Objective Different post-sintering processes are expected to be a reason for alteration in the strength of zirconia. This study evaluated the effect of post-sintering processes on the flexural strength of different types of monolithic zirconia.Materials and Methods A total of 120 classical- (Cz) and high-translucent (Hz) monolithic zirconia discs (1.2 mm thickness and 14 mm in Ø) were prepared, sintered, and randomly divided into four groups to be surface-treated with (1) as-glazed (AG); (2) finished and polished (FP); (3) finished, polished, and overglazed (FPOG); and (4) finished, polished, and heat-treated (FPHT) technique (n = 15). Biaxial flexural strength (σ) was determined on a piston-on-three ball in a universal testing machine at a speed of 0.5 mm/min.Statistical Analysis Analysis of variance, and post hoc Bonferroni multiple comparisons were determined for significant differences (α = 0.05). Weibull analysis was applied for survival probability, Weibull modulus (m), and characteristic strength (σ0). The microstructures were examined with a scanning electron microscope and X-ray diffraction.Results The mean ± standard deviation value of σ (MPa), m, and σ0were 1,626.43 ± 184.38, 9.51, and 1,709.79 for CzAG; 1,734.98 ± 136.15, 12.83, and 1,799.17 for CzFP; 1,636.92 ± 130.11, 14.66, and 1,697.63 for CzFPOG; and 1,590.78 ± 161.74, 10.13, and 1,663.82 for CzFPHT; 643.30 ± 118.59, 5.59, and 695.55 for HzAG; 671.52 ± 96.77, 3.28, and 782.61 for HzFP; 556.33 ± 122.85, 4.76, and 607.01 for HzFPOG; and 598.36 ± 57.96, 11.22, and 624.89 for HzFPHT. The σ was significantly affected by the post-sintering process and type of zirconia (p < 0.05), but not by their interactions (p > 0.05). The Cz indicated a significantly higher σ than Hz. The FP process significantly enhanced σ more than other treatment procedures.Conclusion Post-sintering processes enabled an alteration in σ of zirconia. FP enhanced σ, while FPOG and FPHT resulted in a reduction of σ. Glazing tends to induce defects at the glazing interface, while heat treatment induces a phase change to tetragonal, both resulted in reducing σ. Finishing and polishing for both Cz and Hz monolithic zirconia is recommended, while overglazed or heat-treated is not suggested. When the material is subjected to surface stress and subsequent cracks, high compressive stress can be developed at the crack tips, leading to t- → m-phases transformation with 4 to 4.5% volumetric expansion, rendering crack inhibition phenomenon, known as transformation toughening.2O3. Adding 5% mol. of Y2O3yields a high amount of cubic (c) phase with a smaller grain size of 5-yttrium partially stabilized zirconia (5Y-PSZ). It shows the best enhancement of translucency and aging resistance over the classical 3Y-TZP.Nowadays, all-ceramic restoration has become popular and plays an important role in contemporary restorative dentistry, which is capable of providing a natural esthetic restoration. The ceramic materials must possess high esthetics and be fracture-resistant, especially in the load-bearing area.Post-sintering processes are clinical procedures that clinicians need to perform on the zirconia restorations before delivery to the patients. The restorations need to be ground, adjusted, finished, polished, glazed, or heat-treated.The zirconia blanks were milled into a cylindrical shape with an 18 mm diameter (Ø) from precolored classical translucent monolithic zirconia , and high translucent monolithic zirconia  and sectioned into a disc shape of 1.6 mm in thickness by using a sectioning machine . The dimension of zirconia discs was compensated for sintering shrinkage with the enlargement factor of 1.2302 for Cz and 1.2334 for Hz. The discs were ground flat on both surfaces by silicon carbide abrasive paper with grit 500, 800, and 1,200, respectively, with water coolant on a polishing machine  at a speed of 300 rounds per minute (rpm). Then, the specimens were sintered in a furnace , following the manufacturer's recommendation. A total of 120 zirconia discs of thickness (1.2 ± 0.2 mm) and Ø (14 ± 0.2 mm) were derived. The zirconia discs were sandblasted with 50 µm of alumina oxide powder with a pressure of 30 psi. The specimens were cleaned and allowed to dry at room temperature. The mixing of glazing paste and liquid  was applied over the blasted surface and fired in a ceramic furnace  to produce a glazed surface.All specimens were randomly divided into four groups according to post-sintering surface treatment: AG (as-glazed), FP (finished and polished), FPOG , and FPHT  groups. The specimens in the AG did not receive any surface treatment. The specimens in the FP were ground by cylinder fine diamond finishing bur  by an air-rotor with a speed of 400,000 rpm and water coolant. The contact pressure was exactly 50 g and 30 seconds finishing time for each step in a continuous stroke. The horizontal movement was conducted in one direction with the custom-made load and direction-controlled machine with a fixture for holding the grinding handpiece . The fiThe specimens were tested on the piston-on-three-ball apparatus . The te1is the radius of support circle (mm), r2is the radius of loaded area (mm), and r3is the radius of the specimen (mm).Where P is a load at fracture (N), and b is the specimen thickness (mm), υ is Poisson's ratio = 0.35, ro) as well as the Weibull modulus (m) by using Weibull++statistics  according toThe mean and standard deviation (SD) of σ for each group were compared and analyzed by using ANOVA and post hoc Bonferroni multiple comparisons using statistical software  to determine significant differences in the flexural strength with different post-sintering processes. The result was considered statistically significant at the 95% confidence interval (CI). Weibull analysis was used to determine the reliability of flexural strength and to estimate characteristic strength (σf(σ) is fracture probability, σ is fracture strength, σ0is characteristic strength, and m is Weibull modulus.Where PThe surface topography and fracture surface of the specimens were evaluated with a scanning electron microscope . The crystalline phases of zirconia were determined by their relative proportion of microstructures using an X-ray diffractometer . The crystal structures were examined at a diffraction angle (2θ degree) of 20 to 90 degrees with a 0.02 degrees step size per second intervals by using copper k-alpha radiation. The crystalline phase was analyzed by cross-reference with the standards database of powder diffraction and measured the intensity of the peaks using X'Pert Plus software .o, and m for each group are shown inp < 0.05), but no interaction effect (p > 0.05) was found . The mean ± SD values of flexural strength upon post-sintering surface treatment with AG, FP, FPOG, and FPHT were 1,134.87 ± 523.19; 1,241.23 ± 552.70; 1,116.64 ± 564.06; and 1,111.68 ± 518.99 MPa, respectively (The mean, SD, 95% confidence interval of σ, σs found . The rectively . Post hctively . The poctively .The XRD analysis of the crystalline contents of the Cz and Hz was illustrated inThe SEM photomicrographs revealed the irregularities of the surfaces of the CzAG, CzFPOG, HzAG, and HzFPOG due to small particles of glazing material, and some areas which possibly indicated the incomplete adhesion of the glazing materials as well as several voids inside the glazed layer . The toThis study indicated that post-sintering processes significantly affected the biaxial flexural strength of different types of monolithic zirconia. Therefore, the null hypothesis was rejected for the post-sintering processes and types of zirconia. The grinding, finishing, and polishing procedures were the stepwise method, which was needed to proceed from the coarsest to the finest grit size. The purpose of these procedures was to achieve a smooth, mirror-like surface that provided less susceptibility to bacterial plaque accumulation.Grinding with a coarse diamond bur produced a higher degree of surface roughness than that for the polished surface. The ground zirconia showed significant deterioration in its long-term mechanical properties, which are negatively affected by aging.The multistep zirconia polishing kit can reduce the zirconia surface by a depth of approximately 3 to 4 µm, which is higher than the coarse-diamond ground-induced transformation layer with a thickness of 0.3 µm.The gloss finish was also produced by applying glaze material. Flexural strength results in this study were significantly lowered, possibly because of moisture in the glazing mixture and heat from the glaze firing.Heat treatment can reverse the t- → m-phase transformation when heat is applied at 910°C for 1 minute.o, and survival probability. The m in ceramic was used to determine the reliability of the material and the distribution of flaws. A higher m had higher reliability or homogenous distribution of flaws and greater reliability.owas different in each post-sintering process. The FP had the highest σo, which means that finished and polished zirconia can tolerate more force before it fails and is more durable for long-term use. Moreover, when comparing the σoof the overglazed and heat-treated group to the as-glazed group in both Cz and Hz, the former was found to be lower than the latter.The Weibull analysis provided the m, σobecause of the incompatibility between the glazed layer and the zirconia itself.This study showed that the group subjected to grinding and then glazing exhibited lower σBased on the results of the study, the biaxial flexural strength of the Cz was stronger than that of the Hz. Post-sintering processes affected the flexural strength of monolithic zirconia. The flexural strength of monolithic zirconia can be enhanced through a proper finishing and polishing procedure. Overglazed- and heat-treated processes after finishing and polishing are not necessary because they can reduce the flexural strength of the zirconia. Heat treatment can reverse the phase back to tetragonal but result in a reduction in the flexural strength. However, overglazing shall be done in the esthetic region of restoration to achieve a natural-looking restoration as a gloss surface is also needed for the translucent monolithic zirconia.


