Search Results (99)

The piggyBac+TET-on transposon induction system has a high efficiency in integrating exogenous genes in multiple cell types, can precisely integrate to reduce genomic damage, has a flexible gene expression regulation, and a strong genetic stability. When used in conjunction with somatic cell nuclear transfer experiments, it can precisely and effectively reveal the intrinsic mechanisms of early biological development. This study successfully reprogrammed black-boned sheep fibroblasts (SFs) into induced pluripotent stem cells (iPSCs) using the piggyBac+TET-on transposon system and investigated their impact on early embryonic development. Seven exogenous reprogramming factors (bovine OCT4, SOX2, KLF4, cMyc, porcine NANOG, Lin-28, and SV40 Large T) were delivered into SFs, successfully inducing iPSCs. A growth performance analysis revealed that iPSC clones exhibited a raised or flat morphology with clear edges, positive alkaline phosphatase staining, and normal karyotypes. The transcriptome analysis indicated a significant enrichment of iPSCs in oxidative phosphorylation and cell proliferation pathways, with an up-regulated expression of the ATP5B, SDHB, Bcl-2, CDK1, and Cyclin D1 genes and a down-regulated expression of BAX (p < 0.05). Somatic cell nuclear transfer experiments demonstrated that the cleavage rate (85% ± 2.12) and blastocyst rate (52% ± 2.11) of the iPSCs were significantly higher than those of the SFs (p < 0.05). The detection of trilineage marker genes confirmed that the expression levels of endoderm (DCN, NANOS3, FOXA2, FOXD3, SOX17), mesoderm (KDR, CD34, NFH), and ectoderm (NEUROD) markers in iPSCs were significantly higher than in SFs (p < 0.01). The findings demonstrate that black-boned sheep iPSCs possess pluripotency and the potential to differentiate into all three germ layers, revealing the mechanisms by which reprogrammed iPSCs influence early embryonic development and providing a critical foundation for research on sheep pluripotent stem cells. Full article

Maintaining a Constant False Alarm Rate (CFAR) in the presence of K-distributed sea clutter is vital due to the dynamic and unpredictable nature of maritime environments. However, conventional CFAR detectors suffer significant performance degradation in multi-target scenarios, primarily due to the masking effect caused by interfering targets. To address this challenge, this paper introduces an advanced detection scheme that integrates Linear Density-Based Spatial Clustering for Applications with Noise (Lin-DBSCAN) with CFAR processing. Lin-DBSCAN is specifically tailored to efficiently identify and isolate interfering targets and sea spikes, which typically manifest as outliers in the symmetric reference windows surrounding the Cell Under Test (CUT). By leveraging Lin-DBSCAN, the proposed Lin-DBSCAN-CFAR method effectively filters out anomalous signals from the background clutter, resulting in enhanced detection accuracy and robustness, especially under complex sea clutter conditions. Extensive simulations under varying conditions, including multiple target environments, varying false alarm rates, and different clutter shape parameters, demonstrate that Lin-DBSCAN-CFAR significantly outperforms conventional CFAR approaches. It is noteworthy that the proposed method achieves detection performance comparable to the more computationally intensive DBSCAN-CFAR while significantly reducing computational complexity. Simulation results reveal that Lin-DBSCAN-CFAR requires a 1 to 2 dB lower SNR to reach a detection probability of 0.8 compared with the nearest traditional CFAR techniques, confirming its superiority in both accuracy and efficiency. Full article

