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Written by experts in the field, this text provides a modern introduction to three-dimensional dynamics for multibody systems. It covers rotation matrices, the twist-wrench formalism for multibody dynamics and Lagrangian dynamics, an approach that is often overlooked at the undergraduate level. The only prerequisites are differential equations and linear algebra as covered in a first-year engineering mathematics course. The text focuses on obtaining and understanding the equations of motion, featuring a rich set of examples and exercises that are drawn from real-world scenarios. Readers develop a reliable physical intuition that can then be used to apply dynamic analysis software tools, and to develop simplified approximate models. With this foundation, they will be able to confidently use the equations of motion in a variety of applications, ranging from simulation and design to motion planning and control.
The study aimed to delve into the incidence and risk factors associated with myocarditis and pericarditis following SARS-COV-2-19 vaccination, addressing a notable gap in understanding the safety profile of vaccinations. Through meticulous data selection from the National Health Insurance System (NHIS) database of Korea, the researchers employed both a case-crossover study and a nested case-control design to analyze temporal patterns and risk factors related to carditis occurrences post-immunization. Key findings revealed a significant association between SARS-COV-2-19 vaccination and the occurrence of carditis, with a strong temporal correlation observed within 10 days post-vaccination. Noteworthy factors contributing to carditis risk included the duration between vaccination and carditis, specific comorbidities and medication use. The study concluded by recommending an extended post-vaccination surveillance duration of at least 10 days and underscored the importance of considering individual medical histories and concurrent medication use in assessing vaccine-induced carditis risk. This study might contribute to understanding vaccine safety profiles and emphasizes the significance of comprehensive post-vaccination monitoring protocols.
The choroid plexus produces cerebrospinal fluid, which is crucial for glymphatic system function. Evidence suggests that changes in the volume of the choroid plexus may be associated with glymphatic system function. Therefore, this study aimed to investigate alterations in choroid plexus volume in patients with migraines compared with healthy controls.
Methods:
We enrolled 59 patients with migraines (39 and 20 with episodic and chronic migraines, respectively) and 61 healthy controls. All participants underwent brain magnetic resonance imaging, including three-dimensional T1-weighted imaging. We analyzed and compared choroid plexus volumes between patients with episodic migraines, those with chronic migraines and healthy controls. Additionally, we evaluated the association between choroid plexus volume and the clinical characteristics of patients with migraine.
Results:
The choroid plexus volume in patients with chronic migraines was higher than that in healthy controls (2.018 vs. 1.698%, p = 0.002) and patients with episodic migraines (2.018 vs. 1.680%, p = 0.010). However, no differences were observed in choroid plexus volumes between patients with episodic migraine and healthy controls. Choroid plexus volume was positively correlated with age in patients with migraines (r = 0.301, p = 0.020) and in healthy controls (r = 0.382, p = 0.002).
Conclusion:
We demonstrated significant enlargement of the choroid plexus in patients with chronic migraine compared with healthy controls and those with episodic migraine. This finding suggests that chronic migraine may be associated with glymphatic system dysfunction.
Particle-laden flow through conduits is ubiquitous in both natural and industrial systems. In such flows, particles often migrate across the main fluid stream, resulting in non-uniform spatial distribution owing to particle–fluid and particle–particle interactions. The most relevant lateral particle migration mechanism by particle–fluid interaction is the Segré–Silberberg effect, which is induced by the inertial forces exerted on a particle, as the flow rate increases. However, methods to suppress it have not been suggested yet. Here, we demonstrate that adding a small amount of polymer to the particle-suspending solvent effectively suppresses the Segré–Silberberg effect in a square channel. To accurately determine the position of the particles within the channel cross-sections, we devised a dual-view imaging system applicable to microfluidic systems. Our analyses show that the Segré–Silberberg effect is effectively suppressed in a square microchannel due to the balance between the inertial and elastic forces at an optimal polymer concentration while maintaining nearly constant shear viscosity.
Treatment-resistant depression (TRD) affects 10–30% of patients with major depressive disorder, leading to increased comorbidities, higher mortality, and significant economic and social burdens. This study aimed to compare the efficacy and safety of bupropion and aripiprazole as augmentation therapies for TRD.
