Ppm Group Research Articles

Ultrastructural evaluation of adverse effects on dentine formation from systemic fluoride application in an experimental mouse model.

Fluoride is widely used in dentistry for its caries prevention. To reduce dental caries, the optimal fluoride concentration of public water supplies in the United States is 0.7 ppm. However, excessive systemic fluoride consumption can lead to dental/enamel fluorosis. Numerous studies have explored the effects of fluoride on enamel and enamel-forming cells. However, research on systemic fluoride's impact on dentine is limited, particularly the effect of fluoride on the structure of the dentine-pulp complex. Therefore, this study aimed to identify how excessive fluoride affects dentine microstructure using an experimental mouse model. C57BL6/J male mice (6-9 weeks old) were randomized into four groups (Fluoride at 0, 50, 100, or 125 ppm in drinking water) (n = 4/group). Mice were provided water adlibitum for 6 weeks along with fluoride-free food. Thereafter, mandibular incisors were analysed. Enamel phenotypes were evaluated using light microscopy and quantitative light-induced fluorescence (QLF) to measure fluorosis levels. Dentine morphology was evaluated using micro-CT, scanning electron microscopy (SEM), SEM-EDX (energy-dispersive X-ray), microhardness test and histological imaging. Data were analysed using one-way ANOVA with Dunnett's multiple comparisons as a post hoc test and the Kruskal-Wallis test with Dunn's multiple comparisons post hoc test (p < .05). Mice treated with fluoride at 50-125 ppm developed enamel hypoplasia in their erupting incisors and micro-CT imaging revealed that fluoride 125 ppm caused external resorption of the growing incisor. Dentine mineral density, dentine volume decreased compared with the 0 ppm control, while pulp volume increased compared with the 0 ppm control group. SEM showed wider predentine layer and abnormalities in calcified matrix vesicles derived from odontoblasts in fluoride 100 and 125 ppm groups. Vickers microhardness of dentine significantly decreased in the high-dose group. Fluoride-induced dentine hypoplasia in a dose-dependent manner. Histological evaluation showed excessive fluoride 125 ppm induced micro abscess formation and inflammatory cell infiltration. Fluoride induced dentine dysplasia with a dentine microstructure resembling hypophosphatasia. High doses of systemic fluoride can cause dentine dysplasia. Both three-dimensional and microstructural analyses showed the structural, chemical and mechanical changes in the dentine and the mineralized tissue components, along with external resorption and pulp inflammation.

Nitric Oxide Ameliorates the Effects of Hypoxia in Mice by Regulating Oxygen Transport by Hemoglobin.

Xiaoying Zhou, Wenting Su, Quanwei Bao, Yu Cui, Xiaoxu Li, Yidong Yang, Chengzhong Yang, Chengyuan Wang, Li Jiao, Dewei Chen, and Jian Huang. Nitric oxide ameliorates the effects of hypoxia in mice by regulating oxygen transport by hemoglobin. High Alt Med Biol. 25:174-185, 2024.-Hypoxia is a common pathological and physiological phenomenon in ischemia, cancer, and strenuous exercise. Nitric oxide (NO) acts as an endothelium-derived relaxing factor in hypoxic vasodilation and serves as an allosteric regulator of hemoglobin (Hb). However, the ultimate effects of NO on the hematological system invivo remain unknown, especially in extreme environmental hypoxia. Whether NO regulation of the structure of Hb improves oxygen transport remains unclear. Hence, we examined whether NO altered the oxygen affinity of Hb (Hb-O2 affinity) to protect extremely hypoxic mice. Mice were exposed to severe hypoxia with various concentrations of NO, and the survival time, exercise capacity, and other physical indexes were recorded. The survival time was prolonged in the 5 ppm NO (6.09 ± 1.29 minutes) and 10 ppm NO (6.39 ± 1.58 minutes) groups compared with the 0 ppm group (4.98 ± 1.23 minutes). Hypoxia of the brain was relieved, and the exercise exhaustion time was prolonged when mice inhaled 20 ppm NO (24.70 ± 6.87 minutes vs. 20.23 ± 6.51 minutes). In addition, the differences in arterial oxygen saturation (SO2%) (49.64 ± 7.29% vs. 42.90 ± 4.30%) and arteriovenous SO2% difference (25.14 ± 8.95% vs. 18.10 ± 6.90%) obviously increased. In ex vivo experiments, the oxygen equilibrium curve (OEC) left shifted as P50 decreased from 43.77 ± 2.49 mmHg (0 ppm NO) to 40.97 ± 1.40 mmHg (100 ppm NO) and 38.36 ± 2.78 mmHg (200 ppm NO). Furthermore, the Bohr effect of Hb was enhanced by the introduction of 200 ppm NO (-0.72 ± 0.062 vs.-0.65 ± 0.051), possibly allowing Hb to more easily offload oxygen in tissue at lower pH. The crystal structure reveals a greater distance between Asp94β-His146β in nitrosyl -Hb(NO-Hb), NO-HbβCSO93, and S-NitrosoHb(SNO-Hb) compared to tense Hb(T-Hb, 3.7 Å, 4.3 Å, and 5.8 Å respectively, versus 3.5 Å for T-Hb). Moreover, hydrogen bonds were less likely to form, representing a key limitation of relaxed Hb (R-Hb). Upon NO interaction with Hb, hydrogen bonds and salt bridges were less favored, facilitating relaxation. We speculated that NO ameliorated the effects of hypoxia in mice by promoting erythrocyte oxygen loading in the lung and offloading in tissues.

