Search Results (57)

Background/Objectives: Chronic hyperglycemia drives gingival fibroblast dysfunction, disrupting extracellular matrix homeostasis and the migratory capacity required for wound healing, which contributes to increased periodontal susceptibility in diabetes. Ferulic acid (FA), a dietary phenolic compound with antioxidant and anti-inflammatory properties, has demonstrated wound-healing activity in various tissues; however, its effects on human gingival fibroblasts (HGFs) under hyperglycemic conditions remain unclear. This study investigated the effects of FA on HGF viability, wound closure, and the secreted concentrations of matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and interleukin-6 (IL-6) under normoglycemic and hyperglycemic conditions. Methods: Cultured HGFs (HGF-1 cell line) were pre-conditioned for 72 h under normal glucose (5.5 mM) or high glucose (25 mM), with an iso-osmolar mannitol control to distinguish glucose-specific effects from osmotic changes. Viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and wound closure by scratch assay following FA treatment, while MMP-1, TIMP-1, and IL-6 concentrations in conditioned medium were quantified by enzyme-linked immunosorbent assay (ELISA). Results: FA was non-cytotoxic at all tested concentrations, whereas 50 µM transiently increased viability. FA significantly enhanced wound closure under both conditions, independent of osmotic effects. High glucose increased IL-6 concentration in the conditioned medium (significant at 48 h) and MMP-1 levels. FA partially attenuated MMP-1 levels at specific time points, modestly reduced IL-6 levels, and markedly increased TIMP-1 concentration under both glucose conditions. Conclusions: FA enhanced wound closure in scratch assay and shifted the secreted MMP-1/TIMP-1 concentration ratio toward a more favorable, matrix-protective profile at the protein level, without cytotoxicity, supporting further investigation of its potential role in periodontal tissue repair under high glucose conditions. Full article

Background: Non-proliferative diabetic retinopathy (NPDR) is a common microvascular complication of type 2 diabetes mellitus (T2DM) associated with chronic inflammation, metabolic dysregulation, and multiple interacting systemic factors. This study evaluated the associations of inflammatory biomarkers, serum 25-hydroxyvitamin D [25(OH)D], electrolyte homeostasis, and lipid-derived indices with NPDR. Methods: In this cross-sectional study with prospective recruitment, 98 patients with T2DM were classified into NPDR (n = 55) and non-DR (n = 43) groups. Clinical characteristics, hematological inflammatory indices, serum interleukin-6 (IL-6), C-reactive protein (CRP), 25(OH)D, electrolyte concentrations, lipid profile, and derived biomarkers—including the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic immune–inflammation index (SII), LDL-C/HDL-C ratio, and calculated serum osmolarity—were compared. Receiver operating characteristic (ROC) analysis and multivariable logistic regression were performed to assess discriminatory performance and independent associations with NPDR. Results: Patients with NPDR had significantly lower serum 25(OH)D concentrations than the non-DR group (16.6 ± 7.5 vs. 21.0 ± 5.2 ng/mL, p < 0.001). Serum IL-6 levels (4.2 ± 3.1 vs. 2.8 ± 1.4 pg/mL, p = 0.047), NLR (p = 0.043), LDL-C/HDL-C ratio (p = 0.031), and calculated serum osmolarity (p = 0.033) were significantly higher, whereas lymphocyte count and the lymphocyte-to-monocyte ratio were significantly lower. Among individual biomarkers, serum 25(OH)D showed the best discriminatory performance (AUC = 0.712). A multivariable model including insulin therapy, IL-6, serum 25(OH)D, and the LDL-C/HDL-C ratio demonstrated good discrimination (AUC = 0.813), with 81.8% sensitivity and 81.4% specificity. Bootstrap validation supported the stability of the model. Conclusions: NPDR was associated with lower serum 25(OH)D concentrations, systemic inflammation, and a less favorable LDL-C/HDL-C ratio. A multivariable model integrating inflammatory, metabolic, and nutritional biomarkers showed better discriminatory performance than individual biomarkers. The association between insulin therapy and NPDR likely reflects greater diabetes severity rather than a direct effect of insulin. These findings are exploratory, and prospective studies in larger independent cohorts are needed to determine whether these biomarkers improve risk stratification beyond established clinical factors. Full article

