Clinical utility of gastric fluid cytokine levels in preterm infants for predicting histological chorioamnionitis.

BACKGROUND: The risk of various complications, such as neonatal death, early onset sepsis, and chronic lung disease, is increased in infants born to mothers with chorioamnionitis (CAM). However, predicting the diagnosis of histological CAM (hCAM) in the early postnatal period is challenging for clinicians due to pathological considerations. Therefore, an early diagnostic tool for hCAM is needed. Gastric fluid at birth is considered a suitable biomarker for predicting the intrauterine environment because most of its components are from amniotic fluid, and the sampling technique is less invasive. This study aimed to evaluate the clinical utility of cytokines in the gastric fluid of preterm infants at birth as predictors of hCAM. METHODS: We retrieved gastric fluid and serum from 21 preterm infants with a gestational age of ≤ 32 weeks within 1 h after birth and used cytometric bead array to measure the concentrations of interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor-alpha, and interferon-gamma. We compared the cytokine concentrations in the gastric fluid and serum of the preterm infants born to mothers with or without hCAM. RESULTS: The gastric fluid, serum IL-6, and serum IL-10 concentrations were significantly higher in the hCAM group than that in the non-hCAM group. The best cutoff values for predicting hCAM was > 2,855 pg/mL and > 315 pg/mL for IL-6 in the gastric fluid and serum, respectively. Receiver operating characteristic curves showed that gastric fluid IL-6 concentrations correlated more strongly with the presence of hCAM than serum IL-6 concentrations. CONCLUSION: IL-6 in the gastric fluid at birth may be a more promising biomarker for predicting the presence of hCAM than that in serum. IL-6 concentration analysis in the gastric fluid at birth might help to diagnose hCAM immediately after birth and improve the prognosis of preterm infants.

An approach for live imaging of first cleavage in mouse embryos using fluorescent chemical probes for DNA, microtubules, and microfilaments.

Purpose: Dynamic morphological changes in the chromosome and cytoskeleton occur in mammals and humans during early embryonic development, and abnormalities such as embryonic chromosomal aneuploidy occur when development does not proceed normally. Visualization of the intracellular organelles and cytoskeleton allows elucidation of the development of early mammalian embryos. The behavior of the DNA and cytoskeleton in early mammalian embryos has conventionally been observed by injecting target molecule mRNAs, incorporating a fluorescent substance-expressing gene, into embryos. In this study, we visualized the chronological behavior of male and female chromosome condensation in mouse embryos, beginning in the two-pronuclear zygote, through the first division to the two-cell stage, using fluorescent chemical probes to visualize the behavior of DNA, microtubules, and microfilaments. Method: Mouse two-pronuclear stage embryo were immersed in medium containing fluorescent chemical probes to visualize DNA, microtubules, and microfilaments. Observation was performed with a confocal microscope. Results: This method allowed us to observe how chromosome segregation errors in first somatic cell divisions in mouse embryos and enabled dynamic analysis of a phenomenon called lagging chromosomes. Conclusions: By applying this method, we can observe any stage of embryonic development, which may provide new insights into embryonic development in other mammals.

The first nationwide website survey of the availability and costs of medical and non-medical oocyte cryopreservation in Japan.

Research question: How does the cost-related oocyte cryopreservation (OoC) vary by the facility in Japan, and what data is provided on the websites about OoC procedures? Design: Website survey. The websites of all 621 facilities that provide assistive reproductive technology registered in Japan were surveyed in 2021. Data included the rates of explicit statements regarding the provision of OoC for only medical reasons (medical only group) or non-medical reasons (non-medical group). Based on whether or not facilities that perform OoC clearly stated the cost on their websites, we compared the costs of OoC and annual storage cost between medical only and non-medical groups. Furthermore, we examined the stated number of OoC procedures performed and their clinical outcomes. Results: Of the 621 facilities, 146 (23.5%) clearly stated that they offer OoC on their websites. Of the 88 medical only groups and 58 non-medical groups, 24 (27.3%) and 42 (72.4%) clearly stated the OoC cost, and 27 (30.7%) and 44 (75.9%) clearly states the annual oocyte storage cost, respectively. The OoC costs were significantly higher for the non-medical group than in the medical group. In the medical only group, the annual storage cost remained almost the same regardless of the number of oocytes, while in the non-medical group, the annual storage cost was 2-3 times higher than in the medical only group. Only 16 facilities (16/146, 11.0%) had mentioned the number of OoC procedures, and five facilities (3.4%) provided information on the clinical outcomes after OoC. Conclusion: Costs related to OoC are higher for the non-medical group in Japan. In addition, the websites contain scant information on the costs and clinical outcomes of OoC.

