Comparison of Clinical and Radiological Outcomes after Vertebroplasty and Balloon Kyphoplasty in the Treatment of Osteoporotic Vertebral Compression Fractures.

BACKGROUND: Vertebroplasty (VP) and balloon kyphoplasty (KP) are effective means with which to improve pain and function in osteoporotic vertebral compression fractures. However, the risk of complications after these procedures is poorly understood, with concerns regarding adjacent vertebral fractures. This study retrospectively investigated the clinical and radiological outcomes of these procedures. METHODS: A total of 115 patients who experienced their first vertebral fracture were treated with VP (N=63) or KP (N=52) at the Dankook University Hospital between January 2013 and December 2022. The clinical outcomes were evaluated using the visual analog scale (VAS) preoperative and at 1-year follow-up. Radiological comparisons were performed for kyphosis correction, vertebral height restoration, and postoperative cement leakage. RESULTS: KP was more effective than VP, especially for vertebral body height restoration and kyphotic angle reduction (P<0.05). However, the incidence of cement leakage, new adjacent vertebral fractures, and improvement in pain assessed by VAS did not differ statistically between the 2 groups (P>0.05). CONCLUSIONS: Considering that KP was performed on fractures with severe deformity, no differences were observed in the clinical outcomes and incidence of adjacent vertebral fractures compared Considering that KP was performed for fractures with severe deformity, there was no difference in clinical outcomes and incidence of adjacent vertebral fractures compared to VP. Improvements in radiological measurements were demonstrated. Therefore, KP may be a good treatment option for pain relief and long-term prognosis in patients with high-compressive-rate vertebral fractures.

Antibiofilm and antithrombotic hydrogel coating based on superhydrophilic zwitterionic carboxymethyl chitosan for blood-contacting devices.

Blood-contacting devices must be designed to minimize the risk of bloodstream-associated infections, thrombosis, and intimal lesions caused by surface friction. However, achieving effective prevention of both bloodstream-associated infections and thrombosis poses a challenge due to the conflicting nature of antibacterial and antithrombotic activities, specifically regarding electrostatic interactions. This study introduced a novel biocompatible hydrogel of sodium alginate and zwitterionic carboxymethyl chitosan (ZW@CMC) with antibacterial and antithrombotic activities for use in catheters. The ZW@CMC hydrogel demonstrates a superhydrophilic surface and good hygroscopic properties, which facilitate the formation of a stable hydration layer with low friction. The zwitterionic-functionalized CMC incorporates an additional negative sulfone group and increased negative charge density in the carboxyl group. This augmentation enhances electrostatic repulsion and facilitates the formation of hydration layer. This leads to exceptional prevention of blood clotting factor adhesion and inhibition of biofilm formation. Subsequently, the ZW@CMC hydrogel exhibited biocompatibility with tests of in vitro cytotoxicity, hemolysis, and catheter friction. Furthermore, in vivo tests of antithrombotic and systemic inflammation models with catheterization indicated that ZW@CMC has significant advantages for practical applications in cardiovascular-related and sepsis treatment. This study opens a new avenue for the development of chitosan-based multifunctional hydrogel for applications in blood-contacting devices.

Antimicrobial and Antifouling Effects of Petal-Like Nanostructure by Evaporation-Induced Self-Assembly for Personal Protective Equipment.

Although the personal protective equipment (PPE) used by healthcare workers (HCWs) effectively blocks hazardous substances and pathogens, it does not fully rule out the possibility of infection, as pathogens surviving on the fabric surface pose a substantial risk of cross-infection through unintended means. Therefore, PPE materials that exhibit effective biocidal activity while minimizing contamination by viscous body fluids (e.g., blood and saliva) and pathogen-laden droplets are highly sought. In this study, petal-like nanostructures (PNSs) are synthesized through the vertical rearrangement of colloidal lamellar bilayers via evaporation-induced self-assembly of octadecylamine, silica-alumina sol, and diverse photosensitizer. The developed method is compatible with various fabrics and imparts visible-light-activated antimicrobial and superhydrophobic-based antifouling activities. PNS-coated fabrics could provide a high level of protection and effectively block pathogen transmission as exemplified by their ability to roll off viscous body fluids reducing bacterial droplet adhesion and to inactivate various microorganisms. The combination of antifouling and photobiocidal activities results in the complete inactivation of sprayed pathogen-laden droplets within 30 min. Thus, this study paves the way for effective contagious disease management and the protection of HCWs in general medical environments, inspiring further research on the fabrication of materials that integrate multiple useful functionalities.