Objectives: This study investigated the relationship between interstitial and blood glucose concentrations following ingestion of carbohydrate-containing drinks differing in carbohydrate amount, osmolarity, and glycaemic index. Methods: Ten healthy adults (nine male; age: 22 ± 1 y; height: 177 ± 12 cm; weight: 75 ± 14 kg) completed a double-blind, randomised cross-over study with seven beverage trials varying in carbohydrate (CHO) characteristics. Blood samples were collected at rest and over two hours, while interstitial glucose ([iG]) was recorded using a continuous glucose monitor (Supersapiens, Atlanta, USA). Glycaemic metrics and mean absolute relative difference (MARD) were calculated for hypoglycaemic, euglycaemic, and hyperglycaemic ranges. Data were analysed using repeated-measures ANOVA with Bonferroni correction and paired t-tests (p ≤ 0.05). Results: Interstitial and blood glucose concentrations were similar at baseline but diverged post-ingestion. MARD varied by glucose rate and direction, exceeding 20% during rapid declines (>2 mg/dL/min), where [iG] underestimated [BG] by −7.3 ± 27.1 mg/dL. Accuracy was highest during stable glucose (MARD = 10.5 ± 8.6%). Carbohydrate amount and glycaemic index influenced peak glucose, whereas beverage concentration (5–20%) had minimal effect when CHO amount was fixed, though variation in CGM agreement appeared during post-peak declines. Conclusions: CGM tracked blood glucose overall but showed reduced accuracy during rapid falls or hypoglycaemia. Carbohydrate properties influenced glycaemic response but not sensor agreement when CHO load was constant. Glucose rate and direction of change are key considerations for interpreting CGM data in research and applied settings. Full article

Background/Objectives: Medication errors remain a patient safety concern in neonatal intensive care units (NICU), mainly due to multiple dilution steps, a lack of standardized preparation instructions, and the frequent use of high-alert medications. While standard concentrations (SCs) for intravenous (iv) medication are recommended internationally, a national standard is missing for NICUs in Germany. The aim of this study was to evaluate a proposal for a national list of standardized iv medication concentrations to be used in German NICUs. Methods: In collaboration with the German Society for Neonatology and Pediatric Intensive Care (GNPI) and the Federal Association of German Hospital Pharmacists (ADKA), a multiprofessional expert team, including experts from the medication safety initiatives TELE-KASPER and Kinderformularium.DE and affiliated with seven German university hospitals, evaluated SCs for infusion medication administered to infants weighing 500 g to 5 kg. The evaluation process was based on international SCs lists, clinical practice, stability data, and handling aspects. Medication used in at least four of the seven hospitals was shortlisted. In the first round of the consensus process, an online survey submitted to the German Level-1 NICUs (n = 165) and their affiliated hospital pharmacies identified preferred SCs. In the second round of the consensus process, the expert team further evaluated the results of the survey. Results: The survey response rate was 52%. The consensus process resulted in a list encompassing 50 iv medications and 80 appropriate SCs. Ancillary information on preparation, stability, osmolarity, pH, and practical administration was added. Conclusions: The proposed SCs for infusion medication used in NICUs have the potential to reduce medication errors, simplify electronic prescribing, and improve workflow efficiency. Implementation aligns with international patient safety initiatives to improve medication safety in pediatric patients. Full article

