Klotho is a protein that plays different functions in female fertility. We have previously reported that klotho protein supplementation during in vitro maturation improves porcine embryo development, while klotho knockout for somatic cell cloning completely blocks full-term pregnancy in vivo. However, the effects of the microinjection of klotho protein or klotho knockdown dual vector in porcine embryos at different time points and the specific molecular mechanisms remain largely unknown. In this study, we injected the preassembled cas9 + sgRNA dual vector, for klotho knockdown, into the cytoplasm of the germinal vesicle stage of oocytes and into porcine embryos after 6-h parthenogenetic activation. Similarly, the klotho protein was inserted into the cytoplasm of germinal vesicle stage oocytes and porcine embryos after 6-h parthenogenetic activation. Compared with the controls, the microinjection of klotho dual vector markedly decreased the blastocyst formation rates in germinal vesicle stage oocytes and activated embryos. However, the efficiency of blastocyst formation when klotho protein was inserted before in vitro maturation was significantly higher than that after klotho protein insertion into parthenogenetically activated embryos. These results indicated that klotho knockdown may impair embryo development into blastocyst irrespective of injection timing. In addition, klotho protein injection timing in pig embryos may be an important factor for regulating embryo development.

Birds in flight are prone to collide with various transparent or reflective structures. While bird–window collision has been recognised as a critical conservation issue, collision with other transparent structures has been less understood. Noise barriers made of transparent materials are considered critical hazards for birds; however, little is known about the bird mortality they cause. We conducted the first nationwide-scale estimates of bird-collision mortality caused by transparent noise barriers (TNBs) along roads in the Republic of Korea. The total length of existing roadside transparent noise barriers was estimated at 1,416 km nationwide (as of 2018), and it had been increasing exponentially. Based on carcass surveys at 25 sites, daily mortality at the observed barriers was 0.335 ± 1.132 birds/km on average, and no difference in observed mortality was detected between both sides of a single barrier and between road types (i.e. local roads and motorways). Finally, we estimated that approximately 186,000 birds (95% confidence interval: 162,465–204,812 birds) are killed annually by collisions with roadside TNBs. As privately installed barriers were not considered in this study, the actual mortality is likely be higher than our estimates. Thus, collision with TNBs could become an emerging threat to avian conservation, especially in developing and urbanising regions around the world. As such structures are not formally recognised as conservation issues of importance, more systematic surveys aided by citizen science, both for the status of TNBs and bird-collision mortality, are needed in addition to management and mitigation policies.

Nosocomial transmission of COVID-19 among immunocompromised hosts can have a serious impact on COVID-19 severity, underlying disease progression and SARS-CoV-2 transmission to other patients and healthcare workers within hospitals. We experienced a nosocomial outbreak of COVID-19 in the setting of a daycare unit for paediatric and young adult cancer patients. Between 9 and 18 November 2020, 473 individuals (181 patients, 247 caregivers/siblings and 45 staff members) were exposed to the index case, who was a nursing staff. Among them, three patients and four caregivers were infected. Two 5-year-old cancer patients with COVID-19 were not severely ill, but a 25-year-old cancer patient showed prolonged shedding of SARS-CoV-2 RNA for at least 12 weeks, which probably infected his mother at home approximately 7–8 weeks after the initial diagnosis. Except for this case, no secondary transmission was observed from the confirmed cases in either the hospital or the community. To conclude, in the day care setting of immunocompromised children and young adults, the rate of in-hospital transmission of SARS-CoV-2 was 1.6% when applying the stringent policy of infection prevention and control, including universal mask application and rapid and extensive contact investigation. Severely immunocompromised children/young adults with COVID-19 would have to be carefully managed after the mandatory isolation period while keeping the possibility of prolonged shedding of live virus in mind.

