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1.
Insects ; 15(5)2024 May 16.
Article in English | MEDLINE | ID: mdl-38786917

ABSTRACT

Honey bees (Apis mellifera L.) play vital roles as agricultural pollinators and honey producers. However, global colony losses are increasing due to multiple stressors, including malnutrition. Our study evaluated the effects of four pollen substitute diets (Diet 1, Diet 2, Diet 3, and Control) through field and cage experiments, analyzing 11 parameters and 21 amino acids. Notably, Diet 1 demonstrated significantly superior performance in the field experiment, including the number of honey bees, brood area, consumption, preference, colony weight, and honey production. In the cage experiment, Diet 1 also showed superior performance in dried head and thorax weight and vitellogenin (vg) gene expression levels. Canonical discriminant and principle component analyses highlighted Diet 1's distinctiveness, with histidine, diet digestibility, consumption, vg gene expression levels, and isoleucine identified as key factors. Arginine showed significant correlations with a wide range of parameters, including the number of honey bees, brood area, and consumption, with Diet 1 exhibiting higher levels. Diet 1, containing apple juice, soytide, and Chlorella as additive components, outperformed the other diets, suggesting an enhanced formulation for pollen substitute diets. These findings hold promise for the development of more effective diets, potentially contributing to honey bee health.

2.
Br J Dermatol ; 2024 May 30.
Article in English | MEDLINE | ID: mdl-38815138

ABSTRACT

BACKGROUND: Amelanotic acral melanoma (AAM) is a rare type of acral melanoma associated with poor prognosis. OBJECTIVES: We aimed to investigate the transcriptomic differences between AAM and pigmented acral melanoma (PAM). METHODS: The differences in spatially resolved transcriptome profiles of 9 AAM patients with 29 regions of interest (ROIs) and 11 PAM patients with 46 ROIs were investigated using S100b and CD3 morphology markers. RESULTS: In S100b-positive tumor cell areas, we detected 11 upregulated differentially expressed genes (DEGs), including chaperone/ubiquitin-associated DEGs, and 82 downregulated DEGs, including human leukocyte antigen, in AAMs compared with PAMs. Protein-protein interaction network and pathway analyses revealed significant enrichment of dysregulated translational and nonsense-mediated decay pathways but significant decreases in antigen processing and presentation, interferon signaling, and melanin biosynthesis pathways in S100b-positive ROIs of AAMs compared with those of PAMs. In tumor-associated immune cell areas, the numbers of CD8 T cells (p = 0.044) and M1 macrophages (p = 0.014) were significantly decreased, whereas those of monocytes (p = 0.045) and endothelial cells (p = 0.04) were increased in AAMs compared with those in PAMs. CONCLUSIONS: In conclusion, these findings could widen our understanding of the biological differences between AAMs and PAMs that might result in a different clinical course.

3.
JCO Precis Oncol ; 8: e2300495, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38635931

ABSTRACT

PURPOSE: High-grade neuroendocrine carcinoma (HGNEC) of the lung is an aggressive cancer with a complex biology. We aimed to explore the prognostic value of genetic aberrations and poly(ADP-ribose) polymerase-1 (PARP1) expression in HGNEC and to establish a novel prognostic model. MATERIALS AND METHODS: We retrospectively enrolled 191 patients with histologically confirmed HGNEC of the lung. Tumor tissues were analyzed using PARP1 immunohistochemistry (IHC; N = 191) and comprehensive cancer panel sequencing (n = 102). Clinical and genetic data were used to develop an integrated Cox hazards model. RESULTS: Strong PARP1 IHC expression (intensity 3) was observed in 153 of 191 (80.1%) patients, and the mean PARP1 H-score was 285 (range, 5-300). To develop an integrated Cox hazard model, our data set included information from 357 gene mutations and 19 clinical profiles. When the targeted mutation profiles were combined with clinical profiles, 12 genes (ATRX, CCND2, EXT2, FGFR2, FOXO1, IL21R, MAF, TGM7, TNFAIP3, TP53, TSHR, and DDR2) were identified as prognostic factors for survival. The integrated Cox hazard model, which combines mutation profiles with a baseline model, outperformed the baseline model (incremental area under the curve 0.84 v 0.78; P = 8.79e-12). The integrated model stratified patients into high- and low-risk groups with significantly different disease-free and overall survival (integrated model: hazard ratio, 7.14 [95% CI, 4.07 to 12.54]; P < .01; baseline model: 4.38 [2.56 to 7.51]; P < .01). CONCLUSION: We introduced a new prognostic model for HGNEC that combines genetic and clinical data. The integrated Cox hazard model outperformed the baseline model in predicting the survival of patients with HGNEC.


