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1.
J Chem Inf Model ; 64(9): 3621-3629, 2024 May 13.
Article in English | MEDLINE | ID: mdl-38642039

ABSTRACT

Machine learning (ML) has facilitated property prediction for intricate materials by integrating materials and experimental features such as processing and measurement conditions. However, ML models designed for material properties have often disregarded a common issue of "leakage," resulting in an overestimation of model performance and a decrease in model transferability. This issue can arise from biases inherent in multiple data points obtained from the same experimental group. We provide a critical examination and prevention method of leakage in property prediction for polymer composites. Our proposed method utilizes data partitioning based on the experimental group to ensure that data from the same group are not mixed in both the training and test sets. Evaluation results highlight that the conventional random partitioning unintentionally inflates ML performance through the misuse of experimental features for leaking data bias within the same experimental group rather than explaining the physical causality. In contrast, the proposed method enables the leakage-free utilization of experimental features to improve prediction accuracy while ensuring model transferability. Specifically, when integrating experimental features with polymer and filler features, the conventional method overestimates the prediction performance of electrical conductivity in reducing RMSE by 26% depending on leakage, whereas the proposed method achieves a reduction in RMSE by 5% without leakage. These findings offer valuable guidance for the effective utilization of experimental features in data-driven materials science.


Subject(s)
Machine Learning , Polymers , Polymers/chemistry , Electric Conductivity
2.
Int J Biol Macromol ; 265(Pt 2): 131060, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38521317

ABSTRACT

Various types of hemostatic agents are used to manage bleeding in surgery. Many such agents are animal products, which carry the risk of secondary infection. The aim of this study is to develop a novel hemostatic agent from a non-animal source that quickly stops bleeding, is easy to use, and has no risk of infection. In this study, we synthesized calcium ion-crosslinked sodium alginate (Alg-Na/Ca) by partial substitution of Ca ions for Na ions in sodium alginate. We prepared 12 kinds of Alg-Na/Ca powders with different Ca mass ratios, molecular weights, M/G ratios and particle size distributions and measured their swelling ratio and the burst pressure generated. We found that Alg-Na/Ca began to swell immediately after contact with saline, especially Alg-Na/Ca at Ca mass ratios of 74.1-77.0 % showed a high swelling ratio after 2 min and a high burst pressure, over 200 % and 500 mmHg respectively. Also, there is a correlation between the swelling ratio after 2 min and the burst pressure. Our results suggest that, by optimizing the composition conditions, Alg-Na/Ca may be an effective hemostatic agent that could act as a tamponade by absorbing and swelling at a bleeding site to quickly achieve primary hemostasis.


Subject(s)
Hemostatics , Animals , Hemostatics/pharmacology , Calcium , Hemostasis , Alginates , Ions
3.
Sci Rep ; 14(1): 3082, 2024 02 07.
Article in English | MEDLINE | ID: mdl-38321081

ABSTRACT

Post-stroke gait disorders involve altered lower limb kinematics. Recently, the endpoint of the lower limb has been used as a control variable to understand gait kinematics better. In a cross-sectional study of sixty-seven post-stroke patients, the limb extension angle and effective limb length during gait were used as input variables with a mixed Gaussian model-based probabilistic clustering approach to identify five distinct clusters. Each cluster had unique characteristics related to motor paralysis, spasticity, balance ability, and gait strategy. Cluster 1 exhibited high limb extension angle and length values, indicating increased spasticity. Cluster 2 had moderate extension angles and high limb lengths, indicating increased spasticity and reduced balance ability. Cluster 3 had low limb extension angles and high limb length, indicating reduced balance ability, more severe motor paralysis, and increased spasticity. Cluster 4 demonstrated high extension angles and short limb lengths, with a gait strategy that prioritized stride length in the component of gait speed. Cluster 5 had moderate extension angles and short limb lengths, with a gait strategy that prioritized cadence in the component of gait speed. These findings provide valuable insights into post-stroke gait impairment and can guide the development of personalized and effective rehabilitation strategies.