Background: The framework of measuring effective coverage is conceptually straightforward, yet translation into a single metric is quite intractable. An estimation of a metric linking need, access, utilization, and service quality is imperative for measuring the progress towards Universal Health Coverage. A coverage metric obtained from a household survey alone is not succinct as it only captures the service contact which cannot be considered as actual service delivery as it ignores the comprehensive assessment of provider–client interaction. The study was thus conducted to estimate a one-composite metric of effective coverage by linking varied datasets. Methods: The study was conducted in a rural, remote, and fragile setting in India. Tools encompassing a household survey, health facility assessment, and patient exit survey were administered to ascertain measures of contact coverage and quality. A gamut of techniques linking the varied surveys were employed such as (a) exact match linking and (b) ecological linking using GIS approaches via administrative boundaries, Euclidean buffers, travel time grid, and Kernel density estimates. A composite metric of effective coverage was estimated using linked datasets, adjusting for structural and process quality estimates. Further, the horizontal inequities in effective coverage were computed using Erreygers’ concentration index. The concordance between linkage approaches were examined using Wald tests and Lin’s concordance correlation. Results: A significantly steep decline in measurement estimates was found from crude coverage to effective coverage for an entire slew of linking approaches. The drop was more exacerbated for structural-quality-adjusted measures vis-à-vis process-quality-adjusted measures. Overall, the estimates for effective coverage and inequity-adjusted effective coverage were 36.4% and 33.3%, respectively. The composite metric of effective coverage was lowest for postnatal care (10.1%) and highest for immunization care (78.7%). A significant absolute deflection ranging from −2.1 to −5.5 for structural quality and −1.9 to −8.9 for process quality was exhibited between exact match linking and ecological linking. Conclusions: Poor quality of care was divulged as a major factor of decline in coverage. Policy recommendations such as bolstering the quality via the effective implementation of government flagship programs along with initiatives such as integrated incentive schemes to attract and retain workforce and community-based monitoring are suggested. Full article

Background: Medulloblastoma (MB) is the most aggressive paediatric brain cancer, highlighting the urgent need for new diagnostic and prognostic biomarkers and improved treatments to enhance patient outcomes. Our previous study identified LIN28B, an RNA-binding protein, as a potential diagnostic and prognostic marker for MB and a pharmacological target to inhibit MB cell proliferation and stemness. However, the specific role of LIN28B and its mechanism of action in MB had not been studied. Methods: This study assessed LIN28B’s role in Daoy MB cells using siRNA-mediated silencing. LIN28B silencing was achieved with Dharmacon ON-TARGETplus SMARTpool and confirmed by Western blotting. Proliferation and protein assays evaluated the cell metabolic activity and viability. A proteomics analysis was conducted to examine the effect of LIN28B knockdown on the MB cell protein expression profile. The intracellular lipid droplets were assessed using the Nile Red Staining Kit, and nucleolar B23 protein levels were assessed by immunofluorescence. Both were visualised with a high-content IN Cell Analyser 2200. Results: Effective LIN28B silencing (>80%) was achieved in each experiment. LIN28B knockdown reduced the MB cell viability, impaired ribosome biogenesis, and promoted cellular lipid accumulation, as supported by proteomics and cell-based assays. Conclusions: This study highlights LIN28B as a promising target for regulating MB cell growth, ribosomal biogenesis, and lipid metabolism. Full article

Background: Chronic-Phase Chronic Myeloid Leukemia (C-PCML) is defined by the presence of the BCR-ABL1 fusion gene, which encodes a tyrosine kinase protein that drives the uncontrolled proliferation and survival of leukemic stem cells (LSCs). Nilotinib, a tyrosine kinase inhibitor, targets the activity of BCR-ABL1 by reducing aberrant signaling pathways, which drive the regeneration of LSCs. Despite nilotinib’s action, a population of resilient LSCs persist in the bone marrow (BM) and can indeed drive relapse and progression in CML patients. Methods: Our study investigated the gene expression profiling (GEP) of BM CD34+/lin− cells from 79 CP-CML patients at diagnosis, compared to the BM CD34+/lin− cells from the same patients after 12 months of nilotinib treatment and to the normal counterpart cells from 10 donors (CTRLs). Results: GEP analyses identified 3012 significantly differentially expressed genes across these comparisons. Among these, we focused on certain key genes associated with eight crucial KEGG pathways: CML, cell cycle, JAK-STAT, PI3K-Akt, MAPK, Ras, NF-kB, and ABC transporters. Within these pathways, we observed the up-regulation of several genes at diagnosis compared to both 12 months of nilotinib treatment and the CTRLs. Conclusions: We observed that certain transcriptome features present at diagnosis persisted after 12 months of nilotinib treatment, compared to CTRLs. This suggests that nilotinib may exert selective pressure, potentially supporting the survival and self-renewal of LSCs. Future insights into these pathways could help identify therapeutic targets to improve outcomes in CML. Full article