Subject(s)
Carcinoma, Neuroendocrine , Lung Neoplasms , Humans , Prognosis , Poly (ADP-Ribose) Polymerase-1/genetics , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Retrospective Studies , Carcinoma, Neuroendocrine/genetics , Lung/metabolism , Lung/pathology , Genomics
4.
Bioengineering (Basel) ; 11(4)2024 Mar 29.
Article in English | MEDLINE | ID: mdl-38671758

ABSTRACT

The purposes of this study are to establish and validate a finite element (FE) model using finite element analysis methods and to identify optimal loading conditions to simulate masticatory movement. A three-dimensional FE model of the maxillary and mandibular cortical bone, cancellous bone, and gingiva was constructed based on edentulous cone-beam-computed tomography data. Dental computer-aided design software was used to design the denture base and artificial teeth to produce a complete denture. Mesh convergence was performed to derive the optimal mesh size, and validation was conducted through comparison with mechanical test results. The mandible was rotated step-by-step to induce movements similar to actual mastication. Results showed that there was less than a 6% difference between the mechanical test and the alveolar bone-complete denture. It opened 10° as set in the first stage, confirming that the mouth closed 7° in the second stage. Occlusal contact occurred between the upper and lower artificial teeth as the mouth closed the remaining angle of 3° in the third stage while activating the masseter muscle. These results indicate that the FE model and masticatory loading conditions developed in this study can be applied to analyze biomechanical effects according to the wearing of dentures with various design elements applied.

5.
Int J Mol Sci ; 25(8)2024 Apr 12.
Article in English | MEDLINE | ID: mdl-38673857

ABSTRACT

Honey bees are commonly used to study metabolic processes, yet the molecular mechanisms underlying nutrient transformation, particularly proteins and their effects on development, health, and diseases, still evoke varying opinions among researchers. To address this gap, we investigated the digestibility and transformation of water-soluble proteins from four artificial diets in long-lived honey bee populations (Apis mellifera ligustica), alongside their impact on metabolism and DWV relative expression ratio, using transcriptomic and protein quantification methods. Diet 2, characterized by its high protein content and digestibility, was selected for further analysis from the other studied diets. Subsequently, machine learning was employed to identify six diet-related molecular markers: SOD1, Trxr1, defensin2, JHAMT, TOR1, and vg. The expression levels of these markers were found to resemble those of honey bees who were fed with Diet 2 and bee bread, renowned as the best natural food. Notably, honey bees exhibiting chalkbrood symptoms (Control-N) responded differently to the diet, underscoring the unique nutritional effects on health-deficient bees. Additionally, we proposed a molecular model to elucidate the transition of long-lived honey bees from diapause to development, induced by nutrition. These findings carry implications for nutritional research and beekeeping, underscoring the vital role of honey bees in agriculture.


Subject(s)
Diet , Bees/genetics , Bees/metabolism , Animals , Diet/veterinary , Insect Proteins/genetics , Insect Proteins/metabolism , Animal Feed/analysis , Biomarkers , Transcriptome , Gene Expression Regulation/drug effects
6.
Insects ; 15(3)2024 Mar 18.
Article in English | MEDLINE | ID: mdl-38535396

ABSTRACT

Mosquitoes, the primary vectors of arboviruses, harbor a diverse microbiome that plays a crucial role in their development, immunity, and vector competence. The composition of the mosquito microbiome is heavily influenced by the environment and habitats. Therefore, identifying the relationship between the habitat and the mosquito's microbial community can improve the overall understanding of mosquito biology. However, The microbiome profiles of Culex tritaeniorhynchus and Culex orientalis, known as transmission vectors of the Japanese encephalitis virus, are poorly understood. Using 16S rRNA Illumina sequencing, we hereby investigated the microbial profiles in these two mosquito species collected in several areas in the Republic of Korea. Thirty-six prevalent bacterial families were identified from these mosquito species. The microbial composition variations were primarily influenced by the mosquito collecting sites. Moreover, species biomarkers were identified by utilizing the regional specificity of the mosquito microbiome. Based on the microbiome profiles representing high similarity, Culex orientalis may share an ecological niche with Culex tritaeniorhynchus.