Simultaneous Screening of 322 Residual Pesticides in Fruits and Vegetables Using GC-MS/MS and Deterministic Health Risk Assessments.

The development of efficient methods for evaluating pesticide residues is essential in order to ensure the safety and quality of agricultural products since the Republic of Korea implemented the Positive List System (PLS). The objective of this research was to establish a method for the simultaneous analysis of 322 pesticide residues in fruits and vegetables (such as coffee, potato, corn, and chili pepper), using the quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach in combination with gas chromatography-tandem mass spectrometry (GC-MS/MS). This study introduces a robust, high-throughput GC-MS/MS method for screening the target pesticide residues in agricultural products, achieving the PLS criterion of 0.01 mg/kg LOQ. Despite some compounds not aligning with the CODEX recovery guideline, sufficient reproducibility was confirmed, attesting to the method's applicability in qualitative analyses. A health risk assessment conducted using estimated daily intake/acceptable daily intake ratios indicated low risks associated with product consumption (<0.035391%), thereby confirming their safety. This efficient method holds significant implications for the safe distribution of agricultural products, including during import inspections.

Basic amino acid-mediated cationic amphiphilic surfaces for antimicrobial pH monitoring sensor with wound healing effects.

BACKGROUND: The wound healing process is a complex cascade of physiological events, which are vulnerable to both our body status and external factors and whose impairment could lead to chronic wounds or wound healing impediments. Conventional wound healing materials are widely used in clinical management, however, they do not usually prevent wounds from being infected by bacteria or viruses. Therefore, simultaneous wound status monitoring and prevention of microbial infection are required to promote healing in clinical wound management. METHODS: Basic amino acid-modified surfaces were fabricated in a water-based process via a peptide coupling reaction. Specimens were analyzed and characterized by X-ray photoelectron spectroscopy, Kelvin probe force microscopy, atomic force microscopy, contact angle, and molecular electrostatic potential via Gaussian 09. Antimicrobial and biofilm inhibition tests were conducted on Escherichia coli and Staphylococcus epidermidis. Biocompatibility was determined through cytotoxicity tests on human epithelial keratinocytes and human dermal fibroblasts. Wound healing efficacy was confirmed by mouse wound healing and cell staining tests. Workability of the pH sensor on basic amino acid-modified surfaces was evaluated on normal human skin and Staphylococcus epidermidis suspension, and in vivo conditions. RESULTS: Basic amino acids (lysine and arginine) have pH-dependent zwitterionic functional groups. The basic amino acid-modified surfaces had antifouling and antimicrobial properties similar to those of cationic antimicrobial peptides because zwitterionic functional groups have intrinsic cationic amphiphilic characteristics. Compared with untreated polyimide and modified anionic acid (leucine), basic amino acid-modified polyimide surfaces displayed excellent bactericidal, antifouling (reduction ~ 99.6%) and biofilm inhibition performance. The basic amino acid-modified polyimide surfaces also exhibited wound healing efficacy and excellent biocompatibility, confirmed by cytotoxicity and ICR mouse wound healing tests. The basic amino acid-modified surface-based pH monitoring sensor was workable (sensitivity 20 mV pH-1) under various pH and bacterial contamination conditions. CONCLUSION: Here, we developed a biocompatible and pH-monitorable wound healing dressing with antimicrobial activity via basic amino acid-mediated surface modification, creating cationic amphiphilic surfaces. Basic amino acid-modified polyimide is promising for monitoring wounds, protecting them from microbial infection, and promoting their healing. Our findings are expected to contribute to wound management and could be expanded to various wearable healthcare devices for clinical, biomedical, and healthcare applications.

Transparent, Robust, and Photochemical Antibacterial Surface Based on Hydrogen Bonding between a Si-Al and Cationic Dye.