Episodic ozone exposure in Long-Evans rats has limited effects on cauda sperm motility and non-coding RNA populations

Epidemiological evidence suggests the potential for air pollutants to induce male reproductive toxicity. In experimental studies, exposure to ozone during sensitive windows in the sperm lifecycle has been associated with impaired sperm motility. Subsequently, we sought to investigate the effects of episodic exposure to ozone during sperm maturation in the rat. Long-Evans rats were exposed to either filtered air or ozone (0.4 or 0.8 ppm) for five non-consecutive days over two weeks. Ozone exposure did not impact male reproductive organ weights or sperm motility ∼24 hours following the final exposure. Furthermore, circulating sex hormones remained unchanged despite increased T3 and T4 in the 0.8 ppm group. While there was indication of altered adrenergic signaling attributable to ozone exposure in the testis, there were minimal impacts on small non-coding RNAs detected in cauda sperm. Only two piwi-interacting RNAs (piRNAs) were altered in the mature sperm of ozone-exposed rats (piR-rno-346434 and piR-rno-227431). Data across all rats were next analyzed to identify any non-coding RNAs that may be correlated with reduced sperm motility. A total of 7 microRNAs (miRNAs), 8 RNA fragments, and 1682 piRNAs correlated well with sperm motility. Utilizing our exposure paradigm herein, we were unable to substantiate the relationship between ozone exposure during maturation with sperm motility. However, these approaches served to identify a suite of non-coding RNAs that were associated with sperm motility in rats. With additional investigation, these RNAs may prove to have functional roles in the acquisition of motility or be unique biomarkers for male reproductive toxicity.

Air Pollution, Ozone, and Sulfur Dioxide Can Affect the Blood Serum Lipid Profile and Oxidative Stress of Male Wistar Rats

The majority of the body’s organs are impacted by ambient air pollution (AAP), which is currently a serious environmental global health issue. The objective of this work was to assess the impact of ozone (O3 ), sulfur dioxide (SO2 ), and AAP on oxidative stress (OS) and lipid profile indicators in male Wistar rats. To this end, these rats were exposed for three hours each day for five weeks in control, AAP, O3 (0.6 ppm), and SO2 (10 ppm) groups (each containing 8 animals). Several parameters, such as total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride, and malondialdehyde (MDA), were measured on blood samples. AAP exposure on TAC (P=0.014) and SOD (P=0.05), MDA (P=0.018), and HDL (P=0.003), O3 exposure on TAC (P&lt;0.001), and SO2 exposure on blood serum TAC (P=0.006) all had statistically significant effects. Based on the results, exposure to SO2 and AAP did not significantly alter the lipid profile (P&gt;0.05). According to our research, exposure to O3 , SO2 , and AAP can increase overall antioxidant capacity by stimulating blood serum oxidative defense enzymes. Except for the enhanced effect of O3 exposure on serum HDL, AAP, SO2 , and O3 exposures had no discernible effects on the lipid profile.

Effect of nanoformulation Azadirachta indica on some factors associated with the vectorial capacity and competence of Anopheles aquasalis experimentally infected with Plasmodium vivax

Malaria remains a highly prevalent infectious disease worldwide, particularly in tropical and subtropical regions. Effectively controlling of mosquitoes transmitting of Plasmodium spp. is crucial in to control this disease. A promising strategy involves utilizing plant-derived products, such as the Neem tree (Azadirachta indica), known for its secondary metabolites with biological activity against various insect groups of agricultural and public health importance. This study investigated the effects of a nanoformulation prototype Neem on factors linked to the vector competence of Anopheles aquasalis, a malaria vector in Latin America. Different concentrations of the nanoformulation were supplied through sugar solution and blood feeding, assessing impacts on longevity, fecundity, fertility, and transgenerational survival from larvae to adults. Additionally, the effects of the Neem nanoformulation and NeemAZAL® formulation on the sporogonic cycle of P. vivax were evaluated.Overall, significant impacts were observed at 100 ppm and 1,000 ppm concentrations on adult survival patterns and on survival of the F1 generation. A trend of reduced oviposition and hatching rates was also noted in nanoformulation-consuming groups, with fertility and fecundity declining proportionally to the concentration. Additionally, a significant decrease in the infection rate and intensity of P. vivax was observed in the 1,000 ppm group, with a mean of 3 oocysts per female compared to the control's 27 oocysts per female. In the commercial formulation, the highest tested concentration of 3 ppm yielded 5.36 oocysts per female. Concerning sporozoite numbers, there was a reduction of 52 % and 87 % at the highest concentrations compared to the control group. In conclusion, these findings suggest that the A. indica nanoformulation is a potential as a tool for malaria control through reduction in the vector longevity and reproductive capacity, possibly leading to decreased vector population densities. Moreover, the nanoformulation interfered with the sporogonic development of P. vivax. However, further basic research on Neem formulations, their effects, and mechanisms of action is imperative to gain a more specific perspective for safe field implementation.

