Search Results (95)

The research on cytokine or growth factor (GF) release by leukaemic blasts is a largely unexplored area. This study aimed to evaluate the differential secretory potential of paediatric B-cell precursor and T-cell acute lymphoblastic leukaemia (BCP-ALL and T-ALL, respectively) and acute myeloid leukaemia cells (AMLs) for selected GFs, both basally and upon stimulation with phytohemagglutinin (PHA), lipopolysaccharide (LPS), or phorbol 12-myristate 13-acetate with ionophore A23187 (PMA + I). The concentrations of five GFs: granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) in the supernatants were measured using the Bio-Plex multiplex immunoassay. AML blasts showed the highest basal concentrations of G-CSF, GM-CSF, and VEGF. PHA and LPS stimulation non-selectively enhanced the secretion of G-CSF, GM-CSF, VEGF, and PDGF in BCP-ALL and AML blasts. PMA + I was the strongest GF release inducer, particularly for BCP-ALL and T-ALL blasts, with the latter also showing higher responsiveness to PHA and LPS. Our findings reveal differential, leukaemia-type dependent GF secretion patterns. Lineage-specific responses may be exploitable for targeted therapeutic approaches for distinct AL types. This study is the first to comprehensively assess the extracellular secretion of multiple GFs by paediatric AL cells in cultures using a Bio-Plex multiplex immunoassay. Full article

The agricultural industry faces increasing environmental degradation due to the intensive use of conventional chemical fertilizers, leading to water pollution and alterations in soil composition. In addition, root-knot and cyst nematodes are major constraints to cucumber production, causing severe root damage and yield losses worldwide, underscoring the need for sustainable alternatives to conventional fertilization and pest management. Under greenhouse conditions, a four-month cultivation trial evaluated vegetable oil-, micronutrient-, and activated flavonoid-based biostimulants, applying Key Eco Oil^® (Miami, USA) via soil drench (every 15 days) combined with foliar sprays of CropBioLife^® (Victoria, Australia) and KeyPlex 120^® (Miami, USA) (every 7 days). Results showed reduced parasitic nematodes by 66% in soil and decreased gall formation by 41% in roots. Chlorophyll fluorescence and infrared gas analysis revealed higher oxygen-evolving complex efficiency (38%), increased PSII electron transport, improved the fluorescence decrease ratio, also known as the vitality index (Rfd), and higher CO₂ assimilation compared to conventional treatments. Processed cucumbers showed higher sugar and nearly double ascorbic acid content, with improved flesh consistency and color. Therefore, the application of these bioactive products significantly reduced nematode infestation while enhancing plant growth and physiological performance, underscoring their potential as sustainable tools for crop cultivation and protection. These results provide evidence that sustainable bioactive biostimulants improve plant resilience, productivity, and nutritional quality, offering also an environmentally sound approach to pest management. Full article

Biodegradable magnesium (Mg) alloys are promising materials for temporary orthopaedic implants, combining favourable mechanical properties and superplastic behaviour with in vivo resorption. This enables (i) prolonged implant duration, (ii) fabrication of complex-shaped prostheses via superplastic forming (SPF), (iii) elimination of removal surgery, and (iv) reduced risk of long-term complications. However, rapid corrosion under physiological conditions remains a major limitation, highlighting the need for surface treatments that slow degradation while preserving implant integrity. This study investigates the effects of hydrothermal surface treatment on MgAZ31-SPF alloys, focusing on immunomodulatory responses, antibacterial potential, and degradation behaviour. Hydrothermally treated MgAZ31-SPF (MgAZ31-SPF-HT) extracts released lower Mg²⁺ concentrations (29.2 mg/dL) compared to untreated MgAZ31-SPF (47.5 mg/dL) while maintaining slightly alkaline pH (7–8.7), indicating improved control of early degradation. In vitro assays with human peripheral blood mononuclear cells (hPBMCs) and normal human dermal cells (NHDCs) showed that MgAZ31-SPF-HT extracts maintained higher cell viability over 24–72 h. Gene expression analysis revealed significant downregulation of pro-inflammatory markers CTSE and TNF-α, while protein quantification via ELISA and BioPlex confirmed reduced secretion of TNF-α, TGF-β1, TGF-β2, IL-6, and IL-8, suggesting mitigation of early immune activation. Antibacterial assays demonstrated limited Staphylococcus aureus colonisation on both MgAZ31-SPF and MgAZ31-SPF-HT scaffolds, with CFU counts (~10⁵–10⁶) well below the threshold for mature biofilm formation (~10⁸), and SEM analysis confirmed sparse bacterial distribution without dense EPS-rich layers. Overall, hydrothermal treatment improves Mg alloy biocompatibility by controlling Mg²⁺ release, modulating early immune responses, and limiting bacterial adhesion, highlighting its potential to enhance clinical performance of Mg-based implants. Full article