Healthcare-associated infections can occur and spread through direct contact with contaminated fomites in a hospital, such as mobile phones, tablets, computer keyboards, doorknobs, and other surfaces. Herein, this study shows a transparent, robust, and visible light-activated antibacterial surface based on hydrogen bonds between a transparent silica-alumina (Si-Al) sol-gel and a visible light-activated photosensitizer, such as crystal violet (CV). The study of the bonding mechanisms revealed that hydrogen bonding predominantly occurs between the N of CV and Al-OH. Apart from CV, Si-Al can be combined with a variety of dyes, highlighting its potential for wide application. The Si-Al@CV film selectively generates singlet oxygen using ambient visible light, triggering potent photochemical antibacterial performance against Gram-positive and Gram-negative bacteria. Additionally, the Si-Al@CV film is stable even after mechanical stability tests such as tape adhesion, scratch, bending, and water immersion. In vitro cytotoxicity tests using C2C12 myoblast cells showed that the Si-Al@CV film is a biocompatible material. This work suggests a new approach for designing a transparent and robust touchscreen surface with photochemical antibacterial capability against healthcare-associated infections.

Pathological Evaluation of Porcine Circovirus 2d (PCV2d) Strain and Comparative Evaluation of PCV2d and PCV2b Inactivated Vaccines against PCV2d Infection in a Specific Pathogen-Free (SPF) Yucatan Miniature Pig Model.

Porcine circovirus type 2 (PCV2) is an economically important swine pathogen that causes porcine circovirus-associated diseases (PCVADs). The objective of this study was to evaluate the use of specific pathogen-free Yucatan miniature pigs (YMPs) as an experimental model for PCV2d challenge and vaccine assessment because PCV2-negative pigs are extremely rare in conventional swine herds in Korea. In the first experiment, every three pigs were subjected to PCV2d field isolate or mock challenge. During three weeks of experiments, the PCV2d infection group exhibited clinical outcomes of PCVAD with high viral loads, lymphoid depletion, and detection of PCV2d antigens in lymphoid organs by immunohistochemistry. In the second experiment, three groups of pigs were challenged with PCV2d after immunization for three weeks: a nonvaccinated group (three pigs), a PCV2b-Vac group vaccinated with a commercial PCV2b-based inactivated vaccine SuiShot® Circo-ONE (five pigs), and a PCV2d-Vac group vaccinated with an experimental PCV2d-based inactivated vaccine (five pigs). During the three weeks of the challenge period, nonvaccinated pigs showed similar clinical outcomes to those observed in the PCV2d infection group from the first experiment. In contrast, both the PCV2b and PCV2d vaccinations produced good levels of protection against PCV2d challenge, as evidenced by reduced viral loads, improved growth performance, high virus-neutralizing antibody titers, and less development of PCV2-associated pathological lesions. Taken together, these data suggest that YMPs could be an alternative model for PCV2 challenge experiments, and these animals displayed typical clinical and pathological features and characteristics of protective immunity induced by the vaccines that were consistent with those resulting from PCV2 infections in conventional pigs.

Photobiocidal-triboelectric nanolayer coating of photosensitizer/silica-alumina for reusable and visible-light-driven antibacterial/antiviral air filters.

Outbreaks of airborne pathogens pose a major threat to public health. Here we present a single-step nanocoating process to endow commercial face mask filters with photobiocidal activity, triboelectric filtration capability, and washability. These functions were successfully achieved with a composite nanolayer of silica-alumina (Si-Al) sol-gel, crystal violet (CV) photosensitizer, and hydrophobic electronegative molecules of 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (PFOTES). The transparent Si-Al matrix strongly immobilized the photosensitizer molecules while dispersing them spatially, thus suppressing self-quenching. During nanolayer formation, PFOTES was anisotropically rearranged on the Si-Al matrix, promoting moisture resistance and triboelectric charging of the Si-Al/PFOTES-CV (SAPC)-coated filter. The SAPC nanolayer stabilized the photoexcited state of the photosensitizer and promoted redox reaction. Compared to pure-photosensitizer-coated filters, the SAPC filter showed substantially higher photobiocidal efficiency (∼99.99 % for bacteria and a virus) and photodurability (∼83 % reduction in bactericidal efficiency for the pure-photosensitizer filter but ∼0.34 % for the SAPC filter after 72 h of light irradiation). Moreover, after five washes with detergent, the SAPC filter maintained its photobiocidal and filtration performance, proving its reusability potential. Therefore, this SAPC nanolayer coating provides a practical strategy for manufacturing an antimicrobial and reusable mask filter for use during the ongoing COVID-19 pandemic.