Effects of three disinfection solutions on residual monomers released from resin nanoceramic CAD/CAM blocks

The aim of this study was to evaluate the effects of three disinfection solutions on the amount of monomers released from resin nanoceramic CAD/CAM blocks using high performance liquid chromatography (HPLC). Forty resin nanoceramic CAD/CAM (Cerasmart, GC, Japan) samples (12x14 × 2 mm) were divided into four groups; each group was disinfected using one of four solutions (Group 1: no disinfectant; Group 2: 70 % ethanol; Group 3: 2 % glutaraldehyde; and Group 4: 1 % sodium hypochlorite) for 5 min. Analysis of residual monomers (UDMA and Bis-EMA) amounts was performed using an HPLC instrument (Dionex Ultimate 3000, Thermo Fisher Scientific). After 30 days, the amounts of monomers found were as follows: 14.54 ppm for Group 1; 9.28 ppm for Group 2; 10.60 ppm for Group 3; and 2.76 ppm for Group 4 (the smallest monomer amount) (p < 0.001). Disinfection of indirect restorations prior to cementation can reduce the amount of residual monomers remaining from resin nanoceramic CAD/CAM blocks.

Metabolic equilibrium and reproductive resilience: Freshwater gastropods under nanoplastics exposure

Nanoplastics (NPs) have gained increasing attention due to their widespread presence in aquatic environments and potential adverse effects on organisms. The interaction between NPs and freshwater gastropods can lead to a range of physiological and reproductive disturbances. In this study, we investigated the adverse effects of NPs (two size: 20 nm and 100 nm; three concentrations: 0.5, 50 and 100 ppm) on energy metabolism and reproductive fitness in freshwater gastropods Lymnean stagnalis after 21 days exposure. Briefly, the condition index negatively correlated with increasing NPs concentrations for both sizes. Bioaccumulation revealed a concentration-dependent trend in the 100 nm group, and the highest accumulation appeared in the 100 ppm group, compared to all the rest groups. This phenomenon could be attributed to the larger surface area which facilitates stronger attachment to tissues, while smaller particles could be cleared more readily from body. Carbohydrate and protein reserves remained largely unaffected at all concentrations. However, 100 nm NPs triggered stress responses, increasing lipid production, and 20 nm NPs potentially interfered with mitochondrial function, affecting electron transport system activity. Despite the variations observed in lipid levels and energy cost, the ratio of available energy to energy cost remained stable across for both NPs sizes, and this resilience suggests that cellular energy allocation endured undisturbed, hinting at mechanisms that enable gastropods to maintain their metabolic equilibrium. Reproductively, NPL-exposed groups had fewer clutches, with clutches per collection time decreasing over time for both sizes. In terms of egg development, shell growth and hatching rates remained unaffected, suggesting resilience in aquatic ecosystems.In conclusion, this study underscores the substantial impact of NPs on freshwater gastropods, raising ecological and reproductive concerns. The intricate interplay between nanoparticle size, concentration, and physiological responses highlights the complexity of NPs interactions in aquatic ecosystems, necessitating further research and regulatory measures.

Lactational transfer of sulforaphane-N-acetylcysteine in vivo and in human breast milk

