanism is still unclear. In this study, we identified DATS as a pyroptosis inducer in lung cancer cells. DATS-treated A549 and H460 cells exhibited pyroptotic cell death, with characteristic large bubbles appearing on their plasma membrane and LDH release. DATS induced cell death, arrested the cell cycle at the G2/M phase, and inhibited colony formation in lung cancer cells. Meanwhile, we found that DATS significantly suppressed the malignant features by impairing lung CSC-like properties, including sphere formation ability, CD133 positive cell number, and lung CSCs marker expression. Mechanistically, DATS induced cell pyroptosis via increasing the expression of NLRP3, ASC, Pro Caspase 1, Cleaved Caspase 1, GSDMD, GSDMD-N, and IL-1β. The verification experiments showed that the effects of DATS on pyroptosis and lung CSC-like properties were weakened after Caspase 1 inhibitor VX-765 treatment, indicating that DATS activated NLRP3 inflammasome-mediated pyroptosis by targeting Caspase 1 in lung cancer cells. Moreover, DATS increased ROS overproduction and mitochondrial dysfunction, which contributed to DATS-induced pyroptosis of lung cancer cells. NAC treatment reversed the effects of DATS on pyroptosis and CSC-like properties. In vivo experiment further confirmed that DATS restrained tumor growth. Together, our results suggest that DATS promotes pyroptosis and impairs lung CSC-like properties by activating ROS/Caspase 1 signaling pathway, thereby retarding lung cancer progression.

Artificial intelligence-based model for dose prediction of sertraline in adolescents: a real-world study

ABSTRACT Background Variability exists in sertraline pharmacokinetic parameters in individuals, especially obvious in adolescents. We aimed to establish an individualized dosing model of sertraline for adolescents with depression based on artificial intelligence (AI) techniques. Methods Data were collected from 258 adolescent patients treated at the First Hospital of Hebei Medical University between December 2019 to July 2022. Nine different algorithms were used for modeling to compare the prediction abilities on sertraline daily dose, including XGBoost, LGBM, CatBoost, GBDT, SVM, ANN, TabNet, KNN, and DT. Performance of four dose subgroups (50 mg, 100 mg, 150 mg, and 200 mg) were analyzed. Results CatBoost was chosen to establish the individualized medication model with the best performance. Six important variables were found to be correlated with sertraline dose, including plasma concentration, PLT, MPV, GL, A/G, and LDH. The ROC curve and confusion matrix exhibited the good prediction performance of CatBoost model in four dose subgroups (the AUC of 50 mg, 100 mg, 150 mg, and 200 mg were 0.93, 0.81, 0.93, and 0.93, respectively). Conclusion The AI-based dose prediction model of sertraline in adolescents with depression had a good prediction ability, which provides guidance for clinicians to propose the optimal medication regimen.

Abstract C068: Factors associated with primary resistance to immune checkpoint blockade in early phase clinical trials