This study aimed to evaluate cytokine profiling in a periapical lesion to provide a rationale for future treatment strategies for periapical lesions. Thirteen samples of exudative fluid were collected from such a lesion directly through the root canal. Cytokine profiling was performed using the Bio-Plex system. CXCL10 (C-X-C motif chemokine ligand 10, IP10) was found to be elevated in apical exudates of patients exhibiting favorable healing. To evaluate the role of CXCL10 in cell migration, a Transwell assay was conducted using bone marrow-derived mononuclear cells (BMMCs). Different types of cytokines were detected from the samples of periapical lesion at the initial visit. However, cytokine production varied across patient samples. Release of interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-γ), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, MIP-1β, and tumor necrosis factor (TNF)-α showed differential expression. Comparison of cytokine profiles indicated that cytokine production was variable before and after root canal treatment. In vitro, CXCL10 significantly improved BMMC migration in a dose-dependent manner, supporting clinical findings that elevated CXCL10 levels are associated with favorable healing in apical lesions. Although this study was limited by the small sample size and exploratory design, the cytokine profile of periapical lesions may be useful for assessing the condition of periapical lesions and modulating the immune response to bacterial infection. Full article

Magnesium (Mg) alloys, particularly Mg AZ31, have emerged as promising biomaterials for orthopedic applications due to their biodegradability and favorable mechanical characteristics. Among these, the Mg AZ31+SPF alloy, subjected to hydrothermal (HT) treatment, has demonstrated enhanced bioactivity. Our previous research established that this surface modification supports the osteogenic differentiation of human mesenchymal stem cells (hMSCs) by modulating both canonical and non-canonical signaling pathways, including those implicated in osteogenesis, hypoxic response, exosome biogenesis, and lipid metabolism. In the present study, we extended our investigation to assess the effects of Mg AZ31+SPF+HT and Mg AZ31+SPF extracts on murine pre-osteoclasts (RAW 264.7 cells) over 3- and 6-day treatment periods. The primary objectives were to evaluate biocompatibility and to investigate potential impacts on osteoclastogenesis induction and miRNA expression profiles. Methods: To assess cytocompatibility, metabolic activity, DNA integrity, and morphological alterations in RAW 264.7 cells were evaluated. Osteoclast differentiation was quantified using TRAP staining, alongside the assessment of osteoclastogenic marker expression by qRT-PCR and ELISA. The immunomodulatory properties of the extracts were examined using multiplex BioPlex assays to quantify soluble factors involved in bone healing. Additionally, global miRNA expression profiling was performed using a specialized panel targeting 82 microRNAs implicated in bone remodeling and inflammatory signaling. Results: Mg AZ31+SPF+HT extract exhibited high biocompatibility, with no observable adverse effects on cell viability. Notably, a significant reduction in the number of TRAP-positive and multinucleated cells was observed relative to the Mg AZ31+SPF group. This effect was corroborated by the downregulation of osteoclast-specific gene expression and decreased MMP9 protein levels. Cytokine profiling indicated that Mg AZ31+SPF+HT extract promoted an earlier release of key cytokines involved in maintaining the balance between bone formation and resorption, suggesting a beneficial role in bone healing. Furthermore, miRNA profiling revealed a distinct regulatory signature in Mg AZ31+SPF+HT-treated cells, with differentially expressed miRNAs associated with inflammation, osteoclast differentiation, apoptosis, bone resorption, hypoxic response, and metabolic processes compared to Mg AZ31+SPF-treated cells. Conclusions: Collectively, these findings indicate that hydrothermal treatment of Mg AZ31+SPF (resulting in Mg AZ31+SPF+HT) attenuates pre-osteoclast activation by influencing cellular morphology, gene and protein expression, as well as post-transcriptional regulation via modulation of miRNAs. The preliminary identification of miRNAs and the activation of their regulatory networks in pre-osteoclasts exposed to hydrothermally treated Mg alloy are described herein. In the context of orthopedic surgery—where balanced bone remodeling is imperative—our results emphasize the dual significance of promoting bone formation while modulating bone resorption to achieve optimal implant integration and ensure long-term bone health. Full article