8.
Mol Diagn Ther ; 28(3): 239-248, 2024 May.
Article in English | MEDLINE | ID: mdl-38538969

ABSTRACT

Hemophilia is a genetic disorder that is caused by mutations in coagulation factor VIII (hemophilia A) or IX (hemophilia B) genes resulting in blood clotting disorders. Despite advances in therapies, such as recombinant proteins and products with extended half-lives, the treatment of hemophilia still faces two major limitations: the short duration of therapeutic effect and production of neutralizing antibodies against clotting factors (inhibitor). To overcome these limitations, new hemophilia treatment strategies have been established such as gene therapy, bispecific antibody, and rebalancing therapy. Although these strategies have shown promising results, it is difficult to achieve a permanent therapeutic effect. Advances in the clustered regularly interspaced short palindromic repeat (CRISPR) technology have allowed sustainable treatment by correcting mutated genes. Since genome editing generates irreversible changes in host genome, safety must be ensured by delivering target organs. Therefore, the delivery tool of the CRISPR system is crucial for safe, accurate, and efficient genome editing. Recently, non-viral vector lipid nanoparticles (LNPs) have emerged as safer tools for delivering CRISPR systems than other viral vectors. Several previous hemophilia pre-clinical studies using LNP-CRISPR showed that sufficient and sustainable therapeutic effects, which means that LNP-CRISPR-mediated genome-editing therapy can be a valid option for the treatment of hemophilia. In this paper, we summarize the latest advancements in the successful treatment of hemophilia and the potential of CRISPR-mediated genome-editing therapy using LNPs.


Subject(s)
CRISPR-Cas Systems , Gene Editing , Genetic Therapy , Hemophilia A , Nanoparticles , Humans , Hemophilia A/therapy , Hemophilia A/genetics , Gene Editing/methods , Genetic Therapy/methods , Animals , Hemophilia B/therapy , Hemophilia B/genetics , Factor VIII/genetics , Factor VIII/therapeutic use , Lipids , Genetic Vectors/genetics , Clustered Regularly Interspaced Short Palindromic Repeats/genetics
9.
Insects ; 15(2)2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38392520

ABSTRACT

The nutritional quality of a colony significantly affects its health and strength, particularly because it is required for population growth in the early spring. We investigated the impact of various artificial pollen substitute diets on colony performance in the Republic of Korea during early spring, a critical period for colony health and growth. The colonies were provided with different diets, including the commercial product Megabee (positive control), our developed diet Test A, and four upgraded versions (Diet 1, Diet 2, Diet 3, and Diet 4) of Test A. The negative control group received no supplementary feed. Over 63 days, we observed 24 experimental colonies and assessed various parameters at the colony and individual levels. The results revealed that Diet 2 had the highest consumption and had the most positive impact on population growth, the capped brood area, colony weight, honey bees' weight, and vitellogenin levels. These findings suggested that Diet 2 is most attractive to honey bees and thus holds great promise for improving colony maintenance and development during the crucial early spring period.