Interleukin-33 and Soluble ST2 Levels in Infants with Hypoxic-Ischemic Encephalopathy.

INTRODUCTION: Interleukin (IL)-33 and its receptor ST2L play key roles in the IL-33/ST2 signaling pathway. Soluble ST2 (sST2) inhibits the proper function of IL-33. sST2 levels are increased in patients with several neurological diseases, but in infants with hypoxic-ischemic encephalopathy (HIE), IL-33 and sST2 levels have not been studied. This study aimed to investigate whether serum levels of IL-33 and sST2 are useful as biomarkers of HIE severity and prognostic factors for infants with HIE. METHODS: Twenty-three infants with HIE and 16 controls (gestational age ≥36 weeks and ≥1,800 g birth weight) were enrolled in this study. Serum levels of IL-33 and sST2 were measured at <6 h, 1-2, 3, and 7 days of age. Hydrogen-1 magnetic resonance spectroscopy was performed, and ratios of peak integrals of lactate/N-acetylaspartate (Lac/NAA) were calculated as objective indicators of brain damage. RESULTS: In the moderate and severe HIE, serum sST2 concentrations were increased and there was a good correlation between serum sST2 and HIE severity on days 1-2, whereas no variation was observed in serum IL-33. Serum sST2 levels were positively correlated with Lac/NAA ratios (Kendall's rank correlation coefficient = 0.527, p = 0.024), and both sST2 and Lac/NAA ratios were significantly higher in HIE infants with neurological impairment (p = 0.020 and <0.001, respectively). CONCLUSIONS: sST2 may be a useful predictor of severity and later neurological outcomes in infants with HIE. Further investigation is required to elucidate the relationship between the IL-33/ST2 axis and HIE.

Novel compound heterozygous mutations in the PLEC gene in a neonate with epidermolysis bullosa simplex with pyloric atresia.

Epidermolysis bullosa (EB) is a heterogeneous group of inherited disorders characterized by the blistering of the skin and mucous membranes. Although the molecular basis of EB has been significantly elucidated, the precise phenotypes of the lethal types of EB have not been completely characterized. Herein, we report a severe case of EB with pyloric atresia (PA). The patient was a Japanese boy who not only had skin lesions but also various complications such as PA, dysphagia, hypotonia, infectious keratitis with corneal ulcer, obstructive uropathy and protein-losing enteropathy. Genetic analysis led to the identification of two novel compound heterozygous mutations in the last exon of the plectin (PLEC) gene. Based on this finding, EB simplex with PA was diagnosed. Immunostaining with anti-plectin antibodies revealed truncated plectin proteins lacking the C-terminus in the patient's skin. We also conducted a prenatal diagnosis in subsequent pregnancy. Our report further highlights the crucial role of plectin in many organs and provides valuable information regarding the phenotypes resulting from mutations in the PLEC gene.

Precise size-control and functionalization of gold nanoparticles synthesized by plasma-liquid interactions: using carboxylic, amino, and thiol ligands.

