Search Results (109)

Poloxamer 188 is a polymer that is used as a carrier and stabilizer of pharmacological agents. It has been demonstrated to enhance red blood cell and hemoglobin levels in healthy animals and in select clinical cases. The objective of this study was to assess the efficacy of Poloxamer 188 in CBA mice when administered repeatedly in the carboplatin-induced myelosuppression model. The mice were administered carboplatin once at a dose of 100 mg/kg, and then Poloxamer 188 was orally administered daily at doses of 10 mg/kg, 100 mg/kg, 500 mg/kg, and 1000 mg/kg for 7 and 21 days. Poloxamer 188 at a dose of 1000 mg/kg was found to bring the level of 2,3-bisphosphoglycerate in red blood cells close to control level (p = 0.1331 for the control group compared to Poloxamer at a dose 1000 mg/kg) already from day 8 of the study and in bone marrow resulted in regulation of genes responsible for hematopoiesis. G-GSF at day 8 and TNFα at day 22 gene expression was significantly decreased by 54% (p = 0.012) and 16% (p = 0.024), respectively, with Poloxamer 188 administration at a dose of 100 mg/kg. Additionally, in the bone marrow, the treatment was seen to exert a positive regulatory effect on the genes responsible for hematopoiesis. These findings are consistent with the observed increase in red blood cell by 6.7% (p = 0.001), hemoglobin by 4.7% (p = 0.0053), and reticulocyte percentage by 53.6% (p < 0.0001) following Poloxamer 188 administration at a dose of 1000 mg/kg in CBA mice with myelosuppression. Full article

Background/Objectives: Although the nanocomposite of poly(L-lactic acid) with graphene oxide (PLLA-GO) shows promise for tissue engineering, its specific bioactive interactions with diverse cell lineages during early tissue regeneration remain unclear. This study comprehensively investigated the in vitro multifaceted biocompatibility of PLLA-GO using human fibroblasts (FN1 cells), murine mesenchymal stem cells (mBMSCs), and human umbilical vein endothelial cells (HUVECs). Methods: Morphological analyses were performed using optical and scanning electron microscopy, while proliferation dynamics were assessed via CFSE staining. Cell cycle progression was evaluated using flow cytometry, mitochondrial activity was examined through TMRE staining, and inflammatory cytokine profiling was performed via Cytometric Bead Array (CBA). Results: PLLA-GO exhibited primary biocompatibility across all evaluated cell lines, characterized by efficient adhesion and proliferation. However, significant cell-type-dependent modulations were observed. The FN1 cells exhibited proliferative adaptation but induced accelerated scaffold degradation, as evidenced by a substantial increase in cellular debris (5.93% control vs. 34.38% PLLA-GO; p = 0.03). mBMSCs showed a transient initial proliferative response and a significant 21.66% increase in TNF-α production (179.67 pg/mL vs. 147.68 pg/mL in control; p = 0.03). HUVECs demonstrated heightened mitochondrial sensitivity, exhibiting a 32.19% reduction in mitochondrial electrical potential (97.07% control vs. 65.82% PLLA-GO; p ≤ 0.05), alongside reductions in pro-inflammatory cytokines TNF-α (8.73%) and IL-6 (12.47%). Conclusions: The PLLA-GO processing method is crucial for its properties and subsequent cellular interactions. Therefore, rigorous and specific preclinical evaluations—considering both cellular contexts and fabrication—are indispensable to ensure the safety and therapeutic potential of PLLA-GO in tissue engineering and regenerative medicine. Full article

Antibody–drug conjugates (ADCs) are promising cancer therapeutics, but optimizing their cytotoxic payloads remains challenging. We present DumplingGNN, a novel hybrid Graph Neural Network architecture for predicting ADC payload activity and toxicity. Integrating MPNN, GAT, and GraphSAGE layers, DumplingGNN captures multi-scale molecular features using both 2D and 3D structural information. Evaluated on a comprehensive ADC payload dataset and MoleculeNet benchmarks, DumplingGNN achieves state-of-the-art performance, including BBBP (96.4% ROC-AUC), ToxCast (78.2% ROC-AUC), and PCBA (88.87% ROC-AUC). On our specialized ADC payload dataset, it demonstrates 91.48% accuracy, 95.08% sensitivity, and 97.54% specificity. Ablation studies confirm the hybrid architecture’s synergy and the importance of 3D information. The model’s interpretability provides insights into structure–activity relationships. DumplingGNN’s robust toxicity prediction capabilities make it valuable for early safety evaluation and biomedical regulation. As a research prototype, DumplingGNN is being considered for integration into Omni Medical, an AI-driven drug discovery platform currently under development, demonstrating its potential for future practical applications. This advancement promises to accelerate ADC payload design, particularly for Topoisomerase I inhibitor-based payloads, and improve early-stage drug safety assessment in targeted cancer therapy development. Full article

