Search Results (2,245)

Background/Objectives: Iron deficiency remains a prevalent condition, needing specific laboratory tests for diagnosis. This study aimed to evaluate whether routine complete blood cell count (CBC) parameters can be used within a machine learning framework to predict low ferritin and low transferrin saturation, used as biochemical markers of altered iron status, potentially supporting more targeted laboratory test utilization. Methods: In this single-center retrospective outpatient study, we analyzed 32,437 records from subjects undergoing both complete blood cell count and iron metabolism testing between 2023 and 2026. Low ferritin and low transferrin saturation were defined using sex-specific thresholds. Low ferritin was present in 14,344 subjects (44.2%), whereas low transferrin saturation was present in 7791 subjects (24.0%). After cleaning data and excluding incomplete records, demographic variables and CBC indices were tested as potential predictors. The dataset was split into training and test sets with stratified sampling. Multiple supervised machine learning models, including logistic regression, decision tree, random forest, XGBoost, support vector machine, k-nearest neighbors, and Naive Bayes, were trained. Hyperparameter tuning and model selection were performed using repeated stratified 10-fold cross-validation, optimizing the area under the curve (AUC). Model performance was assessed by AUC, sensitivity, and specificity, and validated on an independent test set. Results: All models showed predictive capability for low ferritin and low transferrin saturation using CBC parameters alone. Ensemble methods, especially random forest and XGBoost, reached the best performance (AUC values of 0.80–0.87 for ferritin and 0.85–0.96 for transferrin saturation). Sensitivity and specificity were balanced, supporting clinical screening applicability. Results were maintained across validation and confirmed in the test set. Prediction of transferrin saturation showed slightly higher accuracy than ferritin. Feature importance analysis identified mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and red blood cell distribution width (RDW) as key predictors. Conclusions: CBC-based machine learning models may help identify subjects with low ferritin or low transferrin saturation, supporting subsequent targeted assessment of iron status. Full article

Background/Objectives: Obesity-related insulin resistance is accompanied by chronic low-grade inflammation, but the extent to which weight loss modifies circulating cytokines in a sex-specific manner remains insufficiently understood. The aim of this study was to assess sex-specific cytokine responses and metabolic adaptation in adults with obesity and insulin resistance following a six-month weight-reduction program (WRP). Methods: Thirty-six participants (24 women and 12 men) with a value of Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) ≥ 2 underwent an individualized low-calorie diet combined with moderate physical activity and health education. Anthropometric, body composition, biochemical, oxidative stress, and cytokine parameters were evaluated before and after the intervention. Results: Both women and men showed significant reductions in body mass, Body Mass Index (BMI), waist circumference, visceral fat area (VFA), body fat mass (BFM), fasting glucose, HOMA-IR, modified Atherogenic Index of Plasma (new-AIP), malondialdehyde (MDA), and Oxidative Stress Index (OSI). Women additionally showed significant decreases in fat-free mass (FFM), skeletal-muscle mass (SMM), total body water (TBW), glycated hemoglobin A_1c (HbA1c), and triacylglycerols, whereas cholesterol in high-density lipoproteins (HDL-C) increased significantly in men. Cytokine changes were selective rather than uniform. Interleukin-1 receptor antagonist (IL-1ra), Interleukin 6 (IL-6), and Tumor Necrosis Factor alpha (TNF-α) decreased in both women and men. In sex-stratified analyses, IL-1β decreased significantly only in women, whereas IL-7 decreased significantly only in men. ClinicalTrials.gov Registration: [NCT07645105] (retrospectively registered on [11 June 2026]). Conclusions: A 6-month lifestyle-based weight-reduction program in adults with overweight or obesity and insulin resistance was associated with metabolic improvement, reduced oxidative stress, and partial attenuation of obesity-related low-grade inflammation. The observed cytokine and metabolic changes suggest sex-related patterns of immunometabolic adaptation to weight reduction. However, these findings should be interpreted cautiously because of the relatively small sex-stratified subgroups and the number of cytokine endpoints analyzed, and they require confirmation in larger, sex-balanced studies. Full article