Liraglutide is known to have much lower weight loss effects in real clinical fields than in randomized clinical trials because of its side effects (SE) and discomfort associated with injections. This study is aimed at determining whether the side effects of liraglutide affect weight reduction and its maintenance in real-world practice.  Endocrinologists conducted a retrospective chart review of data from two tertiary university hospitals. All patients who had been prescribed liraglutide at least once between January 2014 and December 2019 were included. For an average of 3 and 6 months, weight changes due to the presence or absence of SE and discontinuation (MAIN or STOP) of liraglutide were checked. p < 0.001 compared to the baseline). However, there were no significant differences in the weight loss between the MAIN (p = 0.062) and STOP (p = 0.204) groups. At 6 months, the weight reduction was −2.0 ± 0.5% (p < 0.001) in MAIN_SE(-), −2.2 ± 0.7% (p < 0.005) in MAIN_SE(+), −1.7 ± 0.7% (p < 0.01) in STOP_SE(-), and −2.0 ± 0.6% (p = 0.01) in STOP_SE(+), compared to baseline. SEs also caused no significant differences in weight loss between the MAIN (p = 0.787) and STOP (p = 0.694) groups.  Only 40.8% (64/157) of the patients remained on liraglutide for 6 months; 14.7% (23/157) maintained the drug despite SEs (MAIN_SE(+)), and 40.1% (63/157) discontinued the drug despite not having SEs (STOP_SE(-)). At 3 months, there was −5.9 ± 0.6%, −7.9 ± 0.9%, −4.5 ± 0.5%, and −3.4 ± 0.6% weight reduction in the MAIN_SE(-), MAIN_SE(+), STOP_SE(-), and STOP_SE(+) groups, respectively (all  Our study confirmed that the side effects of liraglutide did not affect weight reduction. Moreover, in the real world, the continuous rate of liraglutide use is not high, and the weight gradually increases after 3 months. Therefore, in addition to the side effects of liraglutide, the medical staff should consider various factors that affect drug adherence, consider ways to increase compliance, and continue to ensure management so that patients can maintain their weight. It is important to manage overweight and obesity as they are risk factors for type 2 diabetes , 2. CurrReports on the weight change due to the side effects of liraglutide have been controversial. Some studies have reported that the weight reduction effect is greater than the side effects; however, some have also reported no relationship between the side effects and weight reduction effect of liraglutide , 7. In aLiraglutide is known to have a greater weight reduction effect than other obesity drugs . HoweverIn this study, among the patients who visited Seoul St. Mary's Hospital and Eunpyeong St. Mary's Hospital, from January 2014 to December 2019, those who had been prescribed liraglutide at least once were followed up for an average of 6 months.The index date was designated as the date on which liraglutide was prescribed. Personal information such as age, sex, weight, and body mass index (BMI) of the patients within 2 weeks before the index date was collected. Baseline laboratory results of blood glucose, HbA1c, blood urea nitrogen, and creatinine levels, along with liver function tests , and lipid profiles  were collected. In addition, the history of hypertension, prediabetes, diabetes, dyslipidemia, fatty liver, thyroid disease, and allergies was investigated. All side effects that the patients complained about were confirmed, including gastrointestinal (GI) problems caused by the mechanism of action of liraglutide. Additionally, history of psychiatric disorders or psychiatric treatment was assessed.A visit 2–4 months (average 3 months) from the index date was designated as visit 1, and a visit 5–7 months (average 6 months) later was designated as visit 2. The patient's weight was recorded at each visit, and the change from the baseline weight was calculated. If there were multiple visits within 2–4 months, the lowest weight was selected. If there were multiple visits within 5–7 months of the last visit, the weight closest to 6 months was selected. Additionally, the maximum dosage of liraglutide was determined. If drug discontinuation occurred, the date of discontinuation was estimated through chart review.After liraglutide was prescribed, side effects and discontinuation of the drug were checked for an average of 6 months (up to 7 months) through a direct chart review made by an endocrinologist . Within An endocrinologist with more than 10 years of experience directly reviewed the charts and collected all related data and information for all patients. The patients' charts were reviewed directly by a lead researcher. During statistical analysis, the patients' personal identification numbers were deleted, and a random number was assigned. All personally identifiable data were anonymized and stored in various combinations. Anonymization and security measures were implemented in this study. As this was a retrospective cohort study, there was no possibility of physical or mental harm to the participants; therefore, patient consent was not required. This study was approved by the Clinical Research Ethics Committee of the Catholic Medical Center .t-test considering the Bonferroni correction. The final regression models between weight variation and baseline variables were executed using a linear regression model adjusted for age, sex, and BMI. All statistical analyses were performed using the SAS software , and statistical significance was set at p < 0.05.Baseline variables are expressed as means and standard deviations or numbers, including percentages, whether they were continuous or categorical variables. The association between weight variations during the 3 months after prescription and the degree of discontinuation was assessed using an independent 2; there was no significant difference between the groups. For 6 months, 40.8% patients (64/157) maintained liraglutide, while 59.2% (93/157) stopped taking it. The group that stopped liraglutide stopped it at 3.3 ± 1.2 months. The mean dose of liraglutide was 2.3 ± 0.8 mg, and there was no significant difference between the groups. In the group that maintained liraglutide, 64.1% patients (41/64) had no side effects, and 35.9% (23/64) had side effects. In the group that stopped liraglutide, 67.7% patients (63/93) had no side effects and 32.3% (30/93) had side effects. History of hypertension, prediabetes, and DM did not significantly differ between the groups. In addition, the history of GI problems, such as gastroesophageal reflux disease or ulcers, showed no difference between the groups (p = 0.060). There was no significant difference between the groups regarding bipolar disease, sleep disorders, anxiety disorders, depression, or psychiatric treatment (p = 0.979). Other laboratory findings did not show any significant differences between the groups.From January 1, 2014, to December 31, 2019, 157 patients were prescribed with liraglutide at Seoul St. Mary's Hospital and Eunpyeong St. Mary's Hospital . The meap < 0.001) compared to the baseline in the MAIN group; there was a significant weight reduction of 4.1 ± 0.4%  compared to the baseline in the STOP group (p < 0.001). After 6 months, there was a significant weight reduction of 1.9 ± 0.3%  compared to the baseline in the MAIN group and a significant weight reduction of 1.8 ± 0.5%  compared to the baseline in the STOP group. However, there was no significant difference in weight reduction between the MAIN and STOP groups after 6 months .After 3 months, there was a significant weight reduction of 6.7 ± 0.5%  in the MAIN_SE(-) group, and it was −7.9 ± 0.9%  in the MAIN_SE(+) group. However, there was no significant difference between the two groups (p = 0.062). After an average of 6 months, the weight reduction was −2.0 ± 0.5%  in the MAIN_SE(-) group and −2.2 ± 0.7%  in the MAIN_SE(+) group; there was also no significant difference between the two groups (p = 0.787). After an average of 3 months, in the STOP_SE(-) group, the weight reduction compared to the baseline was −4.5 ± 0.5% , and in the STOP_SE(+) group, it was −3.4 ± 0.6% . There was no significant difference between the two groups (p = 0.204). After 6 months, in the STOP_SE(-) group, the weight reduction compared to the baseline was −1.7 ± 0.7% , and in the STOP_SE(+) group, it was −2.0 ± 0.6% . There was no significant difference between the two groups (p = 0.694)  . Even thp < 0.001) in the MAIN_SE(-) group, 4.6 ± 0.6 kg (p < 0.001) in the MAIN_SE(+) group, 2.2 ± 0.5 kg (p < 0.001) in the STOP_SE(-) group, and 1.2 ± 0.3 kg (p = 0.001) in the STOP_SE(+) group. All groups had a significant weight reduction compared to the baseline at 6 months; however, their weight increased compared to that observed at 3 months with significant results .The greatest weight reduction in all groups was observed at 3 months, regardless of drug maintenance or side effects. However, a weight regain pattern was observed afterward. Regarding the weight difference between 3 and 6 months, the weight regain was 3.4 ± 0.3 kg (p < 0.001). Moreover, the weight loss was 1.7238 times higher in the case of dyslipidemia than in the case of no dyslipidemia (p < 0.01)  group showed greater weight reduction. Drug treatment is important for weight loss, as are diet and exercise . As sideAccording to the results of this study, weight reduction was greatest at around 3 months; however, there was weight regain thereafter. Notably, there was no difference in body weight between the STOP and MAIN groups at 6 months. In our study, the MAIN group lost an average of 5.4 kg (average 6.7%) at 3 months, and after recovering the body weight, it decreased by an average of 1.6 kg (average 2.0%) compared to the baseline at 6 months. There was a study in which 5.0 kg was reduced in 6 months and maintained for 5 years , and theThe reason why our study had a lower weight loss effect at around 6 months compared to other studies is presumed to be a feature of real-world evidence using real-world data , 19. UnlIn our study, the factors affecting weight loss were the presence or absence of drug discontinuation and sex. In a previous study, the effect of the difference in plasma exposure of liraglutide by pharmacokinetics on the weight loss effect was investigated. Weight and sex have been identified as factors affecting the differences in plasma exposure . The samOur study had several limitations owing to its retrospective nature , 29. FirThe major strength of our study is that it evaluated the weight loss effect of liraglutide in an actual clinic, not in a randomized control trial, according to the presence or absence of side effects and discontinuation. All side effects of liraglutide, including those of the GI and nervous systems, were directly checked and included. The effects of these side effects on compliance, such as drug discontinuation at the treatment site, were investigated. Furthermore, this study investigated the effect of liraglutide on weight loss due to drug compliance, which is influenced by side effects.Liraglutide is a representative drug that is maintained continuously despite its side effects because it is often used for cosmetic purposes . Neverth