Comparison of the pathogenicity of porcine reproductive and respiratory syndrome virus (PRRSV)-1 and PRRSV-2 in pregnant sows.

To date, few studies related to the evaluation of the pathogenicity of different PRRSV isolates using a reproductive model have been undertaken, and the main focus has remained on respiratory models using young pigs. This study aimed to evaluate the pathogenicity of two PRRSV-1 isolates (D40 and CBNU0495) and two PRRSV-2 isolates (K07-2273 and K08-1054) in a reproductive model. Pregnant sows were experimentally infected with PRRSV at gestational day 93 or used as an uninfected negative control. Sera were collected at 0, 3, 7, 14, and 19 days post-challenge (dpc) for virological and serological assays. At 19 dpc, all sows were euthanized, and their fetuses were recovered by performing cesarean section and immediately euthanized for sample collection. Here, compared to the other isolates, the CBNU0495 isolate replicated most efficiently in the pregnant sows, and K07-2273 produced the highest rate of reproductive failure even though it did not replicate as efficiently as the other isolates in sows and fetuses, indicating that vertical transmission and reproductive failure due to PRRSV infection do not have any significant correlation with the viral loads in samples from sows and fetuses. Similarly, the viral loads and the histopathological lesions did not show any correlation with each other, as the PRRSV-2-infected groups displayed more prominent and frequent histopathological lesions with lower viral loads than the PRRSV-1-infected groups. However, viral loads in the myometrium/endometrium might be related to the spreading of PRRSV in the fetuses, which affected the birth weight of live fetuses. This study contributes to a better understanding of the pathogenicity of the most prevalent Korean PRRSVs in a reproductive model.

Amino acid-mediated negatively charged surface improve antifouling and tribological characteristics for medical applications.

To prevent infections associated with biomedical catheters, various antimicrobial coatings have been investigated. However, those materials do not provide consistent antibacterial effects or biocompatibility, generally, due to degradation of the coating materials, in vivo. Additionally, biomedical catheters must have low surface friction to reduce tribological damage. In this study, we developed an antifouling surface composed of biocompatible amino acids (leucine, taurine, and aspartic acid) on polyimide, via modification using a series of facile immersion steps with waterborne reactions. The naturally derived amino acid could be formed highly biostable amide bonds on the polyimide surface like peptides. The amino acid-modified surface formed a water layer with antifouling performance through the hydrophilic properties of amino acids. Amino acid-mediated modification reduced adhesion up to 84.45% and 94.81% against Escherichia coli and Staphylococcus epidermidis, respectively, and exhibited an excellent prevention to adhesion against the proteins, albumin and fibrinogen. Evaluation of the surface friction of the catheter revealed a dramatic reduction in the tribological force after amino acid modification on polyimide that of 0.81 N to aspartic acid of 0.44 N. These results clearly demonstrate a reduced occurrence of infections, thrombi and tribological damage following the relatively facile surface modification of catheters. The proposed modification method can be used in a continuous manufacturing process via using the same time of modification steps for the easy producing the product. Moreover, the method uses biocompatible naturally derived materials and can be applied to medical equipment that requires biocompatibility and biofunctionality with polyimide surfaces.

Evaluation of the Cross-Protective Efficacy of a Chimeric PRRSV Vaccine against Two Genetically Diverse PRRSV2 Field Strains in a Reproductive Model.

