Use of an in silico knowledge discovery approach to determine mechanistic studies of silver nanoparticles-induced toxicity from in vitro to in vivo.

BACKGROUND: Silver nanoparticles (AgNPs) are considered a double-edged sword that demonstrates beneficial and harmful effects depending on their dimensions and surface coating types. However, mechanistic understanding of the size- and coating-dependent effects of AgNPs in vitro and in vivo remains elusive. We adopted an in silico decision tree-based knowledge-discovery-in-databases process to prioritize the factors affecting the toxic potential of AgNPs, which included exposure dose, cell type and AgNP type (i.e., size and surface coating), and exposure time. This approach also contributed to effective knowledge integration between cell-based phenomenological observations and in vitro/in vivo mechanistic explorations. RESULTS: The consolidated cell viability assessment results were used to create a tree model for generalizing cytotoxic behavior of the four AgNP types: SCS, LCS, SAS, and LAS. The model ranked the toxicity-related parameters in the following order of importance: exposure dose > cell type > particle size > exposure time ≥ surface coating. Mechanistically, larger AgNPs appeared to provoke greater levels of autophagy in vitro, which occurred during the earlier phase of both subcytotoxic and cytotoxic exposures. Furthermore, apoptosis rather than necrosis majorly accounted for compromised cell survival over the above dosage range. Intriguingly, exposure to non-cytotoxic doses of AgNPs induced G2/M cell cycle arrest and senescence instead. At the organismal level, SCS following a single intraperitoneal injection was found more toxic to BALB/c mice as compared to SAS. Both particles could be deposited in various target organs (e.g., spleen, liver, and kidneys). Morphological observation, along with serum biochemical and histological analyses, indicated that AgNPs could produce pancreatic toxicity, apart from leading to hepatic inflammation. CONCLUSIONS: Our integrated in vitro, in silico, and in vivo study revealed that AgNPs exerted toxicity in dose-, cell/organ type- and particle type-dependent manners. More importantly, a single injection of lethal-dose AgNPs (i.e., SCS and SAS) could incur severe damage to pancreas and raise blood glucose levels at the early phase of exposure.

Using a modified electrical aerosol detector to predict nanoparticle exposures to different regions of the respiratory tract for workers in a carbon black manufacturing industry.

The present study was set out to characterize nanoparticle exposures in three selected workplaces of the packaging, warehouse, and pelletizing in a carbon black manufacturing plant using a newly developed modified electrical aerosol detector (MEAD). For confirmation purposes, the MEAD results were compared with those simultaneously obtained from a nanoparticle surface area monitor (NSAM) and a scanning mobility particle sizer (SMPS). We found that workplace background nanoparticle concentrations were mainly coming from the outdoor environment. Size distributions of nanoparticles for the three selected process areas during the work hours were consistently in the form of bimodel. Unlike nanoparticles of the second mode (simply contributed by the process emissions), particles of the first mode could be also contributed by the forklift exhaust or fugitive emissions of heaters. The percents of nanoparticles deposited on the alveolar (A) region were much higher than the other two regions of the head airway (H), tracheobronchial (TB) for all selected workplaces in both number and surface area concentrations. However, significant differences were found in percents of nanoparticles deposited on each of the three regions while different exposure metrics were adopted. Both NSAM and MEAD obtained quite comparable results. No significant difference can be found between the results obtained from SMPS and MEAD after being normalized. Considering the MEAD is less expensive, less bulky, and easy to use, our results further support the suitability of using MEAD in the field for nanoparticle exposure assessments.

Measurements of respirable dust and nanoparticle concentrations in a titanium dioxide pigment production factory.