Methods
This population-based, retrospective cohort study included adults aged ≥18 years with a diagnosis of depressive disorder who met the criteria for TRD. Data were collected from a nationwide claims database in South Korea. Patients prescribed bupropion were matched 1:1 with those prescribed aripiprazole. Subgroup analyses were performed according to age. An as-treated analysis was performed as the primary analysis, and an intention-to-treat analysis was performed to identify different risk windows. The primary outcome was depression-related hospitalization, and the secondary outcomes were first-time diagnoses of movement disorder and seizure.
Results
A total of 5,619 patients (bupropion: n = 1,568; aripiprazole: n = 4,051) were included in this study. Bupropion was associated with lower risks of hospitalization (hazard ratio [HR]: 0.51; 95% confidence interval [CI] 0.29–0.86) and movement disorders (HR: 0.56; 95% CI 0.36–0.85) than aripiprazole. No significant difference in seizure risk (HR: 0.65; 95% CI 0.30–1.31) was observed between the two treatments. The subgroup analysis of participants aged ≥60 years revealed no significant differences in the three outcomes between the two medications.
Conclusions
Bupropion augmentation is associated with a significantly lower risk of depression-related re-hospitalization and movement disorders in patients with TRD. Therefore, bupropion augmentation can be a comprehensive treatment strategy for TRD.
This study aimed to evaluate factors influencing assessment results in the new health technology assessment (nHTA) system in Korea.
Methods
Publicly available HTA reports obtained from the nHTA website were selected as a data source. A total of 258 nHTA reports including 305 technologies were included in the analysis. The detailed information in the reports was classified into three major categories: technical characteristics, evaluation methods, and publication types. A chi-squared test was used to investigate differences in the levels of evidence (high, medium, or low) and assessment results (pass or fail) according to the three categories. Univariate and multivariate logistic regression analyses were performed to identify factors associated with the levels of evidence and assessment results.
Results
nHTA reports that performed a meta-analysis and included randomized controlled trials for evidence synthesis were associated with higher levels of evidence. The corresponding odds ratios were 5.008 (95% confidence interval [CI]: 1.265, 18.826) and 27.052 (95% CI: 7.802, 103.330), respectively. The analysis showed that as the level of evidence increased, the likelihood of the assessment passing was significantly higher (odds ratio 2.789, 95% CI: 1.284, 6.057). However, univariate analysis indicated that performing a meta-analysis or including randomized controlled trials, both of which affect evidence level, did not have a statistically significant association with assessment results.
Conclusions
This study is the first systematic analysis of the factors influencing the results of nHTA reports in Korea. While higher evidence levels were associated with positive assessment outcomes, factors affecting the evidence level itself did not directly influence assessment results. More efforts are needed to integrate high levels of evidence into assessments.
In 10-minute speaking, N95 respirators significantly decreased SARS-CoV-2 emissions compared with no-mask wearing. However, SARS-CoV-2 was detected in the air even when wearing N95 and surgical masks in patients with high viral loads. Therefore, universal masking of infected and uninfected persons is important for preventing COVID-19 transmission via the air.
Background: Data on antimicrobial use at the national level is crucial to establish domestic antimicrobial stewardship policies and enable medical institutions to benchmark against each other. This study aimed to analyze antimicrobial use in Korean hospitals. Methods: We investigated the antimicrobials prescribed in Korean hospitals between 2018 and 2021, using data from the Health Insurance Review and Assessment. Primary care hospitals (PCHs), secondary care hospitals (SCHs), and tertiary care hospitals (TCHs) were included in this analysis. Antimicrobials were categorized according to the Korea National Antimicrobial Use Analysis System (KONAS) classification, which is suitable for measuring antimicrobial use in Korean hospitals. Results: Out of more than 1,900 hospitals, PCHs and TCHs represented the largest and lowest percentage of hospitals, respectively. The most frequently prescribed antimicrobial in 2021 was piperacillin/β-lactamase inhibitor (9.3%) in TCHs, ceftriaxone (11.0%) in SCHs, and cefazedone (18.9%) in PCHs. Between 2018 and 2021, the most used antimicrobial class according to the KONAS classification was ‘broad-spectrum antibacterial agents predominantly used for community-acquired infections’ in TCHs and SCHs, and 'narrow spectrum beta-lactam agents' in PCH. Total consumption of antimicrobials has decreased from 951.7 to 929.9 days of therapy (DOT)/1,000 patient-days in TCHs and from 817.8 to 752.2 DOT/1,000 patient-days in SCHs during study period, but not in PCHs (from 504.3 to 527.2 DOT/1,000 patient-days). Moreover, in 2021, while use of reserve antimicrobials has decreased from 13.6 to 10.7 DOT/1,000 patient-days in TCHs and from 4.6 to 3.3 DOT/1,000 patient-days in SCHs, it has increased from 0.7 to 0.8 DOT/1,000 patient-days in PCHs. Conclusion: This study confirms that antimicrobial use differs by hospital type in Korea. Recent increases of use of antimicrobials, including reserve antimicrobials, in PCHs reflect the challenges that must be addressed.