Sulforaphane (SFN) is a bioactive phytonutrient found in cruciferous vegetables. There is a lack of detailed information on the lactational transfer of SFN and SFN metabolites, and potential pharmacological effects on breastfeeding infants. We carried out two maternal supplementation studies in a mouse model, wherein lactating dams received either vehicle, 300 or 600 ppm SFN from postnatal day (PND) 1 to 5, or in a second experiment, vehicle or 600 ppm SFN from PND 1 to 14. The parent compound was only detectable in milk and plasma from dams receiving 600 ppm SFN for five days. The predominant metabolite SFN-N-acetylcysteine (SFN-NAC) was readily detected in milk from dams receiving 300 and 600 ppm SFN for five days or 600 ppm for 14 days. Maternal SFN-NAC plasma levels were elevated in both 600 ppm groups. Maternal hepatic and pulmonary expression of NRF2-related genes, Nqo1, Gsta2, Gstm1, and Gstp1, were significantly increased, generally following a dose-response; however, offspring induction varied. PND5 neonates in the 600-ppm group exhibited significantly elevated expression of Nqo1, Gsta2, and Gstp1 in liver, and Gstm1 and Gstp1 in lung. Findings support maternal dietary supplementation with SFN induces NRF2-related gene expression in neonates via lactational transfer of SFN-NAC. However, NQO1 enzyme activity was not significantly elevated, highlighting the need to optimize dosing strategy. Additionally, in a pilot investigation of lactating women consuming a typical diet, without any purified SFN supplementation, 7 out of 8 breast milk samples showed SFN-NAC above the limit of quantification (LOQ). Notably, the one sample below the LOQ was collected from the only participant who reported no consumption of cruciferous vegetables in the past 24 h. The parent compound was not detected in any of the human breast milk samples. Overall, these data indicate lactational transfer of SFN-NAC at dietary relevant levels. Future studies are needed to evaluate pharmacokinetics and pharmacodynamics of lactational transfer for potential preventive or therapeutic effects in breastfeeding children.

From fuzzy-TOPSIS to machine learning: A holistic approach to understanding groundwater fluoride contamination

Fluoride (F−) contamination of groundwater is a prevalent environmental issue threatening public health worldwide and in India. This study targets an investigation into spatial distribution and contamination sources of fluoride in Dhanbad, India, to help develop tailored mitigation strategies. A triad of Multi Criteria Decision Making (MCDM) models (Fuzzy-TOPSIS), machine learning algorithms {logistic regression (LR), classification and regression tree (CART), Random Forest (RF)}, and classical methods has been undertaken here. Groundwater samples (n = 283) were collected for the purpose. Based on permissible limit (1.5 ppm) of fluoride in drinking water as set by the World Health Organization, samples were categorized as Unsafe (n = 67) and Safe (n = 216) groups. Mean fluoride concentration in Safe (0.63 ± 0.02 ppm) and Unsafe (3.69 ± 0.3 ppm) groups differed significantly (t-value = −10.04, p < 0.05). Physicochemical parameters (pH, electrical conductivity, total dissolved solids, total hardness, NO3−, HCO3−, SO42−, Cl−, Ca2+, Mg2+, K+, Na+ and F−) were recorded from samples of each group. The samples from ‘Unsafe group’ showed alkaline pH, the abundance of Na+ and HCO3− ions, prolonged rock water interaction in the aquifer, silicate weathering, carbonate dissolution, lack of Ca2+ and calcite precipitation which together facilitated the F− abundance. Aspatial distribution map of F− contamination was created, pinpointing the “contaminated pockets.” Fuzzy- TOPSIS identified that samples from group Safe were closer to the ideal solution. Among these models, the LR proved superior, achieving the highest AUC score of 95.6 % compared to RF (91.3 %) followed by CART (69.4 %). This study successfully identified the primary contributors to F− contamination in groundwater and the developed models can help predicting fluoride contamination in other areas. The combination of different methodologies (Fuzzy-TOPSIS, machine learning algorithms, and classical methods) results in a synergistic effect where the strengths of each approach compensate for the limitations of the other.

Dose and gender dependence of chlorine inhalation in a conscious ovine model

Characterization of the pathophysiology of ARDS following chlorine gas inhalation in clinically relevant translational large animal models is essential, as the opportunity for clinical trials in this type of trauma is extremely limited. To investigate Cl2 concentration and gender-dependent ARDS severity. Sheep (n = 54) were exposed to air or Cl2 premixed in air at a concentration of 50, 100, 200, and 300 ppm for 30 min under anesthesia/analgesia and monitored for an additional 48 h in a conscious state. Cardiopulmonary variables and survival endpoints were compared between male and female sheep. Overall there were no significant differences in the responses of female and male sheep except pulmonary oxygenation tended to be better in the male sheep (300 ppm group), and the pulmonary arterial pressure was lower (200 ppm group). The onset of mild ARDS (200 < PaO2/FiO2 ≤ 300) was observed at 36 h post exposure in the 50 ppm group, whereas the 100 ppm group developed mild and moderate (100 ≤ PaO2/FiO2 ≤ 200) ARDS by 12 and 36 h after injury, respectively. The 200 ppm and 300 ppm groups developed moderate ARDS within 6 and 3 h after injury, respectively. The 300 ppm group progressed to severe (PaO2/FiO2 ≤ 100) ARDS at 18 h after injury. Increases in pPeak and pPlateau were noted in all injured animals. Compared to sham, inhalation of 200 ppm and 300 ppm Cl2 significantly increased lung extravascular water content. The thoracic cavity fluid accumulation dose-dependently increased with the severity of trauma as compared to sham. At necropsy, the lungs were red, heavy, solidified, and fluid filled; the injury severity grew with increasing Cl2 concentration. The severity of ARDS and mortality rate directly correlated to inhaled Cl2 concentrations. No significant sex-dependent differences were found in measured endpoint variables.