Objective: Head and neck cancer (HNC) is a common cancer represented by nearly 80% oral cavity (OC) and oropharyngeal cancers (OPCs). Seventy percent of OPCs are associated with the Human Papilloma Virus (HPV). Immunotherapy holds the promise of future improvements in treating HNC patients. The study objective was to determine whether durvalumab immunotherapy alone, prior to curative surgery, would significantly impact the oral salivary microbiome in a pilot cohort of HPV negative and positive OC and OPC patients. Methods: Early stage OPC patients with squamous cell carcinoma were recruited: 5 HPV+ and 12 HPV−, and treated with two or three administrations of durvalumab given every two weeks, prior to surgery. Unstimulated saliva was collected and processed for bacterial DNA Isolation and V1–V3 16S rRNA gene next generation sequencing, taxa identification, and determination of relative abundance at four time points: baseline prior to surgery (A) and weekly durvalumab treatment timepoints (B, C, and D). Alpha- and beta-diversity differences for the time series were determined in Primer_v7. MaAsLin2 in R was used to identify potential associations with the time series and/or HPV status. Linear decomposition model (LDM) R-package was used to investigate the relationship of salivary microbiome with HPV status. ROC curves were plotted for significant species in common between MaAsLin2 analysis and FDR-corrected Mann-Whitney U-test using XLSTAT. Results: Longitudinal microbiome data across four timepoints (A, B, C, D) were obtained (HPV+: n = 18 samples; HPV−: n = 46 samples). A total of 416 taxa were detected across all time points, ranging from 336 to 373 per group. There were no differences in α- and β-diversities for all longitudinal comparisons (C vs. BCD, AB vs. CD, or A vs. B, C, or D). However, comparison A vs. D showed a significant increase in Prevotella melaninogenica relative abundance, a potentially pathogenic species able to evade the immune system, after three weeks treatment. Moreover, differences in beta-diversity based on HPV status were found. LDM analysis identified Veillonella atypica, overrepresented in HPV+ group, as the top species accounting for HPV status. Conclusions: The results are consistent with findings from previous studies investigating HNC patients treated with chemoradiotherapy. More research is needed to understand possible impact of immunotherapy on opportunistic bacterial species, although negligible impact from durvalumab treatment on salivary microbiome was observed. Full article

Lin28b and let-7 miRNA regulate mammalian pubertal initiation and Gonadotropin-releasing hormone (GnRH) production. However, it remains unclear which signaling pathways Lin28b regulates to modulate GnRH production. In this study, the mRNA expression levels of Lin28b and let-7 in the pubertal and juvenile goat hypothalamus and pituitary gland were detected, and Lin28b expression in the pubertal hypothalamus decreased significantly compared with that in juvenile tissues. It was predicted that Lin28b might inhibit GnRH1 expression, which was verified in the GnRH-producing cell model GT1-7 cells. Lin28b inhibited GnRH1 expression and promoted Kiss1/Gpr54 signaling. The pyruvate content and the expression of Hif1a and Hk2, which were related to glycolysis, were also promoted by Lin28b overexpression. Additionally, 77 differentially expressed miRNAs (DEMIs) in Lin28b-overexpressed GT1-7 cells were identified. Bioinformatics analysis and mRNA expression of the target genes of DEMIs revealed that the MAPK and PI3K-AKT-mTOR signaling pathways were key pathways that involved the regulatory effect of Lin28b on GnRH. In GT1-7 cells, GnRH1 expression was suppressed by blocking mTOR signaling with rapamycin, which was rescued by Lin28b overexpression. These results indicate that Lin28b-let-7 regulates GnRH1 expression through several pathways, including the Kiss1/Gpr54, MAPK, and mTOR signaling pathways, which are all related to glucose metabolism and provide new insights into the molecular mechanism of the regulatory role of Lin28b on GnRH production. Full article