Using gold nanoparticles (GNPs) in high-standard applications requires GNPs to be fabricated with high-quality size and surface properties. Plasma-liquid interactions (PLIs) have the unique ability to synthesize GNPs without using any reducing agents, and the GNP surface is free of stabilizing agents. It is an extreme advantage that ensures success for the subsequent functionalization processes for GNPs. However, fabricating GNPs via PLIs at the desired size has still been a challenge. Here, we present a simple approach to achieving the precise size-control of GNPs synthesized by PLIs. By adding suitable ligands to the precursor solution, the ligands wrap GNPs which interrupts and slows down the rapid growth of GNPs under PLIs. This way, the size of the GNPs can be precisely controlled by adjusting the ligand concentration. Our results showed that the size of the GNPs in the range of 10-60 nm can be fitted to reciprocal functions of the ligand concentration. The potency of the size-control depends on the type of ligands in the order of thiol > amine > carboxylate. The size-control has been well investigated with four common ligands: l-cysteine, glucosamine, salicylic acid, and terephthalic acid. XPS, FTIR, and zeta potential techniques confirmed the presence of these ligands on GNPs. The results indicated that functionalized ligands could be utilized to control the size and functionalize the GNP surface. Hence our approach could simultaneously achieve two goals: precise size-control and functionalization of GNPs without the ligand-exchange step.

Successful Treatment of Saddle Pulmonary Thromboembolism in 23-Week Preterm Infant.

Saddle pulmonary thromboembolism (PTE) is defined as a thromboembolism straddling the bifurcation of the main pulmonary artery trunk and it is rarely seen in extremely low birth weight infants (ELBWI). Saddle PTE is a critical disease that requires urgent treatment. However, the treatment guidelines for ELBWI are not established. We present the case of a 1-day-old preterm infant (gestational age 23 weeks) who showed sudden desaturation and pulmonary hypertension due to saddle PTE. A thrombus was observed in the bifurcation of the pulmonary artery. The blood flow to the pulmonary artery on the left side was interrupted, and the right side showed severe stenosis. Since the patient was an ELBWI in the acute phase, we decided to use recombinant tissue plasminogen activator (rt-PA) and administered a maintenance dose (0.08 mg/kg/hour), instead of the loading dose. After using rt-PA, the thrombus dissolved in 8 hours without adverse events. This case suggests that starting with a maintenance dose of rt-PA may be an effective treatment option for saddle PTE in ELBWI in the acute phase under the high risk of bleeding.

Demonstration of an intense lithium beam for forward-directed pulsed neutron generation.

As an alternative to research nuclear reactors, a compact accelerator-driven neutron generator that uses a lithium beam driver could be a promising candidate since it produces almost no undesired radiation. However, providing an intense lithium-ion beam has been difficult, and it has been thought that the practical application of such a device would be impossible. The most critical problem of insufficient ion fluxes has been solved by applying a direct plasma injection scheme. In this scheme, a pulsed high-density plasma from a metallic lithium foil generated by laser ablation is efficiently injected and accelerated by a radio-frequency quadrupole linear accelerator (RFQ linac). We have obtained a peak beam current of 35 mA accelerated to 1.43 MeV, which is two orders of magnitude higher than a conventional injector and accelerator system can deliver.

Food Protein-Induced Enterocolitis Syndrome in Children with Down Syndrome: A Pilot Case-Control Study.

Food protein-induced enterocolitis syndrome (FPIES) is a non-immunoglobin E-mediated food hypersensitivity disorder. However, little is known about the clinical features of FPIES in patients with Down syndrome (DS). Medical records of children with DS diagnosed at our hospital between 2000 and 2019 were retrospectively reviewed. Among the 43 children with DS, five (11.6%) were diagnosed with FPIES; all cases were severe. In the FPIES group, the median age at onset and tolerance was 84 days and 37.5 months, respectively. Causative foods were cow's milk formula and wheat. The surgical history of colostomy was significantly higher in the FPIES group than in the non-FPIES group. A colostomy was performed in two children in the FPIES group, both of whom had the most severe symptoms of FPIES, including severe dehydration and metabolic acidosis. The surgical history of colostomy and postoperative nutrition of formula milk feeding may have led to the onset of FPIES. Therefore, an amino acid-based formula should be considered for children who undergo gastrointestinal surgeries, especially colostomy in neonates or early infants. When an acute gastrointestinal disease is suspected in children with DS, FPIES should be considered. This may prevent unnecessary tests and invasive treatments.