Despite the routine use of porcine reproductive and respiratory syndrome (PRRS)-modified live vaccines, serious concerns are currently being raised due to their quick reversion to virulence and limited cross-protection against divergent PRRS virus (PRRSV) strains circulating in the field. Therefore, a PRRS chimeric vaccine (JB1) was produced using a DNA-launched infectious clone by replacing open reading frames (ORFs) 3-6 with those from a mixture of two genetically different PRRSV2 strains (K07-2273 and K08-1054) and ORF1a with that from a mutation-resistant PRRSV strain (RVRp22) exhibiting an attenuated phenotype. To evaluate the safety and cross-protective efficacy of JB1 in a reproductive model, eight PRRS-negative pregnant sows were purchased and divided into four groups. Four sows in two of the groups were vaccinated with JB1, and the other 4 sows were untreated at gestational day 60. At gestational day 93, one vaccinated group and one nonvaccinated group each were challenged with either K07-2273 or K08-1054. All of the sows aborted or delivered until gestation day 115 (24 days post challenge), and the newborn piglets were observed up to the 28th day after birth, which was the end of the experiment. Overall, pregnant sows of the JB1-vaccinated groups showed no meaningful viremia after vaccination and significant reductions in viremia with K07-2273 and K08-1054, exhibiting significantly higher levels of serum virus-neutralizing antibodies than non-vaccinated sows. Moreover, the JB1-vaccinated groups did not exhibit any abortion due to vaccination and showed improved piglet viability and birth weight. The piglets from JB1-vaccinated sows displayed lower viral concentrations in serum and fewer lung lesions compared with those of the piglets from the nonvaccinated sows. Therefore, JB1 is a safe and effective vaccine candidate that confers simultaneous protection against two genetically different PRRSV strains.

Comparing the stability of retinol in liposomes with cholesterol, ß-sitosterol, and stigmasterol.

In this study, cholesterol (CH), ß-sitosterol (SI), and stigmasterol (ST) were explored to improve the stability of retinol in the liposome bilayer. Retinol was incorporated into liposomes composed of soybean-derived L-α-phosphatidylcholine (PC) and 10% sterol (w/w), which were prepared as multilamellar vesicles. Under all conditions, the efficiency of retinol incorporation into liposomes was higher than 99%, and the average particle size of liposomes was similar to that of PC alone. Liposomes were stored at 4 and 25 °C, with and without light, respectively, for 10 days. It was found that during the storage, CH and SI were effective in stabilizing the retinol in liposomes. These results indicate that an appropriate phytosterol could improve the stability of retinol in liposomes.

Defect suppression and photoresponsivity enhancement in methylammonium lead halide perovskites by CdSe/ZnS quantum dots.

Potential strategies such as surface passivation and perovskite material halide mixing may protect material surfaces, improve luminescence, and reduce charge traps for device stability. In this study, we used deep level transient spectroscopy to investigate the effect of CdSe/ZnS core-shell quantum dots (QDs) on defect states and carrier transport in methylammonium (MA) lead halide perovskites (CH3NH3PbX3 where X  = I, Br). In MAPbI3 and MAPbI2Br films with CdSe/ZnS QDs, the density of hole traps located at Ev + 0.37 eV and Ev + 0.56 eV was reduced dramatically. Deep traps at Ev + 0.78 eV and Ev + 1.08 eV were removed, and one broad electron trap signal dominated. Film photoresponsivity under 600-nm wavelength light and a bias voltage of -0.7 V was 10 and 18 mA/W, which is 100 and 27 times larger than the 0.1 and 0.67 mA/W of bare perovskites (PS), respectively. This demonstrates that carrier transport was enhanced due to defect suppression. Our findings on defect suppression and photoresponsivity enhancement provide an important direction for optimizing high-performance PS device fabrication.

Synergetic enhancement in the reactivity and stability of surface-oxide-free fine Al particles covered with a polytetrafluoroethylene nanolayer.

Surface oxide (Al2O3) of reactive fine aluminum (Al) particles for solid fuels, propellants, and brazing materials often restricted oxidative performance, though the passivation film acts to protect Al particles from exploding. Here, we report fine Al particles fully covered with a polytetrafluoroethylene (PTFE) layer instead of an Al2O3 film on the surface. This advance is based on the introduction of strong Al-F bonds, known to be an alternative to the Al-O bonds of surface oxides. The DSC results on the PTFE-coated Al particles exhibit higher reactive-exothermic enthalpy energy (12.26 kJ g-1) than 4.85 kJ g-1 by uncoated Al particles. The artificial aging test of the PTFE layer on the Al particles show long-time stability to the external circumstance compared to those by Al2O3. The activation energy for oxidation was investigated from cyclic voltammetry assessment and the measured peak potentials of the anode curve for PTFE/Al (- 0.45 V) and uncoated Al (- 0.39 V) are achieved, respectively. This means that the PTFE layer is more stable against a sudden explosion of Al particles compared to Al2O3. These results are very useful given its capability to control both the reactivity and stability levels during the oxidation of Al particles for practical applications.