Variable flow emitters are used in subsurface drip irrigation to address challenges in soil moisture transport. This study investigates the impact of flow channel structural parameters on the hydraulic performance and anticlogging ability of emitters using computational fluid dynamics (CFD) simulations and experimental tests. The results show that the realizable k–ε turbulence model can be used to simulate the flow field inside the variable flow emitter flow channel. The nRMSE between the measured (q_m) and simulated (q) values of the flow rate is 11.23%, and the relative error between the measured (x_m) and simulated (x) values of the flow index is 4.66%, which gives a high simulation accuracy. A polar analysis shows that the tooth angle (A) has the smallest effect on the effluent flow rate at 0.1 MPa (q_0.1), x, and particle passage rate (η) of the variable flow emitter. Flow channel depth (D), tooth spacing (B), and tooth height (E) have a different order of precedence in the influence of the three indices, which are D > B > E > A, B > E > D > A and E > B > D > A, respectively. The value of η is positively correlated with the mean flow velocity (v) and the mean turbulent kinetic energy (k) in the flow channel, and η tends to increase and then decrease with the increase of x. The retention time of the particles in the flow channel is closely related to the magnitude of v and k. Three multivariate lin ear regression equations (R² = 0.883–0.995) were constructed for q_0.1, x, and η versus the flow channel structural parameters. The optimal design combination of channel structure parameters for different scenarios was determined using the scipy.optimize.minimize function in Python 3.8.0. The research results provide a reference for the optimal design of variable flow emitters. Full article

Objectives: This study investigated antibiotic resistance and presence of persister cells in Streptococcus agalactiae strains belonging to capsular types Ia/ST-103, III/ST-17, and V/ST-26 in biofilm-like environments. Results: S. agalactiae strains were susceptible to penicillin, clindamycin, and erythromycin. Resistance genes were associated with tetM (80%), tetO (20%), ermB (80%), and linB (40%). Persister cells were detected in bacterial strains exposed to high concentrations of penicillin, clindamycin, and erythromycin. S. agalactiae capsular type III/ST-17 exhibited the highest percentage of persister cells in response to penicillin and clindamycin, while type Ia/ST-103 presented the lowest percentages of persister cells for all antimicrobials tested. Additionally, persister cells were also detected at lower levels for erythromycin, regardless of capsular type or sequence type. Further, all S. agalactiae isolates presented efflux pump activity in ethidium bromide-refractory cell assays. LIVE/DEAD fluorescence microscopy confirmed the presence of >85% viable persister cells after antibiotic treatment. Conclusions: These findings suggest that persister cells play a key role in the persistence of S. agalactiae during antibiotic therapy, interfering with the treatment of invasive infections. Monitoring persister formation is crucial for developing strategies to combat recurrent infections caused by this pathogen. Full article

Prolonged consumption of animal-derived foods containing high levels of lincomycin (LIN) residues can adversely impact human health. Therefore, it is essential to develop specific antibodies and immunoassay methods for LIN. This study utilized computational chemistry to predict the efficacy of LIN haptens prior to chemical synthesis, with subsequent confirmation obtained through an immunization experiment. A hybridoma cell line named LIN/1B11 was established, which is specific to LIN. The optimized indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method exhibited high specificity for detecting LIN residues, with an IC50 value of 0.57 ± 0.03 µg/kg. The method effectively detected LIN residues in pork and milk samples, achieving a limit of detection (LOD) ranging from 0.81 to 1.20 µg/kg and a limit of quantification (LOQ) ranging from 2.09 to 2.29 µg/kg, with recovery rates between 81.9% and 108.8%. This study offers a valuable tool for identifying LIN residues in animal-derived food products. Furthermore, the efficient hapten prediction method presented herein improves antibody preparation efficiency and provides a simple method for researchers in screening haptens. Full article