Subject(s)
Movement Disorders , Stroke Rehabilitation , Stroke , Humans , Biomechanical Phenomena , Cross-Sectional Studies , Gait , Lower Extremity , Muscle Spasticity , Paralysis
4.
J Phys Chem A ; 128(6): 1041-1048, 2024 Feb 15.
Article in English | MEDLINE | ID: mdl-38311924

ABSTRACT

To determine the H-abstraction reaction probabilities of H/O/OH radicals with a polypropylene (PP) surface, a first-principles calculation was performed based on the DLPNO-CCSD(T)/CBS//M06-2X-D3/def-TZVP theory level. The PP chain model used in this study was 2,4,6-trimethylheptane. The rate constants of the H/O/OH radicals with the isolated PP chain model were calculated based on the conventional transition-state theory. By comparing the experimental values and considering the error factors and their compensation, it was concluded that the orders of magnitude of the predicted rate constants were accurate. The resulting rate constants were converted to reaction probabilities between the H/O/OH radicals and the PP surface. The method used in this study is applicable for obtaining theoretical values of surface reaction probabilities based on first-principles calculations. The calculation at the DLPNO-CCSD(T)/CBS theory level has high accuracy but consumes a large amount of computational resources. The study also demonstrated that the double-hybrid functionals, wB97x-2-D3(BJ) and rev-DSD-PBEP86-D3(BJ), with a 3-ζ or 4-ζ basis set, could reproduce the electronic energy values obtained from DLPNO-CCSD(T)/CBS while using only approximately 1/100 of the computational resources required by the latter under our computer configuration.

5.
Opt Lett ; 49(3): 726-729, 2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38300100

ABSTRACT

We experimentally demonstrated the polarization change of femtosecond laser pulses in air by using electric-field-induced second-harmonic generation (E-FISHG) for the first time to our knowledge. The polarization change from linear to elliptical was observed at the laser intensity over the filamentation threshold. These results suggest that the polarization change can occur by the birefringence caused by filamentation. This phenomenon can be used for new applications such as an ultra-fast and precise three-dimensional electric field measurement by E-FISHG. In addition, E-FISHG can be an excellent tool to investigate the characteristics of femtosecond laser propagation such as filamentation.

6.
Ther Apher Dial ; 28(3): 442-452, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38168492

ABSTRACT

INTRODUCTION: A remission induction therapy of granulocyte and monocyte adsorptive apheresis (GMA) was given to patients with Crohn's disease (CD). However, establishing an appropriate treatment strategy for GMA in patients with CD remains unclear. METHODS: This study evaluated the clinical efficacy and subsequent clinical progression after GMA in patients with CD who underwent GMA in seven independent institutions in Japan from 2010 to 2023. RESULTS: Sixteen patients were enrolled. The overall remission and response rates were 25.0% and 68.8%, respectively. All patients responding to GMA received biologics that were continuously used and 36.4% of patients remained on the same biologics 52 weeks after GMA. Notably, all patients who continued the same biologics had previously experienced a loss of response to biologics. CONCLUSION: GMA may exhibit effectiveness even in cases with refractory CD. Moreover, it represents a potential novel therapeutic option for refractory CD with loss of response to biologics.


Subject(s)
Blood Component Removal , Crohn Disease , Granulocytes , Monocytes , Humans , Crohn Disease/therapy , Male , Female , Pilot Projects , Adult , Retrospective Studies , Blood Component Removal/methods , Japan , Treatment Outcome , Middle Aged , Remission Induction/methods , Adsorption , Biological Products/therapeutic use , Young Adult
7.
Int J Mol Sci ; 24(20)2023 Oct 18.
Article in English | MEDLINE | ID: mdl-37894981

ABSTRACT

Germline manipulation at the zygote stage using the CRISPR/Cas9 system has been extensively employed for creating genetically modified animals and maintaining established lines. However, this approach requires a long and laborious task. Recently, many researchers have attempted to overcome these limitations by generating somatic mutations in the adult stage through tail vein injection or local administration of CRISPR reagents, as a new strategy called "in vivo somatic cell genome editing". This approach does not require manipulation of early embryos or strain maintenance, and it can test the results of genome editing in a short period. The newborn is an ideal stage to perform in vivo somatic cell genome editing because it is immune-privileged, easily accessible, and only a small amount of CRISPR reagents is required to achieve somatic cell genome editing throughout the entire body, owing to its small size. In this review, we summarize in vivo genome engineering strategies that have been successfully demonstrated in newborns. We also report successful in vivo genome editing through the neonatal introduction of genome editing reagents into various sites in newborns (as exemplified by intravenous injection via the facial vein), which will be helpful for creating models for genetic diseases or treating many genetic diseases.