According to the UFLC-DAD and LC–ESI–MS results, we found that the complete degradation of TLP can be reached after 60.83 min of UV irradiation using TiO2 as a photocatalyst. Furthermore, we determined that germination, biomass production, and chlorophyll b (Chl b) were not related to the percentage of TLP after irradiation. Chlorophyll a (Chl a) , Chl a+b , and carotenoid (car)  were strongly inversely (negatively) correlated with TLP, while Chl a+b/car  was moderately (positively) related.Environmental pollution is an emerging global issue. Heterogenous photocatalytic degradation, which belongs to the advanced oxidation processes, is a promising sustainable technique for the removal of harmful pollutants  from natural resources (surface and underground waters), as well as wastewaters. In our study, we examined the efficiency of photocatalytic degradation (with TiO Howeverp = 0.15), for SSI, it dropped between 30  and 60 min  only to revert to initial value at 120 min . Further elongation of irradiation beyond 60 min reversed this pattern, likely due to further addition of electromagnetic energy and assuring electrostatic changes of the active site (further release of electrons) towards increased affinity for reactant molecules . The result is fostering photocatalysis of TLP and reversing m(AG/P)  to its initial value. According to the differences between SSI and UV irradiation . However, quadruplication of the initial value for factor loading showed a significant difference for m(AG/P)/[mg] = 229.43 ± 3.57 mg; p ≤ 0.01) from initial loading. Since the sunlight is readily available , where doubling of the initial value yielded an increase in m(AG/P)/[mg] from 229.50 ± 2.90 mg to 251.58 ± 2.90 mg (p ≤ 0.01), only to remain constant at factor loading at 2.0 mg/mL with m(AG/P) = 252.92 ± 2.90 mg (p = 0.42). Connecting with the above-mentioned, the UV irradiation is stronger and more efficient, so the higher m(AG/P) will be already reached at smaller catalyst loading [For SSI to observe the change in the m(AG/P)/[mg], it was not sufficient to increase catalyst loading from 0.5 mg/mL (213.54 ± 3.57 mg) to 1.0 mg/mL (222.17 ± 3.57 mg) where m(AG/P)/[mg] remained constant (s 2112 h ), it is  loading .2 than for ZnO , which represented a smaller increase , while for ZnO m(R/Plant) = 102.83 ± 2.35 mg it was higher for UV than for SSI . Taking into account the earlier statements ) was higher for TiOease 9%; . Plants ease 9%; . Regardlatements , it is op = 0.38).The highest m(R/Plant) = 102.00 ± 2.07 mg was recorded at 1.0 mg/mL, and after that point , remained constant with an additional increase in loadings  = 2.38 ± 0.03 mg  = 2.57 ± 0.03 mg was higher for ZnO than for the TiO 0.03 mg . One sucp = 0.22).Irradiation type had no influence on m(AB/R) . Opposit2O2, and free proline, which indicated the presence of stress-induced reactions in plant tissues. However, since differences were recorded only in concentrations of photosynthetic pigments and biomass, evaluation of other parameters was not reported. Since a variety of factors may influence different shoot and root growth, their ratios were calculated as well.Pigmentation was analyzed to assess whether the treatments applied only during germination affected the subsequent growth of plants. In other words, plant growth relays heavily on photosynthetic apparatus, whose constituents are photosynthetic pigments; hence we measured concentrations of Chlorophyll a (Chl a), Chlorophyll b (Chl b), and carotenoids (car) to gauge the toxicity of TLP. This made sense as carotenoids, besides having the role in broadening the specter of sunlight that can be used in photosynthesis, protect chlorophylls from photooxidation, and thus act also as antioxidants ,48,49,502 /[mg] and m(R/Plant)/[mg], Chl a at 0.5 mg/mL of catalyst loadings (5.69 ± 0.13) increased with an increase to 1.0 mg/mL of loadings (6.18 ± 0.13) where it was the highest, but only to drop to the initial value at 2.0 mg/mL of catalyst loadings (5.83 ± 0.13) or that of 0.5 mg/mL of loadings . These fp = 0.19). For both types of catalyst , UV light had higher values for the Chl a than SSI . For SSI light source, Chl a remained constant regardless of the change in catalyst loadings . However, for the UV light, this was not true; instead, Chl a with SSI at 0.5 mg/mL was 6.43 ± 0.10 and peaked at 6.82 ± 0.10 with 1.0 mg/mL and then dropped to a value no different than the initial value (6.42 ± 0.10) at highest loading of 2.0 mg/mL (p ≤ 0.01). There was no synergistic influence of the type of catalyst, type of light, and catalyst loading on Chl a (p = 0.08). Chl b with ZnO was higher (2.80 ± 0.02) than with TiO2 ; however, this was not true for TiO2 where higher Chl b was found for SSI  than for the UV-light source . An interesting explanation for the higher concentration of pigments under SSI than UV. Similar to the damage that UV irradiation does to human tissues, those are happening in plant tissues [2 has weaker photocatalytic activity than ZnO could be the reasons for higher content of Chl b under SSI than for the UV-light source. More so, the Chl a:Chl b ratio is a reliable indicator of stress in plants. This ratio is the most stable parameter, irrespective of the stress factor plants are exposed to [An interesting pattern was observed for this interaction as type of light source did not modify the Chl b with ZnO  and 7.33 ± 0.13, respectively)  increased with 1.0 mg/mL (8.00 ± 0.16) where it was the highest, only to drop in value with 2.0 mg/mL (7.50 ± 0.16) that was no different from 0.5 mg/mL of loadings (p = 0.12). However, similar to Chl b, Chl a+b was higher for the UV light (8.52 ± 0.16) than for the SSI, where it equaled 6.13 ± 0.16 at p ≤ 0.01. Time of irradiation did not modify the concentration of Chl a+b for different catalyst loadings .The overall concentration of chlorophylls (Chl a+b) with ZnO was higher than with TiOctively)  and it wctively) . These rloadings . The finloadings . One oth2 for both types of light sources. However, for ZnO content of Chl a+b under SSI light increased from initial value of 7.27 ± 0.32 with 0.5 mg/mL of loading to 8.59 ± 0.32 with 1.0 mg/mL and then decreased to initial value of 7.34 ± 0.32 with 2.0 mg/mL (p = 0.02). The optimal catalyst loading is a very important factor in heterogeneous photocatalytic degradation, as mentioned. According to the optimal photocatalytic conditions (p = 0.04). The same conclusions could be taken in this situation. Both catalysts showed no change for Chl a+b value with changing irradiation time under the UV light, and this was also true for samples with ZnO under SSI light. However, TiO2 in combination with SSI light showed significant differences for Chl a+b with different irradiation times (p ≤ 0.01). Here value Chl a+b peaked after 60 min of irradiation (7.92 ± 0.35) from what was initially detected at 30 min (5.16 ± 0.35), then it dropped to the initial value after 120 min of irradiation (4.86 ± 0.35). These findings are in accordance with the weaker photocatalytic activity of TiO2 under SSI caused by the wide band gap . Decreased number of cars under different times of irradiation probably happened because of the negative effect of the present compound on the pigments in wheat. The TLP and its photodegradation intermediates  than with SSI light  than under UV light (4.29 ± 0.06). For ZnO this was inverse, and for samples irradiated with UV light ratio was higher (5.79 ± 0.08) than for the SSI (4.81 ± 0.08) at p ≤ 0.01.Ratio Chl a+b/car for ZnO was higher (5.30 ± 0.05) than for TiOTo find optimal conditions for photocatalytic degradation of TLP and germination of seeds, two mathematical models were created. Predictors for germination of seeds (Seed) and percentage of TLP after irradiation (TLP) were: (i) type of catalyst (CT); (ii) catalyst loading (CL); (iii) germination time (GT); (iv) irradiation time (IT); and (v) source of light (SLG).Firstly, using an adequate model, the degradation of TLP was predicted and calculated by a linear regression model:2 prior adjustment (94.71%) and a good indication that number of predictors had minimal impact on the prediction of variance. It was calculated from the model that complete degradation of TLP (0%) can be achieved by using TiO2 with UV light and loading of 1.0 mg/mL with a minimum length of irradiation of 60.83 min (Here after adjustment (for a number of predictors), the model predicted 94.61% of the variance while this was similar to R0.83 min A.2-adjusted for d.f.) while R2 prior adjustment equaled 65.01% showing that number of predictors had negligible influence on the prediction of variance. Calculated linear regression model was:The germination model explained 64.46% of the variance  after 60.83 min of UV irradiation. LC–MS results also confirmed the successful removal of TLP, as well as the formation of photodegradation intermediates. However, these results have to be further examined, which is going to be the topic of our next study. Additionally, as expected, we proved that the germination time had no influence on the percentage of TLP after irradiation. Our results also suggested that germination time was not related to the percentage of remaining TLP after irradiation. It is important, as germination is one of the most essential processes in plant development, and according to these findings, TLP had no negative effect in this early phase of plant growth. Similarly, a relation with the percentage of TLP after irradiation was not observed in the cases of m(AG/P), m(R/Plant), m(AB/R), and Chl b. On the other hand, a strongly negative effect of the TLP after irradiation was observed in the case of Chl a Chl a+b, and car, while it was moderately (positively) related with Chl a+b/car. Summarizing these data, we successfully proved the toxic effect of TLP and its photodegradation intermediates on the physiology of wheat. Considering the above-mentioned parameters, this strongly implies that photocatalytic degradation is a suitable approach to reduce the toxicity of TLP in the environment and wastewaters. Based on the reported findings, there are many opportunities in the future that can be the topic of further investigation. We suggested a sustainable method for water purification; however, we used TiO2 and ZnO as photocatalysts, which are mostly active only under UV irradiation, as well as TiO2 is not fully safe for the environment. Thus, various nanoparticles and special nanomaterials with less toxic effects and higher photocatalytic activity under natural sunlight could be synthesized by green approaches and investigated in the photodegradation of TLP. In addition, the photodegradation pathways of TLP should also be examined in detail to obtain more information about all the possible degradation products and their effects on the environment. Furthermore, since the toxicity of TLP has not been examined, further research should be conducted in order to determine the possible harmful effect of TLP on other organisms too.In this research, we first determined the optimal conditions for the complete photodegradation of TLP. The findings showed that complete TLP removal can be reached using TiO


This study is the first to investigate how a local government’s humorously framed response strategy on social media to a low-severity crisis influences people’s trust in the local government and their crisis-related behavioral intentions, specifically when considering the government’s responsibility for the crisis. Based on the situational crisis communication theory, we examined the mediating role of experienced positive or negative affect on people’s responses to a local government’s crisis communication strategy. Further, we exploratorily examined the predictive power and moderating role of demographics, sense of humor, disposition to trust, and the respective crisis scenarios. A total of 517 people participated in an online experiment in which they were confronted with three randomly presented fictive crisis scenarios where the local government’s crisis responsibility (high versus low) and the framing of their crisis response strategy  were systematically varied between subjects. First, the results mostly corroborate earlier findings about the degree of crisis responsibility  and about the mediating role of experienced affect. Second, we found that humorously framed strategies negatively influence trust and positive affect . In contrast to earlier findings, the crisis responsibility × framing interaction was not significant. Altogether, the results advise against using humor in crisis communications on social media, even in low-severity crisis. Exploratory analyses indicate that further investigations should focus on specific crisis characteristics and potential moderators. In crisis situations, social media offers local governments a way to communicate directly and quickly with the public and provide up-to-date information. Such crisis communication allows local governments to stay in contact with the public during a crisis, gain their trust, and encourage them to take appropriate actions  depending on the government’s crisis responsibility. We chose trust as the central outcome variable, as the value of information given by governments is strongly reduced if they are not trusted .1Mage= 25.70, SDage = 10.75) were presented with nine different fictive scenarios (randomly ordered) describing crises in local governments’ areas of responsibility. Participants read two versions of each scenario, one where the local government was responsible for the crisis and one where it was not. After reading each scenario, participants rated the attributed crisis responsibility of the local government on a seven-point scale  between 19 and 56 years old  saw 32 fictive Twitter tweets posted by local governments in response to one of nine fictive crisis situations. Each