Background/Objectives: Intrauterine adhesion (IUA) is characterized by endometrial fibrocyte hyperplasia. The LIN28B gene is associated with many proliferative diseases. However, its association with IUA is entirely unknown. We hypothesized that LIN28B gene polymorphisms are responsible for IUA susceptibility after curettage abortion. Methods: In this genetic association study, We genotyped two common polymorphisms (rs369065 C>T and rs314280 A>G) in 107 patients with IUA and 270 controls without IUA after curettage abortion from a Chinese population between July 2022 and May 2023 and analyzed their associations with IUA risk using multiple logistic regression models. Results: The carriers of genotype rs314280 AA of the LIN28B gene showed an increased risk of IUA (AOR [adjusted odds ratio] = 2.12, 95% CI [confidence interval] = 1.151–3.903), compared to GG+GA genotypes. Further stratification analyses showed that the deleterious role of the rs314280 AA genotype was more evident in patients with fewer than four pregnancies (AOR = 2.740, 95% CI = 1.355–5.540), fewer than two births (AOR = 2.676, 95% CI = 1.300–5.509), and fibrous (AOR = 2.082, 95% CI = 1.084–3.997) and muscular adhesions (AOR = 3.887, 95% CI = 1.116–13.540). However, the rs369065 T>C polymorphism of the LIN28B gene was not significantly associated with the occurrence of IUA. Conclusions: The rs314280 AA genotype of the LIN28B gene is associated with an increased risk of IUA in patients after curettage abortion, especially in those with fewer pregnancies or parities and higher disease severity. Our findings implicate a precise choice of clinical counseling and decision-making of IUA, thereby protecting female reproduction. Full article

Oral cancer, particularly oral squamous cell carcinoma (OSCC), is a significant global health challenge because of its high incidence and limited treatment options. Major risk factors, including tobacco use, alcohol consumption, and specific microbiota, contribute to the disease’s prevalence. Recently, a compelling association between diabetes mellitus (DM) and oral cancer has been identified, with metformin, a widely used antidiabetic drug, emerging as a potential therapeutic agent across various cancers, including OSCC. This review explores both preclinical and clinical studies to understand the mechanisms by which metformin may exert its anticancer effects, such as inhibiting cancer cell proliferation, inducing apoptosis, and enhancing the efficacy of existing treatments. Preclinical studies demonstrate that metformin modulates crucial metabolic pathways, reduces inflammation, and impacts cellular proliferation, thereby potentially lowering cancer risk and improving patient outcomes. Additionally, metformin’s ability to reverse epithelial-to-mesenchymal transition (EMT), regulate the LIN28/let-7 axis, and its therapeutic role in head and neck squamous cell carcinoma (HNSCC) are examined through experimental models. In clinical contexts, metformin shows promise in enhancing therapeutic outcomes and reducing recurrence rates, although challenges such as drug interactions, complex dosing regimens, and risks such as vitamin B12 deficiency remain. Future research should focus on optimizing metformin’s application, investigating its synergistic effects with other therapies, and conducting rigorous clinical trials to validate its efficacy in OSCC treatment. This dual exploration underscores metformin’s potential to play a transformative role in both diabetes management and cancer care, potentially revolutionizing oral cancer treatment strategies. Full article