Subject(s)
CRISPR-Cas Systems , Gene Editing , Animals , Gene Editing/methods , CRISPR-Cas Systems/genetics , Animals, Newborn , Zygote
8.
J Immunol Methods ; 521: 113554, 2023 10.
Article in English | MEDLINE | ID: mdl-37661049

ABSTRACT

Antibodies are essential components of the immune system with a wide range of molecular targets. They have been recognized as modalities for treating several diseases and more than 130 approved antibody-based therapeutics are available for clinical use. However, limitations remain associated with its efficacy, tissue permeability, and safety, especially in cancer treatment. Nanoparticles, particularly those responsive to external stimuli, have shown promise in improving the efficacy of antibody-based therapeutics and tissue-selective delivery. In this study, we developed a reliable and accurate method for quantifying the amount of antibody loaded onto lipid nanoparticles modified with Herceptin® (Trastuzumab), an antibody-based therapeutic used to treat HER2-positive cancers, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining. This method proved to be a suitable alternative to commonly used protein quantification techniques, which are limited by lipid interference present in the samples. Furthermore, the amount of Herceptin modified on the liposomes, measured by this method, was confirmed by Herceptin's antibody-dependent cell-mediated cytotoxicity activity. Our results demonstrate the potential of this method as a critical tool for developing tissue-selective antibody delivery systems, leading to improved efficacy and reduced side effects of antibody-based therapeutics.


Subject(s)
Liposomes , Nanoparticles , Trastuzumab , Antibodies
9.
Nihon Shokakibyo Gakkai Zasshi ; 120(7): 590-601, 2023.
Article in Japanese | MEDLINE | ID: mdl-37423730

ABSTRACT

In Japan, establishing a medical cooperation system for patients with inflammatory bowel disease (IBD) between IBD flagship and local care hospitals is a crucial task. Thus, this retrospective multicenter cohort study aims to examine the actual state of medical treatment in patients with IBD via a questionnaire survey administered to eight dependent institutes in Hokkaido, Japan. The present results clarified the clinical disparities of IBD treatment and hospital function between IBD flagship hospitals and local care hospitals. Moreover, the understanding level of IBD treatment in medical staff was significantly lower in local care than in IBD flagship hospitals. Furthermore, an abounding experience of IBD treatment affected the understanding level of IBD treatment of both medical doctors and staff. These findings indicate that selecting patients with IBD corresponding to disease activity, educational system for the current IBD treatment, and promotion of team medicine with multimedical staff can resolve clinical discrepancies between IBD flagship and local care hospitals. The IBD treatment inequities in Japan will be eliminated with the development of an appropriate medical cooperation system between IBD flagship and local care hospitals.


Subject(s)
Inflammatory Bowel Diseases , Humans , Cohort Studies , Inflammatory Bowel Diseases/therapy , Surveys and Questionnaires , Japan
10.
J Clin Med ; 12(9)2023 Apr 24.
Article in English | MEDLINE | ID: mdl-37176545