10.
Sci Rep ; 14(1): 4212, 2024 02 20.
Article in English | MEDLINE | ID: mdl-38378928

ABSTRACT

Although various comorbidities have been noted to be associated with atopic dermatitis (AD) and psoriasis, few studies have compared comorbidities between the two diseases, and little is known about whether these comorbidities vary by the subtypes of psoriasis. In this study of 1:1 age- and sex-matched pair analysis between patients diagnosed with either psoriasis or AD at Asan Medical Center between 1991 and 2020, comorbidities, as determined by the International Classification of Diseases-10 codes, and likelihood ratios of metabolic and neurologic comorbidities in psoriasis compared with AD were studied using a logistic regression model. Among a total of 14,128 patients, the psoriasis group had higher odds of obesity (odds ratio [95% confidence interval]: 1.49 [1.34-1.66]), hypertension (1.14 [1.03-1.26]), diabetes mellitus (1.46 [1.29-1.66]), chronic kidney disease (1.59 [1.22-2.08]), and Parkinson's disease (2.1 [1.15-3.83]) than the AD group. Subgroup analysis revealed that patients with plaque psoriasis had higher odds of obesity (1.18 [1.05-1.33]), hypertension (1.18 [1.06-1.32]), diabetes mellitus (1.53 [1.34-1.75]), chronic kidney disease (1.66 [1.26-2.17]), and Parkinson's disease (2.12 [1.16-3.88]) compared with AD. Meanwhile, guttate psoriasis was associated with higher odds of dementia (3.63 [1.06-12.40]) and patients with generalized pustular psoriasis showed higher odds of diabetes mellitus (5.42 [1.56-18.83]) compared with AD. In conclusion, Asian patients with all types of psoriasis should be closely monitored for the development of metabolic and neurologic diseases, especially men and those aged ≥ 40 years.


Subject(s)
Dermatitis, Atopic , Diabetes Mellitus , Hypertension , Parkinson Disease , Psoriasis , Renal Insufficiency, Chronic , Male , Humans , Dermatitis, Atopic/epidemiology , Psoriasis/complications , Psoriasis/epidemiology , Comorbidity , Hypertension/complications , Hypertension/epidemiology , Diabetes Mellitus/epidemiology , Obesity/epidemiology
11.
Cancers (Basel) ; 16(4)2024 Feb 16.
Article in English | MEDLINE | ID: mdl-38398196

ABSTRACT

Refraction-contrast computed tomography based on X-ray dark-field imaging (XDFI) using synchrotron radiation (SR) has shown superior resolution compared to conventional absorption-based methods and is often comparable to pathologic examination under light microscopy. This study aimed to investigate the potential of the XDFI technique for clinical application in lung cancer diagnosis. Two types of lung specimens, primary and secondary malignancies, were investigated using an XDFI optic system at beamline BL14B of the High-Energy Accelerator Research Organization Photon Factory, Tsukuba, Japan. Three-dimensional reconstruction and segmentation were performed on each specimen. Refraction-contrast computed tomographic images were compared with those obtained from pathological examinations. Pulmonary microstructures including arterioles, venules, bronchioles, alveolar sacs, and interalveolar septa were identified in SR images. Malignant lesions could be distinguished from the borders of normal structures. The lepidic pattern was defined as the invasive component of the same primary lung adenocarcinoma. The SR images of secondary lung adenocarcinomas of colorectal origin were distinct from those of primary lung adenocarcinomas. Refraction-contrast images based on XDFI optics of lung tissues correlated well with those of pathological examinations under light microscopy. This imaging method may have the potential for use in lung cancer diagnosis without tissue damage. Considerable equipment modifications are crucial before implementing them from the lab to the hospital in the near future.

12.
Pathogens ; 13(2)2024 Jan 28.
Article in English | MEDLINE | ID: mdl-38392860

ABSTRACT

Honeybees (Apis mellifera) are pollinating agents of economic importance. The role of the gut microbiome in honeybee health has become increasingly evident due to its relationship with immune function, growth, and development. Although their dynamics at various developmental stages have been documented, their dynamics during the era of colony collapse disorder and immunogenic potential, which are connected to the antagonistic immune response against pathogens, need to be elucidated. Using 16S rRNA gene Illumina sequencing, the results indicated changes in the gut microbiota with the developmental stage. The bacterial diversity of fifth stage larva was significantly different among the other age groups, in which Fructobacillus, Escherichia-Shigella, Bombella, and Tyzzerella were unique bacteria. In addition, the diversity of the worker bee microbiome was distinct from that of the younger microbiome. Lactobacillus and Gilliamella remained conserved throughout the developmental stages, while Bifidobacterium colonized only worker bees. Using an in silico approach, the production potential of lipopolysaccharide-endotoxin was predicted. Forager bees tend to have a higher abundance rate of Gram-negative bacteria. Our results revealed the evolutionary importance of some microbiome from the larval stage to the adult stage, providing insight into the potential dynamics of disease response and susceptibility. This finding provides a theoretical foundation for furthering the understanding of the function of the gut microbiota at various developmental stages related to probiotic development and immunogenic potential.