Effects of adding spermidine carbon quantum dots in feed on growth, intestinal morphology, immunity and disease resistance of Anguilla japonica and Anguilla marmorata

Japanese eel (Anguilla japonica) and giant mottled eel (Anguilla marmorata) are the main eel species reared in recirculating aquaculture system (RAS) in Taiwan. RAS increases the rearing density of eel. On the other hand, it spreads pathogens and diseases more efficiently and uncontrollably. Currently, eel farmers use antibiotics to control disease in RAS, but their misuse can lead to resistance and food-safety issues. Spermidine carbon quantum dots (CQDSpds), a newly developed carbon nanomaterial with positive charges on the surface, can bind to the negatively charged bacterial cell membrane to destroy bacteria and not cause drug resistance. This research investigated the effects of different concentrations of CQDSpds as paste feed additives on the growth performance, intestinal morphology, immune-related gene expression, and disease resistance of Japanese and giant marble eels. The results showed the highest final body weight, weight gain, and specific growth rate of Japanese eel and giant mottled eel fed with 1 ppm and 0.5 ppm CQDSpds added. Compared with the control group, 1 ppm CQDSpds group showed higher lysozyme expression in Japanese eels. In the part of intestinal morphology, there was not a significant difference in intestinal villus height and intestinal morphology between the CQDSpds groups and the control group in both eel species. The survival rate of each group supplemented with CQDSpds was higher than the control group after the E. tarda challenge test, among which the highest were found at 1 ppm group in Japanese eel and 0.5 ppm group in giant mottled eel. By this research, the additive of CQDSpds in eel culturing could improve the growth and immune performance with not caused an impact on the intestine of both eels, which can be a potential feed additive in intensive eel farming.

Clinical impact of permanent pacemaker implantation following surgical ablation of atrial fibrillation concomitant to mitral valve surgery: a multicenter study

Abstract Background Permanent pacemaker (PPM) implantation is a common adverse event following arrhythmia surgery. However, the epidemiology and clinical impact of PPM has been shown in only few studies in heterogeneous population, and the results are controversial. Purpose The present study aimed to investigate the clinical impact of PPM implantation following surgical ablation of atrial fibrillation (AF) concomitant to mitral valve (MV) surgery. Methods We identified 721 patients (aged 60.2±11.0 years, 437 females) who underwent surgical ablation of AF along with MV repair or replacement at five representative institutions in Korea between 2005 and 2017. Of these, 56 had PPM implanted (PPM group) and the remainder (n=665) did not receive a PPM (No-PPM group). The clinical outcomes were compared between the groups, and multivariable Cox proportional hazard analysis with time-varying covariate was conducted to determine the impact of PPM implantation on mortality and stroke. Results The patients in the PPM group were older, and had a higher CHAD2S2-VASc score than those in the No-PPM group (Table). During follow-up of 68.3 months (interquartile range 44.5-105.4 months), no significant survival difference was evident between the groups, however, the PPM group presented a significantly higher incidence of stroke than No-PPM group (p&amp;lt;0.001) (Figure). On time-varying multivariable analysis, the PPM was not a risk factor for mortality (hazard ratio [HR], 0.44; 95% confidence interval [CI], 0.18-1.08; p=0.072), however, old age (HR, 1.10; 95% CI, 1.07-1.13; p&amp;lt;0.001), chronic kidney disease (HR, 1.61; 95% CI, 1.01-2.58; p=0.046) and previous cardiac surgery (HR, 1.86; 95% CI, 1.01-3.21; p=0.027) were independent predictors of mortality in this subset of patients. The only significant risk factor for stroke was CHAD2S2-VASc score (HR, 1.27; 95% CI, 1.01-1.61; p=0.044), and PPM was not associated with risk of stroke (HR, 1.87; 95% CI, 0.71-4.90; p=0.202). Conclusion PPM implantation following surgical ablation of AF concomitant to MV surgery was not associated with an increased risk of mortality or stroke. This study result provides a supporting evidence for an active attempt in surgical ablation despite of inherent risk of PPM implantation.