Introduction: Thyroid cancer and obesity are prevalent conditions with significant global health implications. Differentiated thyroid cancer (DTC) is influenced by various molecular pathways, including those involving Transforming Growth Factor-Beta (TGF-β), a cytokine implicated in cell proliferation, differentiation, immune regulation, and fibrosis. Obesity (BMI > 30) has been associated with thyroid dysfunction and an increased incidence of nodular goiter. However, the relationship between TGF-β levels, thyroid malignancies, and metabolic disturbances remains unclear. This study aimed to analyze TGF-β1, -2, and -3 concentrations in blood serum before and after thyroidectomy in patients with DTC and obese individuals with nodular goiter to evaluate their potential role in thyroid pathology and obesity-related metabolic changes. Methods: A prospective study was conducted at a high-volume surgical center where over 700 thyroidectomy procedures are performed annually. Seventy-six consecutive patients (aged 26–79 years) were included: 21 with DTC and 55 with euthyroid nodular goiter. The latter group was subdivided based on BMI into obese (BMI > 30, n = 26) and non-obese (BMI < 30, n = 29) cohorts. Blood samples were collected preoperatively and on the first postoperative day for TGF-β quantification using the Bio-Plex Pro™ Human Cytokine Assay. Statistical analysis was performed using the Student’s t-test. Results: Postoperatively, patients with DTC exhibited significantly higher TGF-β1 (210,000 pg/mL), TGF-β2 (360 pg/mL), and TGF-β3 (170 pg/mL) levels compared to obese patients with nodular goiter (p < 0.05). In the nodular goiter group, BMI did not significantly influence preoperative TGF-β levels (p > 0.05). However, postoperatively, obese patients showed lower TGF-β1 (100,000 pg/mL) and TGF-β2 (30 pg/mL) levels compared to normal-weight individuals (p = 0.03), while no significant difference was observed for TGF-β3 (p > 0.05). Conclusions: The study highlights distinct alterations in TGF-β isoform levels in thyroid cancer and obesity. Elevated postoperative TGF-β levels in DTC patients suggest a role in tumor progression and response to surgical intervention. In contrast, the reduction of TGF-β1 and TGF-β2 levels in obese patients postoperatively may indicate a complex interplay between obesity, surgical stress, and cytokine regulation. These findings underscore the need for further research into the molecular mechanisms governing TGF-β dynamics in thyroid disorders and obesity, with potential implications for therapeutic interventions. Full article

Wilms tumor (WT), the most common pediatric renal malignancy, shares some clinical and pathological features with chronic kidney disease (CKD). Understanding biomarkers of kidney injury among CKD and WT patients is of high interest due to its potential implications for diagnosis, prognosis, and treatment strategies. This study enrolled twenty pediatric patients with WT (stage I–IV), forty with CKD (stage I–V), and twenty healthy volunteers. Urine samples were collected and six urine biomarkers (calbindin, clusterin, GST-π, IL-18, KIM-1, MCP-1) associated with kidney injury were assessed using the Bio-Plex Pro RBM Human Kidney Toxicity Assays kit (Bio-Plex Manager software 4.0). A comparative analysis of biomarker levels across the three groups revealed distinct patterns. Creatinine levels were notably elevated in CKD (1.32 ± 1.9) compared to WT (0.64 ± 0.26) and the control group. Tested biomarkers were calculated per milligram of urine creatinine, and all the differences among the groups were statistically significant. Pearson’s correlation coefficients showed strong interplay among CKD biomarkers. This study identified variations in biomarker patterns among WT and CKD patients. Understanding biomarker interactions may provide future diagnostic approaches for pediatric kidney conditions. Full article

