BackgroundEnterovirus 71 (EV71) has caused several epidemics of hand, foot and mouth diseases (HFMD) in Asia and now is being recognized as an important neurotropic virus. Effective medications and prophylactic vaccine against EV71 infection are urgently needed. Based on the success of inactivated poliovirus vaccine, a prototype chemically inactivated EV71 vaccine candidate has been developed and currently in human phase 1 clinical trial.Principal FindingIn this report, we present the development of a serum-free cell-based EV71 vaccine. The optimization at each step of the manufacturing process was investigated, characterized and quantified. In the up-stream process development, different commercially available cell culture media either containing serum or serum-free was screened for cell growth and virus yield using the roller-bottle technology. VP-SFM serum-free medium was selected based on the Vero cell growth profile and EV71 virus production. After the up-stream processes (virus harvest, diafiltration and concentration), a combination of gel-filtration liquid chromatography and/or sucrose-gradient ultracentrifugation down-stream purification processes were investigated at a pilot scale of 40 liters each. Although the combination of chromatography and sucrose-gradient ultracentrifugation produced extremely pure EV71 infectious virus particles, the overall yield of vaccine was 7–10% as determined by a VP2-based quantitative ELISA. Using chromatography as the downstream purification, the virus yield was 30–43%. To retain the integrity of virus neutralization epitopes and the stability of the vaccine product, the best virus inactivation was found to be 0.025% formalin-treatment at 37°C for 3 to 6 days. Furthermore, the formalin-inactivated virion vaccine candidate was found to be stable for >18 months at 4°C and a microgram of viral proteins formulated with alum adjuvant could induce strong virus-neutralizing antibody responses in mice, rats, rabbits, and non-human primates.ConclusionThese results provide valuable information supporting the current cell-based serum-free EV71 vaccine candidate going into human Phase I clinical trials.

Background. Hip fractures among older adults are a major public health concern worldwide. This study investigated the potential clinical factors that predict postoperative 1-year activities of daily living (ADL), quality of life (QoL), and mortality in Taiwanese older adults following hip fracture. Methods. This is a prospective cohort study enrolling older adults (≥60 years) who had undergone hip fracture surgery in a single medical center. The comprehensive clinical history of each patient was examined. QoL, ADL, and mortality events were recorded consecutively at 3, 6, and 12 months after operation. The multiple logistic regression model and the generalized estimating equation (GEE) were adopted to identify contributing factors for mortality and postoperative ADL and QoL prognosis, respectively. Results. Among 377 participants with hip fracture, 48 died within 1 year of the index operation. ADL and QoL considerably decreased at 3 months following hip surgery. Old age, high Charlson Comorbidity Index, and American Society of Anesthesiologists grading were crucial predictors for mortality at the 1-year follow-up. The generalized estimating equation analysis indicated that the length of postoperative follow-up time, serum albumin level, patient cognitive status, and handgrip strength were considerably associated with QoL and ADL recovery prognosis in the Taiwanese older adults following hip fracture. Conclusions. Hip fractures have long-lasting effects on the older adults. Our data imply several prognosis predicting parameters that may assist clinicians in accounting for an individual’s personalized risks in order to improve functional outcomes and reduce mortality.

A non-thermal plasma-water system using a microsecond pulsed high-voltage power supply was investigated with air, nitrogen, oxygen, and argon gas feedings individually. Optical emission spectroscopy (OES) was utilized to characterize the primary active species inside the plasmas generated by different gas feedings. The OES method was also employed to estimate the neutral gas and electron temperatures. The pH and the oxidation-reduction potential (ORP) of plasma-activated water (PAW) were measured in the liquid phase. An ion chromatography system (ICS) was employed to present the PAW activity, such as nitrite and nitrate species. Moreover, hydrogen peroxide as a secondary active species inside the activated water, generated by the gases mentioned above, was measured by potassium permanganate titration. It was found that the gas species have a noticeable effect on the pH level as well as the ORP of PAW. In the cases of argon and oxygen plasmas, the pH level of PAW does not change significantly. In contrast, the pH values of PAW generated by air and nitrogen plasmas decline sharply during the treatment time. Moreover, the gas species have a significant impact on the concentrations of nitrite, nitrate, and hydrogen peroxide generated in PAW. The activated water generated by oxygen plasma provides the highest level of hydrogen peroxide. Although the consumed power of argon plasmas was half of the other plasma sources, it provides relatively high hydrogen peroxide contents compared to the nitrogen and air plasmas.