Ozone exposure induces cough hypersensitivity in mice

Purpose: To study the influence of O3 exposure on cough sensitivity, airway barrier function and airway inflammation in mice.&#x0D; Methods: Cough sensitivity was determined in healthy male C57/BL6 mice (aged 8 - 10 weeks) which were exposed to different concentrations of O3 (0.5 - 2 ppm) for 3 h daily for 9 days. Hematoxylin and eosin (H&amp;E) staining of lung tissues, collection of BALF, and cell count were carried out. Inflammatory factor levels in pulmonary tissues were determined by enzyme-linked immunosorbent assay (ELISA), while Western blotting was used to assay TRPA1 and Claudin-1 protein expressions in lung tissues.&#x0D; Results: After 9 days of mice exposure to O3, cough sensitivity increased significantly, and TRPA1 protein was increased in pulmonary tissues, with exposure level of 1 ppm resulting in the highest level of TRPA1 protein expression. Claudin-1 expression in lung tissues of mice decreased after O3 exposure, especially in the groups exposed to O3 levels of 0.5 ppm and 2 ppm. The total cell count in alveolar lavage fluid of mice exposed to O3 was significantly increased (p &lt; 0.05). In addition, O3 exposure increased IL-1α, IL-6 and TNF-α levels, with the most significant increase in the 0.5 ppm group (p &lt; 0.05). Results from histology revealed that all mice had inflammatory reactions and destruction of lung tissues after O3 exposure.&#x0D; Conclusion: Exposure to O3 induces disruption of airway barrier function, infiltration of the airway by inflammatory cells, and increased secretion of inflammatory factors, thereby resulting in enhanced cough sensitivity.

Implementation of analytical quality by design concepts for the optimization of quantitative 1H nuclear magnetic resonance (1H-qNMR) method for quantitation of novel anti-covid drugs (molnupiravir and favipiravir) in their pharmaceutical dosage forms

For the first time in the literature, the Analytical Quality by Design (AQbD) principles were applied to the development of a specific and simple quantitative proton nuclear magnetic resonance (1H-qNMR) method for quantitation of two antiviral drugs, molnupiravir (MOL) and favipiravir (FAV). This 1H-qNMR method was developed within the QbD framework, the new paradigm of quality recently outlined in the ICH Q14 guidelines. Basic principles of AQbD have been adapted starting with early risk assessment and identification of the analytical target profile (ATP), the analytical procedure attributes (APA) and the analytical procedure parameters (APPs). During the scouting phase, dimethyl sulfoxide-d6 (DMSO‑d6) was selected as the deuterated solvent and maleic acid as the internal standard. The quantitative determination was based on the selective proton signals from MOL's pyrimidine ring at 6.82 ppm (doublet) and FAV's hydroxyl group at 8.71 ppm (singlet), as well as a singlet signal from the IS at 6.24 ppm. A Box-Behnken Design was employed in response surface methodology to develop mathematical predictive models relating the APPs to the APAs followed by multicriteria- decision making with the aid of Derringer's desirability function. Finally, the method operable design regions (MODRs) were computed with the aid of Monte Carlo simulations and identified as the multidimensional combination of the APPs where the risk of failure to fulfill the requirements for the APAs was lower than 5 %. The most robust NMR conditions for MOL, with MODR intervals in brackets, were as follows: no. of scans of 72 (16–124.4), a relaxation delay of 19.8 s (1.2–40 s) and an acquisition time of 3.98 s (2.1–6 s). Similarly, for FAV NMR quantitation, the most robust conditions were found to be 72 scans (16–117.3), a relaxation delay of 19.8 s (1.4–40 s) and an acquisition time of 3.98 s (2.2–6 s). The developed method was validated in accordance with the recently endorsed ICH Q2R2 guidelines, and it exhibited detection limits of 1.04 and 0.88 mg mL−1, as well as quantitation limits of 3.45 and 2.95 mg mL−1 for MOL and FAV, respectively. The method was successfully applied to the analysis of the studied analytes in their pharmaceutical products, proving to be simple, fast, non-destructive, and accurate. Additionally, it can be considered an environmentally friendly alternative for quantitation and quality control analysis of pharmaceutical products.

Effect of Different Remineralization Agents on Artificial Caries Lesion: An in-vitro Study

Objective: Remineralization technologies have been shown to arrest or reverse early carious lesions. This study aimed to evaluate and compare the effect of different agents on enamel remineralization in-vitro. Methods: Thirty-five enamel blocks were prepared and divided into the following groups: Group 1; CaGp and Xylitol-containing gel (R.O.C.S Medical Mineral Gel), Group 2; 1100 ppm NaF and 10% CPP-ACP toothpaste (MI Paste One), Group 3; 0.45% SnF2-1150 ppm F toothpaste (Enamelon), Group 4; Positive Control-1450 ppm NaF toothpaste (Colgate Total) and Group 5; Negative Control (deionized water). Microhardness was measured at baseline, after demineralization, and after respective treatments for different treatment groups using a digital Micro Vickers Hardness Tester. For producing demineralized lesions, samples were stored in acidic hydroxyethylcellulose (HEC, pH=4.8) for three days. The Paired Sample t-test, one-way ANOVA, and Tukey were used to compare data and SMH recovery (%SMHR) calculated among treatments. Results: The mean baseline surface microhardness value was statistically non-significant between the groups (p=.378). CaGP and Xylitolcontaining gel demonstrated having the most protective effect against demineralization. The surface remineralization potential of 1100 ppm NaF and 10% CPP-ACP containing novel toothpaste (MI Paste One) was almost similar to the positive control (1450 ppm Fluoride toothpaste)group (p&gt;.05). Conclusions: All treatment groups showed remineralization after respective treatments and these agents can be used as an effective preventive measure for pediatric patients.