The enhancement mechanism of photo-response depending on oxygen pressure for Ga2O3 photo detectors.

We have optimized the responsivity and response speed of a ß-Ga2O3-based photodetector. The ß-Ga2O3 thin films were deposited on a glass substrate under various oxygen partial pressures from 0 to 50 mTorr using pulsed laser deposition. Time-response measurements show that the as-grown ß-Ga2O3 at an oxygen partial pressure of 50 mTorr has the fastest response speed and decay times of 33 and 100 ms, which are better than those prepared at lower oxygen pressures. This sample also showed a high photoresponsivity of 5 A W-1 and detectivity of 1012 cmHz1/2/W. The high performance of the ß-Ga2O3 detector grown at the oxygen partial pressure of 50 mTorr might be due to the reduction of oxygen vacancies caused by the increase in oxygen content during deposition. The results reveal the importance of the oxygen processing gas in promoting photodetector performance.

Generation and protective efficacy of a cold-adapted attenuated genotype 2b porcine epidemic diarrhea virus.

The recent emergence and re-emergence of porcine epidemic diarrhea virus (PEDV) underscore the urgent need for the development of novel, safe, and effective vaccines against the prevailing strain. In this study, we generated a cold-adapted live attenuated vaccine candidate (Aram-P29-CA) by short-term passage of a virulent PEDV isolate at successively lower temperatures in Vero cells. Whole genome sequencing identified 12 amino acid changes in the cold-adapted strain with no insertions and deletions throughout the genome. Animal inoculation experiments confirmed the attenuated phenotype of Aram-P29-CA virus in the natural host. Pregnant sows were orally administered P29-CA live vaccines two doses at 2-week intervals prior to parturition, and the newborn piglets were challenged with the parental virus. The oral homologous prime-boost vaccination of P29-CA significantly improved the survival rate of the piglets and notably mitigated the severity of diarrhea and PEDV fecal shedding after the challenge. Furthermore, strong antibody responses to PEDV were detected in the sera and colostrum of immunized sows and in the sera of their offspring. These results demonstrated that the cold-adapted attenuated virus can be used as a live vaccine in maternal vaccination strategies to provide durable lactogenic immunity and confer passive protection to litters against PEDV.

Long-Term Chemical Aging of Hybrid Halide Perovskites.

Because the power conversion efficiency (PCE) of hybrid halide perovskite solar cells (PSCs) could exceed 24%, extensive research has been focused on improving their long-term stability for commercialization in the near future. In a previous study, we reported that the addition of a number of ionized iodide (triiodide: I3-) ions during perovskite film formation significantly improved the efficiency of PSCs by reducing deep-level defects in the perovskite layer. Understanding the relationship between the concentration of these defects and the long-term chemical aging of PSCs is important not only for obtaining fundamental insight into the perovskite materials but also for studying the long-term chemical stability of PSCs. Herein we aim to identify the origin of the natural decay in PCE during long-term chemical aging of PSCs in the dark based on formamidinium lead triiodide by comparing the performance of control and low-defect (LD) devices. After aging for 200 days, the change in the PCE of the LD devices (1.3%) was found to be half that of the control devices (2.6%). We investigated this difference using grazing incidence wide-angle X-ray scattering, deep-level transient spectroscopy, scanning photoelectron microscopy, and high-resolution photoemission spectroscopy. The addition of I3- was found to reduce the amounts of hydroxide and Ox in the halide perovskites (HPs), affecting the migration of defects and the structural transformation of the HPs.

Molecular characteristics and pathogenic assessment of porcine epidemic diarrhoea virus isolates from the 2018 endemic outbreaks on Jeju Island, South Korea.