ABSTRACT

BACKGROUND: We previously conducted a pilot randomized controlled trial "the MASTER study" and demonstrated that alpha-glucosidase inhibitor miglitol and a dipeptidyl peptidase-4 inhibitor sitagliptin modified postprandial plasma excursions of active glucagon-like peptide-1 (aGLP-1) and active gastric inhibitory polypeptide (aGIP), and miglitol treatment decreased body fat mass in patients with type 2 diabetes (T2D). However, the details regarding the relationships among postprandial plasma aGLP-1 and aGIP excursions, skeletal muscle mass, and body fat mass are unclear. METHODS: We conducted a secondary analysis of the relationships among skeletal muscle mass index (SMI), total body fat mass index (TBFMI), and the incremental area under the curves (iAUC) of plasma aGLP-1 and aGIP excursions following mixed meal ingestion at baseline and after 24-week add-on treatment with either miglitol alone, sitagliptin alone, or their combination in T2D patients. RESULTS: SMI was not changed after the 24-week treatment with miglitol and/or sitagliptin. TBFMI was reduced and the rates of aGIP-iAUC change were lowered in the two groups treated with miglitol, although their correlations did not reach statistical significance. We observed a positive correlation between the rates of aGIP-iAUC and TBFMI changes and a negative correlation between the rates of TBFMI and SMI changes in T2D patients treated with sitagliptin alone whose rates of aGIP-iAUC change were elevated. CONCLUSIONS: Collectively, although T2D patients treated with miglitol and/or sitagliptin did not show altered SMI after 24-week treatment, the current study suggests that there are possible interrelationships among postprandial plasma aGIP excursion modified by sitagliptin, skeletal muscle mass, and body fat mass.

11.
BioTech (Basel) ; 12(2)2023 May 11.
Article in English | MEDLINE | ID: mdl-37218754

ABSTRACT

Genome editing, as exemplified by the CRISPR/Cas9 system, has recently been employed to effectively generate genetically modified animals and cells for the purpose of gene function analysis and disease model creation. There are at least four ways to induce genome editing in individuals: the first is to perform genome editing at the early preimplantation stage, such as fertilized eggs (zygotes), for the creation of whole genetically modified animals; the second is at post-implanted stages, as exemplified by the mid-gestational stages (E9 to E15), for targeting specific cell populations through in utero injection of viral vectors carrying genome-editing components or that of nonviral vectors carrying genome-editing components and subsequent in utero electroporation; the third is at the mid-gestational stages, as exemplified by tail-vein injection of genome-editing components into the pregnant females through which the genome-editing components can be transmitted to fetal cells via a placenta-blood barrier; and the last is at the newborn or adult stage, as exemplified by facial or tail-vein injection of genome-editing components. Here, we focus on the second and third approaches and will review the latest techniques for various methods concerning gene editing in developing fetuses.

12.
Methods Mol Biol ; 2631: 325-340, 2023.
Article in English | MEDLINE | ID: mdl-36995675

ABSTRACT

The clustered regularly interspaced short palindromic repeats (CRISPR) technology has made it possible to produce genome-edited (GE) animals more easily and rapidly than before. In most cases, GE mice are produced by microinjection (MI) or by in vitro electroporation (EP) of CRISPR reagents into fertilized eggs (zygotes). Both of these approaches require ex vivo handling of isolated embryos and their subsequent transfer into another set of mice (called recipient or pseudopregnant mice). Such experiments are performed by highly skilled technicians (especially for MI). We recently developed a novel genome editing method, called "GONAD (Genome-editing via Oviductal Nucleic Acids Delivery)," which can completely eliminate the ex vivo handling of embryos. We also made improvements to the GONAD method, termed "improved-GONAD (i-GONAD)." The i-GONAD method involves injection of CRISPR reagents into the oviduct of an anesthetized pregnant female using a mouthpiece-controlled glass micropipette under a dissecting microscope, followed by EP of the entire oviduct allowing the CRISPR reagents to enter into the zygotes present inside the oviduct, in situ. After the i-GONAD procedure, the mouse recovered from anesthesia is allowed to continue the pregnancy to full term to deliver its pups. The i-GONAD method does not require pseudopregnant female animals for embryo transfer, unlike the methods relying on ex vivo handling of zygotes. Therefore, the i-GONAD method can reduce the number of animals used, compared to the traditional methods. In this chapter, we describe some newer technical tips about the i-GONAD method. Additionally, even though the detailed protocols of GONAD and i-GONAD have been published elsewhere (Gurumurthy et al., Curr Protoc Hum Genet 88:15.8.1-15.8.12, 2016 Nat Protoc 14:2452-2482, 2019), we provide all the protocol steps of i-GONAD in this chapter so that the reader can find most of the information, needed for performing i-GONAD experiments, in one place.