post was preceded by a brief description of the background of the fictive crisis to which the post was related. Information on crisis responsibility was not given to avoid possible interaction effects between the government’s responsibility and the framing of the crisis response strategies  we compared whether humorous tweets were indeed perceived as funnier than rational ones. The three tweets perceived as the funniest, their rational equivalents and their respective scenarios were included in the main study.In pre-study II, 13 participants  between 20 and 58 years old . Participants’ highest completed educational levels were compulsory basic secondary schooling (4.4% of participants), a general certificate of secondary education (13.0%), a general or specialized university entrance qualification (30.4%), a degree from a university or college of higher education (47.8%), or a doctorate (3.3%); the remainder (1.2%) had an unspecified school-leaving qualification. Of the participants, 9.1% pursued a career in local government, 5.4% in crisis management, 34.8% in fields of fire safety, rescue services, or disaster control, and 3.9% in the police force or similar security services; 57.8% worked in none of these areas.Z = 1,660.5, p = 0.048.The a priori power analysis indicated that a sample size of 225 would be sufficient to detect significant effects in the planned parallel mediation models with a power of 0.80, an alpha of 0.05 and a medium effect size of M = 13.35 min, SD = 5.69). This study used a 2 (crisis responsibility: high versus low) × 2  between-subjects design, resulting in four experimental conditions.This study was preregistered on the open science platform As PredictedThe following section provides detailed information on the study materials and measures. Examples for the materials and scales used are given.Fictional crisis scenarios were used to avoid interference from local governments’ crisis histories and negative pre-crisis reputations that could have influenced the effectiveness of crisis communication Coombs . All criIn the respective tweets, the local government informed the public about the situation either in a rational or in a humorous way. All tweets used a subtype of the rebuilding response strategy   than in the low-responsibility ones , F = 495.90, p < 0.001, ηp2 = 0.49, and humorously framed crisis response strategies were perceived as funnier  than the rational ones , F = 356.25, p < 0.001, ηp2 = 0.41.Two ANOVAs suggested that the experimental manipulations worked very well. Local governments were held more responsible for the crisis in the high-responsibility crisis scenarios  = 6.76, p < 0.001, ηp2 = 0.04 (1 – β = 1.00), humorous framing, V = 0.02, F = 3.81, p = 0.010, ηp2 = 0.02 (1 – β = 0.95), and a crisis responsibility × humorous framing interaction effect, V = 0.02, F = 2.97, p = 0.032, ηp2 = 0.02 (1 – β = 0.95).Hypotheses about the main effect of crisis responsibility (H1a−H1c) and humorous framing (H2a−H2c) and their interaction effect (RQ1) on trust, positive affect, and negative affect were tested with a MANOVA. Here, Pillai’s trace indicated significant effects of crisis responsibility, F = 9.57, p = 0.002, ηp2 = 0.02 (1 – β = 0.90), suggesting that trust in the government was higher when the crisis responsibility was low  than when it was high . Furthermore, a significant main effect of framing was found, F = 5.67, p = 0.018, ηp2 = 0.01 (1 – β = 0.63), suggesting that trust in the local government was higher with the rational framing  than the humorous framing . Yet, there was no significant crisis responsibility × framing interaction effect on trust in the local government, F = 3.55, p = 0.060, ηp2 = 0.01.The follow-up ANOVA for trust yielded a significant result for crisis responsibility, F = 12.14, p < 0.001, ηp2 = 0.02 (1 – β = 0.90), suggesting that negative affect was higher when the crisis responsibility was high  than when it was low . Neither the main effect of framing, F = 0.53, p = 0.467, ηp2 < 0.01, nor the interaction effect on negative affect, F =1.80, p = 0.180, ηp2 < 0.01, were significant.The follow-up ANOVA for negative affect showed a significant effect for crisis responsibility, F = 3.41, p = 0.065, ηp2 = 0.01. However, it indicated a significant effect for framing, F = 5.96, p = 0.015, ηp2 = 0.01 (1 – β = 0.63), suggesting that positive affect was lower with the humorous framing  than with the rational framing . The interaction effect on positive affect was not significant either, F = 0.35, p =0.553, ηp2 < 0.01. All effects found in the MANOVA and its follow-up ANOVAs are small according to Cohen . High crisis responsibility led to higher negative affect, resulting in lower trust. The other path coefficients are presented in Fig. Due to missing effects of crisis responsibility on positive affect and humorous framing on negative affect, we subsequently conducted simple instead of parallel mediation analyses. The first mediation analysis tested whether negative affect mediates the relationship between crisis responsibility and trust (H1d). We found a significant indirect effect of crisis responsibility mediated by negative affect, ab = −0.05, 95% CI , p = 0.002 (1 – β = 0.70). Participants seeing the rational framings experienced more positive affect, and, in turn, participants who experienced more positive affect gave higher ratings of trust. The other path coefficients are presented in Fig. The second mediation analysis investigated the mediating role of positive affect on the relationship between framing and trust (H2d). It revealed a significant indirect effect of framing mediated by positive affect, V = 0.04, F = 3.39, p = 0.003, ηp2 = 0.04, but not for framing, V = 0.02, F = 1.43, p = 0.201, ηp2 = 0.02, nor for the crisis responsibility × framing interaction effect, V = 0.01, F = 0.74, p = 0.619, ηp2 = 0.01.The main effect of crisis responsibility (RQ2) and humorous framing (RQ3) on variables measuring behavioral intentions were tested with a MANOVA. Here, Pillai’s trace indicated a significant effect of crisis responsibility, M = 4.90, SD = 1.04; low: M = 5.15, SD = 1.04), they were less willing to follow local government’s advice as a matter of principle , they were less willing to ask family and friends to follow advice , and they were less willing to seek information on the local government’s website . Moreover, the willingness to ask family and friends to follow advice was lower when a humorous  rather than a rational framing was used . All significant effects found in the MANOVA and its follow-up ANOVAs are small according to Cohen’s . Similar to the mediation analyses above, simple instead of parallel mediation analyses were conducted. Mediation analyses were only conducted for the behavioral intention variables that were significantly influenced by crisis responsibility or framing. All results of the mediation analyses can be found in Table F = 8.39, p = 0.004, ηp2 = 0.02, and indicated that participants preferred rational framing  rather than humorous framing .In four exploratory analyses, we examined (1) further predictors of trust; (2) the moderating role of participants’ demographics and characteristics; 3) the moderating effect of the crisis scenarios themselves; and (4) participants’ liking of the tweets depending on both experimental factors. Additionally, four exploratory analyses were examined. the modeInspired by recent propositions that local governments and public authorities may want to use less formal crisis communication strategies on social media, this study offers first insights into the effectiveness of humorous crisis communication on social media by public sector organizations in low-severity crises. It further highlights once again the importance of crisis responsibility in crisis communication.According to our first hypothesis, high attributed crisis responsibility predicted a reduction in trust (H1a) and a rise in negative affect (H1b). This is in line with earlier findings Coombs . We alsoThe lack of correlation between crisis responsibility and positive affect might have occurred because the study’s opposing effects neutralized each other: For example, while positive emotions directed to the organization, such as sympathy, are negatively related with crisis responsibility, the positive emotions associated with one’s own situation in a crisis, such as relief, are partially positively correlated with crisis responsibility  or on experienced emotions (H2b and H2c), such that emotions played no mediating role (H2d), even though the manipulation check showed that humorously framed crisis responses were indeed perceived as humorous. This pattern of results contradicts earlier studies in humor Martin  and adveThe independence of perceived humor and emotional reactions found in our study can be explained by the benign violation theory of humor  could strengthen or reduce the effects found.Finally, one might criticize our use of SCCT Coombs  as a theOur core message for local governments’ crisis communication managers is as follows: If your local government is responsible for a crisis, do not use humorous framing in crisis communication on social media, even in low-severity crises. For humor to have a positive effect, crisis communication managers would have to accurately consider many factors, such as the specific characteristics of the crisis , the local government , and the target group . Additionally, the style of humor  in low-severity crises. We hope that the current study will motivate further investigation of possible moderating and mediating variables, such as quality of emotions and perceived crisis severity. Furthermore, the present research contributes to a growing body of evidence for the SCCT Coombs , suggest


APT-PC was obtained by solvent evaporation with APT and phospholipids, then analyzed by X-ray diffraction, Fourier transform infrared spectroscopy and differential scanning calorimetry. Lipid emulsions are a new formulation that can reduce side effects and improve drug loading.In present, there was no detailed report on the formulation optimization and quality evaluation of aprepitant (APT) injectable lipid emulsion (APT-IE). The aim of the present investigation was to prepare and evaluate its properties of APT-IE loaded with an APT phospholipid complex (APT-PC) APT-IE prepared by High-pressure homogenization and optimized by response surface methodology (RSM). The proportion of sodium oleate, poloxamer 188 and soybean oil were selected as variables for the optimization. The optimal formulation of ATP-IE had the following characteristics: particle size, 82.83 ± 1.89 nm; polydispersity index, 0.243 ± 0.008; zeta potential, −59.0 ± 2.54 mV; encapsulation efficiency, 98.84%±1.43%; drug loading, 7.08 ± 0.16 mg/mL; and osmotic pressure, 301 ± 2.15 mOsmol/kg. Transmission electron microscopy images indicated that the particle diameter of APT-IE was approximately 100 nm, with a morphology of spheroidal or spherical. APT-IE exhibited sufficient stability after storage at 4 ± 2 °C for more than 6 months. The results of the pharmacokinetic study demonstrated that APT-IE had the advantages of better safety, higher bioavailability, and obvious liver targeting than APT solution (APT-SL). The area under the curve (AUC) of APT-IE was 3-fold enhanced compared with APT-SL. The targeted enhancement multiple of APT-IE to liver tissue was greater than that of APT-SL. These results suggested that APT-IE has broad clinical application and industrial production potential. Ltd. . Egg yolk lecithin and sodium oleate were purchased from Shanghai A.V.T. Pharma Co. Ltd. . Poloxamer 188 (F68), sucrose and medium-chain triglyceride (MCT) were purchased from Xi’an Tianzheng Medicinal Materials Co. Ltd. . Ethanol was purchased from Shandong Long Yu Quan Pharmaceutical Excipients Co. Ltd. . All chemicals and reagents were of analytical or HPLC grade.2.1.2.All Sprague-Dawley rats were obtained from the Experimental Animal of North China University of Science and Technology . The animal license number was 2019-0006. Each animal was breaded in a clean environment with enough water and food. The experimental protocols for laboratory animals were evaluated and approved by the University Ethics Committee and complied with the Guide for the Care and Use of Laboratory Animals. All experiments were conducted in accordance with the principles of the European Community Guidelines for the Use of Laboratory Animals.2.2.2.2.1.Excessive amounts of APT were added to 10 mL of