13.
Brain ; 147(6): 2114-2127, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38227798

ABSTRACT

Mutations in the Microrchidia CW-type zinc finger 2 (MORC2) GHKL ATPase module cause a broad range of neuropathies, such as Charcot-Marie-Tooth disease type 2Z; however, the aetiology and therapeutic strategy are not fully understood. Previously, we reported that the Morc2a p.S87L mouse model exhibited neuropathy and muscular dysfunction through DNA damage accumulation. In the present study, we analysed the gene expression of Morc2a p.S87L mice and designated the primary causing factor. We investigated the pathological pathway using Morc2a p.S87L mouse embryonic fibroblasts and human fibroblasts harbouring MORC2 p.R252W. We subsequently assessed the therapeutic effect of gene therapy administered to Morc2a p.S87L mice. This study revealed that Morc2a p.S87L causes a protein synthesis defect, resulting in the loss of function of Morc2a and high cellular apoptosis induced by high hydroxyl radical levels. We considered the Morc2a GHKL ATPase domain as a therapeutic target because it simultaneously complements hydroxyl radical scavenging and ATPase activity. We used the adeno-associated virus (AAV)-PHP.eB serotype, which has a high CNS transduction efficiency, to express Morc2a or Morc2a GHKL ATPase domain protein in vivo. Notably, AAV gene therapy ameliorated neuropathy and muscular dysfunction with a single treatment. Loss-of-function characteristics due to protein synthesis defects in Morc2a p.S87L were also noted in human MORC2 p.S87L or p.R252W variants, indicating the correlation between mouse and human pathogenesis. In summary, CMT2Z is known as an incurable genetic disorder, but the present study demonstrated its mechanisms and treatments based on established animal models. This study demonstrates that the Morc2a p.S87L variant causes hydroxyl radical-mediated neuropathy, which can be rescued through AAV-based gene therapy.


Subject(s)
Genetic Therapy , Animals , Mice , Humans , Genetic Therapy/methods , Hydroxyl Radical/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Charcot-Marie-Tooth Disease/genetics , Charcot-Marie-Tooth Disease/metabolism , Charcot-Marie-Tooth Disease/therapy , Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/metabolism , Fibroblasts/metabolism , Dependovirus/genetics
14.
J Cosmet Dermatol ; 23(3): 931-937, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38169089

ABSTRACT

BACKGROUND: Prurigo nodularis (PN) is an intensively pruritic skin disease that negatively influences quality of life. Cryosim-1 (Intrinsic IB Spot) is a synthetic, selective transient receptor potential melastatin 8 agonist. AIMS: To investigate the efficacy and safety of cryosim-1 in PN patients. PATIENTS/METHODS: A randomized, double-blinded, placebo-controlled clinical trial including 30 patients was conducted. The numerical rating scale (NRS) of pruritus was evaluated before and 2 h after cryosim-1 application at every visit. RESULTS: At week 8, the mean pruritus NRS before serum application (4.7 ± 0.4 treatment, 6.1 ± 0.5 placebo; p = 0.045) and 2 h after serum application (2.8 ± 0.4 treatment, 4.3 ± 0.5 placebo; p = 0.031) were significantly lower in the treatment group, and the mean NRS for sleep disorder was significantly lower in the treatment group (2.2 ± 0.5 treatment, 4.2 ± 0.8 placebo; p = 0.031). The mean satisfaction scales for pruritus improvement were significantly higher in the treatment group (7.2 ± 0.6) than in the placebo group (4.0 ± 0.9; p = 0.005). There was no difference in TEWL between the two groups, and no adverse reactions were reported. CONCLUSIONS: Cryosim-1 is a safe and effective topical treatment for PN patients.