Since the 2013-2014 incursion of the virulent G2b porcine epidemic diarrhoea virus (PEDV) pandemic strains in South Korea, frequent moderate-scale regional outbreaks have recurred. In particular, areas of Jeju Island with extensive swine production have faced repeated epidemics since the re-emergence in 2014. The current study reports the complete genome sequences and molecular characterization of the representative PEDV strains responsible for the 2018 endemic outbreaks on Jeju Island. All isolates were determined to belong genetically to the highly pathogenic pandemic G2b group. Full-length genome sizes of four isolates differed from that of the G2b epidemic field strain due to insertion or deletion (DEL) mutations in the non-structural protein (nsp)- or spike (S) protein-coding regions. The 2018 Jeju isolates shared 96.7%-98.7% and 98.5%-99.4% identity at the S gene and whole-genome levels, respectively, compared to global G2b PEDV strains. Genetic and phylogenetic analyses indicated that the 2018 isolates were closest to the 2014 G2b re-emergent Jeju strains, but appeared to have undergone substantial rapid independent evolution. Among the isolates, a notable nsp3 DEL variant strain, KOR/KNU-1807/2018, was isolated and propagated by continuous passages in Vero cells, and displayed typical PEDV-induced syncytia formation. Genomic sequencing identified a unique 8-nt DEL in the extreme C-terminal region of the S gene at the 4th passage (KNU-1807-P4) compared to its original sample. This DEL resulted in the premature termination of S by nine amino acid residues (EVFEKVHVQ), which contained a KxHxx motif that is a potential endoplasmic reticulum retrieval signal. In vivo animal studies showed that variant strain KNU-1807 had decreased virulence in suckling piglets. These results advance our knowledge regarding the genetic variation and pathogenicity of the G2b PEDV endemic strains prevalent in Jeju swine herds in South Korea.

Assessment of the safety and efficacy of an attenuated live vaccine based on highly virulent genotype 2b porcine epidemic diarrhea virus in nursing piglets.

We have previously reported the generation of the attenuated KNU-141112-S DEL5/ORF3 virus by continuous propagation of highly virulent G2b porcine epidemic diarrhea virus (PEDV) in Vero cells. The present study aimed to assess the safety of S DEL5/ORF3 and to evaluate its effectiveness as a live vaccine for prime-booster vaccinations. Reversion to virulence experiments revealed that the S DEL5/ORF3 strain retains its attenuated phenotype and genetic stability after five successive passages in susceptible piglets. Pregnant sows were primed orally with an S DEL5/ORF3 live vaccine and boosted intramuscularly twice with a commercial killed vaccine at 2-week intervals prior to parturition. This sow vaccination regimen completely protected nursing piglets against virulent G2b challenge, as evidenced by the increase in survival rate from 0% to 100% and the significant reduction in diarrhea intensity, including the amount and duration of PEDV fecal shedding. In addition, despite a 2-3 day period of weight loss in piglets from vaccinated sows after challenge, their daily weight gain was recovered at 7 days post-challenge and became similar to that of unchallenged pigs from unvaccinated sows over the course of the experiment. Furthermore, strong antibody responses to PEDV were verified in the sera and colostrum of immunized sows with the prime-boost treatment and their offspring. Altogether, our data demonstrated that the attenuated S DEL5/ORF3 strain guarantees the safety to host animals with no reversion to virulence and is suitable as an effective primary live vaccine providing durable maternal lactogenic immunity for passive piglet protection.

Differential evolution of antigenic regions of porcine reproductive and respiratory syndrome virus 1 before and after vaccine introduction.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a widespread viral pathogen that has caused tremendous economic losses throughout most pig-producing countries. Nowadays, both PRRSV-1 and PRRSV-2 co-circulate in Korean pig populations, and commercial modified live vaccine (MLV) is predominantly used to control PRRS. Specifically, control strategy using only PRRSV-2 MLV that was used since 1995 cannot prevent the spread of PRRSV-1 and damage from its infection, which led to the first introduction of two additional PRRSV-1 vaccines in 2014. Despite the wide implementation with PRRSV-1 vaccines, there is a lack of knowledge about the currently circulating Korean PRRSV-1 strains. Whole structural genes of PRRSV-1 before (11) and after (17) the introduction of vaccine were compared to determine the genetic evolutionary features of PRRSV. Genetic and phylogenetic analysis indicated that Korean PRRSV-1 shared 91.5 ± 1.7% nucleotide identity but formed a unique clade based on ORF2-7 phylogeny. Bioinformatics showed increased genetic heterogeneity, enhanced diversifying selection, and the emergence of novel glycosylation sites within neutralizing epitopes of minor structural proteins after vaccine introduction. Taken together, our data provide novel insight into the evolution of minor structural proteins of PRRSV-1 in the vaccination era.