Two aphid-transmitted RNA viruses, broad bean wilt virus 2 (BBWV2) and cucumber mosaic virus (CMV), are the most prevalent viruses in Korean pepper fields and cause chronic damage in pepper production. In this study, we employed a screening system for pathotype-specific resistance of pepper germplasm to BBWV2 and CMV by utilizing infectious cDNA clones of different pathotypes of the viruses (two BBWV2 strains and three CMV strains). We first examined pathogenic characteristics of the BBWV2 and CMV strains in various plant species and their phylogenetic positions in the virus population structures. We then screened 34 commercial pepper cultivars and seven accessions for resistance. While 21 pepper cultivars were resistant to CMV Fny strain, only two cultivars were resistant to CMV P1 strain. We also found only one cultivar partially resistant to BBWV2 RP1 strain. However, all tested commercial pepper cultivars were susceptible to the resistance-breaking CMV strain GTN (CMV-GTN) and BBWV2 severe strain PAP1 (BBWV2-PAP1), suggesting that breeding new cultivars resistant to these virus strains is necessary. Fortunately, we identified several pepper accessions that were resistant or partially resistant to CMV-GTN and one symptomless accession despite systemic infection with BBWV2-PAP1. These genetic resources will be useful in pepper breeding programs to deploy resistance to BBWV2 and CMV.

Background: Mumps, a contagious disease, is transmissible by respiratory droplet particles and is preventable by vaccination. In South Korea, mandatory vaccination against mumps has markedly reduced its incidence. However, both the incidence and the number of reported cases of mumps have persistently increased in South Korea since 2007. Despite high vaccination rates, mumps outbreaks continue to occur, and many studies have been conducted on mumps seroprevalence in children and adolescents. In comparison, few reports have been published regarding mumps seroprevalence in healthcare workers (HCWs) in South Korea. Objective: We investigated the seroprevalence of HCWs in South Korea. Methods: This study was conducted at Asan Medical Center, a 2,705-bed tertiary-care hospital in Seoul, South Korea, with 8,329 HCWs. In 2018, we performed mumps antibody testing for HCWs. We administered MMR vaccination to all HCWs whose antibody test yielded equivocal or negative results. However, we did not repeat mumps antibody testing after MMR vaccination. Results: In total, 6,055 HCWs (73%) underwent mumps antibody testing. The overall mumps seropositivity rate was 87% (95% CI, 86%–87%). Seropositivity rates of all birth cohorts ranged from 72% to 92%. Mumps seropositivity rates were 88% in HCWs born before 1970, 87% in those born between 1970 and 1989, and 88% in those born between 1990 and 1995 (P = .59). Mumps seropositivity rates for both women and men HCWs were 87% (3,770 of 4,311 women and 1,517 of 1,744 men); the difference was not statistically significant (P = .62). The overall mumps seropositivity rate was 87%, which was above the herd immunity threshold of 75%–86%. Conclusions: Our results revealed that the overall mumps seropositivity rate in South Korean HCWs was above the herd immunity threshold. On the basis of this finding, we recommend that MMR vaccination after serologic testing may be a more reasonable approach than universal MMR vaccination alone in Korea.

Funding: None

Disclosures: None

Background: Measles is a highly contagious disease that is transmissible by airborne particles but is preventable by vaccination. South Korea has maintained a highly immunized adult population; however, small local outbreaks of measles continued to occur, and there have been some reports of pockets of underimmunity among the young adult population. It is important to know the seroepidemiology of healthcare workers (HCWs) for policy-making process, but data on the seroprevalence of measles in HCWs in South Korea are limited. Methods: We investigated the seroprevalence of HCWs at Asan Medical Center, a 2,705-bed tertiary-care hospital in Seoul, South Korea, with 8,329 HCWs. In 2014, after an outbreak of measles occurred in a university in Seoul, Asan Medical Center required measles IgG tests for all HCWs born in and after 1967 for point-prevalence surveillance. In addition, we have routinely performed measles antibody test for new HCWs since 2014. In 2018, antibody tests were administered to HCWs who were born before 1967 or who had taken a leave of absence in 2014. We provided MMR vaccination to all HCWs whose antibody tests yielded negative results. Results: In total, 7,411 HCWs (89%) underwent measles antibody tests from 2014 to 2018. The overall seropositivity was 73% (95% CI, 72%–74%); seroprevalence was 73% in HCWs born in of after 1967, whereas the seroprevalence in HCWs born before 1967 was 98%. The seroprevalence sharply decreased from 85% in the 1986 birth cohort to 42% in the 1995 birth cohort. Conclusions: In conclusion, the proportion of measles-susceptible individuals was substantially high in HCWs, especially in young adults. Because the impact of measles outbreak in healthcare facilities would be critical, a policy regarding routine serologic screening followed by measles vaccination or routine measles vaccination in healthcare facilities should be considered, especially for young Korean HCWs.