The anxiolytic and circadian regulatory effect of agarwood water extract and its effects on the next generation; zebrafish modelling

Anxiety is one of the most common psychiatric symptoms worldwide. Studies show that there is an increase of >25 % in the prevalence of anxiety with the onset of the COVID-19 pandemic process. Due to the various side effects of drugs used in the treatment of anxiety, interest in natural therapeutic alternatives has increased. Agarwood is a plant used as a natural therapeutic due to its sedative effect as well as many effects such as antioxidant and antibacterial. Although there are many studies with agarwood, comprehensive behavioral studies, including the next generation, are limited. In present study, zebrafish fed with diets containing 10–100 ppm water extract of Agarwood (AWE) for 3 and 8 weeks were exposed to predator stress using Oscar fish in order to test the potential anxiolytic effect of AWE. At the end of the period, zebrafish exposed to predator stress were subjected to anxiety and circadian tests. Histopathological evaluation and immunofluorescent analyzes of BDNF and 5HT4-R proteins were performed in the brains of zebrafish. The effects on the next generation were examined by taking offspring from zebrafish. According to the results, it was observed that AWE had a healing effect on anxiety-like behaviors and on the disrupted circadian rhythm triggered by the predatory stress it applied, especially in the 8 weeks 100 ppm group. Interestingly, it was also found to be effective in offspring of zebrafish fed diets with AWE.

Effects of Mobile-Based Financial Incentive Interventions for Adults at Risk of Developing Hypertension: Feasibility Randomized Controlled Trial.

Hypertension is the leading modifiable risk factor for cardiovascular disease and mortality. Adopting lifestyle modifications, like increasing physical activity (PA), can be an effective strategy in blood pressure (BP) control, but many adults do not meet the PA guidelines. Financial incentive interventions have the power to increase PA levels but are often limited due to cost. Further, mobile health technologies can make these programs more scalable. There is a gap in the literature about the most feasible and effective financial incentive PA framework; thus, pay-per-minute (PPM) and self-funded investment incentive (SFII) frameworks were explored. The aims were to (1) determine the feasibility (recruitment, engagement, and acceptability) of an 8-week mobile-based PPM and SFII hypertension prevention PA program and (2) explore the effects of PPM and SFII interventions relative to a control on the PA levels, BP, and PA motivation. In total, 55 adults aged 40-65 years not meeting the Canadian PA guidelines were recruited from Facebook and randomized into the following groups: financial incentive groups, PPM or SFII, receiving up to CAD $20 each (at the time of writing: CAD $1=US $0.74), or a control group without financial incentive. PPM participants received CAD $0.02 for each minute of moderate-to-vigorous PA (MVPA) per week up to the PA guidelines and the SFII received CAD $2.50 for each week they met the PA guidelines. Feasibility outcome measures (recruitment, engagement, and acceptability) were assessed. Secondary outcomes included changes in PA outcomes (MVPA and daily steps) relative to baseline were compared among PPM, SFII, and control groups at 4 and 8 weeks using linear regressions. Changes in BP and relative autonomy index relative to baseline were compared among the groups at follow-up. Participants were randomized to the PPM (n=19), SFII (n=18), or control (n=18) groups. The recruitment, retention rate, and engagement were 77%, 75%, and 65%, respectively. The intervention received overall positive feedback, with 90% of comments praising the intervention structure, financial incentive, and educational materials. Relative to the control at 4 weeks, the PPM and SFII arms increased their MVPA with medium effect (PPM vs control: η2p=0.06, mean 117.8, SD 514 minutes; SFII vs control: η2p=0.08, mean 145.3, SD 616 minutes). At 8 weeks, PPM maintained a small effect in MVPA relative to the control (η2p=0.01, mean 22.8, SD 249 minutes) and SFII displayed a medium effect size (η2p=0.07, mean 113.8, SD 256 minutes). Small effects were observed for PPM and SFII relative to the control for systolic blood pressure (SBP) and diastolic blood pressure (DBP) (PPM: η2p=0.12, Δmean SBP 7.1, SD 23.61 mm Hg; η2p=0.04, Δmean DBP 3.5, SD 6.2 mm Hg; SFII: η2p=0.01, Δmean SBP -0.4, SD 1.4 mm Hg; η2p=0.02, Δmean DBP -2.3, SD 7.7 mm Hg) and relative autonomy index (PPM: η2p=0.01; SFII: η2p=0.03). The feasibility metrics and preliminary findings suggest that a future full-scale randomized controlled trial examining the efficacy of PPM and SFII relative to a control is feasible, and studies with longer duration are warranted.