Abstract Introduction: Immune checkpoint blockade (ICB) therapies, particularly anti-PD1/PD-L1 monoclonal antibodies, have transformed the field of medical oncology. However, many patients with advanced cancer demonstrate primary resistance to ICB strategies. The identification of objective clinical and biological characteristics associated with treatment resistance is crucial for identifying patients who are most likely not to benefit from conventional or current ICB combinations strategies. Studies and trials dedicated to these patients are essential to overcome such resistance. Methods: We prospectively enrolled patients treated with ICB at Gustave Roussy between 2018 and 2022 in the PREMIS study (NCT03984318). We examined a sub-cohort of patients treated at the early drug development department (DITEP). We identified pre-treatment (baseline) clinical and biological characteristics, including patients’ oncogenic alterations documented via circulating tumor DNA (ctDNA) sequencing in our STING study (NCT04932525), to find a significant difference between patients according to their best objective antitumor response (BOR; complete response [CR], partial response [PR], stable disease [SD], or progressive disease [PD]). Results: We analyzed a total of 227 patients, with paired ctDNA available for 96 patients. Patients were 52% women and 48% men, averaging 59 years old, with 24 different solid tumor types and enrolled in 50 different early phase clinical trials. Eleven percent of patients had received prior ICB treatment. BOR were PD (61%), SD (20%), PR (15%), and CR (4%). We identified clinical characteristics significantly different between patients according to their BOR: performance status (PS), number of previous lines of therapy, number of metastases, presence of liver metastases, absolute neutrophils count (ANC), absolute lymphocyte count (ALC), neutrophil to lymphocyte ratio, CRP, GGT and LDH plasma concentrations, and MLH1 mutations. Acording these results, a score from 0 to 5 was calculated according to the presence of the following characteristics (yes=+1): presence of liver metastases, ALC below the lower limit of the normal (LLN), ANC above the upper limit of the normal (ULN), CRP &amp;gt; ULN, LDH &amp;gt; ULN. Patients with a score ≥ 4 did not develop a response during ICB treatments (n=27, 12%). Among them, 25 (92.6%) had a PS of 0-1 and 9 (33.3%) had a Royal Marsden Hospital (RMH) score of 1. Additionally, both progression-free survival (PFS) and overall survival (OS) decreased significantly with the addition of each point score. Conclusion: Patients with advanced cancer treated by ICB in early phase clinical trials with at least 4 detrimental characteristics (presence of liver metastases, ALC &amp;lt; LLN, ANC, CRP and LDH &amp;gt; ULN) had primary resistance to ICB regardless of tumor type, ICB regimen, PS or RMH. Identification of immunological and oncogenic alterations specific to these patients could support the development of targeted and personalized ICB combination trials to overcome tumor resistance. Citation Format: Stéphane Champiat, Paul Matte, Capucine Baldini, Damien Vasseur, Kaïssa Ouali, Anas Gazzah, Rastilav Bahleda, Arnaud Bayle, Sophie Postel-Vinay, Cristina Smolenschi, Madona Sakkal, Jean-Marie Michot, Antoine Hollebecque, Nathalie Chaput-Gras, Yohann Loriot, Santiago Ponce, Antoine Italiano, Aurélien Marabelle, François-Xavier Danlos. Factors associated with primary resistance to immune checkpoint blockade in early phase clinical trials [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C068.

Using Machine Learning to Identify the Relationships between Demographic, Biochemical, and Lifestyle Parameters and Plasma Vitamin D Concentration in Healthy Premenopausal Chinese Women.

Vitamin D plays a vital role in maintaining homeostasis and enhancing the absorption of calcium, an essential component for strengthening bones and preventing osteoporosis. There are many factors known to relate to plasma vitamin D concentration (PVDC). However, most of these studies were performed with traditional statistical methods. Nowadays, machine learning methods (Mach-L) have become new tools in medical research. In the present study, we used four Mach-L methods to explore the relationships between PVDC and demographic, biochemical, and lifestyle factors in a group of healthy premenopausal Chinese women. Our goals were as follows: (1) to evaluate and compare the predictive accuracy of Mach-L and MLR, and (2) to establish a hierarchy of the significance of the aforementioned factors related to PVDC. Five hundred ninety-three healthy Chinese women were enrolled. In total, there were 35 variables recorded, including demographic, biochemical, and lifestyle information. The dependent variable was 25-OH vitamin D (PVDC), and all other variables were the independent variables. Multiple linear regression (MLR) was regarded as the benchmark for comparison. Four Mach-L methods were applied (random forest (RF), stochastic gradient boosting (SGB), extreme gradient boosting (XGBoost), and elastic net). Each method would produce several estimation errors. The smaller these errors were, the better the model was. Pearson's correlation, age, glycated hemoglobin, HDL-cholesterol, LDL-cholesterol, and hemoglobin were positively correlated to PVDC, whereas eGFR was negatively correlated to PVDC. The Mach-L methods yielded smaller estimation errors for all five parameters, which indicated that they were better methods than the MLR model. After averaging the importance percentage from the four Mach-L methods, a rank of importance could be obtained. Age was the most important factor, followed by plasma insulin level, TSH, spouse status, LDH, and ALP. In a healthy Chinese premenopausal cohort using four different Mach-L methods, age was found to be the most important factor related to PVDC, followed by plasma insulin level, TSH, spouse status, LDH, and ALP.

GPX3 Variant Genotype Affects the Risk of Developing Severe Forms of COVID-19.