Funding: None

Disclosures: None

The experiments reported in this research paper aimed to determine the effect of supplementing different forms of L-methionine (L-Met) and acetate on protein synthesis in immortalized bovine mammary epithelial cell line (MAC-T cells). Treatments were Control, L-Met, conjugated L-Met and acetate (CMA), and non-conjugated L-Met and Acetate (NMA). Protein synthesis mechanism was determined by omics method. NMA group had the highest protein content in the media and CSN2 mRNA expression levels (P < 0.05). The number of upregulated and downregulated proteins observed were 39 and 77 in L-Met group, 62 and 80 in CMA group and 50 and 81 in NMA group from 448 proteins, respectively (P < 0.05). L-Met, NMA and CMA treatments stimulated pathways related to protein and energy metabolism (P < 0.05). Metabolomic analysis also revealed that L-Met, CMA and NMA treatments resulted in increases of several metabolites (P < 0.05). In conclusion, NMA treatment increased protein concentration and expression level of CSN2 mRNA in MAC-T cells compared to control as well as L-Met and CMA treatments through increased expression of milk protein synthesis-related genes and production of the proteins and metabolites involved in energy and protein synthesis pathways.

Due to the lack of an effective prophylactic intervention and diagnosis, human liver fluke Clonorchis sinensis continues to afflict a large human population, causing a chronic inflammatory bile duct disease. With an aim to identify target antigens for sensitive serodiagnosis, adenylate kinase 3 of C. sinensis (CsAK3) was successfully expressed in soluble form in Escherichia coli by fusion to an RNA-interacting domain derived from human Lys-tRNA synthetase and purified by Ni2+-affinity chromatography. Anti-CsAK3 serum was raised by immunization of mice, and Western blotting confirmed that CsAK3 was expressed in adult-stage C. sinensis. Histochemical analysis showed that CsAK3 was localized to the subtegumental tissue of C. sinensis and was excreted into the bile duct of the host. When tested against sera from various parasite-infected patients by enzyme-linked immunosorbent assay, the recombinant CsAK3 elicited a specific response to C. sinensis-infected sera. The results suggest that CsAK3, either alone or in combination with other antigens, could be used for improving the clinical diagnosis of clonorchiasis.

Objectives: Rotator cuff tear is the leading cause of the decline in quality of life for older adults, but comparative evidence on treatment effectiveness is lacking. This study systematically reviewed the effects of various rotator cuff tear treatments through a Bayesian meta-analysis of the related randomized clinical trials (RCTs).

Methods: We searched nine electronic databases for RCTs evaluating rotator cuff tear treatments from their inception through June 2017. A systematic review was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and the National Institute for Health and Care Excellence-Decision Support Unit guidelines (Supplementary Table 1). Outcomes included functional improvement, pain one year after surgical treatment, and tendon structural integrity. The Bayesian network meta-analysis was applied for functional improvement and pain, based on an assumption of consistency and similarity. Tendon integrity was reported descriptively.

Results: Fifteen RCTs were selected. Patients undergoing physiotherapy after open surgery showed statistically significant functional improvements compared with those undergoing physiotherapy only (mean differences, 9.1 [credible interval, 0.9–17.4]). Open surgery with physiotherapy was associated with a decrease in pain 1 year after treatment compared with when physiotherapy was combined with arthroscopic rotator cuff surgery, mini open surgery, platelet-rich plasma therapy, or physiotherapy alone (absolute value of mean difference 1.2 to 1.4). The tendon integrity results were inconsistent.

