RESUMO
Although somatic cell nuclear transfer (SCNT) is a critical component of animal cloning, this approach has several issues. We previously introduced the cytoplasm injection cloning technology (CICT), which significantly improves the quality and quantity of cloned embryos. This study examined the residual status of fused cytoplasmic organelles, such as the endoplasmic reticulum (ER) and lysosomes, in the CICT group during early embryo development. We found that extra-cytoplasmic organelles stained using the ER-Tracker™ Green dye and LysoTracker™ Deep Red probe were fused and dispersed throughout the recipient oocyte and were still visible in day 8 blastocysts. We screened for ER stress, autophagy, and apoptosis-related genes to elucidate the association between the added organelles and improved embryo quality in CICT-cloned embryos. We found that CHOP, ATF4, ATG5, ATG7, and LC3 genes showed non-significantly up- or downregulated expression between CICT- and in vitro fertilization (IVF)-derived embryos but showed significantly (p < 0.05) upregulated expression in SCNT-cloned embryos. Surprisingly, a non-significant difference in the expression of some genes, such as ATF6 and caspase-3, was observed between the CICT- and SCNT-cloned embryos. Our findings imply that compared to conventional SCNT cloning, CICT-derived cloned embryos with additional cytoplasm have much higher organelle activity, lower autophagy, lower rates of apoptosis, and higher embryo development rates.
Assuntos
Clonagem de Organismos , Embrião de Mamíferos , Animais , Bovinos , Clonagem de Organismos/veterinária , Técnicas de Transferência Nuclear/veterinária , Blastocisto , Desenvolvimento Embrionário , Fertilização in vitro/veterinária , Retículo EndoplasmáticoRESUMO
CONTEXT: Telomerase reverse transcriptase is a key factor responsible for structural and cellular alterations in aged oocytes and changes in the structure of the zona pellucida and mitochondria. Telomerase expression is reduced in aged cumulus oocyte complexes, and its activation or enhanced expression would be beneficial for in vitro oocyte maturation and in vitro embryo development. AIMS: This study aimed to investigate telomerase activation by cycloastragenol and its effect on bovine oocyte in vitro maturation, fertilisation, and early embryo development. METHODS: We used qPCR, Western blot, immunofluorescence, reactive oxygen species (ROS) assay,TUNEL assay, JC-1 assay, and invasion assay to analyse the affect of cycloastragenol (CAG) on bovine oocyte maturation, embryo development, embryo quality and implantation potential. KEY RESULTS: Cycloastragenol treatment of oocytes in in vitro maturation (IVM) media significantly (P <0.05) improved oocyte IVM (90.87%), embryo cleavage (90.78%), blastocyst hatching (27.04%), and embryo implantation potential. Telomerase also interacts with mitochondria, and JC-1 staining results showed significantly (P <0.05) higher mitochondrial membrane potential (ΔΨ m) in the CAG-treated group. Furthermore, the inner cell mass (OCT4 and SOX2) and trophoblasts (CDX2) of the control and CAG groups were examined. Moreover, CAG treatment to primary cultured bovine cumulus cells substantially enhanced telomerase activity. CONCLUSIONS: Telomerase activation via cycloastragenol is beneficial for bovine oocyte IVM and for the production of high-quality bovine embryos. IMPLICATIONS: Cycloastragenol is a natural telomerase activator, and could be useful as a permanent component of oocyte maturation media.
Assuntos
Telomerase , Feminino , Animais , Bovinos , Telomerase/genética , Telomerase/metabolismo , Telomerase/farmacologia , Células do Cúmulo/metabolismo , Oócitos/metabolismo , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Implantação do Embrião , Desenvolvimento Embrionário , BlastocistoRESUMO
AIM: We investigated the difference in the relationship between physical activity and low muscle mass resulting from the choice of diagnostic criterion for low muscle mass. METHODS: Our study was cross-sectional, using data from the 2008-2011 National Health and Nutrition Examination Survey. Muscle mass was measured by dual-energy X-ray absorptiometry. Low muscle mass was defined as height-adjusted and weight-adjusted low muscle mass. Odds ratios (ORs) and 95% confidence intervals (CIs) of low muscle mass in relation to physical inactivity were analyzed using multivariate logistic regression analysis. RESULTS: Our study included 3977 older people (1698 men and 2279 women). The prevalence of height-adjusted and weight-adjusted low muscle mass was 4.1% and 11.8%, respectively, in the physically inactive group, and 3.9% and 7.9%, respectively, in the physically active group. The prevalence of weight-adjusted low muscle mass increased by 48% in the physically inactive group in the fully adjusted model (OR, 1.48; 95% CI, 1.13-1.95) and increased by 61% in men and 50% in women (men: OR, 1.61; 95% CI, 1.12-2.31 and women: OR, 1.50; 95% CI, 1.20-2.20) compared with the physically active group. The risk of height-adjusted low muscle mass in men tended to be higher in the physically inactive group than in the physically active group. However, this trend was not observed among women. CONCLUSIONS: Physical inactivity was associated with an increased prevalence of low muscle mass in weight-adjusted measures among elderly adults in Korea. Height-adjusted low muscle mass in women is less useful as an indicator of the relationship between low muscle mass and physical inactivity. Geriatr Gerontol Int 2023; 23: 71-77.