This study compared respirable dust and nanoparticle concentrations measured by different sampling devices at a titanium dioxide pigment factory. Respirable particle mass concentrations, nanoparticle concentrations, particle size distribution and particle metallic content were measured at different sampling locations. The sampling results of the Multi-orifice Uniform Deposit Impactor (MOUDI) showed that the particle size distribution at this titanium dioxide production factory fell in the range of 1-10 mu m. Generally, the higher levels of the respirable particle mass concentrations and nanoparticle number concentrations were near the packing site of the pigment titanium dioxide production factory. Metal analysis results revealed that the titanium dioxide concentrations in respirable dust and nanoparticles were within the limits specified by National Institute for Occupational Safety and Health (NIOSH). During sampling, particle metallic content analysis is essential for identifying the source of particles and for measuring respirable dust and nanoparticle concentrations.

Filter quality of pleated filter cartridges.

The performance of dust cartridge filters commonly used in dust masks and in room ventilation depends both on the collection efficiency of the filter material and the pressure drop across the filter. Currently, the optimization of filter design is based only on minimizing the pressure drop at a set velocity chosen by the manufacturer. The collection efficiency, an equally important factor, is rarely considered in the optimization process. In this work, a filter quality factor, which combines the collection efficiency and the pressure drop, is used as the optimization criterion for filter evaluation. Most respirator manufacturers pleat the filter to various extents to increase the filtration area in the limit space within the dust cartridge. Six sizes of filter holders were fabricated to hold just one pleat of filter, simulating six different pleat counts, ranging from 0.5 to 3.33 pleats cm(-1). The possible electrostatic charges on the filter were removed by dipping in isopropyl alcohol, and the air velocity is fixed at 100 cm s(-1). Liquid dicotylphthalate particles generated by a constant output atomizer were used as challenge aerosols to minimize particle loading effects. A scanning mobility particle sizer was used to measure the challenge aerosol number concentrations and size distributions upstream and downstream of the pleated filter. The pressure drop across the filter was monitored by using a calibrated pressure transducer. The results showed that the performance of pleated filters depend not only on the size of the particle but also on the pleat count of the pleated filter. Based on filter quality factor, the optimal pleat count (OPC) is always higher than that based on pressure drop by about 0.3-0.5 pleats cm(-1). For example, the OPC is 2.15 pleats cm(-1) from the standpoint of pressure drop, but for the highest filter quality factor, the pleated filter needed to have a pleat count of 2.65 pleats cm(-1) at particle diameter of 122 nm. From the aspect of filter quality factor, this study suggests that the respirator manufacturers should add approximately 0.5 pleats cm(-1) to the OPC derived from the generalized correlation curve for pleated filter design based on minimum pressure drop.

Mechanisms of silver nanoparticle-induced toxicity and important role of autophagy.

Safety concerns have been raised over the extensive applications of silver nanoparticles (AgNPs) because nano dimensions make them highly bioactive, being potentially harmful to the exposed humans. Surface physico-chemistry (shape, surface charge, chemical composition, etc.) that mainly dictates nano-bio interactions is relevant for influencing their biocompatibility and toxicity. Although the hazardousness of AgNPs has been demonstrated in vitro and in vivo, mechanistic understanding of the toxicity particularly at the molecular and organismal levels, in addition to oxidative stress and silver ion dissolution, has remained unclear. A growing body of research has elucidated that autophagy, being activated in response to exposure to various nanomaterials, may serve as a cellular defense mechanism against nanotoxicity. Recently, autophagy activation was shown to correlate with AgNPs exposure; however, the subsequent autophagosome-lysosome fusion was defective. As autophagy plays a crucial role in selective removal of stress-mediated protein aggregates and injured organelles, AgNPs-induced autophagic flux defect may consequently lead to aggravated cytotoxic responses. Furthermore, we suggest that p62 accumulation resulting from defective autophagy may also potentially account for AgNPs cytotoxicity. Intriguingly, AgNPs have been shown to interfere with ubiquitin modifications, either via upregulating levels of enzymes participating in ubiquitination, or through impairing the biological reactivity of ubiquitin (due to formation of AgNPs-ubiquitin corona). Ubiquitination both confers selectivity to autophagy as well as modulates stabilization, activation, and trafficking of proteins involved in autophagic clearance pathways. In this regard, we offer a new perspective that interference of AgNPs with ubiquitination may account for AgNPs-induced defective autophagy and cytotoxic effects.