Consistency between chromosomal status analysis of biopsied human blastocyst trophectoderm cells and whole blastocyst cells.

PURPOSE: This study investigated the consistency between results of preimplantation genetic testing for aneuploidy performed on trophectoderm (TE) cells and remaining blastocyst cells. METHODS: TE biopsy was performed on 29 surplus cryopreserved human blastocysts. Biopsy samples and remaining blastocysts were processed using the VeriSeq PGS kit, and chromosomal statuses were compared by next-generation sequencing. RESULTS: Discordance was observed in the chromosomal status of 11 out of 29 blastocysts between the biopsied TE and remaining blastocysts. Concordance was observed in 11 of 12 blastocysts classified as euploid by TE biopsy and in 7 of 17 blastocysts classified as aneuploid. There was 100% concordance (7/7) in cases diagnosed as aneuploid with no mosaicism by TE biopsy. However, discordance was observed in all 10 cases showing mosaicism or partial chromosomal abnormality. CONCLUSION: Chromosomal status analysis based on TE biopsy does not accurately reflect the chromosomal status of the whole blastocyst. The chromosomal status is usually the same between the TE and remaining blastocyst cells in cases diagnosed as euploid or aneuploid with no mosaicism. However, mosaic blastocysts and those with other types of structural rearrangements have a higher risk of inconsistency, warranting caution during embryo selection.

Study protocol for a multicentre, open-label, single-arm phase I/II trial to evaluate the safety and efficacy of ripasudil 0.4% eye drops for retinopathy of prematurity.

INTRODUCTION: Retinopathy of prematurity (ROP) is a vascular proliferative disorder that occurs in preterm infants. Existing treatments are only indicated in severe ROP cases due to the high invasiveness and the potential risk of irreversible side effects. We previously elucidated that ripasudil, a selective inhibitor of the Rho-associated protein kinase, has the ability to inhibit abnormal retinal neovascularisation in animal models. In addition, ripasudil eye drops (Glanatec ophthalmic solution 0.4%) have been already used for the treatment of glaucoma. Since eye drop therapy is less invasive, early intervention for ROP is possible. The purpose of this phase I/II trial is to evaluate the safety and efficacy of ripasudil eye drops for preterm infants with ROP. METHODS AND ANALYSIS: This is a multicentre, open-label, single-arm phase I/II trial. To evaluate the safety and efficacy of ripasudil as much as possible, ripasudil will be administered to all enrolled preterm infants with zone I/II, stage 1, or worse ROP. The safety and efficacy of ripasudil in treated patients will be assessed in comparison to a historical control group. Because this is the first trial of ripasudil in preterm infants, a dose-escalation study (once daily for 1 week, then two times per day for 2 weeks) will be conducted in phase I. After obtaining approval from the independent data and safety monitoring board to continue the trial after the completion of phase I, phase II will be conducted. In phase II, ripasudil eye drops will be administered two times per day for 12 weeks. The primary endpoint in phase II is also safety. Efficacy and pharmacokinetics will be evaluated as secondary endpoints. ETHICS AND DISSEMINATION: This study protocol was approved by the institutional review board at each of the participating centres. Data will be presented at international conferences and published in peer-reviewed journals. TRIAL REGISTRATION NUMBERS: NCT04621136 and jRCT2071200047.

Cell-free DNA in spent culture medium effectively reflects the chromosomal status of embryos following culturing beyond implantation compared to trophectoderm biopsy.