In 2021, the National Research Institute of Cultural Heritage (NRICH) installed a new accelerator mass spectrometer (AMS) to date Korean national heritage. Before conducting radiocarbon dating, the facility was set up and its performance was assessed. AMS system parameters have been optimized. Measurement of standard and blank samples was verified. Intercomparison analyses were also performed using heritage samples. The F14C value of NIST 4990C was 1.3406 and the background level was 0.0024. Both IAEA-C7 and C8 were confirmed to match the reference value within the 1-σ range. The NIST 4990C measurement results confirmed high precision and accuracy, with 1-σ values below 2‰. In the intercomparison, the error for each sample was 2‰. Thus, based on this study, NRICH plans to launch a dating service specializing in cultural heritage.
This study aimed to investigate the diverse clinical manifestations and simple early biomarkers predicting mortality of COVID-19 patients admitted to the emergency department (ED). A total of 710 patients with COVID-19 were enrolled from 6,896 patients presenting to the ED between January 2022 and March 2022. During the study period, a total of 478 patients tested positive for COVID-19, among whom 222 (46.4%) presented with extrapulmonary manifestations of COVID-19; 49 (10.3%) patients displayed gastrointestinal manifestations, followed by neurological (n = 41; 8.6%) and cardiac manifestations (n = 31; 6.5%). In total, 54 (11.3%) patients died. A Cox proportional hazards model revealed that old age, acute kidney injury at presentation, increased total leukocyte counts, low platelet counts, decreased albumin levels, and increased LDH levels were the independent predictors of mortality. The albumin levels exhibited the highest area under the curve in receiver operating characteristic analysis, with a value of 0.860 (95% confidence interval, 0.796–0.875). The study showed the diverse clinical presentations and simple-to-measure prognostic markers in COVID-19 patients presenting to the ED. Serum albumin levels can serve as a novel and simple early biomarker to identify COVID-19 patients at high risk of death.
Climate change poses a major threat to marine ecosystems, with its effects felt worldwide. A major effect of climate change on marine ecosystems is the rise in water temperature, leading to a northward expansion of habitats for marine organisms. Herdmania momus, a species of ascidians (sea squirts), originally found in tropical and subtropical regions, was introduced to the Korean Peninsula. In this study, we examined the habitat of H. momus along the southeastern coast of the Korean Peninsula between 2016 and 2022. We found that H. momus settlements were observed across the entire survey area, with confirmed habitation in Busan in 2016, Ulsan in 2021, and Gyeongju (the northernmost location) in 2022. The observed habitation trend indicates a rapid geographical expansion, occurring approximately 79 years earlier than previously predicted. These observations demonstrate that marine organisms are undergoing a more rapid geographical expansion than previously projected. These unexpected findings should inform government policies related to proactive measures and strategies for managing the impact of climate change on marine ecosystems.
