Mesenchymal Stem Cell-Conditioned Media Modulate HUVEC Response to H2O2: Impact on Gene Expression and Potential for Atherosclerosis Intervention.

Background: We studied the potential of human bone marrow-derived mesenchymal stem cell conditioned media (hBMSC CM) in protecting endothelial cell properties (viability, proliferation, and migrations) from the deleterious effects produced by the inflammatory environment of H2O2. Additionally, we investigated their impact on the endothelial cells' gene expression of some inflammatory-related genes, namely, TGF-ß1, FOS, ATF3, RAF-1, and SMAD3. Methods: Human umbilical vein endothelial cells (HUVECs) were cultured individually under three conditions: alone, with varying concentrations of H2O2, or with varying concentrations of H2O2 and hBMSC CM. HUVEC adhesion, proliferation, and migration were evaluated using the xCELLigence system. The HUVECs' gene expressions were evaluated by real-time polymerase chain reaction (RT-PCR). Results: Generally, we observed enhanced HUVEC viability, proliferation, and migration when cultured in media supplemented with H2O2 and hBMSC CM. Furthermore, the CM modulated the expressions of the studied inflammatory-related genes in HUVECs, promoting a more robust cellular response. Conclusion: This study has illuminated the protective role of hBMSC CM in mitigating the damaging effects of H2O2 on endothelial cell function. Our data demonstrate that hBMSC CM enhances the viability, proliferation, and migration of HUVECs even under oxidative stress conditions. Additionally, the conditioned medium was found to modulate the gene expression of pivotal markers related to inflammation, suggesting a favorable influence on cellular response mechanisms.

Heavy metal levels are positively associated with serum periostin and miRNA-125b levels, but inversely associated with miRNA-26a levels in pediatric asthma cases. A case-control study.

BACKGROUND: The study investigated heavy metals levels [urinary cadmium (U-Cd), erythrocytic cadmium (E-Cd), urinary arsenic (U-As), and whole blood lead (WB-Pb)] in children with bronchial asthma (BA) and tested their associations with serum periostin, miRNA-125b and miRNA-26a levels, and with asthma severity clinically and laboratory [blood eosinophils count (BEC) and serum total immunoglobin E (IgE)]. Also, we tested cut-off points, for the studied parameters, to distinguish BA cases from healthy children. METHODS: This case-control study included 158 children divided into control group; n = 72 and BA group; n = 86. Heavy metals were measured by an inductively coupled plasma-optical emission spectrophotometer. Serum periostin and IgE levels were measured by their corresponding ELISA kits. miRNAs relative expressions were estimated by RT-qPCR using the 2-ΔΔCT method. RESULTS: Heavy metals, serum periostin, and miR-125b levels were significantly high in BA group (p < 0.001). Heavy metals levels correlated positively with serum periostin, miR-125b and IgE levels, BEC, and asthma severity. The reverse was observed regarding serum miR-26a levels. Receiver operating characteristics (ROC) curve analysis showed good to excellent abilities of U-Cd, E-Cd, U-As, WB-Pb, serum periostin, miRNA-125b, and miRNA - 26a, and total IgE levels to distinguish BA cases from healthy children. CONCLUSIONS: Heavy metal toxicity in children is associated with BA severity, increased serum periostin and miRNA-125b levels, and decreased miRNA-26a levels. Specific measures to reduce children's exposure to heavy metals should be taken. Future research should consider blocking miRNA-125b action or enhancing miRNA-26a action to manage BA cases.

Pigment epithelium-derived factor (PEDF) represses the glucose transporter 1 (GLUT1) mRNA expression and may be a potential therapeutic agent in psoriasis: a case-control and experimental study.