Subject(s)
CRISPR-Cas Systems , Gene Editing , Humans , Pregnancy , Female , Mice , Animals , Gene Editing/methods , CRISPR-Cas Systems/genetics , Fallopian Tubes , Oviducts , Clustered Regularly Interspaced Short Palindromic Repeats/genetics , Electroporation/methods , Gonads
13.
Nat Commun ; 14(1): 1795, 2023 Mar 31.
Article in English | MEDLINE | ID: mdl-37002210

ABSTRACT

The ability to drive a spin system to state far from the equilibrium is indispensable for investigating spin structures of antiferromagnets and their functional nonlinearities for spintronics. While optical methods have been considered for spin excitation, terahertz (THz) pulses appear to be a more convenient means of direct spin excitation without requiring coupling between spins and orbitals or phonons. However, room-temperature responses are usually limited to small deviations from the equilibrium state because of the relatively weak THz magnetic fields in common approaches. Here, we studied the magnetization dynamics in a HoFeO3 crystal at room temperature. A custom-made spiral-shaped microstructure was used to locally generate a strong multicycle THz magnetic near field perpendicular to the crystal surface; the maximum magnetic field amplitude of about 2 T was achieved. The observed time-resolved change in the Faraday ellipticity clearly showed second- and third-order harmonics of the magnetization oscillation and an asymmetric oscillation behaviour. Not only the ferromagnetic vector M but also the antiferromagnetic vector L plays an important role in the nonlinear dynamics of spin systems far from equilibrium.

14.
Front Nutr ; 10: 1087471, 2023.
Article in English | MEDLINE | ID: mdl-36819693

ABSTRACT

Background: There are few reports evaluating the relationship between undernutrition and the risk of sarcopenia in type 2 diabetes mellitus (T2DM) patients. Objective: We investigated whether undernutritional status assessed by the geriatric nutritional risk index (GNRI) and controlling nutritional status (CONUT) were associated with the diagnosis of sarcopenia. Methods: This was a cross-sectional study of Japanese individuals with T2DM. Univariate or multivariate logistic regression analysis was performed to assess the association of albumin, GNRI, and CONUT with the diagnosis of sarcopenia. The optimal cut-off values were determined by the receiver operating characteristic (ROC) curve to diagnose sarcopenia. Results: In 479 individuals with T2DM, the median age was 71 years [IQR 62, 77], including 264 (55.1%) men. The median duration of diabetes was 17 [11, 23] years. The prevalence of sarcopenia was 41 (8.6%) in all, 21/264 (8.0%) in men, and 20/215 (9.3%) in women. AUCs were ordered from largest to smallest as follows: GNRI > albumin > CONUT. The cut-off values of GNRI were associated with a diagnosis of sarcopenia in multiple logistic regression analysis (odds ratio 9.91, 95% confidential interval 5.72-17.2), P < 0.001. The superiority of GNRI as compared to albumin and CONUT for detecting sarcopenia was also observed in the subclasses of men, women, body mass index (BMI) < 22, and BMI ≥ 22. Conclusions: Results showed that GNRI shows a superior diagnostic power in the diagnosis of sarcopenia. Additionally, its optimal cut-off points were useful overall or in the subclasses. Future large and prospective studies will be required to confirm the utility of the GNRI cut-off for undernutrition individuals at risk for sarcopenia.

15.
J Environ Manage ; 335: 117557, 2023 Jun 01.
Article in English | MEDLINE | ID: mdl-36842355

ABSTRACT

The application of microalgae to sequester CO2 from flue gases can be an interesting process since it can contribute to mitigate CO2 emission into the atmosphere. One obstacle of such application is the high CO2 concentration in the flue gases, which can lead to low pH in the cultivation medium and hence process failure. This study aims to investigate static CO2 gas supply for microalgae cultivation as a potential alternative that might allow applying different flue gases with different compositions and higher CO2 concentrations. Two sets of experiments were performed. First, the effect of increasing the amount of supplied carbon was tested. In the second experiment, the applicability of such system for different flue gases regarding their oxygen and carbon content was tested. In all experiments, 50 times diluted cow manure digestate was used as a culture medium. By increasing CO2 concentration up to 10% in the supplied air, microalgae growth productivity of 48.7 mg/L/d was achieved. A further improvement of microalgae growth was shown with increasing the gas/culture volume ratio. Microalgae productivity rate increased form 48.7 mg/L/d to 73.5 mg/L/d when the volume of gas increased from 47% to 81% of total volume. Applying CO2 in air (O2 content around 20%) or in N2 (O2 content less than 2%) didn't show any difference regarding inorganic carbon dissolution, pH, ammonium nitrogen removal, CO2 fixation or biomass productivity. Generally, it can be concluded that static gas supply for microalgae cultivation can allow the application of different flue gases from different industries with low or high O2 content and with CO2 concentration as high as 20%. According to our results, a microalgae cultivation system with continuous static gas supply was proposed.