Faecal examinations for helminth eggs were performed on 1869 people from two riverside localities, Vientiane Municipality and Saravane Province, along the Mekong River, Laos. To obtain adult flukes, 42 people positive for small trematode eggs (Opisthorchis viverrini, heterophyid, or lecithodendriid eggs) were treated with a 20–30 mg kg−1 single dose of praziquantel and purged. Diarrhoeic stools were then collected from 36 people (18 in each area) and searched for helminth parasites using stereomicroscopes. Faecal examinations revealed positive rates for small trematode eggs of 53.3% and 70.8% (average 65.2%) in Vientiane and Saravane Province, respectively. Infections with O. viverrini and six species of intestinal flukes were found, namely, Haplorchistaichui, H. pumilio, H. yokogawai, Centrocestus caninus,Prosthodendrium molenkampi, and Phaneropsolus bonnei. The total number of flukes collected and the proportion of fluke species recovered were markedly different in the two localities; in Vientiane, 1041 O. viverrini (57.8 per person) and 615 others (34.2 per person), whereas in Saravane, 395 O. viverrini (21.9 per person) and 155207 others (8622.6 per person). Five people from Saravane harboured no O. viverrini but numerous heterophyid and/or lecithodendriid flukes. The results indicate that O. viverrini and several species of heterophyid and lecithodendriid flukes are endemic in these two riverside localities, and suggest that the intensity of infection and the relative proportion of fluke species vary by locality along the Mekong River basin.
Vascular abnormalities have been frequently reported in elderly adults as a potential risk factor of late-life depression. However, it is still unclear whether stenosis of cerebral arteries may increase risk of depression in the elderly.
Methods:
Study participants were 365 patients 65 years or older with depressive disorder who had undergone brain MRI and angiography (MRA) which were assessed by trained radiologists, and the 15-item Geriatric Depression Scale (GDS-15) and the Mini Mental State Examination (MMSE), and blood glucose and lipid profiles.
Results:
Of the 365 subjects, 108 had at least one location of cerebral artery stenosis (29.59%). Stenosis was associated with age, marital status, infarction, and atherosclerosis. In multivariable linear regression analysis of different locations of stenosis among the whole sample, only bilateral middle cerebral artery (MCA) stenosis was found to have a significant association with higher GDS-15 score (p= 0.0138), and more than 8 scores in the GDS-15 (p= 0.0045), but no significant associations with ACA (anterior cerebral artery), PCA (posterior cerebral artery) or ICA (internal carotid artery). In multivariable linear logistic analysis of different locations among patients with at least one cerebral artery stenosis, left MCA was found to be significantly related to higher GDS-15 scores but not with right MCA (p = 0.0202).
Conclusion:
MCA stenosis is significantly associated with severity of depression in elderly adults with cerebral artery stenosis, especially in those with left MCA stenosis.
Key points
Patients with cerebral artery stenosis found with brain magnetic resonance angiography (MRA) were associated with higher depression severity.
Stenosis of both left and right middle cerebral artery (MCA) was associated with greater depression severity, with left MCA stenosis having a greater influence on depression severity than right MCA stenosis.
Higher depression severity in patients with MCA stenosis suggests that depression in elderly patients is mediated at least in part by vascular pathology of MCA supplied regions and careful investigation and management of cerebral artery stenosis and their risk factors may help reduce the severity of depression in elderly patients who visit psychiatrists.
Previous studies investigating neuropsychological profiles of cognitive impairment people have found a learning curve can be a useful indicator of AD diagnosis or progression. However, the data on the relationship between amyloid β (Aβ) deposition status and the learning curve in amnestic mild cognitive impairment (aMCI) are limited. In this study, we investigate the role of the learning curve in predicting Aβ deposition status in patients with aMCI.
Methods:
This is a cross-sectional study of 67 aMCI patients (N = 67; 33 aMCI with amyloid positive (Aβ-PET (+)), and 34 aMCI with amyloid negative (Aβ-PET (-))). All participants underwent Seoul Neuropsychological Screening Battery for a comprehensive neuropsychological test battery and brain MRI. To determine Aβ deposition status, each participant underwent amyloid PET scans using 18F-florbetaben. The learning curve was obtained using immediate recall of Seoul Verbal Learning Test-learning curve (SVLT- learning curve). The association of cognitive test scores and dichotomized Aβ deposition status was examined using logistic regression models in patients with aMCI. Receiver operating characteristic (ROC) curves were used to examine the predictive ability of cognitive test to detect Aβ deposition status in aMCI.