We investigated the whole blood GLUT1 mRNA expression and serum pigment epithelium-derived factor (PEDF), interleukin-6 (IL-6), fetuin-A, and pentraxin-3 (PTX3) levels in psoriatic patients and tested their correlations with the severity of psoriasis using the psoriasis area and severity index (PASI) score. Also, we tested the GLUT1 mRNA expression after an in vitro treatment of human skin fibroblast (HSF) cell lines with PEDF. The case-control part of the study recruited 74 participants (44 psoriatic patients and 30 healthy volunteers). Whole blood GLUT1 mRNA fold changes were estimated by RT-PCR, and serum PEDF, IL-6, fetuin-A, and PTX3 levels were measured by ELISA kits. In the experimental part, the HSF cell lines were treated with different concentrations of PEDF for different times to test its effect on the GLUT1 mRNA expression. The whole blood GLUT 1 expression significantly increased in psoriatic patients and correlated positively with serum IL-6, fetuin-A, PTX3 levels and with the severity of psoriasis while negatively with serum PEDF levels. The PEDF-treated HSF cell lines showed a time- and dose-dependent decline in the GLUT 1 mRNA expression. The whole blood GLUT 1 mRNA is a non-invasive biomarker that is associated with the severity of psoriasis. PEDF represses GLUT 1 expression and may be a potential therapeutic agent in psoriasis.Trial registration: ClinicalTrials.gov Identifier: NCT04242082.

Exposure to pesticide components causes recurrent pregnancy loss by increasing placental oxidative stress and apoptosis: a case-control study.

We investigated the plasma levels of pesticides components namely polychlorinated biphenyls (PCBs), dieldrin, dichlorodiphenyldichloroethylene (DDE), ethion, malathion, and chlorpyrifos in recurrent pregnancy loss (RPL) cases, and tested their associations with placental oxidative stress (OS) biomarkers [nitric oxide (NO.), thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), and superoxide dismutase (SOD)] and with placental apoptotic/antiapoptotic indices (Bcl-2 and caspase-3), and evaluated their possible cut-off points to distinguish RPL cases. The study recruited 101 pregnant women divided into; G1 [n = 49, control, normal 1st-trimester pregnancy, normal obstetric history with at least one previous normal live birth], G2 [n = 26, cases with missed abortion (< 3 abortions) before 24 weeks of gestation], and G3 [n = 26, cases with missed abortion (≥ 3 abortions) before 24 weeks of gestation]. The plasma pesticide levels were analyzed by gas chromatography-mass spectrometry. Plasma human chorionic gonadotrophin (HCG), placental OS, Bcl-2, and caspase-3, were analyzed by their corresponding methods and kits. Plasma PCBs, DDE, dieldrin, and ethion levels were significantly higher in RPL cases than in normal pregnancies (p ≤ 0.001). These levels correlated positively with placental OS and apoptosis and negatively with plasma HCG levels. Also, these levels were reliable markers of risk to RPL. Malathion and chlorpyrifos were not detected in any of the study's participants. Pesticides may be risk factors in cases of spontaneous RPL cases. They are associated with an increasing placental OS and placental apoptosis. Specific measures should be taken to decrease maternal exposure to these pollutants' sources, especially in underdeveloped and developing countries.

Cytotoxic n-Hexane Fraction of the Egyptian Pteris vittata Functions as Anti-breast Cancer Through Coordinated Actions on Apoptotic and Autophagic Pathways.

We investigated the possible anticancer mechanisms of Pteris vittata [PV] n-hexane extract on MCF-7 [breast cancer cell line]. Cultured cell lines were treated with various concentrations of this extract ± Baf-A1 [autophagic inhibitor]. Cells' viability, apoptotic markers [caspase-7, Bax, and Bcl-2], autophagic markers [light chain 3 [LC-3] and P62/SQSTM1]], and the tumor suppressor P53 and its mRNA were checked by their corresponding methods. Treated cell lines showed significant concentration and time-dependent reductions in cell viability in response to PV-n-hexane extract and also exhibited a concomitant induction of apoptosis [increased chromatin condensation, nuclear fragmentation, and pro-apoptotic Bax, and cleaved caspase-7 levels while decreased Bcl-2 levels] and autophagy [increased autophagosomes vacuoles, and LC3B II levels while decreased P62/SQSTM1 levels]. Moreover, PV-n-hexane extract-treated cells showed significant increases in the P53 and its mRNA levels. The addition of Baf-A1 reversed the PV-n-hexane extract autophagic effects and increased apoptotic cell percentage with a much increase in the cleaved caspase-7 and P53 protein and its mRNA levels. We concluded that the PV-n-hexane extract exhibits cytotoxic effects on the MCF-7 cell line with significant reductions in cell viability and concomitant autophagy and apoptosis induction. Inhibition of autophagy in the PV-treated MCF-7 cells enhances apoptosis via a p35-dependent pathway.