Background/Objectives: Sarcopenic obesity (SO) represents a severe clinical phenotype characterized by the coexistence of reduced skeletal muscle mass and excess adiposity, and is associated with insulin resistance, dyslipidemia, and systemic inflammation. However, easily accessible biomarkers that capture early molecular changes underlying SO are lacking. The aim of this pilot study was to compare circulating microRNA (miRNA) profiles in patients with severe obesity and a sarcopenic obesity phenotype with those of healthy controls and to identify candidate miRNAs suitable for further validation. To the best of our knowledge, this represents one of the first broad screening studies of circulating miRNAs specifically conducted in patients with severe obesity and DXA-confirmed sarcopenic obesity. Methods: In this single-center pilot study conducted in the Czech Republic, fasting plasma samples from 12 adult participants (6 with severe obesity and sarcopenic obesity phenotype, body mass index > 45 kg/m²; 6 healthy controls; age 18–65 years) were analyzed using an RT-qPCR panel comprising 384 assays, including technical controls and 352 target circulating miRNAs. Following predefined quality control and filtering criteria, 224 miRNAs were retained for the final statistical analysis. Six patients with severe obesity were classified according to the ESPEN/EASO 2022 consensus criteria for sarcopenic obesity, while EWGSOP2-based assessment was used for functional evaluation of sarcopenia. Differential expression was evaluated using fold change and exploratory statistical testing. Results: We identified a set of miRNAs with significantly altered expression in SO, including increased muscle-enriched miR-486-5p and hepatocyte-enriched miR-122-5p, and decreased vascular miR-145-5p, as well as several additional miRNAs related to myogenesis, lipid metabolism and inflammatory signaling. miR-451a, a recognized marker of hemolysis, was also increased but was interpreted with caution. Conclusions: Despite the limited sample size, the results of this study suggest that specific circulating miRNAs may reflect key pathophysiological pathways in SO and could serve as promising biomarkers to support risk stratification and monitoring in larger, hypothesis-driven studies. Full article

Background: Ascophyllan, a sulfated polysaccharide extracted from brown seaweed, has shown immunomodulatory and antioxidant effects in preclinical studies, yet human clinical evidence remains scarce. This randomized, double-blind, placebo-controlled pilot trial evaluated the safety and exploratory biological effects of daily ascophyllan supplementation in healthy adults. Methods: Twelve participants were randomized to receive either ascophyllan (n = 6) or placebo (n = 6) for 28 days. Safety was monitored through adverse event reporting and repeated laboratory assessments, including hematology, biochemistry, and inflammatory markers. Immune cell populations were analyzed via serial flow cytometry, serum total antioxidant capacity was measured at multiple time points, and gut microbiome composition was profiled using 16S rRNA gene sequencing. All analyses were exploratory in nature. Results: Ascophyllan supplementation proved well tolerated, with no adverse events observed and stable hematologic, renal, and biochemical parameters throughout the study. Exploratory longitudinal analyses suggested directional modulation of NK-cell-associated phenotypes during ascophyllan supplementation, including directional changes in CD57⁺, NKp46⁺, and NKG2D⁺ NK-cell phenotypes; however, group × time interaction analyses did not remain statistically significant after correction for multiple comparisons. Serum antioxidant capacity showed inter-individual variability with a directional but non-significant increase in the ascophyllan group at intermediate time points. Exploratory microbiome analyses suggested modest directional compositional differences involving members of the Bacteroidaceae and Bifidobacteriaceae families; however, no taxon remained statistically significant after correction for multiple comparisons. Conclusions: These preliminary findings indicate that ascophyllan is safe and well tolerated in healthy adults and may be associated with modulation of innate immune phenotypes, subtle microbiome compositional differences, and directional changes in antioxidant capacity. Larger, adequately powered clinical trials are warranted to confirm these observations and further investigate potential biological and clinical effects. Full article