Subject(s)
Prurigo , TRPM Cation Channels , Humans , Prurigo/drug therapy , Quality of Life , Pruritus/drug therapy , Pruritus/etiology , Administration, Topical , Research Design , TRPM Cation Channels/agonists , Membrane Proteins
15.
Mol Ther Nucleic Acids ; 34: 102050, 2023 Dec 12.
Article in English | MEDLINE | ID: mdl-37916225

ABSTRACT

Gene therapy and rebalancing therapy have emerged as promising approaches for treating hemophilia A, but there are limitations, such as temporary efficacy due to individual differences. Genome editing for hemophilia has shown long-term therapeutic potential in preclinical trials. However, a cautious approach is necessary because genome editing is irreversible. Therefore, we attempted to induce low-level human factor 8 (hF8) gene knockin (KI) using 244-cis lipid nanoparticles and low-dose adeno-associated virus to minimize side effects and achieve a therapeutic threshold in hemophilia A mice. We selected the serpin family C member 1, SerpinC1, locus as a target to enable a combined rebalancing strategy with hF8 KI to augment efficacy. This strategy improved blood coagulation activity and reduced hemophilic complications without adverse effects. Furthermore, hemophilic mice with genome editing exhibit enhanced survival for 40 weeks. Here, we demonstrate an effective, safe, and sustainable treatment for hemophilia A. This study provides valuable information to establish safe and long-term genome-editing-mediated treatment strategies for treating hemophilia and other protein-deficient genetic diseases.

16.
ACS Appl Mater Interfaces ; 15(25): 30975-30984, 2023 Jun 28.
Article in English | MEDLINE | ID: mdl-37310803

ABSTRACT

A series of Al-based isomorphs (CAU-10H, MIL-160, KMF-1, and CAU-10pydc) were synthesized using isophthalic acid (ipa), 2,5-furandicarboxylic acid (fdc), 2,5-pyrrole dicarboxylic acid (pyrdc), and 3,5-pyridinedicarboxylic acid (pydc), respectively. These isomorphs were systematically investigated to identify the best adsorbent for effectively separating C2H6/C2H4. All CAU-10 isomorphs exhibited preferential adsorption of C2H6 over that of C2H4 in mixture. CAU-10pydc exhibited the best C2H6/C2H4 selectivity (1.68) and the highest C2H6 uptake (3.97 mmol g-1) at 298 K and 1 bar. In the breakthrough experiment using CAU-10pydc, 1/1 (v/v) and 1/15 (v/v) C2H6/C2H4 gas mixtures were successfully separated into high-purity C2H4 (>99.95%), with remarkable productivities of 14.0 LSTP kg-1 and 32.0 LSTP kg-1, respectively, at 298 K. Molecular simulations revealed that the exceptional separation performance of CAU-10pydc originated from the increased porosity and reduced electron density of the pyridine ring of pydc, leading to a relatively larger decrease in π-π interactions with C2H4 than in the C-H···π interactions with C2H6. This study demonstrates that the pore size and geometry of the CAU-10 platform are modulated by the inclusion of heteroatom-containing benzene dicarboxylate or heterocyclic rings of dicarboxylate-based organic linkers, thereby fine-tuning the C2H6/C2H4 separation ability. CAU-10pydc was determined to be an optimum adsorbent for this challenging separation.

17.
Sci Rep ; 13(1): 9081, 2023 06 05.
Article in English | MEDLINE | ID: mdl-37277359

ABSTRACT

Microbiota in the mosquito plays an important role in their behavior and vector competence. The composition of their microbiome is strongly influenced by the environment, especially their habitat. The microbiome profiles of adult female Anopheles sinensis mosquitoes from malaria hyperendemic and hypoendemic areas in Republic of Korea were compared using 16S rRNA Illumina sequencing. In different epidemiology groups, the alpha and beta diversity analyses were significant. The major bacterial phylum was Proteobacteria. The most abundant species in the microbiome of hyperendemic mosquitoes were the genera Staphylococcus, Erwinia, Serratia, and Pantoea. Notably, a distinct microbiome profile characterized by the dominance of Pseudomonas synxantha was identified in the hypoendemic area, suggesting a potential correlation between the microbiome profiles and the incidence of malaria cases.