Neonatal heavy metals levels are associated with the severity of neonatal respiratory distress syndrome: a case-control study.

BACKGROUND: This case-control study aimed to compare lead (Pb), cadmium (Cd), and arsenic (As) levels in neonates with respiratory distress syndrome (NRDS) with those levels in normal neonates and tested their associations with the severity of NRDS indicated by the levels of serum surfactant protein D (SP-D) and cord blood cardiac troponin I (CTnI), and high-sensitive C-reactive protein (hs-CRP). METHODS: The study included two groups: G1 (60 healthy neonates) and G2 (100 cases with NRDS). Cord blood Pb, erythrocytic Cd (E-Cd), neonatal scalp hair As (N-As), maternal urinary Cd (U-Cd), and arsenic (U-As) were measured by a Thermo Scientific iCAP 6200, while CTnI, hs-CRP, and SP-D by their corresponding ELISA kits. RESULTS: The levels of cord blood Pb, E-Cd, N-As, U-Cd, U-As, SP-D, CTnI, and hs-CRP were significantly higher in G2 than G1 (p = 0.019, 0.040, 0.003, 0.010, 0.011, < 0.001, 0.004, < 0.001, respectively). While the birth weight, and APGAR score at 1, 5 and 10 min were significantly lower in G2 than G1 (p = 0.002, < 0.001, < 0.001, < 0.001, respectively). The levels of the studied heavy metals correlated positively with the levels of SP-D, CTnI, and hs-CRP. CONCLUSION: Heavy metals toxicity may be accused to be one of the causes of NRDS especially if other apparent causes are not there. Measuring and follow-up of heavy metal levels should be considered during pregnancy.

Modification of SWCNTs with hybrid materials ZnO-Ag and ZnO-Au for enhancing bactericidal activity of phagocytic cells against Escherichia coli through NOX2 pathway.

Zinc oxide-silver (ZnO-Ag), and zinc oxide-gold (ZnO-Au) nano-composites were prepared through wet chemical process and laced into single-walled carbon nanotubes (SWCNTs) to yield ZnO-Ag-SWCNTs, and ZnO-Au-SWCNTs hybrids. These nano-composite-laced SWCNTs hybrids were characterized using Raman spectroscopic, X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) analyses. The hybrids were evaluated for their effects on phagocytic cells and bactericidal activity against the gram-negative bacteria E. coli. Their phagocytic cell activities and intracellular killing actions were found to be significantly increased, as the ZnO-Ag-SWCNTs and ZnO-Au-SWCNTs nano-hybrids induced widespread clearance of Escherichia coli. An increase in the production of reactive oxygen species (ROS) also led to upregulated phagocytosis, which was determined mechanistically to involve the phagocyte NADPH oxidase (NOX2) pathway. The findings emphasized the roles of ZnO-Ag- and ZnO-Au-decorated SWCNTs in the prevention of bacterial infection by inhibiting biofilm formation, showing the potential to be utilized as catheter coatings in the clinic.

Circulating miR-15b, Annexin A1, procalcitonin and interleukin-6 levels differentiate children with metabolically unhealthy obesity from those with metabolically healthy obesity: A case-control study.

