Sleep deprivation boosts O2·- levels in the brains of mice as visualized by a Golgi apparatus-targeted ratiometric fluorescence nanosensor.

Sleep deprivation (SD) is highly prevalent in the modern technological world. Emerging evidence shows that sleep deprivation is associated with oxidative stress. At the organelle level, the Golgi apparatus actively participates in the stress response. In this study, to determine whether SD and Golgi apparatus stress are correlated, we rationally designed and fabricated a novel Golgi apparatus-targeted ratiometric nanoprobe called Golgi dots for O2·- detection. This probe exhibits high sensitivity and selectivity in cells and brain slices of sleep-deprived mice. Golgi dots can be readily synthesized by coprecipitation of Golgi-F127, an amphiphilic polymer F127 modified with a Golgi apparatus targeting moiety, caffeic acid (CA), the responsive unit for O2·-, and red emissive carbon nanodots (CDs), which act as the reference signal. The fluorescence emission spectrum of the developed nanoprobe showed an intense peak at 674 nm, accompanied by a shoulder peak at 485 nm. As O2·- was gradually added, the fluorescence at 485 nm continuously increased; in contrast, the emission intensity at 674 nm assigned to the CDs remained constant, resulting in the ratiometric sensing of O2·-. The present ratiometric nanoprobe showed high selectivity for O2·- monitoring due to the specific recognition of O2·- by CA. Moreover, the Golgi dots exhibited good linearity with respect to the O2·- concentration within 5 to 40 µM, and the limit of detection (LOD) was ~ 0.13 µM. Additionally, the Golgi dots showed low cytotoxicity and an ability to target the Golgi apparatus. Inspired by these excellent properties, we then applied the Golgi dots to successfully monitor exogenous and endogenous O2·- levels within the Golgi apparatus. Importantly, with the help of Golgi dots, we determined that SD substantially elevated O2·- levels in the brain.

Alveolar bone loss and tooth loss contribute to increase in cancer mortality among older patients.

BACKGROUND: Both cancer and periodontitis are more prevalent with age. Information on their relationship in older patients is limited. This study aims to examine whether periodontitis is associated with increased risk of cancer mortality with a ≥ 75-year age group cohort. METHODS: A retrospective cohort study was conducted on 1146 patients who had digital radiographic examinations. Alveolar bone loss and loss of teeth were measured as indicators of periodontitis. Hazard ratio (HR) with 95% confidence interval (CI) were taken as the effect size to summarize the associations between periodontitis and risks of cancer mortality using the multivariate adjusted cox proportional hazards model and competing risk hazard model. RESULTS: Totally, 104 total cancer, 28 lip, oral cavity and pharynx (LOP) cancer, 39 digestive cancer and 13 respiratory cancer cases were documented over 10 years of follow-up. Total cancer (HR 1.27, 95% CI 1.06-1.53) displayed statistically significant associations with alveolar bone loss and tooth loss after adjusting for relevant confounding variables. We also observed borderline significant association between alveolar bone loss and LOP cancer (HR 1.45, 95% CI 0.99-2.12). The above associations were consistent with the results observed from the competing risk hazard models. CONCLUSION: Our results indicate that older patients suffering from tooth loss or alveolar bone loss are at increased risks of cancer mortality, especially for total cancer and LOP cancer.

Occurrence of Microplastics in Fish and Shrimp Feeds.

Plastics with particle sizes of 100 nm to 5 mm are known as microplastics. The contamination of seafood-based feeds by larger microplastics (20 µm to 5 mm) is a growing concern. Here, we analyzed fish and shrimp meals. Microplastics were extracted using density separation methods and characterized using scanning micro Fourier transform infrared spectroscopy (µ-FT-IR). The average microplastic abundance in shrimp meal was 10.7 microplastics·100 g-1. In fish meal, 1.02% of the microplastics were smaller than 1 mm, while most of the microplastics in shrimp meal were 1-5 mm. Eight colors of microplastics were observed; black, red, and orange microplastics have been rarely reported in previous studies. The microplastics found included films, fibers, and fragments, with film-type microplastics being the most common. The main chemical components of fiber-type microplastics were olefins and polyester, while film- and fragment-type microplastics were mainly paraffin and polyethylene. Additional in-depth studies of microplastics in feeds are necessary to provide data support for feed safety assessments.