Background/Objectives: Prognostication in traumatic brain injury (TBI) remains challenging. The urine-to-serum osmolality (U/S) ratio may reflect hypothalamic–pituitary axis integrity, a critical but underexplored prognostic domain. We investigated whether the U/S ratio provides early prognostic value and enhances prediction when combined with conventional severity markers. Methods: This retrospective study included 128 adult TBI patients admitted to a neurosurgical intensive care unit (ICU) with simultaneous osmolality measurements within 6 h of admission. The primary outcome was ICU mortality; the secondary outcome was poor neurological outcomes (Glasgow Outcome Scale 1–3). Results: ICU mortality was 14.1% (18/128), and poor neurological outcome occurred in 41.8% (46/110). Non-survivors had significantly lower U/S ratios than survivors (1.09 ± 0.58 vs. 1.70 ± 0.68, p < 0.001). For ICU mortality, U/S ratios (AUC = 0.803) showed similar discriminative ability to GCS (AUC = 0.806). For poor neurological outcomes, the U/S ratio (AUC = 0.768) significantly outperformed both GCS (AUC = 0.641, p = 0.038) and the Acute Physiology and Chronic Health Evaluation (APACHE) II score (AUC = 0.553, p < 0.001). Combining the U/S ratio with GCS improved mortality prediction (AUC = 0.890), as did combinations with the APACHE II score (AUC = 0.847). The U/S ratio remained independently associated with ICU mortality and poor neurological outcomes after adjusting for GCS or APACHE II scores. Quartile analyses revealed a dose–response relationship, with ICU mortality of 34.4% in Q1 versus 3.1% in Q4 (p for trend < 0.001). Prognostic value was preserved in patients receiving osmotic therapy (n = 86). Conclusions: The U/S ratio is a simple, readily available biomarker that independently predicts mortality and poor neurological outcomes in TBI patients. Particularly for neurological outcome predictions, it outperforms GCS or the APACHE II score alone. Combined with established severity scores, it may serve as a practical bedside tool reflecting hypothalamic–pituitary function in neurocritical care. Full article

Salmonella is one of the main causes of food-borne illness worldwide. In most cases, Salmonella contamination can be traced back to food processing plants and/or to cross-contamination during food preparation. To avoid food-borne diseases, food processing plants use sanitizers and biocidal to reduce bacterial contaminants below acceptable levels. Despite these preventive actions, Salmonella can survive and consequently affect human health. This study investigates the adaptive capacity of the main Salmonella enterica serotypes isolated from the poultry production line, focusing on their replication, antimicrobial resistance, and biofilm formation under stressors such as acidic conditions, oxidative environment, and high osmolarity. Using growth curve analysis, crystal violet staining, and microscopy, we assessed replication, biofilm formation, and antimicrobial resistance under acidic, oxidative, and osmotic stress conditions. Disinfectant tolerance was evaluated by determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of sodium hypochlorite. The antibiotic resistance was assessed using the Kirby–Bauer method. The results indicate that, in general, acidic and osmotic stress reduce the growth of Salmonella. However, no significant differences were observed specifically for serotypes Infantis, Heidelberg, and Corvallis. The S. Infantis isolates were the strongest biofilm producers and showed the highest prevalence of multidrug resistance (71%). Interestingly, S. Infantis forming biofilms required up to 8-fold higher concentrations of sodium hypochlorite for eradication. Furthermore, osmotic and oxidative stress significantly induced biofilm production in industrial S. Infantis isolates compared to a reference strain. Understanding how Salmonella responds to industrial stressors is vital for designing strategies to control the proliferation of these highly adapted, multi-resistant pathogens. Full article

Artificial insemination in goats commonly relies on refrigerated semen doses, yet the optimal energetic substrate to support sperm metabolism remains unclear. This study aimed to evaluate the effects of different energetic substrates on goat buck sperm metabolism and motility when refrigerated at 17 °C. Semen from six Murciano-Granadina male goats were collected and diluted in PBS supplemented with 35 mM of either glucose, fructose, pyruvate, or lactate in the first experiment. In the second experiment, the effects of varying concentrations of pyruvate and/or glucose, NaCl supplementation, and the osmolarity on sperm quality parameters were assessed. Semen was stored at 17 °C for 48 h and evaluated for motility using the CASA system, as well as for viability, mitochondrial membrane potential, and mitochondrial ROS by flow cytometry. The results show that pyruvate and lactate extenders outperformed the others, preserving higher total motility, progressivity, and viability of spermatozoa over 48 h, even at a concentration lower than 35 mM, as in the case of pyruvate. In contrast, glucose had a detrimental effect on sperm quality, reducing viability and healthy population rates while increasing motility, especially at higher concentrations. NaCl supplementation and osmolarity had no significant effect on any of the sperm quality parameters. In conclusion, pyruvate maintains a higher quality and motility of sperm stored at 17 °C in PBS in comparison with a glucose-supplemented extender. Full article