The present study assessed serum miR-15b, Annexin A1, procalcitonin, and interleukin-6 (IL-6) levels in children with metabolically healthy obesity (MHO) and metabolically unhealthy obesity (MUO) and compared them to these levels in a non-obese healthy control group. It also tested the ability of each of these parameters to early differentiate children with MUO from those with MHO. The present study included 620 children [434 males (70%) and 186 females (30%); aged 9-15 years] divided into the following groups: G1, healthy non-obese controls (n=200); G2, MHO (n=246); G3, MUO (n=174). Serum miR-15b, Annexin A1 procalcitonin, IL-6, and other metabolic parameters levels were measured, and clinical examinations were conducted for all of the children. After testing the normality of the variable, Kruskal-Wallis one-way-ANOVA, and Spearman correlation coefficients were used. The area under the receiver operating characteristic curve (AUC) was determined to test the variable's ability to differentiate MUO from MHO. miR-15b, procalcitonin, and IL-6 levels were significantly higher while Annexin A1 levels were significantly lower in G2 and G3 when compared to G1, and in G3 when compared to G2. These levels were positively correlated (Annexin A1 was negatively correlated) with body mass index (BMI) and waist circumference percentiles, and with serum levels of LDL-cholesterol, glucose, HbA1c, insulin, and C-reactive protein (CRP) and with the homeostasis model of insulin resistance (HOMA-IR). The AUC was 0.92, 0.84, 0.82, and 0.67 for miR-15b, Annexin A1, procalcitonin, and IL-6, respectively. In conclusion, determination of serum miR-15b, Annexin A1, and procalcitonin levels could differentiate children with MUO from those with MHO. This may help the early management of these cases and their accompanying complications.

Comparative Phytochemical Profile and Biological Activity of Four Major Medicinal Halophytes from Qassim Flora.

Four halophytic plants, Lycium shawii, Anabasis articulata, Rumex vesicarius, and Zilla spinosa, growing in the central Qassim area, Saudi Arabia, were phytochemically and biologically investigated. Their hydroalcoholic extracts' UPLC-ESIQ-TOF analyses demonstrated the presence of 44 compounds of phenolic acids, flavonoids, saponins, carbohydrates, and fatty acids chemical classes. Among all the plants' extracts, L. shawii showed the highest quantities of total phenolics, and flavonoids contents (52.72 and 13.01 mg/gm of the gallic acid and quercetin equivalents, respectively), along with the antioxidant activity in the TAA (total antioxidant activity), FRAP (ferric reducing antioxidant power), and DPPH-SA (2,2-diphenyl-1-picryl-hydrazyl-scavenging activity) assays with 25.6, 56.68, and 19.76 mg/gm, respectively, as Trolox equivalents. The hydroalcoholic extract of the L. shawii also demonstrated the best chelating activity at 21.84 mg/gm EDTA equivalents. Among all the four halophytes, the hydroalcoholic extract of L. shawii exhibited the highest antiproliferative activity against MCF7 and K562 cell lines with IC50 values at 194.5 µg/mL and 464.9 µg/mL, respectively. The hydroalcoholic extract of A. articulata demonstrated better cytotoxic activity amongst all the tested plants' extracts against the human pancreatic cancer cell lines (PANC1) with an IC50 value of 998.5 µg/mL. The L. shawii induced apoptosis in the MCF7 cell lines, and the percentage of the necrotic cells changed to 28.1% and 36.5% for the IC50 and double-IC50 values at 22.9% compared with the untreated groups. The hydroalcoholic extract of L. shawii showed substantial antibacterial activity against Bacillus cereus ATCC 10876 with a MIC value of 12.5 mg/mL. By contrast, the A. articulata and Z. spinosa exhibited antifungal activities against Aspergillus niger ATCC 6275 with MIC values at 12.5 and 50 mg/mL, respectively. These findings suggested that the L. shawii is a potential halophyte with remarkable biological properties, attributed to its contents of phenolics and flavonoid classes of compounds in its extract.

Investigation of the levels of circulating miR-29a, miR-122, sestrin 2 and inflammatory markers in obese children with/without type 2 diabetes: a case control study.