Background: In rodent models of intracranial tumor development, evaluating the actual burden experienced by animals beyond procedural severity is essential for ethical and legal compliance. This study examined whether voluntary wheel running (VWR) could serve as a sensitive indicator of post-surgical burden following subcutaneous transmitter implantation, tumor cell injection, and tumor resection. It also assessed whether VWR supports the detection of humane endpoints. VWR outcomes were compared with body weight, clinical scores, heart rate, and activity levels recorded via telemetry. Methods: Fourteen male BDIX rats were housed individually in cages equipped with a running wheel. Under general anesthesia, telemetric devices to monitor heart rate and activity were subcutaneously implanted. After recovery, glioblastoma BT4Ca cells were stereotaxically injected into the right frontal cortex. Eight days later, the resulting tumors were microsurgically resected. Body weight, VWR, heart rate, and general activity were continuously monitored until the animals reached humane endpoint criteria, indicated by sudden weight loss and clinical deterioration. Results: On average, body weight and VWR declined significantly after all surgical procedures, with tumor resection causing the most pronounced effect. As animals approached the endpoint, a marked drop in these parameters was observed, along with an increased clinical score (p < 0.05). Activity measures supported these findings, though less consistently than weight and VWR. Conclusions: Monitoring body weight and VWR enables an effective assessment of the actual postoperative burden experienced by rats undergoing surgeries of different procedural complexity. Moreover, VWR is a valuable supplementary tool for identifying humane endpoints alongside body weight and clinical scoring. Full article

Background/Objectives: Phytoestrogens are secondary plant metabolites produced via the phenylpropanoid pathway. They include a broad spectrum of chemical compounds, such as phenolics, flavonoids, isoflavones, coumestans, lignans, and others. Their chemical structures resemble those of estradiol, and they exhibit biological effects similar to those of human estrogens, influencing many physiological processes throughout life in both men and women—including the timing and progression of puberty. Methods: The literature search included databases such as PubMed, Scopus, Web of Science, and Google Scholar with the use of specific keywords. Studies were considered eligible if they reported original findings from observational studies (cohort, case–control, and cross-sectional) or from experimental studies. Results: Phytoestrogens can modulate estrogenic activity and interact with a variety of biological pathways. These compounds may play a role in human development and pubertal processes, contribute to overall health, and potentially help alleviate menopausal symptoms and reduce the risk of certain cancers. Conclusions: Phytoestrogens have numerous positive effects on the human body across various stages of life. Their overall impact and potency, however, seem to be influenced by factors such as intake level, individual genetic variability, and the specific phytoestrogen class consumed. Full article

Background/Objectives: Vitamin D is a nutrient-related secosteroid system with endocrine, paracrine, immunological, and neurodevelopmental actions relevant to nutritional psychiatry. Psychiatric research has often treated vitamin D either as a cross-sectional correlate of depression or as a non-specific supplement expected to act across heterogeneous diagnostic categories. This narrative review aimed to develop a more discriminating framework in which vitamin D is considered a lifespan neuroimmune and immunometabolic signal whose psychiatric relevance depends on developmental timing, biological context, and phenotype. Methods: Evidence was integrated from developmental epidemiology, neonatal dried-blood-spot studies, randomized trials, meta-analyses, Mendelian randomization studies, clinical guidelines, and mechanistic neuroscience. The review focuses on prenatal and neonatal 25-hydroxyvitamin D, vitamin D-binding protein, free and bioavailable vitamin D, vitamin D receptor signaling, immune and microglial pathways, neurotransmitter systems, neurotrophic signaling, mitochondrial function, oxidative stress, hypothalamic–pituitary–adrenal-axis regulation, and the gut–microbiota–immune–brain axis. Results: The available evidence does not support vitamin D as a universal treatment for psychiatric disorders. Instead, vitamin D deficiency and altered vitamin D biology appear most relevant in biologically and clinically defined risk states, including neurodevelopmental vulnerability, inflammatory depression, psychosis liability, severe mental illness with nutritional deprivation, metabolic comorbidity, and cognitive frailty. Mechanistic data support plausible links with cytokine biology, the tryptophan–kynurenine pathway, dopaminergic and serotonergic systems, stress regulation, and neuroimmune homeostasis. Conclusions: Vitamin D should be conceptualized in psychiatry as a context-dependent neuroimmune and immunometabolic signal rather than a generic psychotropic intervention. Future studies should prioritize biomarker-enriched, developmentally timed, nutrition-centered models of precision prevention and adjunctive care. Full article