Purpose: To profile tear cytokine changes in Allogeneic Hematopoietic Stem Cell Transplant (HSCT) patients after instillation of daily topical cyclosporine-A 0.1% cationic emulsion. Methods: Participants in a longitudinal study were given cyclosporine eyedrops daily from 3 to 5 weeks before and 3 months, 6 months, and 12 months post-HSCT. The outcomes included tear cytokine concentration assayed by the Proximity Extension Assay O-linked target 96 platform. The patients were divided into two groups: Group 1 (n = 8 conjunctival CD4 cells responding to cyclosporine) and Group 2 (n = 5 conjunctival CD4 cells not suppressed after cyclosporine, where patients were non-compliant with cyclosporine). All participants had a standardized clinical examination, including meibomian gland evaluation and tear breakup times. Results: The levels of 38 cytokines/chemokines showed significant changes (p < 0.05) over time, and in many, the elevation was marked at one year. These include gamma-interferon, CXCL9, CCL3, and CCL4 (all p < 0.0001). For gamma-interferon, there was significant interaction between group and time at 1 year (p = 0.022), where the cytokine was significantly suppressed in Group 1. Four other cytokines showed significant group and time interaction at 1 year: FGF23, FGF5, LIFR, and Enrage (all p < 0.05). All patients had either withdrawal or a reduction in systemic immunomodulation between 6 months and 1 year. We found several cytokines to be associated with changes in tear osmolarity or symptom scores. Conclusions: HSCT induces significant elevation of 38 tear cytokines/chemokines even without the occurrence of ocular graft-versus-host disease when systemic immunosuppression is reduced within the first year. Topical daily cyclosporine eyedrops can reduce some pro-inflammatory tear cytokines. Full article

The study aimed to evaluate the effectiveness of the cryopreservation protocols for Pseudoplatystoma magdaleniatum semen using dimethyl sulfoxide (DMSO) or methanol (MET) as permeable cryoprotectants at two concentrations (5% and 10%) combined with 12% egg yolk (Y12%) or 5% skimmed milk powder (SMP5%) and glucose (6%), resulting in eight treatments. A semen pool (n = 8) was diluted in a 1:4 ratio, packed in 2.5 mL straws, and frozen in nitrogen vapors. It was thawed at 35 °C for 90 s. Sperm kinetics and motility duration of fresh, prefrozen, and thawed semen were analyzed using a CASA system. The osmolarity of seminal plasma and cryosolutions was estimated. Fertilization (F) and embryo viability (E) rates of thawed semen were evaluated. The osmolarity of seminal plasma was 251.1 ± 3.3 mOsmol/kg and, in the cryosolutions, ranged between 1248.3 ± 19.9 mOsmol/kg (DMSO5% + Y12%) and 3488.2 ± 1.5 mOsmol/kg (MET10% + Y12%). After thawing, total motility ranged from 38.2% to 60.5%, representing a significant reduction compared to fresh semen (95.4 ± 2.1%) (p < 0.05). The best fertilization and embryo viability rates of thawed semen were obtained with DMSO5% + SMP 5% (F = 20.7%, E = 11.7%) and MET10% + SMP5% (F = 20.1%, E = 11.5%) (p < 0.05). A cryopreservation protocol for P. magdaleniatum semen with 5%DMSO or 10%MET combined with SMP5% is possible, but further study is necessary to optimize its fertilizing capacity. Full article

Background/Objectives: A probiotic Streptococcus thermophilus (iHA318) has been demonstrated to alleviate dry eye symptoms in a mouse model. This study investigated its effects on dry eye mitigation in a clinical trial. Methods: A total of 68 volunteers were recruited in the double-blind clinical trial and randomly divided into a probiotic group and a placebo group. The probiotic group received iHA318 capsules daily for 35 days via oral intake, while the placebo group received microcrystalline cellulose capsules. Assessments before and after the intervention were performed for the tear volume (TV), tear break-up time (TBUT), tear osmolarity (Osmo), serum sialic acid (SA) concentrations, and the Ocular Surface Disease Index (OSDI), and an impression cytology analysis was conducted for immunofluorescence detection of NLRP3 expression. Results: The tear volume was significantly increased in the probiotic group, although a placebo effect was observed in the placebo group. The probiotic group showed a significant reduction in tear osmolarity, an extended TBUT, and an improved OSDI score. These parameters were also observed in the placebo group without statistical significance. In addition, the serum SA was significantly increased in the probiotic group in contrast to a slight non-significant increase in the placebo group. Reductions in NLRP3 inflammasome activation and OSDI were found only in the probiotic group. Conclusions: In conclusion, a significant improvement in major dry eye symptoms after iHA318 treatment was observed compared to the placebo group. Full article