Particle Size-Selective Assessment of Protection of European Standard FFP Respirators and Surgical Masks against Particles-Tested with Human Subjects.

This study was conducted to investigate the protection of disposable filtering half-facepiece respirators of different grades against particles between 0.093 and 1.61 µm. A personal sampling system was used to particle size-selectively assess the protection of respirators. The results show that about 10.9% of FFP2 respirators and 28.2% of FFP3 respirators demonstrate assigned protection factors (APFs) below 10 and 20, which are the levels assigned for these respirators by the British Standard. On average, the protection factors of FFP respirators were 11.5 to 15.9 times greater than those of surgical masks. The minimum protection factors (PFs) were observed for particles between 0.263 and 0.384 µm. No significant difference in PF results was found among FFP respirator categories and particle size. A strong association between fit factors and protection factors was found. The study indicates that FFP respirators may not achieve the expected protection level and the APFs may need to be revised for these classes of respirators.

Job insecurity and its association with health among employees in the Taiwanese general population.

As employers respond to intensive global competition through the deregulation of labor, job insecurity has become a widespread problem. It has been shown to have significant health impacts in a growing number of workers, but less is known about its social distribution, the mechanisms through which it may act, and the moderating effects of gender, socioeconomic position, and company size. Utilizing data from a national survey of a representative sample of paid employees in Taiwan, we examined the prevalence of job insecurity and its associations with psychosocial work characteristics and health status. A total of 8705 men and 5986 women aged between 25 and 65 years old were studied. Information on perceived job insecurity, industrial and occupational types, psychosocial work characteristics as assessed by the Job Strain model, and various measures of health status were obtained by a self-administered questionnaire. The overall prevalence of job insecurity was high (50%). Job insecurity was more prevalent among employees with lower education attainment, in blue-collar and construction workers, those employed in smaller companies, and in older women. Insecure employees also reported lower job control, higher job demands, and poor workplace social support, as compared with those who held secure positions. Regression analyses showed that job insecurity was strongly associated with poor health, even with adjustment of age, job control, job demands, and work place social support. The deleterious effects of job insecurity appeared to be stronger in men than women, in women who held managerial or professional jobs than women in other employment grades, and in those working in larger companies than smaller ones. The findings of this study suggest that perceived job insecurity is an important source of stress, and it is accompanied with adverse psychosocial work conditions and poor health. High-risk groups were identified for further investigation.

Prevalence and psychosocial risk factors of upper extremity musculoskeletal pain in industries of Taiwan: a nationwide study.

Prevalence of upper extremity disorders and their associations with psychosocial factors in the workplace have received more attention recently. A national survey of cross-sectional design was performed to determine the prevalence rates of upper extremity disorders among different industries. Trained interviewers administered questionnaires to 17,669 workers and data on musculoskeletal complaints were obtained along with information on risk factors. Overall the 1-year prevalence of neck (14.8%), shoulder (16.6%), and hand (12.4%) disorders were higher than those of the upper back (7.1%) and elbow (8.3%) among those who sought medical treatment due to the complaint. Workers in construction and agriculture-related industries showed a higher prevalence of upper extremity disorders. After multiple logistic regression adjusted for age, education, and employment duration, we found job content, physical working condition, a harmonious interpersonal relationship at the workplace and organizational problems were significant determinants of upper extremity disorders in manufacturing and service industries. Male workers in manufacturing industries showed more concern about physical working conditions while female workers in public administration emphasized problems of job content and interpersonal relationships. We concluded that these factors were major job stressors contributing to musculoskeletal pain of the upper extremity.

Cationic polystyrene nanospheres induce autophagic cell death through the induction of endoplasmic reticulum stress.