soybean oil (SO), corn oil, MCT, SO-MCT mixture, and phosphate buffered solution (PBS) of pH 2–8. The samples were incubated for 48 h at 37 °C in a water bath vibrator after 15 minutes of ultrasonication. The mixture was filtered through a 0.45 μm membrane, with the initial filtrate discarded. Then the concentration of APT was determined by HPLC after diluted appropriately with methanol.2.2.2.n-Octanol and distilled water fully mixed was left to stand for 1 h to ensure separated completely  with methanol, with the water phase directly postmembrane. The APT concentration in both phases were analyzed by HPLC and the partition coefficient was calculated as follows:Co-the concentrations of APT in n-octanol, Cw- the concentrations of APT in water.The classical shake flask method was used for the determination of oil-water partition coefficient of APT. The Solution that 2.3.2.3.1.1w -The concentrations of APT in APT-PC determined by HPLC.w0 -the amount of APT (0.72 g).The APT and EL were dissolved in 2 mL of absolute ethanol with rotary evaporation at 70 °C to obtain dried APT-PC. Moreover, the APT, EL or the dried APT-PC were weighed before and after preparation to ensure anhydrous ethanol completely evaporated. The ratios of APT to EL were settled at ratios of 1:1, 1:2, 1:6, 1:10, 1:15 and 1:17.5 (w/w), and the drug loaded of the phospholipid complex was used to evaluate the optional rate of APT-PC with different proportions. Moreover, the drug loaded in the phospholipid complex was also expressed as the recombination rate (Rer). The calculation of the Rer is as follows:2.3.2.XRD) analysis, samples were acquired at room temperature on a D/max-rA . XRD setup parameters were 40 kV and 30 ma (1.2 KW) Cu Kɑ source. Samples were measured by Fourier transform infrared (FTIR) analysis. The spectral range is set to 4000–400 cm−1. Using differential scanning calorimeter (DSC) analysis, samples were examined using a DSC instrument to test, verify their thermal characteristics and obtain DSC curves. Samples were put into aluminum pans and heated from 25 °C to 300 °C under a nitrogen atmosphere at a rate of rise of 10 °C/min.The properties of APT-PC were characterized by the following three ways. First, samples including free drug (APT), free phospholipid (EL), APT and EL physical mixture (APT-EL-PM) and different mass ratios of APT-PC  were prepared. For X-ray diffraction , poloxamer 188 , and oil  used. The aim of this study was to reduce the size of APT-IE. Due to smaller emulsion droplets undergo reduced Brownian movement, the emulsion is more stable. In addition, the drug absorption can be enhanced with smaller particle sizes and thus realized the greater improvements of bioavailability. The 20 runs designed by CCD were presented in The APT-IE formulation was designed and optimized using the central composite design response surface methodology (CCD-RSM). According to the results of single-factor experiments, three independent variables were designed of the proportion of sodium oleate . The final emulsion was filled into vials under nitrogen protection and sterilized by 0.21 μm Nalgene® nylon filter.2.6.2.6.1.The particle size, PDI and zeta potential of APT-IE were determined by the principle of dynamic light scattering  after APT-IE was diluted with ultrapure water. The pH of APT-IE was obtained by pH meter , and the osmotic pressure was determined with a Model 3250 osmometer .18 column at 30 °C. The mobile phase consisted of acetonitrile-water  at a flow rate of 1.0 mL/min, then the wavelength was set at 210 nm. The DL was calculated by C: concentration of APT, the units of C and DL are both mg/mL.The drug content of APT was determined by HPLC with an Agilent CEE)% of APT-IE was measured by the Sephadex column filtration method. According to the APT-IE elution curve, partial eluent of the emulsion was collected and diluted with methanol. The following equation was used to calculate the EE%:bW: amount of APT in oil; aW: total amount of APT.The encapsulation efficiency (2.6.2.The transmission electron microscopy (TEM)  was used for the observation of morphology and particle size of APT-IE. The sample was diluted 1:50 (v/v) with ultrapure water before being added onto a copper grid. Then the thin liquid film was negatively stained by adding a drop of 2% phosphotungstic acid solution. The sample was placed at room temperature until dried before being TEM observed.2.7.in vitro. The PBS containing 0.5% Tween 80 (pH = 7.4) was used as release medium. APT-IE (1 mL) and APT-SL (1 mL) were taken into dialysis bag respectively, with the water bath at 37 °C. Samples were taken at 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 h, each time 2 mL of release medium was removed and added with 2 mL of fresh release medium. The concentration of APT was determined by HPLC and then calculated the cumulative release rate.Dialysis method was used for the drug release 2.8.The test was performed to measure the hemolytic potential to assess the safety of APT-IE. The red blood cells from rabbits were diluted with normal saline and configured as a 2% red blood cell suspension. The design of hemolysis tests of APT-IE and APT-SL were shown in 2.9.2.9.1.APT-IE is a high-concentration emulsion that needs to be thinned with 0.9% sodium chloride (NaCl) or 5% glucose (G) solution before clinical use. Aseptic withdrawal of 18 mL APT-IE with transfer into 130 mL of 0.9% NaCl or 5% G solution, then observe the appearance and determine the particle size, pH, osmotic pressure and drug concentration of APT-IE at 0, 0.5, 1, 2, 4, and 6h.2.9.2.The long term stability of APT-IE was investigated at 25 ± 2 °C and 4 ± 2 °C for more than 6 months. The samples were evaluated by the changes in appearance, particle size, pH, zeta potential, osmotic pressure and drug content respectively.2.10.2.10.1.n = 6). Both groups were injected with APT-IE or reference APT-SL (11.7 mg/kg) via tail vein. The heparinized tubes were then used to collect blood samples at 0.033, 0.083, 0.25, 0.5, 1, 2, 3, 4, 6, 8, 10 and 12 h by retroorbital vein, respectively. Then the upper plasma was collected from the whole blood by centrifuging it at 12000 rpm for 10 min.The Rats (180 ± 20 g) were divided into two groups  at 40 °C. The mobile phase was methanol-water  at a flow rate of 1.0 mL/min. The wavelength of the UV detector was set at 220 nm.Took 100 μL of plasma or homogenized solution, 400 μL methanol and 20 μL butylparaben  were added. The mixture was vortexed for 10 min, then centrifuged at 14000 rpm for 12 minutes to collect the supernatant. Subsequently, the sample was dried under the protection of nitrogen and reconstituted with methanol. The concentration of APT in plasma or homogenized solution was determined by HPLC equipped with an Agilent C2.11.2.AUC(0–∞) concentration of APT in each organ.The data were analyzed by Winnonlin version 6.4 software  based on a non-compartment model. Moreover, the Winnonlin software was also used to calculate the eR), Targeting efficiency (eT%), Relative targeting efficiency (TeR) and Peak concentration ratio (eC). The parameters were calculated as follows:tAUC refers to AUC of the tissue. t’AUC refers to the total number of tissues, including target tissue.The targeting of each organ was compared based on the following parameters: Relative uptake rate  or too small (<3), the drug transport characteristics may be poor  gradually decreased. From the phospholipid complex sample (1:10), the representative diffraction peak of APT disappeared, which showed that APT was no longer in a crystalline state but in an amorphous state  of APT almost disappeared when the mass ratio was 1:10. We speculated that the phospholipid complex was formed by the interaction of the site with high electronegativity (-C = O) in APT and the polar end in the phospholipid molecule. Furthermore, there was no new absorption peak in the APT-PC spectrum, indicating that the phospholipid complex did not form new compounds.As shown in 3.2.3.The maximum information about compatibility of drugs and excipients can be provided by DSC analysis  of APT-IE. When the amount of EL is 7.2%, the EE (%) of APT-IE can meet the requirements for clinical application with the particle size approximately 100 nm. This is because EL not only can increase the solubility of APT, but also as a emulsifier can reduce the interfacial free energy and form a strong emulsifying membrane to make the emulsion more stable. Therefore, 7.2% EL was added as an emulsifier to ensure sufficient drug loading and stability of APT-IE for further investigation.MCT not only can decrease the viscosity of the oil phase  was performed on the response value. The The equation for calculating the average particle size is as follows:1X2 and X2X3 were significant.The formulation with small particle size tends to be more viscous and could cause pain during intravenous injection  and 7 high-pressure homogenization cycle levels were investigated. As shown in The sterilization method had a significant effect on the characterization and appearance of the APT-IE system. After moist heat autoclaving, the physical properties and appearance of the samples were obviously changed: the color of APT-IE became dark, the liquidity decreased, and some demulsification occurred. Additionally, the drug content and pH value of APT-IE decreased significantly. This result happened may due to the decomposition of phospholipids at high temperature and an increase of free fatty acids. However, no significant changes in appearance, particle size, drug content, and pH value were found after APT-IE sterilized by a 0.21 μm Nalgene® nylon filter. Therefore, the poor compatibility between APT cannot tolerate humid heat autoclaving .The DL of APT-IE was 7.08 ± 0.16 mg/mL, the particle size, PDI, zeta potential, pH value, osmotic pressure and encapsulation efficiency were found to be 82.83 ± 1.89 nm, 0.243 ± 0.008, −59.0 ± 2.54 mV, 8.21 ± 0.07, 301 ± 2.15 mOsmol/kg, and 98.84 ± 1.43% respectively. TEM image of APT-IE was presented in 3.6.2R of Zero order release, first release and Higuchi equations were 0.8410, 0.9121 and 0.9902, respectively. Therefore, the release kinetic of APT-IE was followed the Higuchi equations.The release profile of APT-IE and APT-SL was shown in 3.7.in vitro hemolysis test is considered as a reliable and direct method to predict the safety of injectable preparations. Complete hemolysis was observed in number 7 positive control group, while in number 6 negative control group, the test tube was nearly transparent with the red blood cells sedimentation  in 0.9% NaCl. This result occurred may be related to the presence of cations , the negatively charged droplets adsorb the positive charges and the repulsive force between the droplets is decreased. Therefore, 5% G was suggested for the application in the clinical compatibility of APT-IE.The compatibility stability of APT-IE in combination with 5% G and 0.9% NaCl was investigated within 6 hours. The samples had no color change, precipitation or gas evolution during the 6-hour test period, and there also had no obvious change in drug concentration, pH value or osmotic pressure in both solutions. In addition, the particle size of the samples did not change in 5% G, while slightly changed  of APT in the same tissue after administration of APT-EL and APT-SL. Re>1 manifests the emulsion possess targetability, and Re≤1 shows no targetability, In addition, the larger Re value indicates the better targetability of APT-IE : 55.71%, which are significantly higher than the Re and Te(%) of other organs. The RTe value of liver and heart were larger than 1.00, which indicates enhanced targeting of both tissue by APT-IE compared to APT-SL. At present, APT is widely used as an NK-1 receptor antagonist for the treatment of CINV , soybean oil , egg yolk lecithin , poloxamer 188 , sodium oleate , and sucrose . The emulsion performed good physicochemical properties. Hemolysis tests showed that intravenous injection for APT-IE was safe and suitable. This research confirmed that APT-IE could be stably stored for at least 6 months at 4 ± 2 °C. The results of pharmacokinetic and tissue distribution studies demonstrated that APT-IE expressed increased bioavailability, and obvious liver targeting compared with APT-SL. Consequently, this research suggests that APT-IE loaded with APT-PC has broad prospects for clinical application and provide a data reference for the further study of APT intravenous emulsions.Click here for additional data file.