Soil organic carbon (SOC) and total nitrogen (TN) sequestration are vital for maintaining soil fertility and mitigating climate change. This study aimed to evaluate the effects of different amendments (chemical and biological) and crop rotations on SOC, TN sequestration, and soil aggregate distribution. A six-year field study was conducted, involving five different treatments: a monoculture of peanut (PC), a monoculture of maize (MC), a maize-peanut rotation (M-PR), and peanut continuous cropping with chemical (PCCA) and biological (PCBA) amendments. Soil properties, aggregate size distribution, SOC, TN, and enzyme activities were measured. The results show that the bulk density increased, while the field water−holding capacity and porosity decreased with depth. M-PR had the highest macroaggregate (>0.25 mm) proportion, increasing by 21.6–50.8%. SOC and TN increased with aggregate size and were 23.9–103.6% and 7.0–82.9% higher, than PC and MC, respectively, under the treatments. PCCA showed the highest SOC, TN, and enzyme activities. Structural equation modeling indicated that the C and N contents of aggregates directly influenced SOC and TN sequestration. In conclusion, crop rotation and amendments, especially PCCA, effectively improve soil C and N sequestration, and enhance the soil structure, thereby reducing degradation risks, and potentially decreasing on−farm greenhouse gas emissions. Full article

Background: Elderly patients infected with SARS-CoV-2 are at higher risk of developing cytokine storm and severe outcomes; however, specific immunological and proteomic biomarkers for early prediction remain unclear in this vulnerable group. Methods: We enrolled 182 elderly COVID-19 patients from the Chinese PLA General Hospital between November 2022 and April 2023, categorizing them based on progression to respiratory failure requiring mechanical ventilation (defined as severe progression). Olink proteomic analysis was performed on admission serum from 40 propensity score-matched samples, with differentially expressed proteins (DEPs) validated by cytometric bead array (CBA) in 178 patients. To predict severe progression, a model was developed using a 70% training set and validated on a 30% validation set. LASSO regression screened features followed by logistic regression and receiver operating characteristic (ROC) analysis to optimize the model by incrementally incorporating features ranked by random forest importance. Results: Elderly patients progressing to severe COVID-19 exhibited early immune dysregulation, including neutrophilia, lymphopenia, monocytopenia, elevated procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), neutrophil-to-lymphocyte ratio (NLR), and systemic immune-inflammation index (SII), as well as coagulation dysfunction and multi-organ injury. Proteomics identified a set of biomarkers, including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), and revealed disruptions in signaling pathways, including the mTOR and VEGF signaling pathways. The optimal predictive model, which incorporated PCT, IL-6, monocyte percentage, lymphocyte count, and TRAIL, achieved an area under curve (AUC) of 0.870 (0.729–1.000) during validation. TRAIL levels negatively correlated with fibrinogen (p < 0.05). Conclusions: Elderly COVID-19 patients with severe progression demonstrate early immune dysregulation, hyperinflammation, coagulation dysfunction, and multi-organ injury. The model we proposed effectively predicts disease progression in elderly COVID-19 patients, providing potential biomarkers for early clinical risk stratification in this vulnerable population. Full article

