A New Microsphere-Based Immunoassay for Measuring the Activity of Transcription Factors

There are several traditional and well-developed methods for analyzing the activity of transcription factors, such as EMSA, enzyme-linked immunosorbent assay, and reporter gene activity assays. All of these methods have their own distinct disadvantages, but none can analyze the changes in transcription factors in the few cells that are cultured in the wells of 96-well titer plates. Thus, a new microsphere-based immunoassay to measure the activity of transcription factors (MIA-TF) was developed. In MIA-TF, NeutrAvidin-labeled microspheres were used as the solid phase to capture biotin-labeled double-strand DNA fragments which contain certain transcription factor binding elements. The activity of transcription factors was detected by immunoassay using a transcription factor-specific antibody to monitor the binding with the DNA probe. Next, analysis was performed by flow cytometry. The targets hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-kappa B (NF-κB) were applied and detected in this MIA-TF method; the results that we obtained demonstrated that this method could be used to monitor the changes of NF-κB or HIF within 50 or 100 ng of nuclear extract. Furthermore, MIA-TF could detect the changes in NF-κB or HIF in cells that were cultured in wells of a 96-well plate without purification of the nuclear protein, an important consideration for applying this method to high-throughput assays in the future. The development of MIA-TF would support further progress in clinical analysis and drug screening systems. Overall, MIA-TF is a method with high potential to detect the activity of transcription factors

Optimization and Evaluation of a Chitosan/Hydroxypropyl Methylcellulose Hydrogel Containing Toluidine Blue O for Antimicrobial Photodynamic Inactivation

Photodynamic inactivation (PDI) combined with chitosan has been shown as a promising antimicrobial approach. The purpose of this study was to develop a chitosan hydrogel containing hydroxypropyl methylcellulose (HPMC), chitosan and toluidine blue O (TBO) to improve the bactericidal efficacy for topical application in clinics. The PDI efficacy of hydrogel was examined in vitro against the biofilms of Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa). Confocal scanning laser microscopy (CSLM) was performed to investigate the penetration level of TBO into viable S. aureus biofilms. Incorporation of HMPC could increase the physicochemical properties of chitosan hydrogel including the hardness, viscosity as well as bioadhesion; however, higher HMPC concentration also resulted in reduced antimicrobial effect. CSLM analysis further demonstrated that higher HPMC concentration constrained TBO diffusion into the biofilm. The incubation of biofilm and hydrogel was further performed at an angle of 90 degrees. After light irradiation, compared to the mixture of TBO and chitosan, the hydrogel treated sample showed increased PDI efficacy indicated that incorporation of HPMC did improve antimicrobial effect. Finally, the bactericidal efficacy could be significantly augmented by prolonged retention of hydrogel in the biofilm as well as in the animal model of rat skin burn wounds after light irradiation

Comparison of Genotoxicity and Pulmonary Toxicity Study of Modified SiO₂ Nanomaterials

Surface-modified nano-SiO2 is a common additive in many products. However, the safety of nano-SiO2 products under various modifications is still unclear. In this study, we investigated the genotoxicity and acute pulmonary toxicity of nano-SiO2 with or without modification. The samples used in this study included: sample A (SA, 55.16 nm, 411.3 mg/mL), modified sample A (mSA, 82.29 nm, 37.7 mg/mL), sample B (SB, 22 nm, 358.0 mg/mL), and modified sample B (mSB, 86.64 nm, 37.7 mg/mL). In the genotoxicity study, we conducted an Ames test, chromosomal aberration test (CA), and a micronucleus (MN) test. The SA, mSA, and mSB groups showed negative results in all these genotoxicity tests. Only SB showed a weakly positive reaction in these assays, but the genotoxicity could be reversed after S9 metabolism or modification. In the acute pulmonary toxicity test, the rats were given an intratracheal instillation (IT) (0.5 mL/kg) of diluted samples and sacrificed after 1 or 14 days. The mortality rate, number of leukocytes and cytokines of TNF-α in the bronchoalveolar lavage fluid (BALF), and the pathology in the lungs were determined. The results revealed that mSA posed acute toxicity in rats. After modification, the pulmonary toxicity was increased in mSA but decreased in mSB on Day 1, and no significant difference was observed on Day 14. In conclusion, there was no observed genotoxicity in either SA or SB, while mSA posed acute inhalation toxicity to rats that decreased in mSB after modification. This indicates that the decrease in pH level in SA and decrease in the solid content in SB are considered after the trifluorosilane surface-modified amorphous nano-silica