Assuntos
Músculo Esquelético , Comportamento Sedentário , Masculino , Humanos , Feminino , Idoso , Músculo Esquelético/diagnóstico por imagem , Inquéritos Nutricionais , Estudos Transversais , População do Leste Asiático , PrevalênciaRESUMO
The lack of drugs that target both disease progression and tissue preservation makes it difficult to effectively manage rheumatoid arthritis (RA). Here, we report a porous silicon-based nanomedicine that efficiently delivers an antirheumatic drug to inflamed synovium while degrading into bone-remodeling products. Methotrexate (MTX) is loaded into the porous silicon nanoparticles using a calcium silicate based condenser chemistry. The calcium silicate-porous silicon nanoparticle constructs (pCaSiNPs) degrade and release the drug preferentially in an inflammatory environment. The biodegradation products of the pCaSiNP drug carrier are orthosilicic acid and calcium ions, which exhibit immunomodulatory and antiresorptive effects. In a mouse model of collagen-induced arthritis, systemically administered MTX-loaded pCaSiNPs accumulate in the inflamed joints and ameliorate the progression of RA at both early and established stages of the disease. The disease state readouts show that the combination is more effective than the monotherapies.
Assuntos
Antirreumáticos , Artrite Reumatoide , Camundongos , Animais , Metotrexato/farmacologia , Metotrexato/uso terapêutico , Nanomedicina , Silício , Porosidade , Cálcio , Artrite Reumatoide/tratamento farmacológico , Antirreumáticos/uso terapêutico , Portadores de Fármacos/uso terapêutico , Inflamação/tratamento farmacológicoRESUMO
BACKGROUND: Isolated anterior cerebral artery territory (ACA) infarction is a rare phenomenon, and is known to have distinctive clinical features. Little is known regarding the clinical prognosis of isolated ACA territory infarction with associated factors, and its impact on dwelling and job status. We investigated the short- and long-term outcomes of anterior cerebral artery (ACA) territory infarction, and the associated factors involved in the development of the distinctive symptoms. METHODS: This retrospective study in a prospective cohort of acute ischaemic stroke patients included consecutively enrolled patients with isolated ACA territory infarction. We investigated the functional status using the modified Rankin scale (mRS) score at discharge, three months' post-discharge, and one-year post-discharge. We also investigated the occlusion site of the ACA (proximal vs. distal); presence of distinctive symptoms of ACA territory infarction including behaviour changes, indifference, aphasia, and urinary incontinence; and the effect of these symptoms on dwelling and job status one year after discharge. RESULTS: Between April 2014 and March 2019, 47 patients with isolated ACA territory infarction were included. Twenty-nine patients (61.7 %) had good outcomes (mRS ≤ 2) at discharge; however, the mRS score increased at three months (40; 85.1 %, p < 0.001) and one year (41; 87.2 %) post-discharge. Occlusion of the ACA proximal segment was independently associated with the development of distinctive symptoms (adjusted odds ratio, 17.68; 95 % confidence interval: 2.55-122.56, p < 0.05). Twenty-one (48.8 %) patients with good outcomes at one year experienced a change in dwelling status and job loss; 20 (95.2 %) of them had distinctive ACA territory symptoms with proximal ACA occlusion. CONCLUSIONS: Short- and long-term outcomes of isolated ACA territory infarction were favourable. However, proximal segment occlusion was associated with the development of distinctive symptoms, possibly related to future dwelling and job status.
Assuntos
Infarto da Artéria Cerebral Anterior , Recuperação de Função Fisiológica , Idoso , Feminino , Humanos , Infarto da Artéria Cerebral Anterior/complicações , Infarto da Artéria Cerebral Anterior/patologia , Infarto da Artéria Cerebral Anterior/terapia , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Estudos Retrospectivos , Fatores de RiscoRESUMO
A hybrid composite of silver nanoparticles (AgNPs) and porous silicon microparticles (pSiMPs) was developed and applied for the computed tomography (CT) scanning of the lungs as an image-guided localization agent. We confirmed the grafting of AgNPs on oxidized pSiMPs template using various analytical equipment, including a scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). The hybrid composite showed a high CT contrast intensity (>1000 HU) that enabled us to produce and view images of the lungs. In addition, it showed the ability to maintain a strong CT signal at the injected area of the rabbit's lungs, up to an hour, without spreading. The lack of toxicity and immune response indicated that the composite could be fully utilized as a new image-guided localization agent of CT scans for lung cancer surgery.