Objectives: Atrial fibrillation (AF) is the most common sustained arrhythmia associated with stroke, heart failure, and increased mortality. Due to its efficacy and safety, cryoballoon ablation (CBA) is widely accepted for rhythm control; however, long-term AF recurrence remains a challenge. Continuous monitoring with implantable loop recorders (ILRs) enhanced by artificial intelligence (AI) can detect both symptomatic and asymptomatic episodes, potentially optimizing patient management. This analysis assessed the long-term effectiveness of CBA in maintaining sinus rhythm and investigated the role of ILR-guided monitoring in enhancing therapeutic decisions. Methods: Data from 91 patients with paroxysmal or persistent atrial fibrillation (AF) who underwent pulmonary vein isolation using cryoballoon ablation at four Italian centers between April 2022 and April 2024 were analyzed. All patients received an insertable loop recorder (ILR) before or during hospitalization for ablation, allowing for the continuous remote monitoring of arrhythmias. Baseline demographics, procedural details, AF occurrence, AF burden (calculated as the total duration of all AF episodes occurring within a day and categorized by episode duration), therapeutic adjustments, and the effect of artificial intelligence (AI) on data processing were evaluated. Results: The cohort’s average age was 62.4 years, with 24.2% of participants being female. Physician-confirmed AF recurrence was noted in 26.7% of patients at 12 months and 49.5% at 24 months. The device data indicated a daily AF burden of ≥6 min in 47.2% at 12 months, with 25.9% surpassing 1 h. AI algorithms decreased false-positive alerts by 21%, resulting in an estimated saving of 19 clinician hours. In patients with pre-ablation ILR data, the median AF burden significantly decreased from 7% to 0.2% (p = 0.017). ILR-guided monitoring affected treatment adjustments, leading to the discontinuation of antiarrhythmic therapy in 36 patients and redo ablations in 8. Conclusions: Continuous ILR monitoring, combined with AI-driven analysis, enables the detection of AF recurrences and burden, thereby facilitating timely therapeutic adjustments. Full article

Malaria, the deadliest parasitic disease in the world, is sexually dimorphic, inflammatory, and oxidative. Males experience more severe symptoms and mortality than females do; therefore, the roles of 17β-estradiol and testosterone in this phenomenon have been studied. Both hormones affect oxidative stress, the primary mechanism of Plasmodium elimination. Estradiol has antioxidant activity, but the role of testosterone is controversial. Testosterone increases oxidative stress by reducing superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities, which increase lipoperoxidation in the testis. However, the antioxidant properties of testosterone in prostate and nervous tissue have also been reported. The discrepancies are probably because when testosterone levels increase, the aromatase enzyme transforms testosterone into estrogens that possess antioxidant activity, which masks the results. Therefore, it is unknown whether testosterone is involved in the sexual dimorphism that occurs in oxidative stress in malaria. In this work, we administered testosterone and simultaneously inhibited aromatase with letrozole to evaluate the role of testosterone in the sexually dimorphic pattern of oxidative stress that occurs in the blood, spleen, and brain of male and female CBA/Ca mice infected with Plasmodium berghei ANKA (P. berghei ANKA). Testosterone triggers parasitemia in males, who also display more oxidative stress than females in the absence of infection, leading to sexually dimorphic patterns. Interestingly, increasing testosterone levels in infected mice reduced oxidative stress in males and increased oxidative stress in females, reversing or eliminating the dimorphic patterns observed. Oxidative stress varies in each tissue; the brain was the most protected, while the blood was the greatest damaged. Our findings highlight the role of testosterone as a regulator of oxidative stress in a tissue and sex-specific manner; therefore, understanding the role of testosterone in malaria may contribute to the development of sex-specific personalized antimalarial therapies. Full article

Approximately 10–35% of people living with HIV (PLHIV) on antiretroviral therapy (ART) fail to restore CD4⁺ T cell counts, a state known as immunological non-responder (INR) characterized by persistent immune activation and elevated cytokine levels. Objective: This study aimed to identify cytokines that can serve as biomarkers for immune activation and inflammation in INR patients. Methods: We conducted a cross-sectional study comparing two groups: INRs (PLHIV on ART with viral suppression) and immunological responders (IRs). We analyzed 40 samples of virologically suppressed PLHIV, measuring CD4⁺ T cell counts, viral load via RT-PCR, and cytokine levels through cytometric bead array (CBA). Results: The INR group exhibited significantly higher median serum levels of IL-6 (1.74 pg/mL vs. 0.94 pg/mL, p = 0.016) and IL-10 (1.65 pg/mL vs. 0.92 pg/mL, p = 0.03) compared to the IR group. Conclusions: Elevated IL-6 and IL-10 levels may serve as potential markers to distinguish INR from IR patients with areas under the curve (AUC) of 0.731 and 0.707, respectively. Full article