In SARS-CoV-2 infection, excessive activation of the immune system intensively increases reactive oxygen species levels, causing harmful hyperinflammatory and oxidative state cumulative effects which may contribute to COVID-19 severity. Therefore, we assumed that antioxidant genetic profile, independently and complemented with laboratory markers, modulates COVID-19 severity. The study included 265 COVID-19 patients. Polymorphism of GSTM1, GSTT1, Nrf2 rs6721961, GSTM3 rs1332018, GPX3 rs8177412, GSTP1 rs1695, GSTO1 rs4925, GSTO2 rs156697, SOD2 rs4880 and GPX1 rs1050450 genes was determined with appropriate PCR-based methods. Inflammation (interleukin-6, CRP, fibrinogen, ferritin) and organ damage (urea, creatinine, transaminases and LDH) markers, complete blood count and coagulation status (d-dimer, fibrinogen) were measured. We found significant association for COVID-19 progression for patients with lymphocytes below 1.0 × 109/L (OR = 2.97, p = 0.002). Increased IL-6 and CRP were also associated with disease progression (OR = 8.52, p = 0.001, and OR = 10.97, p < 0.001, respectively), as well as elevated plasma AST and LDH (OR = 2.25, p = 0.021, and OR = 4.76, p < 0.001, respectively). Of all the examined polymorphisms, we found significant association with the risk of developing severe forms of COVID-19 for GPX3 rs8177412 variant genotype (OR = 2.42, p = 0.032). This finding could be of particular importance in the future, complementing other diagnostic tools for prediction of COVID-19 disease course.

Continuous Dosing of Apohemoglobin-Haptoglobin in Beta Thalassemic Mice to Establish Safety and Efficacy

Thalassemia is a blood disorder that affects the production of hemoglobin (Hb), the crucial protein responsible for carrying and delivering oxygen to tissues. It is caused by mutations in the genes that control the production of hemoglobin, leading to decreased or absent production of one of the globin chains (alpha or beta) that make up hemoglobin. Individuals with thalassemia often need regular blood transfusions to maintain adequate Hb levels. However, frequent transfusions can cause iron overload in the body, necessitating chelation therapy to remove excess iron and prevent organ damage. The iron overload also results in higher levels of hemolysis, leading to the release of free acellular Hb and heme, which the body's natural scavenging proteins (hemopexin and haptoglobin) cannot effectively handle. To address this problem, our research group has recently explored a promising solution using a bioengineered Hb and heme scavenger called apohemoglobin-haptoglobin (apoHb-Hp) complex. This complex works as a scavenger for heme and hemoglobin from the circulation, helping to eliminate vasoconstriction, enhance perfusion, and reduce inflammation. We conducted a study to test the safety and effectiveness of apoHb-Hp in beta-thalassemic mice, and the results were quite encouraging. Initially, we treated twenty beta-thalassemic mice with different dosing regimens of apoHb-Hp, ranging from 25 to 100 mg/kg. The mice received the complex every other day for a total of three doses. After the final dose, we measured various parameters, including total bilirubin concentration, plasma LDH, and plasma iron. As the dosing amount increased, there was a progressive decrease in all three measurements, reaching a therapeutic plateau at 75 mg/kg. In he second part of the study, we treated eight beta-thalassemic mice with a dose of 80 mg/kg of apoHb-Hp for six weeks, every other day. Again, we measured total bilirubin concentration, plasma LDH, and plasma iron, and observed a significant drop compared to baseline levels at the start of the study. These promising results suggest that apoHb-Hp could be a safe and effective treatment option for beta-thalassemia. Its ability to scavenge heme and Hb, along with its positive effects on various markers, makes it a potential game-changer in the management of this genetic blood disorder. Further research and testing will be necessary to fully explore the potential of apoHb-Hp as a therapeutic option for individuals with beta-thalassemia.

Integrating serum pharmacochemistry and network pharmacology to explore the molecular mechanisms of Acanthopanax senticosus (Rupr. & Maxim.) Harms on attenuating doxorubicin-induced myocardial injury