Subject(s)
Gases , Microalgae , Cattle , Animals , Gases/chemistry , Carbon Dioxide/chemistry , Manure , Carbon , Biomass
16.
Spine Surg Relat Res ; 6(5): 540-544, 2022 Sep 27.
Article in English | MEDLINE | ID: mdl-36348674

ABSTRACT

Introduction: Lumbar spondylolysis is common in pediatric athletes, and many athletes can return to sports with conservative treatment. There are two initial treatment strategies: bony union or pain management, but the outcomes of these strategies have not been clarified. The purpose of this study is to investigate the rates of return to sports (RTS) and recurrence in pediatric athletes after conservative treatment for lumbar spondylolysis and to compare both treatment strategies. Methods: A total of 180 patients with lumbar spondylolysis were managed with a trunk brace and cessation of sports activity (bone union [BU] group, n=95) or treated for pain only (pain management [PM] group, n=85). RTS and recurrence rates according to type of conservative treatment were compared. Results: The RTS rate was 98.9% in the BU group and 97.6% in the PM group at 4.7±1.9 and 1.8±1.7 months, respectively. Recurrence occurred in 7.4% of patients in the BU group at 19.0±16.0 months and in 4.8% of the PM group at 17.8±5.2 months. Conclusions: The RTS rate in pediatric athletes with lumbar spondylolysis was high at more than 95%, regardless of type of conservative treatment. The mean time to RTS was longer in the BU group than in the PM group (4.7 vs. 1.8 months) because of the time required for bone healing. There were several cases of recurrence after RTS. Strategies to prevent recurrence of lumbar spondylolysis in pediatric athletes are discussed.

17.
Int J Mol Sci ; 23(19)2022 Oct 06.
Article in English | MEDLINE | ID: mdl-36233155

ABSTRACT

Fibroblast growth factor 5 (FGF5) is an important molecule required for the transition from anagen to catagen phase of the mammalian hair cycle. We previously reported that Syrian hamsters harboring a 1-bp deletion in the Fgf5 gene exhibit excessive hair growth in males. Herein, we generated Fgf5 mutant mice using genome editing via oviductal nucleic acid delivery (GONAD)/improved GONAD (i-GONAD), an in vivo genome editing system used to target early embryos present in the oviductal lumen, to study gender differences in hair length in mutant mice. The two lines (Fgf5go-malc), one with a 2-bp deletion (c.552_553del) and the other with a 1-bp insertion (c.552_553insA) in exon 3 of Fgf5, were successfully established. Each mutation was predicted to disrupt a part of the FGF domain through frameshift mutation (p.Glu184ValfsX128 or p.Glu184ArgfsX128). Fgf5go-malc1 mice had heterogeneously distributed longer hairs than wild-type mice (C57BL/6J). Notably, this change was more evident in males than in females (p < 0.0001). Immunohistochemical analysis revealed the presence of FGF5 protein in the dermal papilla and outer root sheath of the hair follicles from C57BL/6J and Fgf5go-malc1 mice. Histological analysis revealed that the prolonged anagen phase might be the cause of accelerated hair growth in Fgf5go-malc1 mice.