AIM: The present work investigated serum levels of miR-29a, miR-122 and sestrin2 in obese children with/without type-2-diabetes mellitus (T2DM), and their correlations with inflammatory, metabolic and anthropometric parameters. METHODS: The study included 298 children, divided into: G1 (control, n = 136), G2 (obese without diabetes, n = 90) and G3 (obese with T2DM, n = 72). Metabolic and anthropometric parameters, miR-29a, miR-122 relative expressions, and sestrin2, high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were measured by their specific methods. The data was processed and analyzed by SPSS V.26 using the corresponding tests. After testing the variables' normality, Kruskal-Wallis one-way-ANOVA, Spearman correlations coefficient were used. RESULTS: Significant higher serum miR-29a, miR-122, IL-6, hsCRP and TNF-α and lower sestrin2 levels were found in G2 and G3 than G1 and in G3 than G2 (p= > 0.001 for all). Especially in G3, miR-29a and miR-122 levels correlated positively while sestrin2 levels correlated negatively with waist circumference and BMI percentiles, serum levels of LDL-cholesterol, triacylglycerol, total cholesterol, HbA1c%, glucose, insulin, c-peptide, homeostatic model assessment-insulin resistance (HOMA-IR), IL-6, hsCRP and TNF-α. CONCLUSION: The change in the serum miR-29a, miR-122 and sestrin2 levels in obese children with/without T2DM may suggest a possible role of these biomarkers in the pathogenesis of childhood obesity and their accompanied complications e.g. inflammations and T2DM. Also, further studies are required to test drugs that antagonize the action miR-29a and miR-122 or upregulate sestrin2 in the management of these cases.

Thymoquinone Lowers Blood Glucose and Reduces Oxidative Stress in a Rat Model of Diabetes.

The aim of the present study was to assess the short-term effects of Thymoquinone (TQ) on oxidative stress, glycaemic control, and renal functions in diabetic rats. DM was induced in groups II and III with a single dose of streptozotocin (STZ), while group I received no medication (control). The rats in groups I and II were then given distilled water, while the rats in group III were given TQ at a dose of 50 mg/kg body weight/day for 4 weeks. Lipid peroxidase, nitric oxide (NO), total antioxidant capacity (TAC), glycated haemoglobin (HbA1c), lipid profiles, and renal function were assessed. Moreover, the renal tissues were used for histopathological examination. STZ increased the levels of HbA1c, lipid peroxidase, NO, and creatinine in STZ-induced diabetic rats in comparison to control rats. TAC was lower in STZ-induced diabetic rats than in the control group. Furthermore, rats treated with TQ exhibited significantly lower levels of HbA1c, lipid peroxidase, and NO than did untreated diabetic rats. TAC was higher in diabetic rats treated with TQ than in untreated diabetic rats. The histopathological results showed that treatment with TQ greatly attenuated the effect of STZ-induced diabetic nephropathy. TQ effectively adjusts glycaemic control and reduces oxidative stress in STZ-induced diabetic rats without significant damaging effects on the renal function.

Association between circulating microRNAs 486, 146b and 15b and serum betatrophin levels in obese; type 2 diabetic and non-diabetic children.