Ethnopharmacological relevanceAcanthopanax senticosus (Rupr. & Maxim.) Harms (AS), also known as Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. or Siberian ginseng, has a rich history of use as an adaptogen, a substance believed to increase the body's resistance to stress, fatigue, and infectious diseases. As a traditional Chinese medicine, AS is popular for its cardioprotective effects which can protect the cardiovascular system from hazardous conditions. Doxorubicin (DOX), on the other hand, is a first-line chemotherapeutic agent against a variety of cancers, including breast cancer, lung cancer, gastric cancer, and leukemia, etc. Despite its effectiveness, the clinical use of DOX is limited by its side effects, the most serious of which is cardiotoxicity. Considering AS could be applied as an adjuvant to anticancer agents, the combination of AS and DOX might exert synergistic effects on certain malignancies with mitigated cardiotoxicity. Given this, it is necessary and meaningful to confirm whether AS would neutralize the DOX-induced cardiotoxicity and its underlying molecular mechanisms. Aim of the studyThis paper aims to validate the cardioprotective effects of AS against DOX-induced myocardial injury (MI) while deciphering the molecular mechanisms underlying such effects. Materials and methodsFirstly, the cardioprotective effects of AS against DOX-induced MI were confirmed both in vitro and in vivo. Secondly, serum pharmacochemistry and network pharmacology were orchestrated to explore the in vivo active compounds of AS and predict their ways of functioning in the treatment of DOX-induced MI. Finally, the predicted mechanisms were validated by Western blot analysis during in vivo experiments. ResultsThe results demonstrated that AS possessed excellent antioxidative ability, and could alleviate the apoptosis of H9C2 cells and the damage to mitochondria induced by DOX. In vivo experiments indicated that AS could restore the conduction abnormalities and ameliorate histopathological changes according to the electrocardiogram and cardiac morphology. Meanwhile, it markedly downregulated the inflammatory factors (TNF-α, IL-6, and IL-1β), decreased plasma ALT, AST, LDH, CK, CK-MB, and MDA levels, as well as increased SOD and GSH levels compared to the model group, which collectively substantiate the effectiveness of AS. Afterward, 14 compounds were identified from different batches of AS-dosed serum and selected for mechanism prediction through HPLC-HRMS analysis and network pharmacology. Consequently, the MAPKs and caspase cascade were confirmed as primary targets among which the interplay between the JNK/Caspase 3 feedback loop and the phosphorylation of ERK1/2 were highlighted. ConclusionsIn conclusion, the integrated approach employed in this paper illuminated the molecular mechanism of AS against DOX-induced MI, whilst providing a valuable strategy to elucidate the therapeutic effects of complicated TCM systems more reliably and efficiently.

Resolution of fibrosis in mdx dystrophic mouse after oral consumption of N-163 strain of Aureobasidium pullulans produced β-glucan

Recent advances in the management of Duchenne muscular dystrophy (DMD), such as exon skipping and gene therapy, though have reached a clinical stage, the outcome at its best is still considered suboptimal. In this study, we evaluated a novel N-163 strain of Aureobasidium pullulans produced β-glucan (Neu-REFIX) for its potential as an adjuvant to slow down the progression of the disease by anti-inflammatory and anti-fibrotic effects. In this study, 45 mice in the three groups, 15 each in a group; Gr. 1 normal mice, Gr.2 mdx mice as vehicle, and Gr.3 mdx mice administered the N-163 β-glucan for 45 days. The N-163 β-glucan group showed a significant decrease in the plasma ALT, AST, and LDH levels (126 ± 69 U/l, 634 ± 371 U/l, 3335 ± 1258 U/l) compared with the vehicle group (177 ± 27 U/l, 912 ± 126 U/l, 4186 ± 398 U/l). Plasma TGF-β levels increased, and plasma IL-13 levels decreased in the N-163 group. The inflammation score of HE-stained muscle sections in the N-163 group (1.5 ± 0.8) was lower than that in the vehicle group (2.0 ± 0.8). The N-163 strain β-glucan group (24.22 ± 4.80) showed a significant decrease in the fibrosis area (Masson’s Trichrome-positive area) compared with the vehicle group (36.78 ± 5.74). The percentage of centrally nucleated fibres evaluated by Masson’s trichrome staining was 0 in the normal group, while it increased to 80% in the vehicle group but remained at 76.8% in the N-163 group. The N-163 β-glucan group showed a significant decrease in the fibrosis area. Considering their safety and easy oral consumption, Neu-REFIX β-glucan could be worth large multicentre clinical studies as adjuvant in slowing down the progress of DMD.

Manipulated sperm motility via soot nanoparticles-induced biochemical alterations in human seminal plasma

Obtaining spermatozoa with progressive motility, via postejaculatory activation with pharmacological agents such as theophylline and pentoxifylline, is crucial for the success rate of assisted reproduction in couples with severe male factor infertility. Regrettably, the possibility of premature acrosome reactions and impared oocyte function questions the practical applicability of phosphodiesterase inhibitors. The rapid development of nanotechnologies promotes the use of hydrophobic rapeseed oil soot as a non-cytotoxic biomaterial for sperm motility activation, but the scarcity of knowledge regarding the interactions of soot with components from the seminal plasma hinders the eventual commercialization of this cutting-edge approach. Aiming to eliminate this shortcoming, the current study shows for the first time how the soot nanomaterials alter the biochemistry of human seminal plasma. Upon 270 min incubation with soot nanoparticles, the activity of AST, ALT, CK, LDH and GGT enzymes in the seminal plasma of ten patients changes inversely to the registered sperm motility (i.e., lower enzyme activity, higher sperm motility and vice versa). This phenomenon is primarily related to termination of the enzymes-substrate binding or extraction of enzymes from the gametes via chemical bonding with the soot. These novel mechanisms depend on the physicochemical features of used carbon nanomaterials, revealing opportunities for predictable tuning of the sperm reproductive potential.