The corpus luteum is the principal progesterone-secreting gland, while primary cilia function as pivotal organelles in intercellular signal transduction. Together, they play an essential role in the establishment and maintenance of pregnancy. However, the mechanisms underlying the role of primary cilia in granulosa cell luteinization in mouse ovaries remain poorly understood. This study discovered the regularity of primary cilia in mouse ovaries and revealed the role of primary cilia in regulating progesterone synthesis in luteinized granulosa cells. In vivo test results showed that the expression of primary cilia was obvious in the corpus luteum. The secretion of P4 in mice was significantly increased at 6, 12, 24, 48, and 72 h. The secretion of P4 and the expressions of luteinization markers (STAR, 3β-HSD) and primary ciliate proteins (IFT88, Arl13B) were significantly up-regulated at different time points (6, 12, 24 h and 24, 48, 72 h), and the INS group was significantly higher than the LH group and the control. In vitro test results showed that the follicular granulosa cells were luteinized under INS, the length and number of primary cilia increased, and the secretion of progesterone increased. The expression levels of STAR and 3β-HSD of the primary cilia marker Arl13B and luteinization markers were increased, while the expression levels of CYP19A1 were decreased. Ciliobrevin A (CBA) and Y-27632 2HCl were used to regulate the expression of primary cilia. The results showed that after CBA treatment, the expression level of cilia protein Arl13B decreased, and the secretion level of P4 and the expression levels of STAR and 3β-HSD decreased, indicating that the level of luteinization decreased. Conversely, after inducing ciliogenesis with Y-27632 2HCl, the results were the opposite of those observed with CBA treatment. In conclusion, our study demonstrates that primary cilia regulate the expression of steroidogenic enzymes, thereby promoting progesterone secretion by granulosa cells in mice and ensuring proper luteinization. Full article

Escherichia coli (E. coli) has the ability to induce clinical and subclinical mastitis in dairy cows, causing a huge loss for the dairy industry. In this study, 51 subclinical mastitis isolates and 36 clinical mastitis isolates from eight provinces of China between 2019 and 2021 were used to investigate the differences in their biological characteristics. The results showed that B1 (52.9%) and A (39.1%) were the predominant phylogroups; R1 (50.6%) was the predominant lipopolysaccharide (LPS) core type; and 44 STs (ST10 and ST58 were the most sequence-prevalent STs) and 2 new STs (ST14828 and ST14829) were identified; however, no significant difference was observed between the clinical and subclinical group strains. To compare the virulence gene differences between the clinical and subclinical mastitis-related isolates, 18 common virulence genes (including afaE, eaeA, papC, saa, sfa, ompA, aer, irp2, iucD, escV, sepD, east1, estB, stx2e, CNF1, cba, hlyA and traT) were determined using the PCR method. The results showed that the detection rates of traT, irp2 and iucD in clinical mastitis isolates were significantly higher than those in subclinical mastitis isolates (p ˂ 0.05). Meanwhile, subclinical-group E. coli had stronger biofilm formation abilities than the clinical group (p < 0.05) in 78 (89.7%) mastitis-related E. coli that could form biofilms. Furthermore, 87 mastitis-related E. coli showed severe resistance against tetracycline (37.9%), ampicillin (36.8%), streptomycin (34.5%) and cotrimoxazole (28.7%); their most prevalent resistance genes were blaCTX-M (33.3%), tetA (27.6%), sul2 (18.4%) and strB (28.7%). It was noteworthy that the clinical-group strains had a higher resistance against ampicillin and possessed higher amounts of the resistance gene blaCTX-M (p < 0.05) compared to the subclinical group. This study aims to provide references for preventing the E. coli isolates from inducing different types of mastitis. Full article