Preeclampsia, one of the leading causes of maternal and fetal morbidity and mortality, affects approximately 3–5% of pregnancies worldwide. However, its etiology remains poorly understood. The aim of this study was to identify molecular markers of preeclampsia. Protein concentrations in blood and urine were determined using the Bio-Plex Kidney Toxicity 1 assay Bio-Rad, Hercules, CA, USA followed by magnetic separation and flow cytometry. This study included 51 patients with preeclampsia and 25 healthy pregnant women. The results revealed that five out of the six serum biomarkers of kidney injury were elevated in the preeclampsia group compared to the control group (calbindin 1, clusterin, glutathione transferase pi (GSTP1), monocyte chemotactic protein 1 (MCP-1), and kidney injury molecule type 1 (KIM-1)). Additionally, the serum concentrations of calbindin 1, clusterin, GSTP1, and KIM-1 were significantly higher in both early-onset and late-onset preeclampsia compared to the control group. The analysis of urinary proteins showed that only the KIM-1 concentration was elevated in late-onset preeclampsia compared to the control group. These findings suggest that the calbindin 1, clusterin, GSTP1, KIM-1, and MCP-1 concentrations in maternal plasma could serve as potential biomarkers for monitoring kidney injury in preeclamptic women. This study provides a foundation for future research to explore novel biomarkers of preeclampsia and renal injury in pregnant women. Full article

Infections with the Hepatitis B (HBV) and Hepatitis C (HCV) viruses share some transmission routes, which is why coinfection with these viruses becomes common, especially in endemic areas. This study evaluated the immunological response profile, viral load, and liver damage in groups monoinfected with HBV or HCV and in those co-infected with HBV/HCV. The groups were composed of 22 patients monoinfected by HCV, 22 patients monoinfected by HBV, and 34 co-infected by HBV/HCV, according to serological markers and molecular biology tests. The study was carried out from December 2017 to October 2019. Virus detection employed enzyme immunoassay, Enzyme-Linked Immunosorbent Assay (ELISA), and real-time PCR, while liver function and fibrosis were assessed using biochemical tests and Fibroscan. To research the immunological profile, cytokines were quantified using the BIO-Plex Pro Human Cytokine. Comparing the groups, both mono- and co-infected patients exhibited a Th1 immune response profile. HCV monoinfection notably showed significantly elevated serum levels of INF-γ (p < 0.01) and TNF-α (p < 0.01). The viral load was significantly higher in the HCV monoinfected group when compared to the other groups. Regarding liver damage, patients with a high level of fibrosis (F4) presented significant levels of cytokines INF-γ (p < 0.001), IL-17 (p < 0.0001), and TNF-α (p < 0.0001). Full article

Foods such as Curcuma longa L. and propolis can attenuate inflammation in patients with chronic kidney disease (CKD) undergoing hemodialysis (HD). This study aimed to evaluate the effects of microcapsules loaded with Curcuma longa L. and propolis on inflammatory markers and uremic toxins in patients undergoing HD. In this randomized, double-blind clinical trial, 40 patients were divided into two groups: an intervention group (137 mg/day of Curcuma and 500 mg/day of green propolis) in the form of microcapsules, and a placebo group, both administered for 8 weeks. Cytokines were analyzed using a multiplex assay (Bio-Plex Magpix^®). Malondialdehyde was evaluated as a marker of lipid peroxidation. Uremic toxins were analyzed by reversed-phase high-performance liquid chromatography. Demographic and clinical data were obtained from medical records. A total of 38 patients completed the study: 18 were in the intervention group (49 ± 16.2 years; 8 men) and 20 were in the control group (49 ± 18.7 years; 10 men). There was a reduction in levels of C-reactive protein (p = 0.026) and MIP-1 (p = 0.019) in the intervention group. No change in uremic toxins was observed. In conclusion, the intervention with microcapsules containing Curcuma longa L. and green propolis showed potential anti-inflammatory effects in patients with CKD undergoing HD. These findings warrant investigation in larger, long-term trials. Full article