Helicobacter pylori infection worldwide. This study investigates the potential bactericidal effects of fosfomycin combinations with clarithromycin, metronidazole, ciprofloxacin, amoxicillin, rifampicin, and doxycycline against thirty-six H. pylori strains using the checkerboard and time-kill assay methods. The results showed that ≥ 50% of the strains were resistant to the six antibiotics. Remarkably, only six strains exerted resistance to these antibiotics, with the minimum inhibitory concentrations (MICs) ranges of (3.2–12.8 mg/l), (32–256 mg/l), (3.2–51.2 mg/l), (3.2–25.6 mg/l), (1.6–3.2 mg/l), and (25.6 > 51.2 mg/l), respectively. The seven antibiotics were evaluated through in silico studies for their permeability and ability to bind UDP-N-acetylglucosamine1-carboxyvinyltransferase (MurA) of H. pylori. The results indicated that fosfomycin exhibited the highest predicted membrane permeability  and binding affinity  for H. pylori MurA, compared to other tested antibiotics. The combinations of fosfomycin with these antibiotics exerted synergistic interactions  against the six strains. Importantly, the combinations of fosfomycin with clarithromycin, doxycycline and rifampicin achieved bactericidal effects (reduction ≥ 3.0 Log10 cfu/ml) against the most resistant H. pylori strain. Notably, these effects increased with presence of metronidazole, which enhanced the activity of the fosfomycin combination with amoxicillin from a weak inhibition to bactericidal effect. This study provides evidence that the combination of fosfomycin with either clarithromycin, amoxicillin, doxycycline, or rifampicin  could be a promising option for treating MDR H. pylori infection.Antibiotic resistance represents the main challenge of  Helicobacter pylori is a Gram negative, microaerophilic, motile, and spiral-shaped bacterium. It represents one of the most frequent bacterial human infections worldwide , metronidazole (MET) and ciprofloxacin (CIP) exceeded 15%  in all developed and developing countries  is a broad-spectrum antibiotic, with putative activity against multidrug resistant (MDR) Gram-positive and Gram-negative pathogens. It inhibits the early stage of the bacterial cell wall synthesis. Several studies have investigated the synergistic effects of FOS when combined with other antibiotics that act via a different mechanism of action, thereby allowing for decreased dosages and lower toxicity  used in this study were previously isolated from gastric biopsy specimens from patients with gastric and peptic ulcer. The strains were previously identified based on standard biochemical and molecular (16S rRNA) approaches  glycerol at − 70 °C until use. All the experiments of this study were carried out under microaerophilic conditions using an anaerobe jar  with microaerophilic gas-generating kit .The H. pylori strains to FOS, CLA, MET, CIP, AM, RIF and DO (European pharmacopoeia reference standards) was evaluated through the determination of MIC using agar plate dilution method according to European Committee on Antimicrobial Susceptibility Testing guidelines . The final concentrations of FOS and MET were ranging from 0.5 to 512 mg/l, whereas the final concentrations of CLA, CIP, AM, RIF and DO were ranging from 0.00315 to 51.2 mg/l. Following the inoculation, the plates were dried at room temperature and then incubated at 37 ℃ for 4 days. The MIC was defined as the lowest concentration that inhibited visible growth of bacteria.The susceptibility of thirty six  EUCAST, . Brieflyhttps://swissmodel.expasy.org/). A template search with BLAST and HHblits has been performed against the SWISS-MODEL template library based on the ProMod3 target template alignment and evaluated using the QMEAN score function  were used to build the three dimensional (3D) model with template sequence of high sequence identity according to  sequence was performed using the SWISS-MODEL prediction tools (The docking experiment was performed with Glide’s Extra Precision (XP) program from Schrödinger 16.4  were considered as positive and negative controls, respectively. Following the inoculation, the plates were incubated for 48 h at 37 °C. Additionally, FICs of triple combinations of FOS/MET with CLA, CIP, AM, RIF and DO were determined by the same above-mentioned method. FIC index of the combinations was calculated by the sum of the FIC of each antibiotic alone . FIC index of antibiotic combinations defined as synergy (ΣFIC ≤ 1), indifference (1.0 < ∑FIC ≤ 4) or antagonism (ΣFIC > 4)  and serial tenfold dilutions were prepared in sterile sodium chloride solution  as needed. Three replicates of each diluent were spotted on MHA supplemented with 5% defibrinated sheep blood, dried at room temperature and then incubated 4 days at 37 °C (Inoculated and un-inoculated MHB were considered as positive and negative controls). The data were analyzed by using mean colony counts (Log10 cfu/ml) from the replicates of each diluent at each time interval. The limit of quantification was 2 Log10 cfu/ml. The synergy of the combination was defined as a 2 Log10 cfu/ml decrease compared with the most active antibiotic in this combination, whereas the bacteriostatic and bactericidal effects were defined as 2 and 3 Log10 cfu/ml decease relative to the initial inoculum, respectively.The bactericidal activities of antibiotics  and their respective combinations with FOS (in the presence/absence of MET) against the representative strain were evaluated by performing time-kill assay  using the agar plate dilution method. As shown in Fig. H. pylori strains were found resistant to the six tested antibiotics.The susceptibility of 36 H. pylori strains are summarized in Table H. pylori in either the CLSI or the EUCAST. The MICs ranges of the six antibiotics against the test strains were (3.2–12.8 mg/l), (32–256 mg/l), (3.2–51.2 mg/l), (3.2–25.6 mg/l), (1.6–3.2 mg/l), and (25.6–> 51.2 mg/l), respectively. The MIC of FOS against these strains was ranging from 128 to 256 mg/l. Additionally, all the test strains were classified as MDR since they exhibited resistance to more than two antibiotics related to different antibiotic categories. Notably, H. pylori HP-1 exhibited the highest MIC values of all tested antibiotics. Thus, this strain was selected as a representative strain for time kill studies.The MICs of the seven different antibiotics against the six MDR H. pylori MurA was determined based on the principle of homology modeling, using of a templet (PDB:5UJS) from Campylobacter jejuni ATCC 700,819. The structure analysis indicated that H. pylori MurA showed 60.05% sequence identity with crystal structure of MurA protein from Campylobacter jejuni ATCC 700,819. Additionally, the InterPro protein families and domains of H. pylori MurA demonstrated that the Mur A possesses one active site contain Cys117 and 3 binding sites contain Arg93, Asp308 and Leu330, respectively  bonding interactions in the best docking are also described in Table 234 and Thr329. Under neutral condition (pH 7), the docking score of FOS was − 5.708 kcal/mol, possessing one negative charge and forming 3H bonds with Arg234 and Thr329. Oppositely, under acidic conditions, the docking score of FOS deceased to 2.945 kcal/mol in the pH values ranging from 6 to 3, and it was found in the 0 state with very low binding affinity, forming 3H bonds with Arg234 and Asp308.The effect of different pH levels on the docking score of FOS are summarized in Table H. pylori membranes was evaluated and the results indicated that FOS had greater membrane permeability compared to other tested antibiotics, with ∆G insert a value of − 37.537630 kcal/mol, followed by AM and DO, with ∆G insert values of − 31.255440 and − 29.848466 kcal/mol, respectively. Whereas, RIF and CIP showed moderate membrane permeability, with ∆G insert values of − 17.08 and − 13.84 kcal/mol, respectively. The lower membrane permeability was observed with MET and CLA, with ∆G insert values of − 6.04 and − 5.67 kcal/mol  and their respective combinations with FOS against most resistant strain (HP-1) are presented in Fig. 10 reduction in bacterial count, respectively. Moreover, these combinations exhibited bactericidal effects after 24 h post-treatment, with 3.2, 3.8 and 3.18 Log10 cfu/ml reduction in the initial inoculum, respectively. Notably, MET improved the activity of the combination of FOS with AM against HP-1 from a weak inhibition to a bacteriostatic effect within 3 h of treatment, with a reduction of 2.28 log10 cfu/ml, followed by bactericidal effects after 6 h of treatment and lasted up to 24 h, with a reduction of 3.28 and 3.56 log10 cfu/ml, respectively. Additionally, MET enhanced the bactericidal activity of FOS combinations with CLA, RIF and DO against the representative strain 24 h post-treatment, with the reduction in bacterial count increasing from 3.2, 3.18 and 3.6 to 4.04, 4.09 and 4.02 log10 cfu/ml, respectively. On the other hand, the presence of MET did not influence the activity of FOS/CIP combination against HP-1  defined it as critically important due to its potential efficacy against MDR Gram-positive and Gram-negative bacteria , which used as the first and second lines for the treatment of H. pylori infections. Furthermore, FOS binds the protein active site of H. pylori MurA by forming 4 H bonds with Glu190, and 2H bond with Arg236 and Thr307. The obtained results are consistent with previous studies, which revealed that the bactericidal effects of FOS are due to its binding to the protein active site of the Mur A transferase, rendering it inactive. FOS inhibits the peptidoglycan biosynthesis by preventing the formation of UDP-N-acetylglucosamine-enolpyruvate from UDP-N-acetylglucosamine and phosphoenolpyruvate, resulting in the first step of bacterial cell wall synthesis is disrupted, which ultimately led to the destruction of the bacterial cell  for 234, and Thr329 and two salt bridges with Arg234 as it possess two negative charge, on the other hand at pH value of 7, the binding affinity was − 5.708 kcal/mol with the formation of 3 H-bonds with Arg234, and Thr329 and one salt bridges with Arg234 as it possess one negative charge, However at pH value range from 6 to 3, the docking score was 2.945 kcal/mol which mean very low binding affinity with the formation of 3 H-bonds with Arg234, and Asp308 under non ionized state.Based on the State Penalty, the smaller value under alkaline and neutral conditions 0.6255, and 0.2534 kcal/mol, respectively showed better effect in comparison to low value under acidic condition  compared to other tested antibiotics, which exhibited low membrane permeabilities, with ∆G Insert ranging from − 5.67 to − 31.26 