Results:
Logistic regression models showed that SVLT-learning curve and Rey Complex Figure Test- delayed recall (RCFT-delayed recall) scores were significantly associated with Aβ deposition status. In ROC analysis to assess the predictive power, SVLT-learning curve (area under the curve (AUC) = 0.734, P = 0.001) and RCFT-delayed recall (AUC = 0.739, P = 0.001) independently discriminated Aβ-PET (+) and Aβ-PET (-). The combination of these clinical markers (SVLT-learning curve and RCFT-delayed recall) improved the predictive accuracy of Aβ-PET (+) (AUC = 0.833, P < 0.001).
Conclusions:
Our findings of association of Aβ deposition status with SVLT-learning curve and RCFT- delayed recall suggest that these cognitive tests could be a useful screening tool for Aβ deposition status among aMCI patients in resource-limited clinics.
Although, attempts to apply virtual reality (VR) in mental healthcare are rapidly increasing, it is still unclear whether VR relaxation can reduce stress more than conventional biofeedback.
Methods:
Participants consisted of 83 healthy adult volunteers with high stress, which was defined as a score of 20 or more on the Perceived Stress Scale-10 (PSS-10). This study used an open, randomized, crossover design with baseline, stress, and relaxation phases. During the stress phase, participants experienced an intentionally generated shaking VR and serial-7 subtraction. For the relaxation phase, participants underwent a randomly assigned relaxation session on day 1 among VR relaxation and biofeedack, and the other type of relaxation session was applied on day 2. We compared the StateTrait Anxiety Inventory-X1 (STAI-X1), STAI-X2, the Numeric Rating Scale (NRS), and physiological parameters including heart rate variability (HRV) indexes in the stress and relaxation phases.
Results:
A total of 74 participants were included in the analyses. The median age of participants was 39 years, STAI-X1 was 47.27 (SD = 9.92), and NRS was 55.51 (SD = 24.48) at baseline. VR and biofeedback significantly decreased STAI-X1 and NRS from the stress phase to the relaxation phase, while the difference of effect between VR and biofeedback was not significant. However, there was a significant difference in electromyography, LF/HF ratio, LF total, and NN50 between VR relaxation and biofeedback
Conclusion:
VR relaxation was effective in reducing subjectively reported stress in individuals with high stress.
Titanium dioxide (TiO2, rutile) nanoparticles, inorganic ultraviolet absorbers, are used extensively in sunscreen cosmetics as an inorganic ultraviolet (UV) absorber to prevent skin damage; because of their nanotoxicity, use in combination with a support, such as montmorillonite (Mnt), rather than alone, is suggested. Mnt-supported TiO2 composites (Mnt-TiO2) for sunscreens are most suitable when the particles are spherical and of relatively uniform size, which are normally accomplished by spray drying, but this is difficult to achieve because of the naturally layered structure of Mnt. The objective of the present study was, therefore, to find the ideal characteristics of spray-drying nozzles to produce the desired spherical shape and size distribution of the Mnt-TiO2 composite particles. The starting Mnt was extracted from natural bentonite by particle-size separation. An ultrasonic nozzle in the spray dryer was selected for use in the synthesis of Mnt-TiO2 composites based on the particle-size distribution (PSD) of Mnt prepared using a two-fluid nozzle and an ultrasonic nozzle at 453 K. The incorporation of TiO2 in the final Mnt-TiO2 composites was examined by X-ray powder diffraction (XRD) and elemental analysis. With increasing TiO2 concentration, the TiO2 content and average particle size of the Mnt-TiO2 composites increased. Scanning electron microscopy (SEM) images showed that all samples prepared had uniform and nearly spherical shapes. Absorbance of UV by Mnt-TiO2 (5:1) composites was greater than that by either purified Mnts or pure TiO2. The present study demonstrated a simple method, using a spray dryer with an ultrasonic nozzle, to synthesize Mnt-TiO2 composites of uniform size and shape suitable for cosmetic application.