Owing to their favorable stability and safety profile, postbiotics—defined as functional metabolites originating from probiotic microbes—have attracted growing interest in the field of companion animal health care. The present work sought to evaluate how postbiotics prepared from Lactiplantibacillus plantarum and Pediococcus lactis influence lipid metabolism, blood biochemistry, and the composition of fecal microbiota in felines. Eighteen clinically healthy adult cats were assigned at random to three equal-sized groups: a control treatment (n = 6, a basal diet), a L. plantarum postbiotic L-27-2 group (n = 6, a basal diet with 10⁹ CFU equivalent/kg/day), and a P. lactis L-14-1 postbiotic group (n = 6, a basal diet with 10⁹ CFU equivalent/kg/day). The supplementation lasted four weeks. Both postbiotics significantly lowered serum total cholesterol (TC) and triglycerides (TG) concentrations (p < 0.05), demonstrating a modulatory action on lipid metabolism in cats. The L-27-2 postbiotic selectively enriched beneficial taxa, notably Bifidobacterium (p < 0.05). Furthermore, fecal levels of malodorous compounds were significantly decreased by both treatments (p < 0.05). The results suggested both postbiotics have potential positive effects in cats. These findings are preliminary and hypothesis-generating. Full article

Introduction: Lung cancer, the leading cause of cancer-related mortality worldwide, is a heterogeneous malignancy comprising distinct histological and molecular subtypes, with non-small cell lung cancer (NSCLC) accounting for approximately 85% of cases and adenocarcinoma (ADC) representing the most prevalent histotype. An emerging pathological feature of NSCLC, spread through air spaces (STAS)—defined as the extension of tumor cells into the lung parenchyma beyond the main tumor margin—has been associated with worse disease-free and overall survival and has been proposed as a possible predictor of recurrence to guide surgical extent. Concurrently, recent comprehensive genomic profiling of early-stage NSCLC has highlighted the need to interpret multi-omics data and their relationship with pathological variables, including IASLC histological subtypes, to better personalize treatment strategies. In this context, we investigated the overall distribution of STAS and its association with tumor mutational profiles and IASLC histological subtypes in a large real-world cohort of lung adenocarcinoma patients from the LANTERN project. Materials and Methods: In a prospective, multicenter observational study (March 2023–December 2024), 271 NSCLC patients were enrolled, and clinicopathological, immunohistochemical, and genomic data were collected; comprehensive genomic profiling was performed using the TruSight Oncology 500 assay to analyze 523 cancer-related genes, tumor mutational burden (TMB), and microsatellite instability; and STAS was assessed according to IASLC criteria. Adenocarcinoma accounted for roughly 90% of the cases, with a median age of 69 years and a predominance of stage IV disease (49.5%). STAS was evaluable in 162 cases and was detected in 17.9% of tumors. Results: STAS-positive tumors showed a higher trend towards locally advanced and advanced disease; no differences were observed in sex, age, smoking status, tumor mutational burden, or PD-L1 expression. Additionally, STAS-positive tumors showed a higher association with micropapillary, mucinous, and papillary patterns, whereas the acinar pattern was more frequent in STAS-negative tumors. The most frequently mutated genes were TP53, KRAS, EGFR, and STK11, with no significant differences between groups; ROS1 alterations were absent in STAS-negative tumors but detected more frequently in STAS-positive cases. Conclusions: Overall, these findings indicate that STAS positivity is associated with high-risk histological subtypes and advanced disease, suggesting its importance as a marker of tumor aggressiveness and emphasizing the need for its systematic evaluation in lung adenocarcinoma to better guide surgical planning and patient risk assessment. Full article

Newborn screening in England is a national program with laboratories adhering to common screening algorithms. Until recently, screening for inherited metabolic disorders was provided by ten laboratories using laboratory-developed tests (LDTs) and three using commercial assays: harmonization of results proved challenging. Introduction of hereditary tyrosinemia type 1 screening meant LDTs required modification to include the measurement of succinylacetone, and subsequent re-validation. This provided an opportunity to implement a single commercial reagent kit in all laboratories. It was anticipated that this would improve analytical performance and harmonization. This study aimed to determine whether these goals were achieved. Verification across the 13 laboratories revealed that the commercial kit reduced inter-laboratory variation for all analytes demonstrating improved harmonization. However, this was achieved by applying instrument-specific correction factors to all analytes, the magnitude of which were significant, indicating a lack of standardization. Performance of succinylacetone was limited by instrument-dependent background interference from the methionine stable isotope label, underscoring the need to establish evidence-based screening cut-off values (COV) rather than adopting published thresholds. This study emphasizes the need for traceable reference materials to improve laboratory quality and the value of screening outcome data. Full article