kcal/mol. From these findings, which agree with previous studies  showed good synergistic effects (FIC index < 1) against all The data of time-kill curves of the single and combinations of used antibiotics were consistent with those of the checkerboard experiments. Time-kill curves of single antibiotics  against representative strain (HP-1) showed a considerable regrowth similar to control after 24 h of post-treatment. Additionally, FOS combinations with AM, CIP and MET exhibited an initial reduction within 3–6 h post-treatment followed by a considerable regrowth similar to control after 24 h of post-treatment. These findings were in agreement with those mentioned by previous studies, which revealed that the regrowth phenomenon might be due to that the total bacterial burden contained two particular subpopulations with different susceptibility in which the selective amplification of resistant sub-population take over the preferential killing of the susceptible sub-population at a specified time of interaction  against MDR H. pylori MurA, compared to other tested antibiotics, which used as the first and second lines for the treatment of H. pylori infections. Hence, FOS is potentially a promising antibiotic against H. pylori infection. Additionally, this antibiotic enhances the activity of CLA, DO, RIF and AM against MDR H. pylori by decreasing their MICs to the susceptible breakpoints. Moreover, the combinations of FOS with these antibiotics exert bactericidal effects against MDR H. pylori, especially with the presence of metronidazole. Thus, the combinations of FOS with CLA, DO, RIF and AM could be a promising option for treating MDR H. pylori infection, especially with the presence of metronidazole.Based on in silico analysis, we found that FOS exhibited the highest predicted membrane permeability and binding affinity for


Cucumis melo L. called “Carosello leccese” was grown in a greenhouse with a soilless technique under light-emitting diodes (LEDs) used as supplementary light system. The obtained results showed that “Carosello leccese” contains up to 71.0 mg·g−1 dried weight (DW) of potassium and several bioactive compounds important for human health such as methyl gallate (35.58 µg·g−1 DW), α-tocopherol (10.12 µg·g−1 DW), and β-carotene (up to 9.29 µg·g−1 DW under LEDs). In fact, methyl gallate has antioxidative and antiviral effects in vitro and in vivo, tocopherols are well recognized for their effective inhibition of lipid oxidation in foods and biological systems and carotenoids are known to be very efficient physical and chemical quenchers of singlet oxygen. Finally, it was demonstrated that the LEDs’ supplementary light did not negatively influence the biochemical profile of the peponids, confirming that it can be considered a valid technique to enhance horticultural production without reducing the content of the bioactive compounds of the fruits.The world population is projected to increase to 9.9 billion by 2050 and, to ensure food security and quality, agriculture must sustainably multiply production, increase the nutritional value of fruit and vegetables, and preserve genetic variability. In this work, an Apulian landrace of  Cucumis melo L. (melon) of the Cucurbitaceae family  in an unheated polymethacrylate greenhouse with a maximum height of 4.5 m. Plants were cultivated on a surface of 80 m−2·s−1 of extra photosynthetic photon flux density (PPFD). Two light spectra were tested: red (660 nm) + blue (440 nm) (R + B) and red + blue + far red (730 nm) , the system measured incoming sunlight in the greenhouse at four-second intervals and turned on the LED light bars whenever the ambient PPFD dropped below 200 µmol·m−2·s−1. More detailed information on SL management and LEDs fixtures characteristics have been reported by Palmitessa et al. [LED interlight fixtures , Italy) were used as a supplementary light (SL) system to supply 170 µmol·ma et al. .Cucumis melo L. subsp. melo conv. flexuosus) was grown for 81 days after transplant (DAT) in an extra-seasonal growth cycle with a soilless technique with close-cycle nutrient solution (NS) management. The seedlings were grown until the second true leaf stage at the plant nursery  and transplanted in 10 L pots filled with a mixture composed of peat  and perlite  in a 1:1 (v/v) ratio. Ten pots were placed on each aluminum trough bench . Distance between plants on the bench was 0.30 cm with a stem density of 2.78 plants per m−2. The experimental layout consisted of eleven trough benches . The principal stem of each plant was trained vertically, and the lateral stems were topped after the second node. Periodically, the oldest leaves were removed. The greenhouse temperature was controlled by natural ventilation through ridge openings set to open automatically above 20 °C. Pollination was guaranteed by the introduction of bumblebees (Bombus terrestris L.) into the greenhouse. Temperature and relative humidity were recorded by a CR1000 datalogger . NS composition and management during the crop cycle have been extensively described by Palmitessa et al. [A landrace of the of Cucurbitaceae family called “Carosello leccese” . Carotenoid standards  were purchased from Cayman chemicals ; antheraxanthin was tentatively identified by comparison with retention times and literature UV spectra and quantified by means of lutein standard. Chlorophylls in acetone were purchased from DHI Water & Environment v/v) was added to 100 mg of dried weight (DW) samples. They were vortexed , sonicated for 30 min in a Labsonic177 LBS1-10 ultrasonic bath , and shaken with a magnetic stirrer for 5 h. The samples were centrifuged at 4500× g for 10 min and the supernatants were dried through an evaporator at 40 °C. The dry residue was suspended in 100 µL of methanol 80% (v/v), filtered with 0.2 µm filters, and analyzed with HPLC as reported by Laddomada et al. [v/v) H3PO4 in water (solution B) as follows: isocratic elution, 100% B, 0–30 min; linear gradient from 100% B to 85% B, 30–55 min; linear gradient from 85% B to 50% B, 55–80 min; linear gradient from 50% B to 30% B, 80–82 min; post time, 10 min before the next injection. The flow rate of the mobile phase was 1.0 mL/min, the injection volume was 20 µL and the temperature of the column was set at 30 °C. The wavelength used for the quantification of gallic acid, methyl gallate, and 3,4 dihydroxybenzoic acid was 280 nm; of p-coumaric acid was 295 nm; of clorogenic acid was 320 nm; and of rutin was 350 nm.Polyphenols were extracted with the methodology of Abu-Reidah et al.  and bared 1 min, 00 rpm, sv/v) (B), and tert-methyl butyl ether (C). The gradient elution was as follows: 0 min, 95% A and 5% B; 0–12 min, 80% A, 5% B and 15% C; 12–42 min, 30% A, 5% B and 65% C; 42–60 min, 30% A, 5% B and 65% C; 60–62 min, 95% A, and 5% B. The column was re-equilibrated for 10 min between runs. The flow rate was 1.0 mL/min, and the column temperature was maintained at 25 °C. The injection volume was 10 µL. Wavelengths used for quantification of tocopherols, carotenoids, and chlorophylls were 290 nm, 450 nm, and 665 nm, respectively.Triplicate aliquots of 500 mg DW samples were resuspended in 5 mL distilled water, thus obtaining a homogeneous suspension. Isoprenoid  extraction was performed on 500 mg of the homogeneous suspension through the method of Sadler et al.  as modif−1 NaOH. Results were expressed as mg g−1 DW.“Carosello leccese” fruit samples  were freeze-dried and milled as described in −1 FW) of that sugar in fruits. The formula suggested by Renna et al. [The sweetness index (SI) was calculated based on the content and sweetness properties of individual carbohydrates by multiplying the sweetness coefficient of each sugar  by the concentration  concentrations were determined according to D’Imperio et al. [3 at 65%  in a microwave digestion system . The digestion procedure was performed in two steps: 15 min to reach 200 °C and 10 min maintained at 200 °C . The digested solutions were cooled, diluted, and filtered using a 0.45 µm filter. Samples were analyzed with an inductively coupled plasma-optical emission spectrometry  to measure Ca2+, K+, Mg2+, and Na+ in radial mode and B3+, Fe2+, Mn2+, Zn2+, and Ni2+ in axial mode [Macro and microelement  using the general linear model . The experimental factors were fixed by two-way analysis of variance (ANOVA), and the orthogonal contrasts technique was used to establish differences between means (two contrasts): (1) NL vs. LEDs; (2) (R + B) vs. (R + B + FR).−1 vs. 3021 g∙plant−1) [−1 DW [Supplementary light management and light effects on “Carosello leccese” fruit yield and morphology have already been discussed by Palmitessa et al. . Brieflyplant−1) . However) [−1 DW , respectp-coumaric > chlorogenic acid > rutin. Methyl gallate represented almost 50% of the total polyphenol content, with an average of 35.6 ± 0.7 µg·g−1 DW  and β-tocopherol (β-T) . α-T wascis-β-carotene > violaxanthin > zeaxanthin . Generally, the total sugar content in “Carosello leccese” fruits was higher when the plants were cultivated with LED modules  > phosphorus (P5+) > calcium (Ca2+) > magnesium (Mg2+) > sodium (Na+) > iron (Fe2+) > zinc (Zn2+) > boron (B3+) > manganese (Mn2+) > nickel (Ni2+)  . Potassid 50 DAT . Like pod 50 DAT . Moreovereatment . Differe·kg−1 DW , respect+ B + FR .Recent studies have suggested that LEDs are the emerging technology to use as supplementary light in greenhouse cultivation or as a single source of light energy in indoor farming ,37,38. UCucumis melo L. subsp. melo conv. flexuous , which are widespread in Apulia region [Plants are a remarkable source of antioxidants, such as polyphenols, carotenoids, and vitamins, and they play an important role in the human diet . For thia region ,9. The rCucumis melo L. have a rich polyphenols profile [p-coumaric, chlorogenic acid, and rutin are the main phenolic compounds identified (Hordeum vulgare L.) by Kowalczewski et al. [p-coumaric acid, and chlorogenic acid have antioxidant and antiradical activity [Carosello leccese fruits. The results obtained showed that polyphenol profile was not influenced by supplementary light of the LEDs, thus encouraging the use of this technology in out-of-season cultivation cycles.Polyphenols are known for their ability to scavenge free radicals through hydrogen atom transfer, single electron transfer, and/or chelation of metal cations to contribute to the plant defense system against biotic and abiotic stresses and are also important for human health . Our exp profile , and in entified . Our resentified . Howeveri et al. . Methyl i et al. ,46. Galli et al.  and is ri et al. . Both gai et al. ,50,51. Mi et al. . 