Conclusions: Some surgical treatments were associated with significant improvement in function and pain, but evidence regarding their comparative effectiveness is still lacking. A well-designed RCT discussing functional and structural treatment outcomes is needed in future.

Allicin (AL) regulates the cellular redox, proliferation, viability, and cell cycle of different cells against extracellular-derived stress. This study investigated the effects of allicin treatment on porcine oocyte maturation and developmental competence. Porcine oocytes were cultured in medium supplemented with 0 (control), 0.01, 0.1, 1, 10 or 100 μM AL, respectively, during in vitro maturation (IVM). The rate of polar body emission was higher in the 0.1 AL-treated group (74.5% ± 2.3%) than in the control (68.0% ± 2.6%) (P < 0.1). After parthenogenetic activation, the rates of cleavage and blastocyst formation were significantly higher in the 0.1 AL-treated group than in the control (P < 0.05). The reactive oxygen species level at metaphase II did not significantly differ among all groups. In matured oocytes, the expression of both BAK and CASP3, and BIRC5 was significantly lower and higher, respectively, in the 0.1 AL-treated group than in the control. Similarly, the expression of BMP15 and CCNB1, and the activity of phospho-p44/42 mitogen-activated protein kinase (MAPK), significantly increased. These results indicate that supplementation of oocyte maturation medium with allicin during IVM improves the maturation of oocytes and the subsequent developmental competence of porcine oocytes.

Echinochloa species are among the most troublesome weeds in rice cultivation, and grow in a broad habitat range in Korea. Although various ecotypes of Echinochloa have been collected as germplasm for future studies, it has been difficult to classify them due to their high level of morphological similarity. This study was thus conducted to develop and investigate the phylogenetic relationships between 77 Echinochloa accessions with the use of 23 simple sequence repeat (SSR) markers and 24 morphological traits. Of 77 Echinochloa accessions, including 57 accessions from Korea and 5 reference species, late watergrass was clearly clustered as a distinctive group from barnyardgrass and other Echinochloa species. In this analysis, we also identified core genetic and morphological markers that can be used for the future identification and classification of Echinochloa species. Five out of 23 SSR makers produced distinctive bands that discriminate late watergrass from barnyardgrass and other Echinochloa species. Four morphological traits of the reproductive organs were the most influential contributors for classifying Echinochloa species. Although there was no clear consensus generated in this study between SSR markers and morphological trait analyses, our results support the potential use of the selected SSR markers and morphological traits in future studies of Echinochloa.

Many transgenic domestic animals have been developed to produce therapeutic proteins in the mammary gland, and this approach is one of the most important methods for agricultural and biomedical applications. However, expression and secretion of a protein varies because transgenes are integrated at random sites in the genome. In addition, distal enhancers are very important for transcriptional gene regulation and tissue-specific gene expression. Development of a vector system regulated accurately in the genome is needed to improve production of therapeutic proteins. The objective of this study was to develop a knock-in system for expression of human fibroblast growth factor 2 (FGF2) in the bovine β-casein gene locus. The F2A sequence was fused to the human FGF2 gene and inserted into exon 3 of the β-casein gene. We detected expression of human FGF2 mRNA in the HC11 mouse mammary epithelial cells by RT-PCR and human FGF2 protein in the culture media using western blot analysis when the knock-in vector was introduced. We transfected the knock-in vector into bovine ear fibroblasts and produced knock-in fibroblasts using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system. Moreover, the CRISPR/Cas9 system was more efficient than conventional methods. In addition, we produced knock-in blastocysts by somatic cell nuclear transfer using the knock-in fibroblasts. Our knock-in fibroblasts may help to create cloned embryos for development of transgenic dairy cattle expressing human FGF2 protein in the mammary gland via the expression system of the bovine β-casein gene.