RAW264.7 murine macrophages (American Type Culture Collection [ATCC], Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µ g/mL streptomycin. All reagents were purchased from Euroclone (Milan, Italy). Cells were maintained in humidified air with 5% CO 2 at 37 ° C. For all experiments, the cells were grown to approximately 70%–90% confluence and culture medium with 2% heat- inactivated fetal bovine serum was used. 37
was lower than that in nonlymph metastatic primary cervi- cal cancer tissues (LN-) (p<0.05, t-test, Figure 6B) and lower in LN- than that in lymph metastatic cervical cancer tissues (LN+) (p<0.05, t-test, Figure 6B). All cervical tis- sues exhibited lower GPX2 expression than distal metastatic lymph nodes (LNM) (p<0.001, t-test, Figure 6B). We also analyzed the association between the pathological stage and GPX2 protein expression in 100 cervical cancer patients according to the standard of International Federation of Gynaecology and Obstetrics (FIGO) stage. The expression of GPX2 protein was associated with greater lymph node metastasis (p<0.001, x 2 -test, Table 1), although it was not associated with age, tumor size or histological type in cervical cancer patients. Taken together, these results sug- gest that GPX2 is closely related to tumor metastasis and the pathological stage in cervical cancer.
process of NDDs have received significant attention. Generally, excessive production of free radicals, ROS, and reactive nitrogen species or the deregulation of detoxifying and/or repairing systems causes OS, either individually or together. Therefore, it would be very practical to develop drugs for the clearance of oxidative free radicals or activa- tion of the antioxidant defense system for NDD treatment. This study, for the first time, provides in vitro results show- ing that through the clearance of H2O2 -induced ROS and reduced activation of ROS-dependent Src-MAPK/AKT signaling pathways, orientin protected PC12 cells against H2O2 -induced cell apoptosis and oxidative damage.
tion using a McFarland nephelometer. Sterile Müller–Hinton Agar No. 2 (20 ml) was poured into petri dishes and allowed to cool in aseptic conditions. 100 µ l of the test inoculum was uniformly spread onto the plates and 5 mm diameter cups were made in the plates using a sterile borer. Different concentrations of cystone and gentamicin were prepared in sterile distilled water and 50 µ l of each dilution was intro- duced in triplicate wells, along with the test drug dilutions, and solvent controls. All petri dishes were refrigerated at 4 ° C for 60 minutes for drug diffusion to occur before being incubated at 37 ° C for 24 hours. Further, the minimum inhibitory concentration (MIC) of the above test com- pounds was determined by the liquid broth dilution method described earlier and the least concentration of both cystone and gentamicin, which inhibited the microbial growth, was taken as the MIC. 27
Previous studies have demonstrated a signi ﬁ cant asso- ciation between in ﬂ ammasome and IDD progression. 29,30 In consistence with previous researches, the results in this study demonstrated that NLRP3, which is a key factor in the in ﬂ ammasome pathway, was up-regulated in IVD sam- ples from IDD cases. Advanced analyses demonstrated the correlation between cGAS and Sting expression and NLRP3 levels in IDD cases and that provide us evidence of the potential regulation effect of cGAS/Sting on NLRP3. Even no previous studies focused on the effect of the cGAS/Sting pathway on the in ﬂ ammasome in IDD, the cGAS/Sting pathway was reported to play a key role in the in ﬂ ammasome activation. 31 The in ﬂ ammatory corpus- cles of human bone marrow cells can be activated by the cGAS/Sting recognition mechanism, which can also acti- vate the innate immune system ’ s response to viral DNA. The researchers found that the activation of the cGAS/ Sting pathway would lead to the programmed cell death unrelated to the antiviral response. When the activation of the cGAS/Sting pathway exceeds a certain threshold, the sting protein will induce the cleavage of a kind of cell inner membrane vesicles called lysosomes and induce in ﬂ ammasome. 32 Considering that both cGAS/Sting and in ﬂ ammasome are commonly reported biological pro- gresses in various diseases, we propose that the cGAS/ Sting pathway could trigger NLRP3 activation and con- tribute in the development of IDD. More in-vitro and in- vivo studies are required to con ﬁ rm this hypothesis.