Background/Objectives: Menopause is accompanied by substantial changes in endogenous sex hormones that influence metabolic regulation. However, the associations of specific hormones with type 2 diabetes (T2D) risk in postmenopausal women remain inconsistent. This study aimed to quantify the relationships between incident T2D and follicle-stimulating hormone (FSH), oestradiol, testosterone, and sex hormone-binding globulin (SHBG), and to examine cross-sectional differences in hormone concentrations between postmenopausal women with and without T2D. Methods: MEDLINE, Embase and Cochrane CENTRAL were searched from database inception to 21 June 2024. Eligible studies included prospective cohort, nested case–control and case–control designs. Associations with incident T2D were pooled using Hartung–Knapp–Sidik–Jonkman random-effects meta-analysis. Both categorical and continuous estimates were extracted, prioritising maximally adjusted models. Risk of bias was assessed using ROBINS-E and the Newcastle–Ottawa Scale. Results: Sixteen studies (18 articles; n = 16,180) were included. Higher SHBG was consistently associated with lower T2D risk in cohort analyses (RR 0.55; 95% CI 0.38–0.72; I² ≈ 0%). Higher FSH was also associated with lower risk (high vs. low: HR 0.55, 95% CI 0.29–0.81), although continuous estimates showed heterogeneity. Higher oestradiol was associated with increased T2D risk (RR 1.61, 95% CI 1.18–2.03; I² ≈ 6%), while testosterone was not significantly associated with incident T2D (RR 1.11, 95% CI 0.73–1.50). Cross-sectional analyses indicated lower SHBG and higher testosterone in women with T2D. Conclusions: Endogenous hormone profiles and SHBG concentrations are associated with T2D in postmenopausal women, with the most consistent evidence for an inverse association between SHBG and incident T2D. Because the available evidence is observational and partly heterogeneous, these findings should be interpreted as associations rather than causal or clinically predictive effects. Standardised measurement, repeated pre-diagnostic sampling and external validation are required before these biomarkers can be considered for routine risk stratification. Full article

Background: Phenylketonuria (PKU) is a rare genetic disorder caused by mutations in the phenylalanine hydroxylase (PAH) gene, resulting in a deficiency of the enzyme responsible for metabolizing phenylalanine (Phe) and its accumulation. PKU can be identified through newborn screening (NBS) or genetic sequencing; however, both approaches have limitations, including high false-discovery rates and variants of uncertain significance (VUS). This study aims to identify a PKU metabolomic profile using unique biomarkers to enhance early diagnosis and improve treatment outcomes. Methods: Dried blood spot (DBS) samples from 65 patients diagnosed with PKU and matched healthy controls were collected through the NBS program. An untargeted metabolomics analysis was conducted using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to profile metabolites and investigate altered metabolic pathways in patients with PKU. Results: A total of 418 significantly dysregulated metabolites were identified in PKU patients. Among them, 90 metabolites were identified as endogenous human metabolites. The most significantly affected pathways were those related to the metabolism of aromatic amino acids and polysaccharides. Moreover, lipid metabolic pathways were dysregulated, including those involved in fatty acid and phospholipid biosynthesis. In addition to phenylalanine (AUC = 0.994), 1,11-Undecanedicarboxylic acid (UDCA) (AUC = 0.969) was significantly elevated in patients with PKU, suggesting it is a promising potential biomarker for PKU. Conclusions: Untargeted metabolomics revealed distinct metabolic alterations in patients with PKU, providing insights into disease pathophysiology. The identification of UDCA as a consistently elevated metabolite supports its potential utility as a supplementary biomarker for PKU diagnosis and monitoring. Further validation in larger cohorts, using a targeted metabolomics approach, is warranted. Full article