Subject(s)
Anopheles , Malaria , Microbiota , Animals , Female , RNA, Ribosomal, 16S/genetics , Mosquito Vectors , Malaria/epidemiology , Anopheles/genetics , Republic of Korea/epidemiology
18.
Small ; 19(39): e2302023, 2023 09.
Article in English | MEDLINE | ID: mdl-37246275

ABSTRACT

Deoxyribonuclease-I (DNase-I), a representative endonuclease, is an important biomarker for the diagnosis of infectious diseases and cancer progression. However, enzymatic activity decreases rapidly ex vivo, which highlights the need for precise on-site detection of DNase-I. Here, a localized surface plasmon resonance (LSPR) biosensor that enables the simple and rapid detection of DNase-I is reported. Moreover, a novel technique named electrochemical deposition and mild thermal annealing (EDMIT) is applied to overcome signal variations. By taking advantage of the low adhesion of gold clusters on indium tin oxide substrates, both the uniformity and sphericity of gold nanoparticles are increased under mild thermal annealing conditions via coalescence and Ostwald ripening. This ultimately results in an approximately 15-fold decrease in LSPR signal variations. The linear range of the fabricated sensor is 20-1000 ng mL-1 with a limit of detection (LOD) of 127.25 pg mL-1 , as demonstrated by spectral absorbance analyses. The fabricated LSPR sensor stably measured DNase-I concentrations from samples collected from both an inflammatory bowel disease (IBD) mouse model, as well as human patients with severe COVID-19 symptoms. Therefore, the proposed LSPR sensor fabricated via the EDMIT method can be used for early diagnosis of other infectious diseases.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Animals , Mice , Humans , Surface Plasmon Resonance/methods , Gold/chemistry , Metal Nanoparticles/chemistry , Biosensing Techniques/methods , Deoxyribonucleases
19.
Mol Ther Nucleic Acids ; 32: 161-172, 2023 Jun 13.
Article in English | MEDLINE | ID: mdl-37064777

ABSTRACT

Recent therapeutic strategies for hemophilia include long-term therapeutic gene expression using adeno-associated virus (AAV) and rebalancing therapy via the downregulation of anticoagulant pathways. However, these approaches have limitations in immune responses or insufficiency to control acute bleeding. Thus, we developed a therapeutic strategy for hemophilia B by a combined rebalancing and human factor 9 (hF9) gene knockin (KI) using a lipid nanoparticle (LNP) and AAV. Antithrombin (AT; Serpin Family C Member 1 [Serpinc1]) was selected as the target anticoagulation pathway for the gene KI. First, the combined use of LNP-clustered regularly interspaced short palindromic repeats (CRISPR) and AAV donor resulted in 20% insertions or deletions (indels) in Serpinc1 and 67% reduction of blood mouse AT concentration. Second, hF9 coding sequences were integrated into approximately 3% of the target locus. hF9 KI yielded approximately 1,000 ng/mL human factor IX (hFIX) and restored coagulation activity to a normal level. LNP-CRISPR injection caused sustained AT downregulation and hFIX production up to 63 weeks. AT inhibition and hFIX protein-production ability could be maintained by the proliferation of genetically edited hepatocytes in the case of partial hepatectomy. The co-administration of AAV and LNP showed no severe side effects except random integrations. Our results demonstrate hemophilia B therapy by a combination of rebalancing and hF9 KI using LNP and AAV.

20.
Small ; 19(1): e2204336, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36403243

ABSTRACT

This study focuses on the applicability of single-atom Mo-doped graphitic carbon nitride (GCN) nanosheets which are specifically engineered with high surface area (exfoliated GCN), NH2 rich edges, and maximum utilization of isolated atomic Mo for propylene carbonate (PC) production through CO2 cycloaddition of propylene oxide (PO). Various operational parameters are optimized, for example, temperature (130 °C), pressure (20 bar), catalyst (Mo2 GCN), and catalyst mass (0.1 g). Under optimal conditions, 2% Mo-doped GCN (Mo2 GCN) has the highest catalytic performance, especially the turnover frequency (TOF) obtained, 36.4 h-1 is higher than most reported studies. DFT simulations prove the catalytic performance of Mo2 GCN significantly decreases the activation energy barrier for PO ring-opening from 50-60 to 4.903 kcal mol-1 . Coexistence of Lewis acid/base group improves the CO2 cycloaddition performance by the formation of coordination bond between electron-deficient Mo atom with O atom of PO, while NH2 surface group disrupts the stability of CO2 bond by donating electrons into its low-level empty orbital. Steady-state process simulation of the industrial-scale consumes 4.4 ton h-1 of CO2 with PC production of 10.2 ton h-1 . Techno-economic assessment profit from Mo2 GCN is estimated to be 60.39 million USD year-1 at a catalyst loss rate of 0.01 wt% h-1 .

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