Background: Hepatitis B virus (HBV) is a major cause of morbidity and mortality globally, and chronic infections are associated with cirrhosis and hepatocellular carcinoma. Issues with conventional treatments and vaccines mean there is a need for new therapeutic vaccines, which must elicit a strong and sustainable immune response. Here, we evaluated the immunogenicity of dual-antigen vaccines containing hybrid surface (hy-LHBs) and core (HBc) antigens, combined with a carboxyl-vinyl polymer (CVP) as a mucoadhesive excipient, following intranasal administration in mice. Methods: Mice were intranasally administered a mixed vaccine (10 µg of hy-LHBs and 2.5 or 10 µg of HBc) with or without a CVP excipient, and they were assessed for their immune response (levels of IgGs or IgA antibodies in an ELISA, IFN-γ level in splenocytes in an ELISpot assay, and cytokine/chemokine levels in a BioPlex assay). A protein stability assay was also conducted for vaccine formulations with and without excipients. Results: Significantly enhanced IgG production was noted targeting hy-LHBs and (less markedly) HBc at 10 µg/antigen, but only a non-significant elevation was noted with the vaccine containing 2.5 µg HBc. The BioPlex assay showed a significant increase in IL-2 (#00-07, 0B), IL-12(p40)(#00), eotaxin (#00), MIP1α (#00, #00-07, 0B), and MCP-1 (#00-07, 0B) in mice that received treatment compared to those of untreated mice. The endpoint titers of IgG1 and IgG2a were measured, which were higher with CVP excipients than without. From the IgG2a/IgG1 ratio, a higher IgG1 response was induced by CVPs to hy-LHBs and a higher IgG2a response was induced to HBc. Th2-dominant phenotype to hy-LHBs was induced with CVP#00 in an ELISpot assay. The highest anti-hy-LHBs antibody titer was noted with the conventional CVP#00 excipient. Consistent with these results, a higher amount of neutralizing antibodies of HBV was induced with CVP#00 treatment and followed by #00-03 and #14-00. Conclusions: We consider that the addition of CVP excipients to vaccine formulation enhances immunogenicity and HBV antigen stability for intranasal vaccines. This effect was seen for both humoral and cell-mediated immune responses, indicating the potential of CVPs as excipients in intranasal HBV vaccines. Full article

Background: Reliable biomarkers are urgently needed to aid in the differential diagnosis, prognosis, disease progression monitoring, and prediction of therapeutic response in patients with systemic sclerosis (SSc). This study aimed to evaluate a panel of potentially pathogenic circulating cytokines and chemokines in a cohort of SSc patients. Methods: Serum samples were obtained from 35 SSc patients and 40 age- and sex-matched healthy controls. Twenty-three cytokines/chemokines were quantified using a Luminex^® multiplex immunoassay (BioRad-BioPlex 200 System-Lumine x-Map technology R&D Systems, USA) following the manufacturer’s instructions and customized procedures. Data were acquired using Bioplex manager v 6.1. Data analysis was performed using GraphPad Prism v.8 (GraphPad Software, Inc.), with significance defined as p ≤ 0.05. V.8 Results: Levels of TNFRII and MMP-8 were significantly higher in SSc patients compared to healthy controls, while IL-1RII levels were significantly elevated in healthy individuals (p < 0.001 for all comparisons). Patients with elevated ESR at baseline (>30 mm/h) showed higher IL-15 levels (p = 0.019). A strong positive correlation was found between MCP-1 and the modified Rodnan skin score (mRSS) (p = 0.009, r = 0.740), and a moderate correlation between TNFRII and diffusing capacity for carbon monoxide (p = 0.046, r = 0.339). PLS regression identified MMP-8, MCP-1, TNFRII, IL-15, and IL-1RII as key predictors of SSc, with MMP-8 having the strongest influence. The logistic regression model showed high performance (AUC = 0.93, accuracy = 87.5%). Conclusions: TNFRII, MMP-8, and IL-1RII may play a pathogenic role in SSc. TNFRII, in particular, may serve as a biomarker for pulmonary involvement, aligning with its known role in pro-fibrotic pathways. These findings support their utility in diagnosis and disease profiling. Full article