Introduction: Obesity is an important risk factor for atrial fibrillation (AF) development. Data on cryoballoon ablation (CBA) outcomes in obese patients have so far been scarce. We reviewed the existing literature to compare the efficacy and safety of CBA in obese versus non-obese AF patients. Methods: A systematic literature search was conducted for studies comparing clinical outcomes (arrhythmia recurrence and/or procedural data and/or safety outcomes) between obese and non-obese patients undergoing CBA for AF. Statistical pooling was performed according to a random-effects model with generic inverse-variance weighting of relative risks (RRs) and standardised mean differences (SMDs) computing risk estimates with 95% confidence intervals (CIs). Results: Obese and non-obese patients had comparable arrhythmia recurrence rates (normal versus overweight, RR = 0.95, 95% CI: 0.82–1.11, p = 0.55, I²% = 91%; normal versus class I obesity, RR = 0.97, 95% CI: 0.82–1.13, p = 0.68, I²% = 87%; normal versus class II obesity, RR = 0.98, 95% CI: 0.91–1.07, p = 0.29, I²% = 65%). Procedure time was marginally increased in obese patients compared to non-obese counterparts (normal versus overweight, SMD = 0.05, 95% CI: −0.15–0.26, p = 0.62, I²% = 74%; normal versus class I obesity, SMD = 0.10, 95% CI: −0.00–0.19, p = 0.06, I²% = 2%; overweight versus class I obesity, SMD = 0.11, 95% CI: 0.01–0.21, p = 0.048, I²% = 25%). Regarding radiation exposure, fluoroscopy time was increased in patients with class I obesity compared to normal-weight or overweight patients and dose area product was also increased in obese patients compared to non-obese patients. Lastly, the risk of complications did not differ between obese and non-obese patients. Statistical heterogeneity and the small number of patients included are the main limitations of this study. Conclusion: CBA seems to be effective for obese patients suffering from AF, featuring also similar safety outcomes with non-obese individuals. Radiation exposure was increased in obese patients. Full article

Extensive fertilization and irrigation in commercial plant nurseries generates runoff containing high levels of nutrients. Biochar-amended bioretention systems with internal water storage zones (IWSZs) have been shown to enhance total nitrogen removal from urban runoff. However, their effectiveness for treatment of nursery runoff and the role of biochar in the IWSZ remain understudied. The goal of this research was to investigate nitrogen transformations in pilot-scale bioretention systems treating nursery runoff with varying biochar-amendment strategies: (a) throughout both the unsaturated zone and the IWSZ (CBA), (b) only in the unsaturated zone (PBA). Variables investigated included hydraulic loading rate (HLR; 0.11, 0.22, and 0.55 cm/min), IWSZ depth (44 and 69 cm), and the presence of plants (Muhlenbergia capillaris). The presence of biochar in the IWSZ (CBA) enabled significantly greater nitrogen removal (p = 0.031) compared to PBA. CBA had improved hydraulic efficiency by mitigating short-circuiting (34% increase in mean retention time) and likely enhanced performance by promoting nutrient uptake and microbial activity. Three times the above ground plant biomass was observed in CBA vs. PBA (0.73 kg in CBA vs. 0.23 kg in PBA). The highest nitrogen removal efficiency (84%) was achieved in the planted CBA unit at an HLR of 0.22 cm/min and IWSZ depth of 69 cm. A spreadsheet-based tool, utilizing a logarithmic regression model for CBA (R² = 0.88 for TIN, 0.86 for NO_x) and PBA (R² = 0.50 for TIN, 0.60 for NO_x), was developed for system design to achieve nitrogen removal targets. The greater variability in the PBA-fitted model (lower R²) compared to CBA (higher R², better fit) suggests biochar’s ability to mitigate short-circuiting and improve hydraulic performance. Full article