Comparison of Genotoxicity and Pulmonary Toxicity Study of Modified SiO2 Nanomaterials

Surface-modified nano-SiO2 is a common additive in many products. However, the safety of nano-SiO2 products under various modifications is still unclear. In this study, we investigated the genotoxicity and acute pulmonary toxicity of nano-SiO2 with or without modification. The samples used in this study included: sample A (SA, 55.16 nm, 411.3 mg/mL), modified sample A (mSA, 82.29 nm, 37.7 mg/mL), sample B (SB, 22 nm, 358.0 mg/mL), and modified sample B (mSB, 86.64 nm, 37.7 mg/mL). In the genotoxicity study, we conducted an Ames test, chromosomal aberration test (CA), and a micronucleus (MN) test. The SA, mSA, and mSB groups showed negative results in all these genotoxicity tests. Only SB showed a weakly positive reaction in these assays, but the genotoxicity could be reversed after S9 metabolism or modification. In the acute pulmonary toxicity test, the rats were given an intratracheal instillation (IT) (0.5 mL/kg) of diluted samples and sacrificed after 1 or 14 days. The mortality rate, number of leukocytes and cytokines of TNF-α in the bronchoalveolar lavage fluid (BALF), and the pathology in the lungs were determined. The results revealed that mSA posed acute toxicity in rats. After modification, the pulmonary toxicity was increased in mSA but decreased in mSB on Day 1, and no significant difference was observed on Day 14. In conclusion, there was no observed genotoxicity in either SA or SB, while mSA posed acute inhalation toxicity to rats that decreased in mSB after modification. This indicates that the decrease in pH level in SA and decrease in the solid content in SB are considered after the trifluorosilane surface-modified amorphous nano-silica

A New Microsphere-Based Immunoassay for Measuring the Activity of Transcription Factors

<p>Abstract</p> <p>There are several traditional and well-developed methods for analyzing the activity of transcription factors, such as EMSA, enzyme-linked immunosorbent assay, and reporter gene activity assays. All of these methods have their own distinct disadvantages, but none can analyze the changes in transcription factors in the few cells that are cultured in the wells of 96-well titer plates. Thus, a new microsphere-based immunoassay to measure the activity of transcription factors (MIA-TF) was developed. In MIA-TF, NeutrAvidin-labeled microspheres were used as the solid phase to capture biotin-labeled double-strand DNA fragments which contain certain transcription factor binding elements. The activity of transcription factors was detected by immunoassay using a transcription factor-specific antibody to monitor the binding with the DNA probe. Next, analysis was performed by flow cytometry. The targets hypoxia-inducible factor-1&#945; (HIF-1&#945;) and nuclear factor-kappa B (NF-&#954;B) were applied and detected in this MIA-TF method; the results that we obtained demonstrated that this method could be used to monitor the changes of NF-&#954;B or HIF within 50 or 100 ng of nuclear extract. Furthermore, MIA-TF could detect the changes in NF-&#954;B or HIF in cells that were cultured in wells of a 96-well plate without purification of the nuclear protein, an important consideration for applying this method to high-throughput assays in the future. The development of MIA-TF would support further progress in clinical analysis and drug screening systems. Overall, MIA-TF is a method with high potential to detect the activity of transcription factors.</p