Subject(s)
Fibroblast Growth Factor 5 , Hair , Sex Characteristics , Animals , Female , Fibroblast Growth Factor 5/genetics , Fibroblast Growth Factor 5/metabolism , Hair/growth & development , Hair Follicle/metabolism , Male , Mice , Mice, Inbred C57BL , Mutation , Nucleic Acids/metabolism , Sex Factors
18.
J Clin Med ; 11(20)2022 Oct 15.
Article in English | MEDLINE | ID: mdl-36294410

ABSTRACT

Tissue-specific stem cells exist in tissues and organs, such as skin and bone marrow. However, their pluripotency is limited compared to embryonic stem cells. Culturing primary cells on plastic tissue culture dishes can result in the loss of multipotency, because of the inability of tissue-specific stem cells to survive in feeder-less dishes. Recent findings suggest that culturing primary cells in medium containing feeder cells, particularly genetically modified feeder cells expressing growth factors, may be beneficial for their survival and proliferation. Therefore, the aim of this study was to elucidate the role of genetically modified human feeder cells expressing growth factors in maintaining the integrity of primary cultured human deciduous dental pulp cells. Feeder cells expressing leukemia inhibitory factor, bone morphogenetic protein 4, and basic fibroblast growth factor were successfully engineered, as evidenced by PCR. Co-culturing with mitomycin-C-treated feeder cells enhanced the proliferation of newly isolated human deciduous dental pulp cells, promoted their differentiation into adipocytes and neurons, and maintained their stemness properties. Our findings suggest that genetically modified human feeder cells may be used to maintain the integrity of primary cultured human deciduous dental pulp cells.

19.
Int J Mol Sci ; 23(18)2022 Sep 14.
Article in English | MEDLINE | ID: mdl-36142589

ABSTRACT

Improved genome editing via oviductal nucleic acids delivery (i-GONAD) is a new technology enabling in situ genome editing of mammalian zygotes exiting the oviductal lumen, which is now available in mice, rats, and hamsters. In this method, CRISPR/Cas9 genome-editing reagents are delivered directly to the oviducts of pregnant animals (corresponding to late zygote stage). After intraoviductal instillation, electric shock to the entire oviduct was provided with a specialized electroporation (EP) device to drive the genome editing reagents into the zygotes present in the oviductal lumen. i-GONAD toward early zygotes has been recognized as difficult, because they are tightly surrounded by a cumulus cell layer, which often hampers effective transfer of nucleic acids to zygotes. However, in vivo EP three min after intraoviductal instillation of the genome-editing reagents enabled genome editing of early zygotes with an efficiency of 70%, which was in contrast with the rate of 18% when in vivo EP was performed immediately after intraoviductal instillation at Day 0.5 of pregnancy (corresponding to 13:00-13:30 p.m. on the day when vaginal plug was recognized after natural mating). We also found that addition of hyaluronidase, an enzyme capable of removing cumulus cells from a zygote, slightly enhanced the efficiency of genome editing in early zygotes. These findings suggest that cumulus cells surrounding a zygote can be a barrier for efficient generation of genome-edited mouse embryos and indicate that a three-minute interval before in vivo EP is effective for achieving i-GONAD-mediated genome editing at the early zygote stage. These results are particularly beneficial for researchers who want to perform genome editing experiments targeting early zygotes.


Subject(s)
Gene Editing , Nucleic Acids , Animals , CRISPR-Cas Systems , Electroporation/methods , Female , Gene Editing/methods , Gonads , Humans , Hyaluronoglucosaminidase/genetics , Mammals/genetics , Mice , Oviducts , Pregnancy , Rats , Ribonucleoproteins/genetics , Zygote
20.
Phys Rev Lett ; 129(10): 107201, 2022 Sep 02.
Article in English | MEDLINE | ID: mdl-36112457

ABSTRACT

Spin current is a key to realizing various phenomena and functionalities related to spintronics. Recently, the possibility of generating spin current through a photogalvanic effect of magnons was pointed out theoretically. However, neither a candidate material nor a general formula for calculating the photogalvanic spin current in materials is known so far. In this Letter, we develop a general formula for the photogalvanic spin current through a magnetic resonance process. This mechanism involves a one-magnon excitation process in contrast to the two-particle processes studied in earlier works. Using the formula, we show that GHz and THz waves create a large photogalvanic spin current in the antiferromagnetic phase of bilayer CrI_{3} and CrBr_{3}. The large spin current arises from an optical process involving two magnon bands, which is a contribution unknown to date. This spin current appears only in the antiferromagnetic ordered phase and is reversible by controlling the order parameter. These results open a route to material design for the photogalvanic effect of magnetic excitations.

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