Cerebrospinal fluid (CSF) is a fluid critical to brain development, function, and health. It is actively secreted by the choroid plexus, and it emanates from brain tissue due to osmolar exchange and the constant contribution of brain metabolism and astroglial fluid output to interstitial fluid into the ventricles of the brain. CSF acts as a growth medium for the developing cerebral cortex and a source of nutrients and signalling throughout life. Together with perivascular glymphatic and interstitial fluid movement through the brain and into CSF, it also acts to remove toxins and maintain metabolic balance. In this study, we focused on cerebral folate status, measuring CSF concentrations of folate receptor alpha (FOLR1); aldehyde dehydrogenase 1L1, also known as 10-formyl tetrahydrofolate dehydrogenase (ALDH1L1 and FDH); and total folate. These demonstrate the transport of folate from blood across the blood–CSF barrier and into CSF (FOLR1 + folate), and the transport of folate through the primary FDH pathway from CSF into brain FDH + ve astrocytes. Based on our hypothesis that CSF flow, drainage issues, or osmotic forces, resulting in fluid accumulation, would have an associated cerebral folate imbalance, we investigated folate status in CSF from neurological conditions that have a severity association with enlarged ventricles. We found that all the conditions we examined had a folate imbalance, but these folate imbalances were not all the same. Given that folate is essential for key cellular processes, including DNA/RNA synthesis, methylation, nitric oxide, and neurotransmitter synthesis, we conclude that ageing or some form of trauma in life can lead to CSF accumulation and ventricular enlargement and result in a specific folate imbalance/deficiency associated with the specific neurological condition. We believe that addressing cerebral folate imbalance may therefore alleviate many of the underlying deficits and symptoms in these conditions. Full article

Ceftolozane-tazobactam (CT) is used for the treatment of complicated infections and for multidrug-resistant strains of Pseudomonas aeruginosa and extended-spectrum beta-lactamase-producing enterobacteria. In certain cases, simultaneous administration of CT and parenteral nutrition (PN) may be required, but compatibility of Y-site co-administration is unknown. The aim of this study was to analyse the physicochemical compatibility of CT Y-site administered with PN. We evaluated a protocolized PN approach for critical patients in our center. We studied both bolus infusion (2 g ceftolozane/1 g tazobactam in 1 h) and continuous infusion (CI) (6 g ceftolozane/3 g tazobactam) strategies. Samples were visually observed against light, microscopically inspected, and pH was analysed using a pH meter. The mean lipid droplet diameter (MDD) was determined via dynamic light scattering. CT concentration was quantified using HPLC–HRMS. No alterations were observed through visual or microscopic inspection. Changes in pH were ≤0.2, and changes in osmolarity were less than 5%. MDD remained below 500 nm (284.5 ± 2.1 for bolus CT and 286.8 ± 7.5 for CI CT). CT concentrations at t = 0 h and t = 24 h remained within prespecified parameters in both infusion strategies. CT is physiochemically compatible with PN during simulated Y-site administration at the tested concentration and infusion rates. Full article