Nanoparticles (NPs) have been used to produce a wide range of products that have applications in imaging and drug delivery in medicine. Due to their chemical stability, well-controlled sizes and surface charges, polystyrene (PS) NPs have been developed as biosensors and drug delivery carriers. However, the possible adverse biological effects and underlying mechanisms are still unclear. Recently, autophagy has been implicated in the regulation of cell death. In this study, we evaluated a library of PS NPs with different surface charges. We found that NH2-labeled polystyrene (NH2-PS) nanospheres were highly toxic with enhanced uptake in macrophage (RAW 264.7) and lung epithelial (BEAS-2B) cells. Furthermore, NH2-PS could induce autophagic cell death. NH2-PS increased autophagic flux due to reactive oxygen species (ROS) generation and endoplasmic reticulum (ER) stress caused by misfolded protein aggregation. The inhibition of ER stress decreased cytotoxicity and autophagy in the NH2-PS-treated cells. In addition, the Akt/mTOR and AMPK signaling pathways were involved in the regulation of NH2-PS-triggered autophagic cell death. These results suggest an important role of autophagy in cationic NP-induced cell death and provide mechanistic insights into the inhibition of the toxicity and safe material design.

Prevalence of musculoskeletal disorder among workers in Taiwan: a nationwide study.

Musculoskeletal disorder (MSD) is a common problem among workers. In spite of the numerous reports on MSD in various specific groups of workers, few data on the prevalence in the general working population are available except for back pain. We analyzed the information collected through a nationwide survey in Taiwan in 1994 to estimate the prevalence of MSD by age, gender, and education level and identify high-risk industries. In the survey, a standard questionnaire was distributed to a representative sample of 22,475 non-self-employed workers in Taiwan. National estimates were obtained by applying a weight to each participant. Among the sampled workers, 18,942 (84.3%) participated, and 37.0% (standard error=0.4%) had MSD. Female workers had a significantly higher overall prevalence than male workers (39.5% vs. 35.2%, p<0.05). Education and age also had significant associations with MSD (p<0.001 in both genders). "Lower back and waist" were the most frequently affected body parts (18.3% among males and 19.7% among females), but the prevalence of MSDs of the neck, shoulders, hands and wrists were also above 10%. The top ten high-risk major industries for MSD of various body parts for each gender were identified, and some industries, including "Basic Metal Industries" and "Buildings Construction," were among the top ten for multiple body parts. Our study showed that MSDs of body parts other than the back are also prevalent, especially in the neck, shoulders, hands and wrists. We also identified high-risk industries for further research and intervention.

Cytotoxicity, oxidative stress, apoptosis and the autophagic effects of silver nanoparticles in mouse embryonic fibroblasts.

With the advancement of nanotechnology, nanomaterials have been comprehensively applied in our modern society. However, the hazardous impacts of nanoscale particles on organisms have not yet been thoroughly clarified. Currently, there exist numerous approaches to perform toxicity tests, but common and reasonable bio-indicators for toxicity evaluations are lacking. In this study, we investigated the effects of silver nanoparticles (AgNPs) on NIH 3T3 cells to explore the potential application of these nanoparticles in consumer products. Our results demonstrated that AgNPs were taken up by NIH 3T3 cells and localized within the intracellular endosomal compartments. Exposure to AgNPs is a potential source of oxidative stress, which leads to the induction of reactive oxygen species (ROS), the up-regulation of Heme oxygenase 1 (HO-1) expression, apoptosis and autophagy. Interestingly, AgNPs induced morphological and biochemical markers of autophagy in NIH 3T3 cells and induced autophagosome formation, as evidenced by transmission electron microscopic analysis, the formation of microtubule-associated protein-1 light chain-3 (LC3) puncta and the expression of LC3-II protein. Thus, autophagy activation may be a key player in the cellular response against nano-toxicity.

Particle size concentration distribution and influences on exhaled breath particles in mechanically ventilated patients.