This prospective study evaluated the accuracy of non-invasive preimplantation genetic testing for aneuploidy (niPGT-A) using cell-free DNA in spent culture medium, as well as that of preimplantation genetic testing for aneuploidy (PGT-A) using trophectoderm (TE) biopsy after culturing beyond implantation. Twenty frozen blastocysts donated by 12 patients who underwent IVF at our institution were investigated. Of these, 10 were frozen on day 5 and 10 on day 6. Spent culture medium and TE cells were collected from each blastocyst after thawing, and the embryos were cultured in vitro for up to 10 days. The outgrowths after culturing beyond implantation were sampled and subjected to chromosome analysis using next-generation sequencing. Chromosomal concordance rate, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), false-positive rate (FPR), and false-negative rate (FNR) of niPGT-A and PGT-A against each outgrowth were analyzed. The concordance rate between the niPGT-A and outgrowth samples was 9/16 (56.3%), and the concordance rate between the PGT-A and outgrowth samples was 7/16 (43.8%). NiPGT-A exhibited 100% sensitivity, 87.5% specificity, 88.9% PPV, 100% NPV, 12.5% FPR, and 0% FNR. PGT-A exhibited 87.5% sensitivity, 77.8% specificity, 87.5% PPV, 75% NPV, 14.3% FPR, and 22.2% FNR. NiPGT-A may be more accurate than PGT-A in terms of ploidy diagnostic accuracy in outgrowths.

Live visualisation of electrolytes during mouse embryonic development using electrolyte indicators.

Studies have shown that some electrolytes, including Na+ and K+, play important roles in embryonic development. However, these studies evaluated these electrolytes by using inhibitors or knockout mice, with no mention on the changes in the intracellular electrolyte concentrations during embryogenesis. In this study, we used the electrolyte indicators CoroNa Green AM and ION Potassium Green-2 AM to directly visualise intracellular concentrations of Na+ and K+, respectively, at each embryonic developmental stage in mouse embryos. We directly observed intracellular electrolyte concentrations at the morula, blastocyst, and hatching stages. Our results revealed dynamic changes in intracellular electrolyte concentrations; we found that the intracellular Na+ concentration decreased, while K+ concentration increased during blastocoel formation. The degree of change in intensity in response to ouabain, an inhibitor of Na+/K+ ATPase, was considered to correspond to the degree of Na+/K+ ATPase activity at each developmental stage. Additionally, after the blastocyst stage, trophectoderm cells in direct contact with the blastocoel showed higher K+ concentrations than in direct contact with inner cell mass, indicating that Na+/K+ ATPase activity differs depending on the location in the trophectoderm. This is the first study to use CoroNa Green AM and ION Potassium Green-2 AM in mouse embryos and visualise electrolytes during embryonic development. The changes in electrolyte concentration observed in this study were consistent with the activity of Na+/K+ ATPase reported previously, and it was possible to image more detailed electrolyte behaviour in embryo cells. This method can be used to improve the understanding of cell physiology and is useful for future embryonic development studies.

Cooperation between single third-party logistics provider and government for emergency disaster relief operations in Thailand.

The planning of disaster relief operations is important for governments. These strategies are expected to reduce the impact on human lives and the economy during disasters and can be executed with the cooperation of third-party logistics to quicken disaster relief responses. In this study, we focus on investigating the importance of communication and the cooperation of government with third-party logistics providers to reduce the response time during disaster relief operations. In Thailand, although there are several third-party logistics providers, a large single third-party stands out owing to its availability of resources and distribution networks. We analyzed the effect of the disaster relief operations performed by the government in coordination with a single third-party provider for two sections of the disaster-relief logistics process: pretransport and transport. In our investigation, the inclusion of a third-party logistics provider during the transport process showed a reduction in response time irrespective of the scale of the disaster, along with a reduction in the supply saturation time for the required demand. The findings of this study suggest that the emergency response time during a disaster can be reduced in Thailand through the inclusion of a single third-party logistics provider. In addition, this study suggests an emergency response model for Thailand with the included third-party logistics to improve disaster relief operations.

Control of current waveform of laser ion source using pulsed magnetic field.