Investigation of the potential mutagenicity of ethyl tertiary-butyl ether in the tumor target tissue using transgenic Big Blue Fischer 344 rats following whole body inhalation exposure.

Ethyl tertiary-butyl ether (ETBE) is a fuel oxygenate used for the efficiency of motor vehicle fuels and their octane ratings. ETBE has been reported to induce liver adenomas in male rats in a 2-year bioassay at the highest inhalation concentration tested of 5000 ppm. To investigate the potential mutagenicity of ETBE in the liver, male Big Blue Fischer 344 rats were exposed for 28 consecutive days (6h/day) to 0, 500, 1500, and 5000 ppm ETBE. The treated rats were sacrificed 3 days post-exposure and the frequencies of cII mutants were evaluated in the liver and bone marrow tissues. The mutant frequency (MF) of the liver in the negative control group was 36.3 × 10-6 and this value was not significantly differentin ETBE-exposed animals (39.4, 37.3, and 45.9 × 10-6 in 500, 1500, and 5000 ppm groups, respectively). In the bone marrow, the mean MF in the negative control was 32.9 × 10-6 which was not different from the means of the exposed groups (33.8, 22.6, and 32.0 × 10-6 for groups exposed to 500, 1500 and 5000 ppm, respectively). These data, along with consistent negative response reported in the literature for other apical genotoxicity endpoints informs that mutagenicity is not likely the initial key event in the mode of action for ETBE-induced hepatocarcinogenesis in the rat.

Effects of Fluoride Exposure on Thiol/ Disulphide Homeostasis, Testicular Oxidative Stress and Histopathological Changes in Rats

Objective: This study investigated the effects of fluoride-induced toxicity on the reproductive system. For this purpose, testicular oxidative stress, thiol/disulpide homeostasis, sperm motility, density, and histopathology of testicular tissue were evaluated. Material and Methods: In the experiment, 4 groups were formed, with eight animals (n=8) in each group. Group 1, control/saline, groups 2, 3, and 4 were administered orally sodium fluoride (NaF) at doses of 25, 50, and 100 ppm, respectively, for 30 days. Then the rats were sacrificed and testicular tissue samples were evaluated by sperm examination and histopathologically. Biochemical parameters malondialdehyde (MDA), catalase (CAT), advanced oxidation, and protein products (AOPP) were measured spectrometrically. Thiol/disulphide homeostasis was studied with a fully automated analysis method. Results: NaF exposure increased MDA and AOPP levels and decreased CAT levels. Disrupted thiol/disulphide homeostasis. Histopathological examination revealed integrity between germinal cells and tubular atrophy. NaF showed reduced sperm motility and density and increased abnormal sperm count. The findings of the NaF 100 ppm group were statistically significant when compared with the Control and other NaF groups (p˂0.05). Conclusion: The findings concluded that fluoride might cause pathological conditions and infertility in testicular tissue.

Effect of Prosthesis-Patient Mismatch on Long-Term Clinical Outcomes After Bioprosthetic Aortic Valve Replacement

BackgroundProsthesis-patient mismatch (PPM) is common following surgical aortic valve replacement (SAVR). ObjectivesThe purpose of this study was to quantify the impact of PPM on all-cause mortality, heart failure hospitalization, and reintervention following bioprosthetic SAVR. MethodsThis observational nationwide cohort study from SWEDEHEART (Swedish Web system for Enhancement and Development of Evidence based care in Heart disease Evaluated According to Recommended Therapies) and other national registers included all patients who underwent primary bioprosthetic SAVR in Sweden from 2003 to 2018. PPM was defined according to the Valve Academic Research Consortium 3 criteria. Outcomes were all-cause mortality, heart failure hospitalization, and aortic valve reintervention. Regression standardization was used to account for intergroup differences and to estimate cumulative incidence differences. ResultsWe included 16,423 patients (no PPM: 7,377 [45%]; moderate PPM: 8,502 [52%]; and severe PPM: 544 [3%]). After regression standardization, the cumulative incidence of all-cause mortality at 10 years was 43% (95% CI: 24%-44%) in the no PPM group compared with 45% (95% CI: 43%-46%) and 48% (95% CI: 44%-51%) in the moderate and severe PPM groups, respectively. The survival difference at 10 years was 4.6% (95% CI: 0.7%-8.5%) and 1.7% (95% CI: 0.1%-3.3%) in no vs severe PPM and no vs moderate PPM, respectively. The difference in heart failure hospitalization at 10 years was 6.0% (95% CI: 2.2%-9.7%) in severe vs no PPM. There was no difference in aortic valve reintervention in patients with or without PPM. ConclusionsIncreasing grades of PPM were associated with long-term mortality, and severe PPM was associated with increased heart failure. Moderate PPM was common, but the clinical significance may be negligible because the absolute risk differences in clinical outcomes were small.