Humans produce exhaled breath particles (EBPs) during various breath activities, such as normal breathing, coughing, talking, and sneezing. Airborne transmission risk exists when EBPs have attached pathogens. Until recently, few investigations had evaluated the size and concentration distributions of EBPs from mechanically ventilated patients with different ventilation mode settings. This study thus broke new ground by not only evaluating the size concentration distributions of EBPs in mechanically ventilated patients, but also investigating the relationship between EBP level and positive expiratory end airway pressure (PEEP), tidal volume, and pneumonia. This investigation recruited mechanically ventilated patients, with and without pneumonia, aged 20 years old and above, from the respiratory intensive care unit of a medical center. Concentration distributions of EBPs from mechanically ventilated patients were analyzed with an optical particle analyzer. This study finds that EBP concentrations from mechanically ventilated patients during normal breathing were in the range 0.47-2,554.04 particles/breath (0.001-4.644 particles/mL). EBP concentrations did not differ significantly between the volume control and pressure control modes of the ventilation settings in the mechanically ventilated patients. The patient EBPs were sized below 5 µm, and 80% of them ranged from 0.3 to 1.0 µm. The EBPs concentrations in patients with high PEEP (> 5 cmH2O) clearly exceeded those in patients with low PEEP (≤ 5 cmH2O). Additionally, a significant negative association existed between pneumonia duration and EBPs concentration. However, tidal volume was not related to EBPs concentration.

Size distributions and exposure concentrations of nanoparticles associated with the emissions of oil mists from fastener manufacturing processes.

The aims of the present study were set out to measure size distributions and estimate workers' exposure concentrations of oil mist nanoparticles in three selected workplaces of the forming, threading, and heat treating areas in a fastener manufacturing plant by using a modified electrical aerosol detector (MEAD). The results were further compared with those simultaneously obtained from a nanoparticle surface area monitor (NSAM) and a scanning mobility particle sizer (SMPS) for the validation purpose. Results show that oil mist nanoparticles in the three selected process areas were formed mainly through the evaporation and condensation processes. The measured size distributions of nanoparticles were consistently in the form of uni-modal. The estimated fraction of nanoparticles deposited on the alveolar (AV) region was consistently much higher than that on the head airway (HD) and tracheobronchial (TB) regions in both number and surface area concentration bases. However, a significant difference was found in the estimated fraction of nanoparticles deposited on each individual region while different exposure metrics were used. Comparable results were found between results obtained from both NSAM and MEAD. After normalization, no significant difference can be found between the results obtained from SMPS and MEAD. It is concluded that the obtained MEAD results are suitable for assessing oil mist nanoparticle exposures.

Evaluation of exhalation valves.

Certification tests currently employed by some industrialized nations to certify respirators require that when challenged with air flow at a constant suction head of 25 mmH2O, the leakage into the facepiece from the exhalation valve(s) should not exceed 30 ml min(-1). However, the test alone might not reflect the leakage rate under different levels of vacuum. To study the characteristics of leakage through exhalation valves, a leakage meter was built to measure the leakage rate of four brands of exhalation valves. Seven valves of each brand were tested for leakage rate at pressure drops ranging from 15 to 45 mmH2O (or extended to 75 mmH2O for some valves). Two types of leaks, i.e. fiber insertion and arch dent on the valve seat, were used to investigate the effect of leak shape on the characteristic leak flow as a function of pressure drop. A leakage data point represented a mean value of five measurements. The results showed that the leakage rate could increase or decrease with increasing pressure drop inside the respirators, indicating that the current method of conducting leakage tests, at a constant suction head of 25 mmH2O, does not guarantee better protection than when the respirators are used at a lower suction head. Moreover, based on the sample valves tested in this study, it was found that valve leakage was mostly caused by defects in the valve seats. Both fiber-insertion and arch-dent leaks had either increasing or decreasing characteristic curves, depending on the size of the leak and the material of the valve membrane.