Factors associated with registration for organ donation among clinical nurses

[[abstract]]Purpose Healthcare professionals play an important role in the organ donation process. The aim of this study was to examine the organ donation registration rate and related factors among clinical nurses. Material and methods In this cross-sectional, correlational study, we used mailed questionnaires to collect data from four geographical areas and three hospital levels in Taiwan from June 6 to August 31, 2018. Two thousand and thirty-three clinical nurses participated in this study. Results Participants’ mean age was 34.47 years, and 95.7% were women. Of them, 78.3% were willing to donate their organs and 20.6% had registered for organ donation after death. The results of logistic regression showed that in the personal domain, higher age (odds ratio (OR) = 1.03, p < 0.001), better knowledge of organ donation (OR = 1.09, p < 0.001), and a positive attitude toward organ donation (OR = 2.91, p < 0.001) were positively associated with organ donation registration, while cultural myths (OR = 0.69, p < 0.001) were negatively correlated. In the policy domain, the convenience of the registration procedure (OR = 1.45, p < 0.001) was positively associated with registration. A gap between willingness to donate and actual registration was observed. Conclusions Personal factors played an important role in organ donation registration. Therefore, efforts to improve knowledge and inculcate positive cultural beliefs about organ donation among clinical nurses are recommended. There is also a need to cooperate with government policies to provide appropriate in-service training and policy incentives and establish an efficient registration process

Factors associated with the intention to recommend pertussis vaccination for postpartum women: A survey in Taiwan of obstetrician–gynecologists' knowledge, beliefs, and attitudes

AbstractObjectiveTo examine obstetrician–gynecologists' knowledge, beliefs, and attitudes associated with the intention to recommend adult tetanus, reduced diphtheria, and acellular pertussis (Tdap) vaccination to postpartum women.Materials and MethodsA survey instrument was mailed to a total of 2686 members of the Taiwan Association of Obstetrics and Gynecology to assess provider demographic characteristics, occupational information, pertussis knowledge, and beliefs and attitudes about vaccination. The intention to recommend pertussis vaccination to postpartum women was evaluated. Trend chi-square statistics and multivariate logistic models were used to determine variables that were significantly associated with intention to recommend vaccination.ResultsOf the 676 surveys returned (25.2%), 510 participants were active in obstetric practice. A statistically significant difference was found in mean ± SD knowledge scores for pertussis disease and pertussis vaccination between obstetrician–gynecologists who intended to recommend and those who did not intend to recommend postpartum Tdap vaccination (disease: 2.99 ± 2.2 vs. 2.31 ± 1.9, respectively, p < .001; vaccination: 2.64 ± 2.2 vs. 1.36 ± 1.8, respectively, p < .001). Obstetrician–gynecologists who were in favor of postpartum Tdap vaccination were more likely to: (1) rate the change in pertussis incidence among adults as increased; (2) rate pertussis disease among newborn infants as highly severe; (3) rate pertussis as highly contagious; and (4) understand the current recommendation of important organizations for protecting infants against pertussis.ConclusionOur study of obstetrician–gynecologists' attitudes and intentions concerning postpartum Tdap vaccination may be useful in guiding the design of effective perinatal Tdap immunization programs nationwide

Identification of a c.544C>T mutation in WDR34 as a deleterious recessive allele of short rib-polydactyly syndrome

Objective: Single-nucleotide polymorphism (SNP) microarrays and whole-exome sequencing (WES) are tools to precisely diagnose rare autosomal recessive (AR) diseases. In this study, SNP chip and WES were used to identify a mutated location in WDR34 in a baby born to consanguineous parents. Case report: The baby, born at 36 gestational weeks had a small thoracic cage, symmetric short proximal bones, and polydactyly. Radiography showed short ribs with reduced lung volume and pulmonary opacities, compatible with asphyxiating thoracic dystrophy or short rib-polydactyly syndrome (SRPS). At 4 months of age, she died of pulmonary hypoplasia and sepsis. SNP microarray and evaluation tool confirmed WDR34 as the candidate gene. WES detected an AR mutation at c.554C > T [p.Arg182Trp] in WDR34. Conclusion: This study was the first to identify c.544C > T [p.Arg182Trp] mutation in WDR34 in a patient with SRPS. According to the database, the homozygous mutation of c.544C > T in WDR34 was deleterious and the prevalence of heterozygous mutation was relatively higher in Asian population. More studies of this mutation in patients with SRPS are required