D-Limonene complexed with cyclodextrin attenuates cardiac arrhythmias in an experimental model of doxorubicin-induced cardiotoxicity: Possible involvement of calcium/calmodulin-dependent protein kinase type II

BackgroundArrhythmias are one manifestation of the cardiotoxicity that results from doxorubicin (Doxo) administration. Although cardiotoxicity is an anticipated outcome in anticancer therapies, there is still a lack of treatment options available for its effective management. This study sought to evaluate the possible cardioprotective effect of complex d-limonene (DL) plus hydroxypropyl-β-cyclodextrin (HβDL) during treatment with Doxo, focusing on the arrhythmic feature. MethodsCardiotoxicity was induced in Swiss mice with Doxo 20 mg/kg, with 10 mg/kg of HβDL being administered 30 min before the Doxo. Plasma CK-MB and LDH levels were analyzed. Cellular excitability and susceptibility to cardiac and cardiomyocyte arrhythmias were evaluated using in vivo (pharmacological cardiac stress) and in vitro (burst pacing) ECG protocols. Ca2+ dynamics were also investigated. The expression of CaMKII and its activation by phosphorylation and oxidation were evaluated by western blot, and molecular docking was used to analyze the possible interaction between DL and CaMKII. ResultsElectrocardiograms showed that administration of 10 mg/kg of HβDL prevented Doxo-induced widening of the QRS complex and QT interval. HβDL also prevented cardiomyocyte electrophysiological changes that trigger cellular arrhythmias, such as increases in action potential duration and variability; decreased the occurrence of delayed afterdepolarizations (DADs) and triggered activities (TAs), and reduced the incidence of arrhythmia in vivo. Ca2+ waves and CaMKII overactivation caused by phosphorylation and oxidation were also decreased. In the in silico study, DL showed potential inhibitory interaction with CaMKII. ConclusionOur results show that 10 mg/kg of βDL protects the heart against Doxo-induced cardiotoxicity arrhythmias, and that this is probably due to its inhibitory effect on CaMKII hyperactivation.

Mailuoning oral liquid attenuates convalescent cerebral ischemia by inhibiting AMPK/mTOR-associated apoptosis and promoting CREB/BDNF-mediated neuroprotection

Ethnopharmacological relevanceIschemic stroke is divided into acute, subacute and convalescent phases according to the time of onset. Clinically, Mailuoning oral liquid (MLN O) is a traditional Chinese patent medicine for treating ischemic stroke. Previous studies have shown that MLN O could prevent acute cerebral ischemia-reperfusion. However, its underlying mechanism remains unclear. Aim of the studyTo investigate the relationship between neuroprotection and apoptosis for clarifying MLN O mechanism in the recovery phase of ischemic stroke. Materials and methodsWe imitated stroke using middle cerebral artery occlusion/reperfusion (MCAO/R) in vivo and oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro models. The infarct volume, neurological deficit scores, HE staining, Nissl staining, TUNEL staining, immunohistochemistry, and Western blot were correspondingly performed to find pathological changes and detect neuronal apoptosis in rat cerebral cortex. The contents of LDH, Cyt-c, c-AMP and BDNF in rat plasma and cerebral cortex were detected by ELISA. Cell viability was measured by CCK8 assay. Cell morphology, Hoechst 33342 staining and Annexin-V-Alexa Fluor 647/PI staining were performed to assess neuronal apoptosis. The expression levels of proteins were evaluated by western blotting. ResultsMLN O obviously reduced brain infarct volume and neurological deficit scores in MCAO rats. MLN O inhibited inflammatory cell infiltration and neuronal apoptosis, but promoted gliosis, neuronal survival, and neuroprotection in the cortical region of MCAO rats. Additionally, MLN O decreased the amount of LDH and cytochrome c, while increasing the expression of c-AMP in the plasma and ischemic cerebral cortex of MCAO rats, and promoting the expression of BDNF in the cortical tissue of MCAO rats. Besides, MLN O improved cell viability, restored cell morphology, while attenuating cell damage, inhibiting neuronal apoptosis following OGD/R in PC-12 cells. Moreover, MLN O inhibited apoptosis by suppressing the expression of pro-apoptotic-associated proteins, including Bax, cytochrome c, Cleaved caspase 3 and HIF-1α, whereas accelerating the expression of Bcl-2 in vivo and in vitro. Furthermore, MLN O inhibited the activity of AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR), but activated the signaling pathway of cAMP-response element binding protein (CREB)/brain-derived neurotrophic factor (BDNF) in MCAO rats and OGD/R-stimulated PC-12 cells. ConclusionsThese results demonstrated that MLN O inhibited AMPK/mTOR to affect apoptosis associated with mitochondria, leading to improve CREB/BDNF-mediated neuroprotection in the recovery period of ischemic stroke in vivo and in vitro.