Polystyrene nanospheres-induced hepatotoxicity in swamp eel (Monopterus albus): From biochemical, pathological and transcriptomic perspectives.

As ubiquitous emerging pollutants, microplastics (MPs) in aquatic environments have aroused critical global concerns. Despite the occurrence and characteristics of MPs in freshwater agroecosystems well-described by our previous study, their ecotoxicological implications in Monopterus albus remains unfathomed. Herein, we dissected toxic effects and mechanisms of PS-NPs exposure against M. albus hepatic tissues at concentrations of 0.5 (L), 5 (M), 10 (H) mg/L for 28 days using physiochemical measurements, histopathological analysis and transcriptomic sequencing. Results showed that upon PS-NPs treatments, levels of ROS, MDA, 8-OHdG and MFO activity were significantly enhanced relative to the control (C) group, while SP content and T-AOC activity were dramatically suppressed, suggesting ROS burst, lipid peroxidation and DNA damage may occur in liver tissues. This oxidative damage further triggered impaired hepatic function and histopathology, disordered lipid metabolism and hepatocyte apoptosis, as reflected by significantly diminished activities of GPT, GOT, ACP, AKP and LDH, paralleled with augmented levels of TG, TC and HSI as well as Cytc and Caspase-3,8,9 activities. Noticeably, concentration-dependent rises of apoptotic rate, vacuolar degeneration and lipid deposition were manifest in TUNEL, H&E and ORO staining. In addition, a total of 375/475/981 up-regulated as well as 260/611/1422 down-regulated DEGs in C vs L, C vs M and C vs H categories were identified based on RNA-seq, respectively. These DEGs were significantly annotated and enriched into GO terms (membrane, cytoplasm, response to stimuli, oxidation-reduction process) as well as KEGG pathways (ether lipid metabolism, apoptosis, chemical carcinogenesis-reactive oxygen species, non-alcoholic fatty liver disease). Moreover, signaling cascades Keap1-Nrf2, p53 and PPAR were either substantially initiated or dysregulated to orchestrate PS-NPs hepatotoxicity featuring oxidative damage, hepatocyte apoptosis and lipid steatosis. Collectively, this study not only expounded on toxicological mechanisms whereby PS-MPs exerted deleterious effects on M. albus, but also pointed to ecological risks of PS-MPs-induced hepatoxicity and lipid steatosis in this commercially-important species.

[A study on decreasing the instrument detection limit of atomic fluorescence spectrometry (AFS-930) for Hg].

In the present study, the detection limit of atomic fluorescence spectrometry (AFS-930) was decreased to 2 ng x L(-1) (n=6) based on several optimizing modifications, including that the sub-high voltage of photomultiplier tube and the current of hollow-cathode lamp were elevated to 280 V and to 30 mA, respectively, and the height of atomization cell was set as 10 mm; In addition, the concentration of KBH4 was decreased to 0.5% (KOH 0.2%). With the optimized parameters, a good standard curve of Hg concentration versus intensity of fluorescence (If) could been obtained readily, after that, a 4-ng x L(-1)-Hg water samples was measured accurately with a little relative standard deviation (RSD) of <5%, while for approximately 2-ng x L(-1)-Hg waters the RSD varied within a wide range of 10.9%-27.2%, likely due to the absorption of Hg by polyethylene vessels used in this study and/or due to the contamination by analysis grade reagents used in this study. By using low-absorption polytetrafluoroethylene (PTFE) materials and the guaranteed reagents, the instrument detection limit was further decreased to 1 ng x L(-1) (n=10).

Injectable long-acting in situ forming systems for Radix Ophiopogonis polysaccharide.

In the area of injectable long-acting formulations, the in situ forming system (ISFS) is an attractive alternative for its various superiorities. In this study, both hydrophilic and hydrophobic in situ forming systems, using Poloxamer and sucrose acetate isobutyrate (SAIB) or poly(D,L-lactide-co-glycolide) copolymer (PLGA) as carrier, respectively, were investigated for Radix Ophiopogonis polysaccharide (ROP), a natural anti-myocardial ischemic fructan. A reasonable and applicable range of formulations were selected from each carrier for in vivo study by investigating their rheological property. The results from in vivo evaluation show that relatively promising sustained behaviors were achieved by formulations 24% P407/10% P188, 40% PLGA30k/NMP, and 30% PLGA50k/NMP. Significant differences of drug release kinetics were observed between in situ thermally-induced Poloxamer-based hydrogels and in situ solvent exchange-induced hydrophobic PLGA depots. This suggests that different ISFS could be chosen to provide different application purpose for polysaccharide drugs. In the case of ROP, Poloxamer-based ISFS is promising for short-term acute therapies; however, PLGA-based ISFS might be promising for long-term precaution or/and cure of myocardial ischemia.