BACKGROUND: This study tested the association between serum levels of microRNA-486, -146b and -15b and betatrophin in normal and obese children with/without type 2 diabetes mellitus (T2DM). METHODS: the study included 120 children; divided into three groups: G1 (50 healthy), G2 (35 obese) and G3 (35 obese with T2DM). The levels of microRNA-486, 146b and 15b and serum betatrophin were measured by their corresponding methods. RESULTS: serum microRNA-486, -146b, -15b and betatrophin levels were significantly high in G3 followed by G2 then G1 (p = 0.002, > 0.001, > 0.001, and > 0.001, respectively). Especially in G3, these levels correlated positively with the BMI percentile (r = 0.44, 0.58, 0.38, and 0.46, p = 0.007, > 0.001, 0.021, and 0.005, respectively), serum glucose (r = 0.56, 0.49, 0.82, 0.60, and 0.42, p > 0.001, 0.003, > 0.001, and > 0.001, respectively) and HbA1c% (r = 0.56, 0.39, 0.66, and 0.42, p > 0.001, 0.019, > 0.001, and 0.032, respectively) while, showed negative correlations with correlated with serum insulin levels (r = - 0.37, - 0.42, - 0.58, and - 0.41, p = 0.021, 0.012, > 0.001 and 0.013, respectively) and with serum C-peptide levels (r = - 0.76, - 0.50, - 0.35 and - 0.42, p > 0.001, 0.002, 0.036 and 0.011, respectively). Serum betatrophin levels correlated positively with microRNA-486, -146b and -15b levels in G2 (r = 0.35, 0.80, and 0.67, p = 0.036, > 0.001, and,> 0.001, respectively), and in G3 (r = 0.57, 0.36, and 0.38, p > 0.001, 0.029 and, 0.023, respectively). CONCLUSIONS: Circulating microRNA-486, 146b and 15b increase significantly in obese children with T2DM and these levels correlate positively with serum betatrophin levels. Further studies are required to test the role of targeting of these microRNAs and betatrophin in the timely management of obesity and/or T2DM in children.

Association of serum sestrin 2 and betatrophin with serum neutrophil gelatinase associated lipocalin levels in type 2 diabetic patients with diabetic nephropathy.

PURPOSE: Understanding the pathogenesis and the molecular mechanisms of diabetic nephropathy (DN) helps its timely detection and prevention. The current work aims tomeasure serum sestrin 2 and betatrophin levels in healthy and type diabetic (T2DM)subjects with/or without diabetic nephropathy (DN) and also to test their correlation with serum neutrophil gelatinase associated lipocalin (sNGAL); indicator of DN. METHODS: This study included 96 subjects; 20 healthy (G1) and 76 T2DM [22 normoalbuminuric (G2), 35 microalbuminuric (G3) and 19 macroalbuminuric (G4)]. Serum sestrin 2, betatrophin and NGAL were measured by their corresponding kits. RESULTS: Significant low levels of serum sestrin 2 andhigh levels of serum betatrophin were found in T2DM group when compared to G1 (p = 0.002,p > 0.001, respectively) and this difference is manifested in G4 followed, in order, by G3, G2 then G1 (p= > 0.001 for both). Also, serum sestrin2 levels showed significant negative correlations with sNGAL in G1 (r = -0.497, p = 0.026), G2 (r = -0.784, p > 0.001), G3 (r = -0.894, p > 0.001) and G4 (r = -0.896, pp. > 0.001) while serum betatrophin levels showed significant positive correlations with sNGAL in G2 (r = 0.681, p > 0.001), G3 (r = 0.518, p > 0.001) and G4 (r = 0.727, p > 0.001). CONCLUSION: Serum sestrin 2 levels decrease significantly while betatrophin levels increase significantly in T2DM patients with DN especially those with macroalbuminuria. These levels have significant effect strengths on the indicator of diabetic nephropathy; sNGAL which might indicate theirvaluablerole in the timely detection and prevention of the development of DN.

Environmental factors and apoptotic indices in patients with intrauterine growth retardation: a nested case-control study.

BACKGROUND: Egypt has one of the highest incidences of IUGR. The current study investigates the effect of heavy metals toxicity as risk factors of IUGR and determines the possible role of increased apoptosis in their pathogenesis. METHODS: This study was conducted in Assiut, Egypt, included 60 women diagnosed to have IUGR. We measured lead and cadmium levels in blood besides arsenic and cadmium levels in urine. Neonatal scalp hair sample were analyzed for arsenic content. Quantitative determination of human placental Bcl-2 and caspase-3 were performed. RESULTS: There are significantly higher levels of heavy metals and caspase-3 and lower levels of placental Bcl-2 in the IUGR group. The levels of heavy metals were positively correlated with caspase-3 while negatively correlated (except cadmium) with Bcl-2 levels. CONCLUSIONS: There is an alarming high level of heavy metals toxicity in Egypt that was positively correlated to IUGR. Increased placental apoptosis may be one of the possible mechanisms behind the effect.