Large bilateral adrenal masses: interest of plasma LDH measurement to detect adrenal lymphoma

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Prognosis of Hyperviscosity Syndrome in Newly Diagnosed Multiple Myeloma in Modern-Era Therapy: A Real-Life Monocentric Study

Introduction Hyperviscosity syndrome (HVS) is a rare and life-threatening complication of multiple myeloma (MM). HVS was a criterion of MM treatment in the previous international recommendation, but it was withdrawn in the IMWG 2014 guidelines. Moreover, studies about the prognosis of HVS in modern-era therapy are missing. Our objective is to describe MM patients with HVS and to evaluate its prognosis impact. Methods Inclusion criteria for the case were patients aged ≥18 years newly diagnosed with MM with HVS in Saint Louis Hospital, Paris, France, between 2011 and 2021. HVS diagnosis was based on presence of neurological signs and/or evaluation by ophthalmologic examination for symptoms or high spike. Two controls were selected for each case in the same period with the best fit set on age, sex, year of diagnosis, and front-line treatment. The primary outcome was overall survival. Results Thirty-nine MM patients with HVS (7 severe forms and 32 ophthalmologic confirmation) were included with a median age of 59 years (IQR 51-68). The main isotype of immunoglobulin was IgG (69%, median 61 g/L), followed by IgA (28%, median 41 g/L). Translocation t(11;14), presented in 37% of the cohort (13 IgG and 1 IgA), was associated with a higher spike level than other patients (for IgG: 71 versus 59 g/L, p=0.0001). HVS clinical presentation was heterogenous: 23% asymptomatic (median spike at 60 and 44 g/L for IgG and IgA), 59% mild (epistaxis, headache, blurry vision) and 18% severe with neurological signs, respectively. Fifteen (38%) patients required ICU admission: severe HVS for 6, association with other complications for 6 and only for therapeutic plasma exchange (TPE) realization for 3. No major ischemic or bleeding was observed at diagnosis. For HVS management, in addition to myeloma therapy, TPE was performed for 35 patients (92%), which was well tolerated, and 3 patients received only steroids. One month after HVS diagnosis and care, no rebound effect of monoclonal spike was observed and 33 (84%) patients had complete clinical recovery. Three patients (8%) died, and three (16%) had an ophthalmic sequela of HVS. All patients had at least one CRAB criterion except one who had two focal lesions on MRI. Most of the patients received bortezomib and high-dose steroids (95%) associated with an immunomodulatory drug (43%) or alkylating agents (42%). Two patients (5%) received Daratumumab. For younger patients (< 65 years), 21 patients (84%) received autologous stem cell transplantation upfront. After the exclusion of one patient with an early loss in follow-up (< 3 months), the global response rate (≥PR) was 92%. The median time to the subsequent treatment was 2.1 years (95%IC 1.4-3.4). At subsequent relapse, 6 patients (3 IgG, 2 IgA and 1 IgM) had HVS recurrence. With a median follow-up of 5.2 years, the median OS for HVS patients was 3.6 years (95%CI 2.7-not reached). Among patients older than 65 years, the early mortality rate (< 3 months) was 21%. Presence of > 5% of blood plasma cell (HR 6.06, p=<.01), high-risk cytogenetics (HR 3, p=0.03), R-ISS3 score (HR 3.3, p=0.04), and LDH above normal level (HR 3.6, p=0.01) shortened the OS. Compared to controls, HVS MM patients had a higher spike level at diagnosis, higher ISS3 rate, higher bone marrow plasma cells and lower fibrinogen levels (Table). HVS myeloma had dismal OS compared to controls (median: 3.6 vs. 7.7 years, p=0.01, Figure). In a multivariate analysis of pooled HVS and control patients, HVS (HR 2.7, 95%CI 1.4-5.4, p=0.004) and high-risk cytogenetics (HR 2.8, 95%CI 1.4-5.7, p=0.005) were two independent adverse factors for OS. HVS remained an independent factor for OS for myeloma with the addition of the ISS score or R-ISS score in the model. Conclusion HVS at myeloma diagnosis is a rare but urgent complication associated with high lethality in older patients. HVS was associated with high tumor burden and dismal survival, independent of principal prognosis scores in modern treatment era. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Correlation of Certain Biochemical Constituents of Seminal Plasma with Semen Characteristics in Pantja Buck