Targeting delivery of Radix Ophiopogonis polysaccharide to ischemic/reperfused rat myocardium by long-circulating macromolecular and liposomal carriers.

Drug delivery to ischemic myocardium is an enormous challenge. This work aimed to characterize cardiac delivery behaviors of mono-polyethylene glycosylated (PEGylated) conjugates and long-circulating liposomes (L-Lps) with Radix Ophiopogonis polysaccharide (ROP) as drug. The results showed that compared to native ROP, 32-, 52-, and 45-fold increases in blood half-life were achieved by 20-kDa PEG mono-modified ROP (P(20k)-R), 40-kDa PEG mono-modified ROP (P(40k)-R), and ROP-loaded L-Lp, respectively. With comparable blood pharmacokinetics, ROP-loaded L-Lp showed both significantly higher targeting efficacy and drug exposure in infarcted myocardium than P(40k)-R. With regard to P(20k)-R, both its targeting efficacy and its level in infarcted myocardium at 3 hours postdose were comparable to P(40k)-R, but its level in blood and myocardium reduced obviously faster. As a whole, the results indicate that both loading in L-Lps and mono-PEGylation are effective in targeting drug to ischemic myocardium, but the former appears to induce stronger effects.

Increased cardiac distribution of mono-PEGylated Radix Ophiopogonis polysaccharide in both myocardial infarction and ischemia/reperfusion rats.

Although PEGylation plays an important role in drug delivery, knowledge about the distribution behavior of PEGylated drugs in ischemic myocardia is rather limited compared to nanoparticles. This work therefore aims to characterize the targeting behavior of the anti-myocardial ischemic mono-PEGylated conjugates of Radix Ophiopogonis polysaccharide (ROP) in two clinically relevant animal models, ie, the myocardial infarction (MI) model and the ischemia/reperfusion (IR) model. To determine the effect of the molecular size of conjugates, two representative conjugates (20- and 40-kDa polyethylene glycol mono-modified ROPs), with hydrodynamic size being approximately and somewhat beyond 10 nm, respectively, were studied in parallel at three time points postdose after a method for determining them quantitatively in biosamples was established. The results showed that the cardiac distribution of the two conjugates was significantly enhanced in both MI and IR rats due to the enhanced permeability and retention effect induced by ischemia. In general, the cardiac targeting efficacy of the conjugates in MI and IR rats was approximately 2; however, different changing in targeting efficacy with time was observed between MI and IR rats and also between the conjugates. Although the enhanced permeability and retention effect-based targeting efficacy for mono-PEGylated ROPs was not high, they, as dissolved macromolecules, are prone to diffusion in the cardiac interstitium space, and thus, facilitate the drug to reach perfusion-deficient and nonperfused areas. These findings are helpful in choosing the cardiac targeting strategy.

Poloxamer-based in situ hydrogels for controlled delivery of hydrophilic macromolecules after intramuscular injection in rats.

This study is aimed to investigate the applicability of poloxamer 407 (P407) and 188 (P188)-based temperature-sensitive in situ hydrogel (TSHG) in sustained delivery of hydrophilic macromolecules following intramuscular administration. Polyethylene glycols (PEGs) with molecular weight of 5-, 20-, and 40-kDa were used as model drugs, which can represent the common size range of hydrophilic macromolecular drugs using TSHG. The correlation between the level of poloxamers and thermogelling transition temperatures (Tsol-gel) was established and two formulations "20% P407/10% P188" and "24% P407/10% P188" were chosen for further study. The results showed that the release kinetics of PEGs was close to zero order. Sustained in vivo behaviors were achieved by both of the two formulations for all the PEGs though variations were seen. Lower molecular weight PEG showed more remarkable pharmacokinetic improvements. No significant differences in pharmacokinetics were observed between the two formulations for the same PEG. This suggested that 20-24% P407/10% P188 formulations, with accordingly Tsol-gel in the range of 24.6 °C-31.7 °C, might be freely chosen to achieve comparable pharmacokinetics for hydrophilic macromolecular drugs after intramuscular injection.