Abstract. Selective oxidation of phenols is of great interest in terms of practically valuable hydroquinone and pyrocatechin. The purpose of this paper is to investigate the kinetics of the oxidation reaction of 4-tert-butylphenol by an aqueous solution of hydrogen peroxide in the presence of titanosilicates. Macrokinetic model of experimental data was obtained. To minimize the relative error at different temperatures we use genetic algorithm and simulated annealing methods to solve the optimization problems. To perform the calculations a separate software module the mathematical chemistry Web-laboratory was developed. A sub module “The inverse problem” for the solving the inverse kinetic problem with the use of Java-interface for Octave was designed.
many reactions because of their thermal and chemical stability and the possibilities for modification.They bear many similarities to metal complexes of macrocyclic ligands and metalloporphyrins because they possess rigid co-ordination sites surrounding a metal centre . With d 0 electronic configuration of tungsten(VI) species, they do not cause excessive catalytic dismutation of hydrogen peroxide . They have been used as oxidative catalysts in many reactions , . Tetrabutylammonium (TBA) salts of polyoxotungstates are soluble in organic solvent, they can be transferred from aqueous phase into weak polar or non-polar organic phase allowing good contact between the organic substrates with the catalytic active species . Balulu et al. reported catalytic activities of the (TBA) 4 [PW 11 M(H2O)O 39 ].2H 2O (M = Fe, Mn, and V) in
Precursor solutions (0.50 M) were prepared by dissolving appropriate amounts of tin (II) 2-ethylhexanoate (Aldrich, 95%) used as Sn in xylene (Carlo Erba, 98.5%). The precursor mixture was fed into a nozzle at a constant feed rate of 5 ml/min using a syringe pump. At the end of the nozzle the precursor solution was dispersed by 4.30 l/min oxygen forming a spray with a pressure drop at the capillary tip kept constant at 1.5 bars by adjusting the orifice gap area. A sheath gas flow of 3.92 l/min of O2 was supplied concentrically
curve shape which are depicted in Fig. 5a-c. This method (BET) leads to the identification of the isotherm profile as type IV in the BDDT system which is typical for mesoporous material. The BJH pore size distribution demonstrates that all the samples obtained have narrow pore diameter range. The BET surface area (S BET ), pore volume (Vp), pore
used to determine the total protein concentrations (Sangon Biotech, Shanghai, China). A 40 μ g total protein was loaded onto 8% sodium dodecyl sulfate (SDS) polyacrylamide gel, separated, and transferred to a 0.45 μ m PVDF membrane (Pall, Gelman Laboratory, USA); then, membranes were blocked with 5% BSA in buffer for 2h at room temperature. Followed by incuba- tion with antibodies against speci ﬁ c primary antibodies: anti-StAR (12225-1-AP, diluted at 1:1500, Proteintech, USA), anti-CYP11A1 (13363-1-AP, diluted at 1:2000, Proteintech, USA), anti-CYP17A1 (ab125022, diluted at 1:1000, Abcam, Cambridge, UK), anti-LHR (19968- 1-AP, diluted at 1:500, Proteintech, USA), anti-Keap-1 (60027-1-Ig, diluted at 1:500, Proteintech, USA), anti- Nrf-2 (16396-1-AP, diluted at 1:5000, Proteintech, USA), anti-NQO1 (ab80588, diluted at 1:1000, Abcam, Cambridge, UK), anti-HO-1 (27282-1-AP, diluted at 1:1500, Proteintech, USA), anti-GSH-Px (sc-166120, diluted at 1:500, Santa Crus, USA), anti-TNF- α (17,590-1-AP, diluted at 1:1000; Proteintech, USA), anti-IL-1 β (16806-1-AP, diluted at 1:300, Proteintech, USA), anti-i-NOS (18985-1-AP, diluted at 1:600, Proteintech, USA), anti-COX-2 (BA0738, diluted at 1:400, BOSTER, China), anti-BAX (diluted at 1:1000, Abcam, Cambridge, UK), anti-Bcl-2 (diluted at 1:1000, Abcam, Cambridge, UK), and rabbit diluted 1:10,000; anti- β -actin (ARH4149, Antibody Revolution, USA) overnight at 4°C, washed 3 times in TBST for 10 min, and incubated with horseradish peroxidase-conjugated goat anti-rabbit IgG or goat anti-mouse IgG antibody (1:1000; Abgent, San Diego, CA, USA) for 1 h. Densitometric analysis was normalised using β -actin as an internal control. Bands were visualised using an enhanced chemiluminescence (ECL) kit (Sangon Biotech). Quantity One software was used to quantify each band area and density on the blots. Quanti ﬁ ed band intensities are presented as the fold-change from the control. The experiments were repeated three times and were performed independently.