Background: Pantja is a dual purpose indigenous goat breed, reared in Tarai region of Uttarakhand and Uttar Pradesh. There is a lack of thorough study about the seminal characteristics of this goat breed. Hence, the present study was conducted to evaluate the level of certain biochemical constituents and their relationship with macroscopical and microscopical semen characteristics in Pantja buck. Methods: A total thirty two ejaculates from four sexually mature Pantja buck were analysed. The ejaculates were evaluated for volume (0.36±0.06 ml), mass activity (4.47±0.22), concentration (3.54±0.13 x 109/ml), individual motility (85.28±1.17%), viability (88.28±0.78%), morphological abnormalities (4.70±0.17%), plasma membrane integrity (86.31±0.62%) and acrosome integrity (94.31±1.05%) by standard semen evaluation protocol. Seminal plasma ALT (18.10±0.45 U/L), AST (124.04±0.86 U/L), ALP (49.98±0.38 U/L), LDH (216.36±1.78 U/L), GSH-Px (10.90±0.21 U/ml), MDA (2.14±0.05 nmol/ml), albumin (2.29±0.03 gm/dl), globulin (2.52±0.02 gm/dl), total proteins (4.81±0.05 gm/dl), calcium (11.07±0.18 mg/dl) and phosphorus (12.34±0.20 mg/dl) were determined spectrophotometrically. Two tailed Pearson’s correlation was applied to check the relationship among the above attributes. Result: A significant correlation was observed among the biochemical constituents of seminal plasma and seminal attributes of Pantja bucks. The selected biochemical constituents also showed significant correlation with each other.

Impact of Elevated LDH on Cystatin C-Based Glomerular Filtration Rate Estimates in Patients with Cancer.

Background: The determination of renal function is crucial for the clinical management of patients with cancer. The glomerular filtration rate (GFR) serves as a key parameter, estimated by creatinine clearance determination in 24-h collected urine (CrCl) as well as equation-based approaches (eGFR) relying on serum creatinine (eGFR CKD EPIcrea) or serum cystatin C (eGFR cystatin C). Serum creatinine and serum cystatin C levels differentially depend on muscle and tumor mass, respectively. Although muscle and tumor mass may thus represent confounding factors, comparative studies for eGFR estimate approaches in cancer patients are lacking. Methods: The present study retrospectively analyzed GFR estimates based on equations of creatinine (eGFRcr), cystatin C (eGFRcys) and combined creatinine-cystatin C levels (eGFRcr-cys) in a subset of patients. The associations of LDH with cystatin C or LDH with eGFRcr, eGFRcys and GFRcr-cys were explored. Results: The laboratory values of 123 consecutive patients were included. The median age was 59 (24–87) and 47.2% were female. There was a statistically significant difference in the mean of CKD EPIcrea (85.17 ± 21.63 mL/min/1.73 m2), CKD EPIcys (61.16 ± 26.03 mL/min/1.73 m2) and CKD EPIcrea-cys (70.42 ± 23.89 mL/min/1.73 m2) (p < 0.0001). Spearman’s correlation analysis revealed a significant correlation of elevated plasma LDH >1.5 UNV and cystatin C values (r = 0.270, p < 0.01, n = 123). LDH values >1.5 UNV were associated with significantly lower CKD EPIcys (r = 0.184, p < 0.01) or CKD EPIcrea-cys (r = 0.226, p < 0.05) estimates compared to CKD EPIcrea. Conclusions: The inclusion of cystatin C as a biomarker led to a lower eGFR estimates compared to creatinine alone or in a combination of both cystatin C and creatinine. The level of cystatin C correlated with the level of LDH, suggesting that the use of cystatin C-based calculations of GFR in cancer patients with elevated LDH should be used with caution.