Background/Objectives: The literature suggests that the cranial base angle is considered one of the contributing factors to sagittal jaw malpositions when its relationship with the viscerocranium is examined. Our study aims to compare and evaluate the outcomes of fixed functional orthopedic treatment in patients with mandibular retrognathia across different cranial base groups. Methods: Participants were treated at Erciyes University with fixed functional appliances and categorized by CBA into low (<130°), medium (130°–134°), and high (>134°) groups. A total of 39 patients were included: 13 in the low CBA group (7 males, 6 females; mean age 14.62 ± 1.12 years), 13 in the medium CBA group (3 males, 9 females; mean age 14.38 ± 0.96 years), and 13 in the high CBA group (4 males, 9 females; mean age 14.08 ± 1.04 years). Results: In the low CBA group, Ar-Go-N (p = 0.005) and SNA (p = 0.023) angles significantly decreased, while the ANB angle and Wits appraisal significantly decreased across all groups (p < 0.05). The high CBA group showed increases in ANS-Me, N-Me, N-ANS, and N-Gn lengths (p < 0.05). The medium and high CBA groups had significant increases in S-Go and ANS-Gn lengths, while Co-Gn length increased significantly in the low and high CBA groups (p < 0.05). Incisor measurements (IMPA, L1-APog, L1/NB, L1-NB) increased in all groups, with overjet and overbite reduced (p < 0.05). U1/PP (p = 0.039), U1/SN (p = 0.043), U1-NA (p = 0.030), and U1/NA (p = 0.025) parameters increased in the low CBA group, with the Upper Lip–E distance decreasing significantly in the low and high CBA groups (p < 0.05). A comparison between groups showed significant differences in U1/PP, U1-NA, and U1/NA parameters, which increased in the low and medium CBA groups but decreased in the high CBA group. Conclusions: CBA influences treatment outcomes. The low CBA group experienced decreases in Ar-Go-N and SNA angles, while the high CBA group showed increases in certain vertical facial dimensions. Incisor parameters rose in the low and medium CBA groups but decreased in the high CBA group, suggesting limited CBA effects on treatment results. Full article

Giant cell arteritis (GCA) is an autoimmune/autoinflammatory disease affecting large vessels in patients over 50 years old. The disease presents as an acute inflammatory response with two phenotypes, cranial GCA and large-vessel vasculitis (LV)-GCA, involving the thoracic aorta and its branches. 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET-CT) is among the imaging techniques contributing to diagnosing patients with systemic disease. However, its association with soluble inflammatory markers is still elusive. This proof-of-concept study aims to identify novel soluble serum biomarkers in PET/CT-positive patients with LV-GCA and associate them with active (0 months) and inactive disease (6 months following treatment), in sequential samples. The most-diseased-segment target-to-background ratio (TBR_MDS) was calculated for 13 LV-GCA patients, while 14 cranial GCA and 14 Polymyalgia Rheumatica patients with negative initial PET/CT scans served as disease controls. Serum macrophage-related cytokines were evaluated by cytometric bead array (CBA). Finally, previously published NMR/metabolomics data acquired from the same blood sampling were analyzed along with PET/CT findings. TBR_MDS was significantly increased in active versus inactive disease (3.32 vs. 2.65, p = 0.006). The analysis identified nine serum metabolites as more sensitive to change from the active to inactive state. Among them, choline levels were exclusively altered in the LV-GCA group but not in the disease controls. Cytokine levels were not associated with PET/CT activity. Combining CRP, ESR, and TBR_MDS with choline levels, a composite index was generated to distinguish active and inactive LV-GCA (20.4 vs. 11.62, p = 0.001). These preliminary results could pave the way for more extensive studies integrating serum metabolomic parameters with PET/CT imaging data to extract sensitive composite disease indexes useful for everyday clinical practice. Full article

Batch heterogeneous catalytic ozonation experiments were performed using commercial and synthesized nanoparticles as catalysts in aqueous ozone. The transferred ozone dose (TOD) ranged from 0 to 150 μM, and nanoparticles were added in concentrations between 0 and 1.5 g L⁻¹, with all experiments conducted at 20 °C and a total volume of 240 mL. A Ce-doped TiO₂ catalyst (1% molar ratio of Ce/Ti) was synthesized via the sol–gel method. Response surface methodology (RSM) was applied to identify the most significant factors affecting the removal of selected pharmaceuticals, with TOD emerging as the most critical variable. Higher TOD resulted in greater removal efficiencies. Furthermore, it was found that the commercially available metal oxides α-Al₂O₃, Mn₂O₃, TiO₂, and CeO₂, as well as the synthesized CeTiO_x, did not increase the catalytic activity of ozone during the degradation of ibuprofen (IBF) and para-chlorobenzoic acid (pCBA). Carbamazepine (CBZ) and diclofenac (DCF) are compounds susceptible to ozone oxidation, thus their complete degradation at 150 μM transferred ozone dose was attained. The limited catalytic effect was attributed to the rapid consumption of ozone within the first minute of reaction, as well as the saturation of catalyst active sites by water molecules, which inhibited effective ozone adsorption and subsequent hydroxyl radical generation (^●OH). Full article