Abstract: Over the last decade, magnetic iron oxide nanoparticles (IONPs) have drawn much attention for their potential biomedical applications. However, serious in vitro and in vivo safety concerns continue to exist. In this study, the effects of uncoated, Fe m O n -SiO 2 composite flake-like, and SiO 2 -Fe m O n core-shell IONPs on cell viability, function, and mor- phology were tested 48 h postincubation in human umbilical vein endothelial cell culture. Cell viability and apoptosis/necrosis rate were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay and annexin V-phycoerythrin kit, respectively. Cell mor- phology was evaluated using bright-field microscopy and forward and lateral light scattering profiles obtained with flow cytometry analysis. All tested IONP types were used at three different doses, that is, 0.7, 7.0, and 70.0 µg. Dose-dependent changes in cell morphology, viability, and apoptosis rate were shown. At higher doses, all types of IONPs caused formation of binucle- ated cells suggesting impaired cytokinesis. Fe m O n -SiO 2 composite flake-like and SiO 2 -Fe m O n core-shell IONPs were characterized by similar profile of cytotoxicity, whereas bare IONPs were shown to be less toxic. The presence of either silica core or silica nanoflakes in composite IONPs can promote cytotoxic effects.
may not be observed. In fact, the TPR peak locations are affected by reduction kinetics. The kinetics of reduction can be affected by a wide range of variables, including crystallite size, support interaction, and reduction gas composition [17, 22]. The effects of crystallite size and support interaction can be superimposed on each other. Thus, while a decrease in metal oxide crystallite size can result in faster reduction due to greater surface area/volume ratio, smaller particles may interact more with the support, slowing reduction. As seen from Fig. 4, the amounts of H2 consumed during TPR related to the
Thereafter, two experienced cardiologists, without knowl- edge of the patients’ prognosis, calculated the CHADS 2 and R 2 CHADS 2 scores. The CHADS 2 nomenclature represents congestive heart failure (C), HT (H), age (A), DM (D), and stroke (S). The CHADS 2 score was calculated by assigning one point each for the presence of congestive heart failure, HT, and DM and by assigning two points for a history of stroke or TIA. The glomerular filtration rate (GFR) was calculated using the Chronic Kidney Disease Epidemiology (CKD-EPI) equation:
of bacterial colonies in the control and treated groups of different microstructures after 12-h cultivation. The control (absence of particles) and Fe 3 O 4 @SiO 2 microsphere-treated solutions did not inhibit the bacterial growth. The Fe 3 O 4 @ SiO 2 –Ag seed showed higher antibacterial efficiency compared with its precursor Fe 3 O 4 @SiO 2 microspheres. After the decoration of Ag seeds and flower-like Ag pet- als, the number of E. coli and MRSA colonies gradually decreased. These results prove that Ag-coated magnetic microcomposites exhibit antibacterial activities against both Gram-negative and Gram-positive bacterial patho- gens. The bactericidal effect also increased significantly with increasing Ag coating components surrounding the magnetic core. Few colonies of E. coli were found during treatment with the Fe 3 O 4 @SiO 2 @Ag microflowers, whereas bacterial colonies of MRSA were still observed in Ag microflower-treated group. MRSA showed less suscepti- bility to the Fe 3 O 4 @SiO 2 @Ag microflowers than E. coli. The Fe 3 O 4 @SiO 2 @Ag microflowers cannot effectively kill MRSA at 20 μ g mL −1 within 60 min. However, almost no