The Daitokuji Old Collection’s 500 Luohans Paintings 五百羅漢圖, painted by Southern Song Mingzhou 明州 artists Lin Tinggui 林庭珪 and Zhou Jichang 周季常, have become a focal point in recent studies on the Chinese Buddhist material culture of the Song Dynasty. Among the 500 Luohans series, five paintings are related to the ancient Indian legend of Emperor Aśoka’s creation of 84,000 stupas. These paintings include “Building a Stupa” (No.78), “King Aśoka Stupa Emitting Light” (No.79), “Precious Stupa on the Rock” (No.80), and “Miracle of Light-Emitting Relics” (No.81), which are currently housed in the Daitokuji 大德寺 in Kyoto, Japan, and “Luohans Watching the Relics’ Light” (B5), which is housed in the Museum of Fine Arts in Boston, the US. However, the way in which the “King Aśoka Stupa” 阿育王塔 and “Relics’ Light” 舍利光 series were integrated into the overall visual narrative of the 500 Luohans in the Daitokuji and Boston collections, as well as the profound meanings and social-cultural contexts embedded in these images, have been rarely studied in depth. The introduction of the miraculous relics theme into the Daitokuji Old Collection’s 500 Luohans Paintings originates from an earlier version by the monk Fa Neng. However, Fa Neng’s version recorded by the Northern Song literati Qin Guan 秦觀 does not mention the King Aśoka Stupa. The artists had considerable freedom in depicting miraculous relic phenomena and King Aśoka Stupa. The specific details of King Aśoka Stupa’s background in Tiantai Mountain 天台山, such as rock bridges, waterfalls, and rock caves, as well as the craftsmanship of King Aśoka Stupa, reflect particular contemporary ideas. The vivid depictions of the “King Aśoka Stupa” and “Relics’ Light” in the Daitokuji Present Collection and the Boston Collection of the 500 Luohans may indicate a close connection between the creation of these images and the fervent Relics Worship at King Aśoka Temple 阿育王寺 in Mingzhou during Southern Song. This paper synthesizes these images, ancient Chinese and Japanese manuscripts, and fieldwork insights to interpret the sources and significance of these images. Full article

Background: Single-nucleotide polymorphisms in LIN28B, critical regulators of female growth and puberty, have been linked to age at menarche. Methods: We assessed the association of rs7759938, rs314280, and rs314276 with menarcheal age in girls of Greek descent. We reviewed the records of 248 girls who had their first menstruation before 18 years and who attended the Greek Departments of Pediatric Endocrinology from January 2021 to July 2023. Genotyping was performed by standard DNA-based methods. Association analyses involved both parametric and non-parametric tests. Results: The average age of breast and pubic hair development was 9.95 years, and the age at menarche was 11.55 years. Menarche occurred ≤11 years (mean 10.24 years) in 108 girls (43.5%) and >11 years (mean 12.55 years) in 140 (56.5%). The girls’ menarcheal age correlated significantly with that of their mothers (average 12.1 years, p-value < 0.0001, Spearman’s r 0.350). The dominant rs7759938(TT) genotype was the most common (55.2%), followed by the dominant rs314276(CC) (53.2%) and dominant rs314280(TT) (14.5%) genotypes. Conclusions: There was no association between age at menarche and any of the polymorphism genotypes/alleles or between genotypes/alleles and birth weight, gestational week, mode of delivery, and maternal age at menarche. Future large sample studies are warranted to confirm these results. Full article

The delay in wound healing caused by chronic wounds or pathological scars is a pressing issue in clinical practice, imposing significant economic and psychological burdens on patients. In particular, with the aging of the population and the increasing incidence of diseases such as diabetes, impaired wound healing is one of the growing health problems. MicroRNA (miRNA) plays a crucial role in wound healing and regulates various biological processes. Our results show that miR–618 was significantly upregulated during the inflammatory phase of wound healing.Subsequently, miR–618 promotes the secretion of pro–inflammatory cytokines and regulates the proliferation and migration of keratinocytes. Mechanistically, miR–618 binds to the target gene–Atp11b and inhibits the PI3K–Akt signaling pathway, inhibiting the epithelial–mesenchymal transition (EMT) of keratinocytes. In addition, the PI3K–Akt signaling pathway induces the enrichment of nuclear miR–618, and miR–618 binds to the promoter of Lin7a to regulate gene transcription. Intradermal injection of miR–618 antagomir around full–thickness wounds in peridermal mice effectively accelerates wound closure compared to control. In conclusion, miR–618 antagomir can be a potential therapeutic agent for wound healing. Full article