Assuntos
Nanopartículas Metálicas , Prata , Animais , Pulmão/diagnóstico por imagem , Porosidade , Coelhos , Silício , Tomografia , Tomografia Computadorizada por Raios XRESUMO
Fluorescence-based in vivo imaging is one of the most important tools for monitoring of biological processes in cells and tissues of live animal models. Fluorescence imaging agents have also been used to monitor the microcirculation. Tracking microcirculation of the blood is vital to gain further insight into various vascular disease-related anomalies within the human body. As monitoring of vascular circulation is performed with visualization of both immune cells and pathogens, which are mainly labelled with red and green, the favorable color option for blood vessels could be blue. However, currently available blueish color-labeled agents for vascular monitoring is generally confronted with quick bleaching, because of its short excitation and emission wavelengths. Hereby, what we propose in this report is a newly generated bright blue fluorescent dextran, named HCD-70K that monitors the blood vessels using blue and inter-compatible typical fluorescent materials. DBCO-functionalized dextran-70K was fabricated with hydroxy-coumarin dye via metal-free bioorthogonal click chemistry, and generated HCD-70K, which can flow within the blood vessel and decipher the whole structure of the blood vessel successfully. The synthesis, spectroscopic analysis, and quantum chemical calculations were conducted. Using two-photon microscopy, efficient deep in vivo blood vessel imaging of a mouse model revealed exceptional bio-imaging capabilities of the HCD-70K and consequently it provided a promising opportunity for efficient vascular visualization in various research areas.
Assuntos
Vasos Sanguíneos/diagnóstico por imagem , Dextranos/química , Corantes Fluorescentes/química , Imagem Óptica , Fótons , Animais , Teoria da Densidade Funcional , Dextranos/administração & dosagem , Dextranos/síntese química , Corantes Fluorescentes/administração & dosagem , Corantes Fluorescentes/síntese química , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Estrutura MolecularRESUMO
Silencing of aberrantly expressed microRNAs (miRNAs or miRs) has emerged as one of the strategies for molecular targeted cancer therapeutics. In particular, miR-21 is an oncogenic miRNA overexpressed in many tumors, including ovarian cancer. To achieve efficient administration of anti-miR therapeutics, delivery systems are needed that can ensure local accumulation in the tumor environment, low systemic toxicity, and reduced adverse side effects. In order to develop an improved anti-miR therapeutic agent for the treatment of ovarian cancer, a nanoformulation is engineered that leverages biodegradable porous silicon nanoparticles (pSiNPs) encapsulating an anti-miR-21 locked nucleic acid payload and displaying a tumor-homing peptide for targeted distribution. Targeting efficacy, miR-21 silencing, and anticancer activity are optimized in vitro on a panel of ovarian cancer cell lines, and a formulation of anti-miR-21 in a pSiNP displaying the targeting peptide CGKRK is identified for in vivo evaluation. When this nanoparticulate agent is delivered to mice bearing tumor xenografts, a substantial inhibition of tumor growth is achieved through silencing of miR-21. This study presents the first successful application of tumor-targeted anti-miR porous silicon nanoparticles for the treatment of ovarian cancer in a mouse xenograft model.
Assuntos
Portadores de Fármacos , MicroRNAs , Nanopartículas , Neoplasias Ovarianas , Silício , Animais , Linhagem Celular Tumoral , Portadores de Fármacos/química , Portadores de Fármacos/farmacologia , Feminino , Humanos , Camundongos , Camundongos Nus , MicroRNAs/química , MicroRNAs/genética , MicroRNAs/farmacologia , Nanopartículas/química , Nanopartículas/uso terapêutico , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Porosidade , Silício/química , Silício/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Porous silicon nanoparticles (pSiNPs) have been utilized within a wide spectrum of biological studies, as well as in chemistry, chemical biology, and biomedical fields. Recently, pSiNPs have been constantly coming under the spotlight, mostly in biomedical applications, due to their advantages, such as controlled-release drug delivery in vivo by hydrolysis-induced degradation, self-reporting property through long life-time photoluminescence, high loading efficiency of substrate into pore, and the homing to specific cells/organ/bacteria by surface functionalization. However, the systematic degradation rate analysis of surface-functionalized pSiNPs in different biological media has not been conducted yet. In this paper, we prepared four different surface-functionalized pSiNPs samples and analyzed the degradation rate in six different media (DI H2O (deionized water), PBS (phosphate-buffered saline), HS (human serum), DMEM (Dulbecco's modified Eagle's medium), LB (lysogeny broth), and BHI (brain heart infusion)). The obtained results will now contribute to understanding the correlation between surface functionalization in the pSiNPs and the degradation rate in different biological media. The characterized data with the author's suggestions will provide useful insights in designing the new pSiNPs formulation for biomedical applications.
RESUMO
Porous silicon has been utilized within a wide spectrum of industries, as well as being used in basic research for engineering and biomedical fields. Recently, surface modification methods have been constantly coming under the spotlight, mostly in regard to maximizing its purpose of use. Within this review, we will introduce porous silicon, the experimentation preparatory methods, the properties of the surface of porous silicon, and both more conventional as well as newly developed surface modification methods that have assisted in attempting to overcome the many drawbacks we see in the existing methods. The main aim of this review is to highlight and give useful insight into improving the properties of porous silicon, and create a focused description of the surface modification methods.