Background: The coronavirus–caused disease (COVID-19), first identified in China in December 2019, has spread worldwide becoming a global pandemic. Although people infected with the SARS-CoV-2 virus presented mainly respiratory and gastrointestinal symptoms, an increase in cardiovascular incidents was observed in several scientific studies. SARS-CoV-2 virus has been shown to disrupt the normal immune response leading to a dysregulation of immune system function and massive production of inflammatory cytokines commonly known as “cytokine storm”. Methods: 57 patients eventually participated in the study, assigned to non–COVID (24 patients) and COVID (33 patients) groups. After signing consent to participate in the study, each patient was given a self–administered questionnaire to fill out prior to specimen collection, anthropometric measurements were taken and venous blood was collected for the following determinations: pro- and anti–inflammatory cytokines using a Bio–Plex 200 system, oxidative stress markers and basic hematological blood parameters. Results: showed statistically significant higher values of IL–1Ra and IL–1β in the COVID-19 group. Of the oxidative stress markers, only MDA levels were higher in the COVID-19 group. Conclusions: the results of our study provide evidence and support the occurrence of elevated levels of IL–1Ra, IL–1β and MDA in the COVID-19 group of patients, which are associated with a worse course and prognosis of COVID-19. A better understanding of the pathophysiology and dysregulation of the immune system associated with the cytokine storm is essential to select patients at risk and develop effective drugs and vaccines. Full article

An Mg-based alloy device manufactured via a superplastic forming process (Mg-AZ31+SPF) and coated using a hydrothermal method (Mg AZ31+SPF+HT) was investigated as a method to increase mechanical and osteointegration capability. The cell viability and osteointegrative properties of alloy-derived Mg AZ31+SPF and Mg AZ31+SPF+HT extracts were investigated regarding their effect on human mesenchymal stem cells (hMSCs) (maintained in basal (BM) and osteogenic medium (OM)) after 7 and 14 days of treatment. The viability was analyzed through metabolic activity and double-strand DNA quantification, while the osteoinductive effects were evaluated through qRT-PCR, osteoimage, and BioPlex investigations. Finally, a preliminary liquid mass spectrometry analysis was conducted on the secretome of hMSCs. Biocompatibility analysis revealed no toxic effect on cells’ viability or proliferation during the experimental period. A modulation effect was observed on the osteoblast pre-commitment genes of hMSCs treated with Mg-AZ31+SPF+HT in OM, which was supported by mineralization nodule analysis. A preliminary mass spectrometry investigation highlighted the modulation of protein clusters involved in extracellular exosomes, Hippo, and the lipid metabolism process. In conclusion, our results revealed that the Mg AZ31+SPF+HT extracts can modulate the canonical and non-canonical osteogenic process in vitro, suggesting their possible application in bone tissue engineering. Full article

Background: Psoriasis is a chronic, multi-system inflammatory disease frequently associated with metabolic syndrome and lipid disturbances. Apolipoproteins, as essential regulators of lipid metabolism, may play a critical role in these metabolic abnormalities, potentially influencing disease severity and systemic inflammation. The aim of this study was to compare serum concentrations of chosen apolipoproteins in patients with psoriasis before and after treatment with acitretin or narrowband UVB (NB-UVB). Methods: This study was conducted on 39 patients with psoriasis. The concentration of nine apolipoproteins and C-reactive protein was quantified using the Bio-Plex Immunoassay Kit. Results: The serum concentrations of ApoA2, ApoC1, ApoD, ApoE, and ApoJ were higher in the acitretin group compared to the NB-UVB group before treatment, while the ApoA1/ApoA2 ratio was lower. We also observed a negative association between the Psoriasis Area and Severity Index (PASI) and ApoA1/ApoA2 ratio in the patients before the treatment. Conclusions: The results of this study confirm the presence of metabolic disturbances in psoriatic patients. The treatment with NB-UVB or acitretin did not cause any significant changes in the apolipoproteins profile. Thus, we found no detrimental impact of acitretin on the apolipoproteins profile, despite the observed rise in total cholesterol concentration after the treatment. Further research is needed to explore whether specific therapeutic approaches can modify these disturbances and potentially improve long-term cardiovascular outcomes in this population. Full article