Background Use of appropriate child passenger safety restraints reduces injury in infants, with rear facing restraints favored over forward facing. In 2011, the American Academy of Pediatrics (AAP) began recommending that infants and children under the age of 2 years be restrained in a rear-facing seat installed in the vehicle’s rear seat. This study examines the practice of rear-facing restraints pre- and post-AAP recommendations for children under 2 years. Methods Data from the Fatality Analysis Reporting System (FARS) from 2008 to 2015 were used to examine restraint status and injuries in rear-seated infants and toddlers aged 0 to less than 2 years involved in fatal collisions ( n = 4966). Subpopulation analyses were conducted on 1557 children with seat facing direction recorded. Multivariable logistic regression was used to generate odds ratios (OR) with 95% confidence intervals (CI). Covariates considered for inclusion in the multivariable model included passenger characteristics (age, gender, seating position), driver characteristics (age, gender, seat belt status, alcohol status, drug status, previous traffic violations), vehicle characteristics (vehicle type), and crash-level characteristics (day/night, weekday/weekend, rush hour, expressway/surface street, urban/rural). Results Approximately 6.7% (330 of 4996) of infants and toddlers were unrestrained with mortality that was approximately triple that of restrained infants (40.0% vs 13.7%, P < 0.0001). In multivariable adjusted models, predictors of an infant being unrestrained included unrestrained driver (OR: 3.17, 95% CI: 2.38–4.21), driver aged less than 20 years (OR: 2.18, 95% CI: 1.42–3.34), driver alcohol use (OR: 2.21, 95% CI: 1.42–3.44), center-seated infant (OR: 1.55, 95% CI: 1.19–2.03) and weekday crash (OR: 1.52, 95% CI: 1.12–2.01). Of all rear-seated children whose restraint status were reported (4966), rear-facing restraint use increased from 5.0% to 23.2% between 2008 and 2015 ( P < 0.0001). The odds of rear-facing restraint use increased after introduction of the AAP guideline among infants aged 0 to < 1 year old (OR: 2.12, 95% CI: 1.46–3.10) and among toddlers aged 1 to < 2 years old (OR: 1.97, 95% CI: 1.03–3.79). Conclusion Trends in the use of rear-facing child restraints improved over the timeframe of this study, but remain low despite the introduction of AAP guidelines and the strengthening of child restraint laws.

PURPOSE:The purpose of the study was to evaluate the clinical features, treatment, and outcome of patients with conjunctival papilloma.MATERIALS AND METHODS:Twenty-two patients (22 eyes) with biopsy-proven conjunctival papilloma between January 2005 and January 2015 in a tertiary medical center were retrospectively reviewed. Clinical profiles, treatment, outcome, and factors related to recurrence were evaluated.RESULTS:There were 16 males (73%) and 6 females (27%), with a mean age of 47 years. The most common location of conjunctival papilloma was the caruncle (43%), followed by palpebral conjunctiva (29%), bulbar conjunctiva (14%), and fornix (14%). Recurrence developed in five patients (22.7%). The risk of postoperative recurrence was significantly related to the presence of bulbar conjunctival papilloma with corneal involvement (P = 0.043) and surgical excision alone (P = 0.039). One case with multiple recurrences developed nonkeratinizing carcinoma. Two young females developed conjunctival papilloma even after receiving human papillomavirus vaccinations.CONCLUSION:The recurrence of conjunctival papilloma is not uncommon, especially for those patients underwent surgical excision alone. Surgical excision with adjunctive therapy and long-term follow-up is rational for the treatment of conjunctival papilloma.