Cold Water Immersion Improves the Recovery of Both Central and Peripheral Fatigue Following Simulated Soccer Match-Play.

The present study aimed to investigate the effect of cold water immersion (CWI) on the recovery of neuromuscular fatigue following simulated soccer match-play. In a randomized design, twelve soccer players completed a 90-min simulated soccer match followed by either CWI or thermoneutral water immersion (TWI, sham condition). Before and after match (immediately after CWI/TWI through 72 h recovery), neuromuscular and performance assessments were performed. Maximal voluntary contraction (MVC) and twitch responses, delivered through electrical femoral nerve stimulation, were used to assess peripheral fatigue (quadriceps resting twitch force, Qtw,pot) and central fatigue (voluntary activation, VA). Performance was assessed via squat jump (SJ), countermovement jump (CMJ), and 20 m sprint tests. Biomarkers of muscle damages (creatine kinase, CK; Lactate dehydrogenase, LDH) were also collected. Smaller reductions in CWI than TWI were found in MVC (-9.9 ± 3%vs-23.7 ± 14.7%), VA (-3.7 ± 4.9%vs-15.4 ± 5.6%) and Qtw,pot (-15.7 ± 5.9% vs. -24.8 ± 9.5%) following post-match intervention (p < 0.05). On the other hand, smaller reductions in CWI than TWI were found only in Qtw,pot (-0.2 ± 7.7% vs. -8.8 ± 9.6%) at 72 h post-match. Afterwards, these parameters remained lower compared to baseline up to 48–72 h in TWI while they all recovered within 24 h in CWI. The 20 m sprint performance was less impaired in CWI than TWI (+11.1 ± 3.2% vs. +18 ± 3.6%, p < 0.05) while SJ and CMJ were not affected by the recovery strategy. Plasma LDH, yet no CK, were less increased during recovery in CWI compared to TWI. This study showed that CWI reduced both central and peripheral components of fatigue, which in turn led to earlier full recovery of the neuromuscular function and performance indices. Therefore, CWI might be an interesting recovery strategy for soccer players.

Abstract P1041: Redox Regulator MnTnBuOE-PyP5+ Protects Cardiomyocyte Against Oxidative Stress Mediated Myocardial Ischemia/reperfusion Injury

Background: Much evidence suggests that oxidative stress plays a major role in the pathogenesis of myocardial ischemia/reperfusion (I/R) injury and is a potential target for therapeutic interventions. We tested the hypothesis that the novel redox regulator, MnTnBUOE-2-PyP 5+ (BMX-001), mediates cardioprotection through reduction of oxidative stress in myocardial tissue following I/R injury. Methods and Results: BMX-001 (2mg/Kg, IP) or vehicle was administered in mice 4 hours before myocardial ischemia. Mice were then subjected to I/R injury by ligating the left anterior descending artery for 1 hour followed by 24 hours of reperfusion. Following 24 hours of reperfusion, infarct size, cardiomyocyte apoptosis, and SOD2 level were determined, and echocardiography was performed. Our study revealed that the redox regulator provided protection against I/R-induced injury in mice as evident by a significant reduction in infarct size, plasma LDH level, and oxidative stress. In addition, BMX-001 administration in mice markedly increased FoxO3a and SOD2 levels. BMX-001 treatment attenuated I/R induced cardiomyocyte apoptosis in-vivo and in-vitro as evident by a reduction in the number of TUNEL positive cells and improved cell viability. Echocardiography at 24 hours of reperfusion revealed improved ejection fraction and fractional shortening in mice treated with BMX-001. Furthermore, in-vitro treatment of BMX-001 (10μM) 24 hours before hypoxia/reoxygenation in H9c2 cells improved oxidative stress as evident by a reduction in cardiolipin peroxidation, mitochondrial superoxide level, and 4-HNE adducted proteins. Conclusion: Pre-treatment with redox regulator BMX-001 showed a beneficial effect in attenuating cardiomyocyte death, mitochondrial superoxide, and cardiolipin peroxidation following I/R injury in H9c2 cells. Pre-treatment with SOD2 mimetic improved heart function, decreased infarct size, and cardiomyocyte apoptosis in mice following I/R injury. Therefore, BMX-001 can be a novel cardioprotective agent against oxidative stress-induced myocardial I/R injury.