We investigated plasma behavior with a solenoid-generated static and pulsed magnetic field by measuring time evolution in a transverse ion current profile to control the ion current waveform of a laser ion source. The results showed that static magnetic fields cannot enhance the ion current in the slow region of time of flight (TOF). However, a pulsed magnetic field whose magnetic flux density is rising while a plasma passes through the solenoid can enhance the ion current in the same TOF region. The results showed that applying a pulsed magnetic field to a laser-produced plasma is an effective way to control an ion current waveform in a laser ion source to produce an ion beam with a flat-top-shaped pulse. By using a pulsed magnetic field, the ion current waveform peak was held for ∼50 µs.

Neutron generator based on intense lithium beam driver.

We are proposing a compact neutron generator based on a Li beam driver. The proposed neutron generator comprises a laser ion source, a radio-frequency quadrupole linear accelerator (RFQ linac), a drift tube linac, and a target containing protons. In the generator, the lithium ion is used as a projectile instead of protons to utilize the kinematic focusing technique. The technique enables us to enhance the neutron flux without increasing the beam energy, which is important to develop a clean compact neutron generator. Moreover, the combination of a laser ion source and a RFQ linac with the direct plasma injection scheme will provide several tens of mA of a fully ionized lithium beam, which is much higher than that of conventional heavy ion sources comparable with proton drivers. Neutrons are generated by the nuclear reaction of the lithium ions and protons in the beam target. In this paper, we reported the current status of the development. For RFQ, we designed the RFQ rods to accelerate 40 mA of 7Li3+. We fabricated and installed the rods into a cavity, and, as a first test, accelerated 10 mA of C6+ successfully.

Low charge state lithium beam production from chemical compounds with laser ion source.

In recent years, the primary ion source for the Brookhaven National Laboratory has been the laser ion source, which provides many types of ions within a short switching time of several seconds. The task is difficult for other ion sources. In the previous work, we tested metallic lithium as a target material of the laser irradiation. Although an intense lithium beam was demonstrated, some operational difficulties were observed due to its reactiveness to oxygen. For accelerator applications, a more robust and reliable target material has been demanded. For this purpose, we tested lithium niobate, LiNbO3. Our study investigated the optimization of power density to produce low charge state lithium ions. We struck LiNbO3 with the laser and found lithium ion quantities for five different power densities. Based on the data obtained, we can conclude that the most efficient production of Li1+ occurs when the laser power density is 5 × 108 W/cm2.

Kinetics of meiotic maturation in oocytes from unstimulated ovaries and duration of pronucleus presence and preimplantation development.

OBJECTIVE: To evaluate the meaning of meiotic maturation kinetics and duration of pronucleus presence (DPP) for parthenogenetic activation outcome. DESIGN: Retrospective study. SETTING: University hospital. PATIENT(S): Eight patients with endometrioid adenocarcinoma and 65 patients who underwent in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI). INTERVENTION(S): After collection of oocytes from nonstimulated ovaries of patients with endometrioid adenocarcinoma, in vitro maturation (IVM) and parthenogenetic activation performed with time-lapse imaging; after ICSI, embryos similarly incubated with time-lapse imaging. MAIN OUTCOME MEASURE(S): Timing of the release of the first polar body (fPB), DPP, and developmental stage with IVM and parthenogenetic activation; after ICSI, assessment of DPP and preimplantation developmental stage. RESULT(S): With IVM, 55.2% of oocytes matured; 53.1% of fPBs were released within 24 hours, and 46.9% of fPBs were released after 24 hours. Regarding developmental stage, oocytes that released fPB later during IVM tended to develop more than oocytes that released the fPB within 24 hours. For embryos from parthenogenetic activation the DPP was statistically significantly shorter than the DPP of embryos from ICSI. With ICSI, the DPP was statistically significantly shorter in embryos that developed to ≥8 cells than embryos whose final development included ≤7 cells. The development rate in parthenogenetic activation was statistically significantly lower than that in ICSI. CONCLUSION(S): Embryo development is negatively affected by DPP that is too short or too long. When the DPP was short with parthenogenetic activation, embryo development did not proceed, indicating that DPP is an important determinant of parthenogenetic activation outcomes as with the timing of fPB release.