Honey is a natural sweetener made by bees that exhibits antimicrobial activity, mainly related to its H₂O₂ content. The aim of this work was to research the H₂O₂ concentration of 24 Spanish honeys from different botanical origins, studying their possible correlation with glucose oxidase (GOx), catalase (CAT), and anti-Staphylococcus aureus activities (minimal inhibition concentration (MIC), minimal bactericidal concentration (MBC), and percentage of inhibition at 5% (w/v) honey against Staphylococcus aureus), as well as possible correlations among all the analyzed parameters. The results showed that the H₂O₂ concentration did not depend on the botanical origin of the honeys. There were neither correlations between the H₂O₂ concentration and the activities of GOx and CAT, nor between GOx and antimicrobial activity. However, CAT and antimicrobial activities were positively correlated. Therefore, CAT could be successfully used as a possible marker of the antimicrobial activity of honeys against Staphylococcus aureus. Furthermore, a linear regression model has been fitted to explain the antimicrobial activity from CAT and GOx activity and H₂O₂ concentration. Although H₂O₂ is one of the compounds involved in honey’s antibacterial activity, this capacity also strongly depends on other honey components (such as low water activity, acidity, osmolarity, and phenolic compounds). The very high anti-Staphylococcus aureus activity exhibited by all samples could be interesting for commercial honey-based formulations also helping to promote local beekeeping. Full article

Absorption spectra of red blood cell (RBC) suspensions are investigated in an osmolarity range in the medium from 200 mOsm to 900 mOsm. Three spectral parameters are used to characterize the process of swelling or shrinkage of RBC—the absorbance at 700 nm, the Soret peak height relative to the spectrum background, and the Soret peak wavelength. We show that with an increase in the osmolarity, the absorbance at 700 nm increases and the Soret peak relative height decreases. These changes are related to the changes in the RBC volume and the resulting increase in the hemoglobin intracellular concentration and index of refraction. Confocal microscopy and flow cytometry measurements supported these conclusions. The maximum wavelength of the Soret peak increases with increasing osmolarity due to changes in the oxygenation state of hemoglobin. Using these spectrum parameters, the process of osmosis in RBCs can be followed in real time, but it can also be applied to various processes, leading to changes in the volume and shape of RBCs. Therefore, we conclude that UV–Vis absorption spectrophotometry offers a convenient, easily accessible, and cost-effective method to monitor changes in RBC, which can find applications in the field of drug discovery and diagnostics of RBC and hemoglobin disorders. Full article

Endogenous positively charged organic substances, including neurotransmitters and cationic uremic toxins, as well as exogenous organic cations such as the anti-diabetic medication metformin, serve as substrates for organic cation transporters (OCTs) and multidrug and toxin extrusion proteins (MATEs). These proteins facilitate their transport across cell membranes. Vectorial transport through the OCT/MATE axis mediates the hepatic and renal excretion of organic cations, regulating their systemic and local concentrations. Organic cation transporters are part of the remote sensing and signaling system, whose activity can be regulated to cope with changes in the composition of extra- and intracellular fluids. Glucose, as a source of energy, can also function as a crucial signaling molecule, regulating gene expression in various organs and tissues. Its concentration in the blood may fluctuate in specific physiological and pathophysiological conditions. In this work, the regulation of the activity of organic cation transporters was measured by incubating human embryonic kidney cells stably expressing human OCT1 (hOCT1), hOCT2, or hMATE1 with high glucose concentrations (16.7 mM). Incubation with this high glucose concentration for 48 h significantly stimulated the activity of hOCT1, hOCT2, and hMATE1 by increasing their maximal velocity (V_max), but without significantly changing their affinity for the substrates. These effects were independent of changes in osmolarity, as the addition of equimolar concentrations of mannitol did not alter transporter activity. The stimulation of transporter activity was associated with a significant increase in transporter mRNA expression. Inhibition of the mechanistic target of rapamycin (mTOR) kinase with Torin-1 suppressed the transporter stimulation induced by incubation with 16.7 mM glucose. Focusing on hOCT2, it was shown that incubation with 16.7 mM glucose increased hOCT2 protein expression in the plasma membrane. Interestingly, an apparent trend towards higher hOCT2 mRNA expression was observed in kidneys from diabetic patients, a pathology characterized by high serum glucose levels. Due to the small number of samples from diabetic patients (three), this observation must be interpreted with caution. In conclusion, incubation for 48 h with a high glucose concentration of 16.7 mM stimulated the activity and expression of organic cation transporters compared to those measured in the presence of 5.6 mM glucose. This stimulation by a diabetic environment could increase cellular uptake of the anti-diabetic drug metformin and increase renal tubular secretion of organic cations in an early stage of diabetes. Full article