titanium isopropoxide and Gold (III) chloride starting materials  as starting materials. This solution was added drop wise into a dilute ethanol solution under stirring material. The dopant concentrations in at% fraction of Au were set as 0.25%, 0.5% and 0.75%, respectively. In a typical run, the precursor is fed into a FSP reactor by a syringe pump with a rate of 5 ml/min while 5 l/min O2 was being dispersed (5/5
The knowledge of medical use of oxygen has been broadly expanding since 1929, when a book titled “ Ozone and Its Therapeutic Actions ” was published. This publication covered more than hundred diseases that could be treated by the ozone therapy. Ozone ’ s hemodynamic and anti-in ﬂ ammatory properties are now well documen- ted. Through proper administration (never via inhalation) of precise and high doses (in comparison to the environ- mental contamination), ozone brings bene ﬁ ts as it decreases blood cholesterol, improves glycemic index, as well as stimulates antioxidant defence. 12 The therapeutic dosage ranges from 10 to 80 µg/mL. 15 It has been shown that the immune system is altered by the ozone. The
The procedures for generation and drug loading of TfDMP are shown in Figure 1. By coupling GA with amino- immobilized Fe 3 O 4 @SiO 2 , the surface of the magnetic nanoparticle can be modified with both carboxyl and protected amino groups. The dual function magnetic nanoparticles produced were used as carriers. As a multi-armed crosslinker, PLGA has large numbers of carboxyl functional groups, as well as an amino end-group. Thus, PLGA can couple with the carboxyl of DMP, in the presence of NHS and EDC, through an amide bond. Thereby, large numbers of carboxyl functional groups can be introduced onto the surface of DMP. It is known that DOX has an amino group, which is not the active site for DOX effectiveness. Thus, DOX can be immobilized (under vacuum) onto the PLGA-coated layer of the DMP, in the presence of NHS and EDC, through amide bonding. Finally, after deprotection of DMP’s amino groups, Tf can be coupled, through amide bonding.
catalytic activity of carbon dioxide hydrogenation over all copper based catalysts exhibits in Table 2 and Fig. 4. The catalytic activities were consistent for 4 hours. The carbon dioxide hydrogenation can occur despite no hydrogen reduction prior to the hydrogenation took place. The hydrogen chemisorptions were carried out for Cu/ZrO 2 650, Cu/CaO and Cu/ZrO 2 _CaO300 catalysts after hydrogen reduction at 250 o C for 3 h.
growth period were incubated with different concentrations of Gd-NPs (2 × phosphate buffered saline [PBS], 10 µ M, 1 µ M, and 100 nM) in DMEM (Dulbecco’s Modified Eagle’s Medium) at 37 ° C, 5% CO 2 . The L-929 cells treated only with culture media served as negative controls, lipopolysaccharide (500 ng/mL) was used as the positive control. All groups were cultured for 24 and 48 hours post-treatment. We then added 20 µ L of MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) to each well for another 4 hours of incubation, then removed the culture medium and replaced with 100 µ L DMSO (dimethyl sulfoxide), followed by 10 minutes of incubation. The absorbance at 490 nm was measured by a microplate reader (Bio-Rad Laboratories Inc., Hercules, CA, USA).
CH 4 produced in the intestinal lumen, the results indicated that 11.7% of the patients were diag- nosed “positive” for hypolactasia, differently from what was expected. Conversely, taking into consideration the sum of H2 and CH 4 , the percentage increased to 62.8%, closer to the expected one. No significant differences were found when comparing the 2 groups for age, gender, or symptoms. The sizable difference between the 2 approaches is likely linked to gut microbiome variability, and consequently the different production of the 2 gases, in the population. Conclusion: The threshold normally used for lactose breath test should be reconsidered and changed, merging H2 and CH 4 stoichiometric values to increase sensitivity.