The development of nanoparticles incorporating Gd3+ has attracted interest in the field of contrasting nanomedicine for magnetic resonance (MR) and computed tomography (CT). In order to achieve an effective contrasting performance, the amount and stability of Gd incorporated as well as particle-size control of the nanoparticles should be considered simultaneously. In the current study, Gd-diethylenetriaminepentaacetate(Gd-DTPA) was incorporated into layered double hydroxide (LDH) to meet the physicochemical properties required for MR-CT dual contrasting nanomedicine. Strategically, the particle size and nanometer incorporation of Gd-DTPA into LDH (GL-R hybrid) were controlled homogeneously using the reverse micelle method. X-ray diffraction showed that the hybrid obtained possessed a hydrotalcite phase. Dynamic light scattering and electron microscopic analyses showed that the hybrid had a controlled size of <200 nm with significant homogeneity. Fast Fourier-transform of transmission electron microscopy confirmed that the platelets of the GL-R hybrids were oriented randomly to form inter-particle space, enabling the Gd-DTPA moiety to be encapsulated stably. The encapsulation efficiency of Gd-DTPA was 20.8%, which was sufficiently high compared with other Gd-DTPA-incorporatedLDH. According to X-ray photoelectron spectroscopy and Fourier-transform infrared spectroscopy, Gd-DTPA incorporated in the hybrid preserved its structure intact. Its potential as a dual modal contrast agent was demonstrated by measuring the concentration-dependent Hounsfield unit and magnetic resonance relaxivity, which were determined to be 230 at 2 mg/mL and 5.8 in the range ~0.2–1 mM-Gd, respectively.
Critical congenital heart disease (CCHD) refers to a group of heart defects that cause serious, life-threatening symptoms in the neonatal period and requires timely surgical or catheter interventions. We tried to explore current status of CCHD burden and the effect of early diagnosis of CCHD to mortality using the Korean national health insurance (NHI) data.
Methods
We analyzed the national health insurance (NHI) data from 2014 to 2018. We identified CCHD patients using the diagnosis codes and intervention codes from the claim data and the prevalence, mortality and medical expenditure of CCHD were analyzed. We linked neonatal data with their mother’s medical claim data and developed retrospective cohort data set for analyzing the effect of early diagnosis to mortality and related outcomes of CCHD treatment.
Results
The annual prevalence of neonatal CCHD in Korea was 0.144% percent. A total of 2,241 CCHD neonates, 1,546 (69.0%) underwent cardiac ultrasound within three days after birth, and mothers of 419 neonates had a record of prenatal fetal ultrasound (18.7%). In our comparison of neonates diagnosed with CCHD within three days of birth with those diagnosed with CCHD on or after day 4 of birth, the probability of early diagnosis increased for preterm infants and infants with low birth rate. Regarding mortality rate, most types of CCHD showed a significantly higher mortality rate in the early diagnosis group.
Conclusions
The reason for the high mortality rate despite a high early diagnosis rate pertains to the high percentage of patients with severe conditions that induce a serious heart rate within three days of birth. More than half of the neonates with CCHD were found to have not undergone a prenatal fetal ultrasound, rendering this an important policy target.
Critical congenital heart disease (CCHD) refers to a group of heart defects that cause serious, life-threatening symptoms in the neonatal period and requires timely surgical or catheter interventions. We reviewed evidence for incorporating a mandatory neonatal CCHD screening test as a national public health project for all neonates born in Korea by analyzing the validity and cost-effectiveness of neonatal CCHD screening using pulse oximetry in Korea.
Methods
We performed a rapid literature review to establish models for the diagnostic accuracy and economic evaluation of pulse oximetry. Also, we analyzed the prevalence, mortality, and medical expenditure for different types of CCHD using the national health insurance (NHI) data. We analyzed the cost-effectiveness of pulse oximetry by comparing the group of neonates who received a combination of a physical examination and pulse oximetry, and group of neonates who only received a physical examination. For the cost-effectiveness analysis for the CCHD screening test in this study, we used a duration of one year, diagnostic accuracy as the clinical endpoint, and Life Year Gain (LYG) as the effectiveness indicator.
Results
We used recent systematic review he pooled sensitivity can be enhanced from 76.5 percent (pulse oximetry alone) to 92 percent (combined with physical examination). We used a total of 2,334 neonates with CCHD data for the economic model. Our analysis revealed that adding pulse oximetry to the routine neonatal physical examination leads to 2.34 of LYG and a cost difference of USD1,080,602, showing a ICER of KRW610,063,240 (USD461,857)/LYG.
Conclusions
Considering the benefit of LYG and cost of reducing the complications and after effects of newborns with CCHD who survived early diagnosis, it is considered to be worthwhile in Korea for a mandatory screening test.
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.