Background and Objectives: Oxidative stress is recognized as an important contributor to heart failure (HF) pathophysiology, but the relationships of individual oxidative and antioxidant biomarkers with symptomatic severity and systolic dysfunction remain insufficiently defined. This study examined circulating oxidative and nitrosative stress markers across New York Heart Association (NYHA) classes and left ventricular ejection fraction (LVEF) categories in HF and their associations with HF severity. Materials and Methods: In this case–control study, 85 patients with HF and 33 healthy controls were included. Malondialdehyde (MDA), nitrates and nitrites (NOx), superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), sulfhydryl (SH) groups, and NT-proBNP were measured. Group differences were analyzed using the Kruskal–Wallis test with post hoc comparisons. Adjusted ordinal logistic regression models examined associations with NYHA class and LVEF category, and receiver operating characteristic (ROC) analysis evaluated discriminatory performance. Results: Compared with controls, all biomarkers differed significantly across NYHA classes and LVEF categories (all p < 0.001). In separate adjusted models, higher NOx, MDA, and NT-proBNP were associated with worse NYHA class and more impaired LVEF, whereas higher antioxidant marker levels were associated with lower odds of severe HF. In combined models, NOx remained independently associated with worse NYHA class (OR 1.07, 95% CI 1.04–1.11; p < 0.001), while MDA remained independently associated with more impaired LVEF (OR 1.02, 95% CI 1.00–1.03; p = 0.022). NT-proBNP showed the best discrimination for NYHA III/IV versus I/II (AUC 0.966), while among oxidative biomarkers NOx performed best for symptomatic severity (AUC 0.782) and MDA for LVEF ≤ 40% (AUC 0.751). Conclusions: HF is characterized by increased oxidative and nitrosative stress together with reduced antioxidant defense. NOx appears more closely related to symptomatic severity, whereas MDA appears more closely related to systolic dysfunction. However, NT-proBNP remained the strongest overall discriminator. NOx and MDA may provide complementary mechanistic information on redox imbalance across HF severity categories. Full article

Background: Oxidative stress plays a key role in the pathogenesis of complications in preterm infants, including bronchopulmonary dysplasia (BPD). Thymosin β4 (Tβ4) and thymosin β10 (Tβ10) are proteins involved in tissue repair and responses to oxidative stress, but their role in extremely preterm neonates remains poorly understood. Methods: A total of 149 saliva samples from 18 infants with gestational age < 30 weeks were analyzed. Salivary proteins and their proteoforms were characterized using an integrated proteomic platform based on nano-HPLC-ESI-MS. Relative quantification was performed using extracted ion current (XIC) peak areas. Associations with postmenstrual age, oxygen requirement, and BPD development were assessed, including longitudinal analysis using generalized estimating equation (GEE) models. Results: Significant correlations were found between postmenstrual age and total Tβ4 (p = 0.001), oxidized Tβ4 percentage (p = 0.025), and total Tβ10 (p = 0.043). Higher oxygen requirement was associated with lower levels and percentages of oxidized Tβ10 (p = 0.005; p < 0.001). No significant differences were observed during the first week of life between neonates who later developed BPD and those who did not. However, longitudinal analysis showed that in neonates without BPD, total and oxidized Tβ10 and total Tβ4 increased over time, whereas in neonates with BPD, these biomarkers remained stable or decreased. The increase in oxidized Tβ10 percentage was slower in infants with BPD. Conclusions: Although no early differences were detected, longitudinal trajectories of Tβ4 and Tβ10 differed between infants with and without BPD. Postnatal changes in these proteins may be associated with differences in clinical course and exposure to postnatal oxidative stress. Full article

Objective: The development of nanobubble technology has expanded to various gas molecules, such as nitric oxide (NO). NO donors have been frequently used for both therapeutic and diagnostic purposes. However, the use of NO in nanoformulations has received little attention thus far. This research aimed to assess the subchronic toxicity of NO nanobubbles in a preclinical study design. Methods: Ninety days of subchronic toxicity testing based on modified OECD guidelines in Sprague Dawley rats was used in this study to assess potential adverse effects of intravenous administration of NO nanobubbles (NONBs). Hematological factors, serum biochemistry, and histology of the liver, kidney, heart, lung, and spleen were investigated. Results: The intravenous NONB injection trial using graded doses of 0.01 mL, 0.04 mL, and 0.06 mL resulted in no deaths during the 90-day treatment period. However, liver disturbance and electrolyte imbalance emerged. This condition was supported by histological findings in the liver and kidney, which showed potential reversible damage, and persistent hemorrhage of the spleen. Conclusions: Intravenous administration of NONBs at a dose of up to 0.06 mL in this subchronic toxicity test remains safe. Further studies with adjustments to the test formulation are highly encouraged before the clinical trial stage in humans. Full article