3,4-dihactivity . Rutin hactivity ,54. Due Tocopherols usually occur in one of four forms, namely α-, β-, γ-, and δ-tocopherol, differing only in the number and position of methylation on the chromanol ring. The number and position of methyl groups of tocopherols influence their ease of donating hydrogen, hence their antioxidant effectiveness. They are well recognized for their effective inhibition of lipid oxidation in foods and biological systems . Since vCarotenoids are known to be very efficient physical and chemical quenchers of singlet oxygen, as well as potent scavengers of other reactive oxygen species, enhancing the immune response, suppressing cancer development, and protecting eye tissues . β-carotC. melo L.) [−1 of FW of glucose and 7.5 g∙kg−1 FW of fructose [−1 FW of glucose, 18.7 g∙kg−1 FW of fructose [−1 of FW [−1 FW, while under NL it was 8.61 g∙kg−1 FW (−1 FW (−1 FW) and was not influenced by supplementary light treatment. This was another confirmation that the fruits of “Carosello leccese” were harvested at an immature stage. [Sweetness Index (SI) was calculated to determine the sweetness perception of fruits . In thismelo L.) , “Carosefructose , while mfructose . Conside−1 of FW , the glu∙kg−1 FW . TherefoW  and its content 32 DAT was 35.5 mg∙kg−1 FW ; 32 DAT, it was 17.7 mg∙kg−1 FW  was another important element contained in “Carosello leccese” fruit tissues, and it was on average 4.96 mg∙kg−1 FW . In addition, given that the global population is expected to grow from 7 to 9 billion by 2050, there will be rapidly increasing demand for vegetable products. Doing more with less is imperative. Our results indicate that light technology could be used as an innovative method for increasing the yield [Considering the mineral element content, the highest values were found in the fruits harvested 32 DAT and generally light treatments did not affect their concentrations  despite −1 FW . This wakg−1 FW; ), in cuc [−1 FW; ), and in [−1 FW; ). The se; [−1 FW . This wakg−1 FW; ) and in  [−1 FW; ). The th∙kg−1 FW . This wakg−1 FW; ), in cuc [−1 FW; ), and in [−1 FW; ). The fo∙kg−1 FW . This wakg−1 FW; ), in cuc [−1 FW; ), and in [−1 FW; ). Finall∙kg−1 FW . This wa , but it [−1 FW; ) and “Ba [−1 FW; ) cultivaCucumis melo L. subsp. melo conv. Flexuosus, is well suited to greenhouse cultivation. To increase the cultivation of “Carosello leccese”, LEDs have been proposed as a system to obtain high yield through out-of-season production [“Carosello leccese”, an Apulian landrace of oduction . The res


In recent years, questions concerning its effect on the gastrointestinal microbiota have been raised. In the present study, we examined interactions between bacteria and TiO2. The study involved six pathogenic/opportunistic bacterial strains and four different-sized TiO2 types: three types of food-grade E171 compounds and TiO2 NPs (21 nm). Each bacterial strain was exposed to four concentrations of TiO2 . The differences in the growth of the analyzed strains, caused by the type and concentration of TiO2, were observed. The growth of a majority of the strains was shown to be inhibited after exposure to 300 and 600 mg/L of the food-grade E171 and TiO2 NPs.Food-grade titanium dioxide (TiO Since it contains nano-fractions [Titanium dioxide  bacteria, which may also come into contact with TiOntestine , 7. Thisal cells , 9 and a2 NPs applied in food production on selected pathogenic and opportunistic intestinal bacteria (Table 2 concentrations) in which intestinal bacteria come into contact with this food additive.In the present study, we focused on the effect of unmodified TiOia Table . The aim2 were used in the study. Food-grade TiO2 (E171) was purchased from three Polish suppliers: Warchem Sp z o.o., Marki; Biomus, Lublin; and Food Colors, Piotrków Trybunalski . For comparison purposes, TiO2 NPs were purchased from Sigma-Aldrich (CAS Number: 718467-100G. Titanium (IV) oxide, nanopowder, 21 nm) (no. 4). The characteristics of the studied TiO2 are presented in our earlier work [Four types of TiO2 type were prepared daily, before each experiment, in deionized water . Next, each sample was subjected to 30-min sonication in an ultrasonic bath .Aqueous suspensions of each TiOPure MRS media, as well as media with the addition of nanoparticles at four concentrations , were prepared. The growth of bacteria Table  was cont2  . The growth curve for the bacteria cultured on the medium supplemented with these two concentrations of all types of nanoparticles  was similar to that in the control. An exception was the M. luteus strain, whose growth was inhibited already at the lowest concentration (60 mg/L)  Fig. , and sig2 applied at the concentrations of 300 mg/L and 600 mg/L inhibited the growth of four bacterial strains. Interestingly, at the 300 mg/L concentration, there were evident differences in the bacterial responses to the different E171/NPs TiO2 forms used in the experiment  for L. monocytogenes , E. coli O157:H7 , and S. enteritidis  were 2, 3, and 3 mg/mL, respectively. Bacillus subtilis (wild type 3610 strain) growth was slightly decreased by 13 μg/mL of TiO2 NPs , after 72 h of cultivation (zones of inhibition in soft-solid agar LB medium) [2 NPs and Aeroxyde P25  were added at 32–320 μg/mL to batch cultures in non-irradiated conditions induced dose-dependent inhibition of the growth of E. coli K12 MG1655. In the case of food-grade E171 at 320 μg/mL, TiO2 NPs caused complete inhibition of the growth of E. coli after 20 h of the cultivation [2 NPs (D = 13 nm) inhibited the growth of E. coli and Enterobacter cloacae, as shown using the disc diffusion method, but no sophisticated calculations using statistical tools were performed in that study [This study demonstrated that the four different types of TiOi et al. , the min medium) . Both fo2 NPs  were tested against S. typhimurium and E. coli. At 50 μg/mL, the 40 nm, 60 nm, and 80 nm TiO2 NPs induced 3.96, 4.45, and 7.15% cell death, respectively. At the concentration of 250 μg/mL, TiO2 NPs caused a 10% increase in the killing rate, compared to 50 μg TiO2/mL, but the trend was similar at both concentrations applied. Similar results were obtained for the S. typhimurium population, where the killing percentage was nearly 3% at the 50 μg/mL concentration and varied to 2 decimal places only. However, in the S. typhimurium population treated with TiO2 NPs with mean diameters of 60 nm and 80 nm, the killing rate increased 2–threefold along with the higher dose of NPs applied (250 vs. 50 μg NPs/mL) [2 NPs with an average size of ~ 40 nm. The isolated TiO2 NPs decreased the growth of probiotic bacteria  over a concentration range of 125, 250, and 500 μg/mL in vitro (plate method). At the concentration of 500 μg TiO2/mL, NPs inhibited the growth of the probiotic formulation by 66 ± 6.1% and 71 ± 5.6% in anaerobic and aerobic conditions, respectively. Significant reduction in the count of E. coli (ATCC 8739), Salmonella paratyphi (ATCC 9150), and L. monocytogenes   by TiO2 NPs  was observed by Anaya-Esparza et al. [2/mL of nutrient broth significantly reduced the growth of S. enterica var. enteridis, E. coli, S. aureus, and B. cereus in a dose-dependent manner. Sroila et al. [2 NPs and showed that, in the dark, 1–6 nm TiO2 NPs  were efficient inhibitors of the growth of E. coli after 4 h of incubation. The authors concluded that TiO2 NPs can diffuse into the cell membrane of E. coli and disorganize the cell membrane, leading to deformation of the cell and disorganization of intracellular structures. The minimum inhibition concentrations of TiO2 NPs  for the growth of E. coli, S. enteritidis, and L. monocytogenes were 2.00 ± 0.33, 2.50 ± 0.17, and 1.00 ± 0.14 mg/mL, respectively [2 NPs (with particle size < 20 nm), incorporated in sago-based starch film on the growth of E. coli and Salmonella typhimurium. However, in their experiments, the film was complex and cannot be directly compared with our studies.In another study, TiO NPs/mL) . Khan et NPs/mL)  isolateda et al. . Ripollea et al.  reporteda et al.  preparedectively  reported2. In selected cases, we obtained similar results as other authors, concerning the effective concentrations of TiO2 that may have an influence on bacterial growth [2 NPs with a well-defined size in a very narrow range). Thirdly, although we and the cited authors have tested bacteria of the same genus, the outcome may be species/strain dependent. Due to all these factors, the comparison of the results must be made with the awareness of their existence. Another issue is the analysis of how TiO2 affects the growth of bacteria. Unfortunately, we do not have more detailed information yet, what factors determine the reduction of growth and propagation of bacteria in the presence of TiO2, this knowledge is also not included in the cited works.As it can be seen, the above-cited authors reported on the reduction in the growth of bacteria in the presence of TiOl growth , 17, 20,l growth , 19, 21.2 NPs had no influence on the growth of discussed microorganisms. Escherichia coli (ATCC8099) was not inhibited by P25 TiO2 NPs  at 0.15 mg NPs/plate [E. coli by pure TiO2 NPs at 0.5 g/L in the dark and under the light. Rokicka-Konieczna et al. [E. coli K12  in the dark in the presence of 0.1 g/L of TiO2 NPs . Sethi and Sakthivel [E. coli by pure anatase (titania) in the dark in the presence of 0.5 mg/mL of TiO2 NPs. Aguas et al. [E. coli by commercial TiO2  suspended in the solution at the concentration of 50 mg TiO2 NPs/L. No inhibition of the growth of E. coli and S. paratyphi A in the dark was observed in the presence of TiO2 NPs [Last but not least, some studies demonstrated that TiOPs/plate . Similara et al.  showed nakthivel  showed ns et al.  showed nK12 ATCC ,425 in tTiO2 NPs .2 NPs in most of the analyzed strains, at concentrations similar to those reported by a number of other authors. However, there are also discrepancies between works, concerning the effective concentrations of TiO2. The methods of detection of microorganisms, the type of TiO2 (food grade or NPs), or species/strain variability may influence the outcome of the study. Due to these factors, it is necessary to check the mechanism of action of TiO2 on the bacterial cell; therefore, further investigations in this field are indispensable for elucidation of the potential toxicity of NPs to the human microbiome.The complexity and variability of microbiota species in each human impede the assessment of the effect of food additives on this ecosystem. The present study has demonstrated inhibition of bacterial growth caused by both food-grade E171 and TiOESM1(ZIP 8.20 MB)ESM2(DOCX 438 KB)