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1.
Int J Mol Sci ; 24(3)2023 Jan 17.
Article in English | MEDLINE | ID: mdl-36768155

ABSTRACT

Irreversible cell-cycle-arrested cells not undergoing cell divisions have been thought to be metabolically less active because of the unnecessary consumption of energy for cell division. On the other hand, they might be actively involved in the tissue microenvironment through an inflammatory response. In this study, we examined the mitochondria-dependent metabolism in human cells irreversibly arrested in response to ionizing radiation to confirm this possibility. Human primary WI-38 fibroblast cells and the BJ-5ta fibroblast-like cell line were exposed to 20 Gy X-rays and cultured for up to 9 days after irradiation. The mitochondrial morphology and membrane potential were evaluated in the cells using the mitochondrial-specific fluorescent reagents MitoTracker Green (MTG) and 5,5',6,6'-tetraethyl-benzimidazolylcarbocyanine iodide (JC-1), respectively. The ratio of the mean MTG-stained total mitochondrial area per unit cell area decreased for up to 9 days after X-irradiation. The fraction of the high mitochondrial membrane potential area visualized by JC-1 staining reached its minimum 2 days after irradiation and then increased (particularly, WI-38 cells increased 1.8-fold the value of the control). Their chronological changes indicate that the mitochondrial volume in the irreversible cell-cycle-arrested cells showed significant increase concurrently with cellular volume expansion, indicating that the mitochondria-dependent energy metabolism was still active. These results indicate that the energy metabolism in X-ray-induced senescent-like cells is active compared to nonirradiated normal cells, even though they do not undergo cell divisions.


Subject(s)
Fluorescent Dyes , Mitochondria , Humans , X-Rays , Mitochondria/metabolism , Cell Cycle Checkpoints , Membrane Potential, Mitochondrial/radiation effects , Fluorescent Dyes/metabolism
2.
J Radiat Res ; 64(2): 300-303, 2023 Mar 23.
Article in English | MEDLINE | ID: mdl-36617210

ABSTRACT

We examined here normal human cells with large deletions encompassing the hypoxanthine-phosphoribosyltransferase 1 (HPRT1) gene on X chromosome. Expression levels of genes on X chromosome were analyzed by microarray and RT-qPCR method, and differentially expressed genes (DEGs) were extracted. We found that DEGs were not limited to the genes flanking deleted regions but spread over the entire X chromosome. Interestingly, the gene regulation patterns were similar to a large extent among independent clones that have similar-sized large deletions involving the HPRT1 gene. Thus, it is indicated that an impact of large deletion on possible epigenetic transcriptional regulation is not limited to the regions proximal to the deletion region.


Subject(s)
Chromosomes, Human, X , Hypoxanthine Phosphoribosyltransferase , Humans , X-Rays , Chromosomes, Human, X/genetics , Hypoxanthine Phosphoribosyltransferase/genetics , Clone Cells
3.
Int J Mol Sci ; 22(23)2021 Nov 26.
Article in English | MEDLINE | ID: mdl-34884589

ABSTRACT

The non-targeted effects of radiation have been known to induce significant alternations in cell survival. Although the effects might govern the progression of tumor sites following advanced radiotherapy, the impacts on the intercellular control of the cell cycle following radiation exposure with a modified field, remain to be determined. Recently, a fluorescent ubiquitination-based cell-cycle indicator (FUCCI), which can visualize the cell-cycle phases with fluorescence microscopy in real time, was developed for biological cell research. In this study, we investigated the non-targeted effects on the regulation of the cell cycle of human cervical carcinoma (HeLa) cells with imperfect p53 function that express the FUCCI (HeLa-FUCCI cells). The possible effects on the cell-cycle phases via soluble factors were analyzed following exposure to different field configurations, which were delivered using a 150 kVp X-ray irradiator. In addition, using synchrotron-generated, 5.35 keV monochromatic X-ray microbeams, high-precision 200 µm-slit microbeam irradiation was performed to investigate the possible impacts on the cell-cycle phases via cell-cell contacts. Collectively, we could not detect the intercellular regulation of the cell cycle in HeLa-FUCCI cells, which suggested that the unregulated cell growth was a malignant tumor. Our findings indicated that there was no significant intercellular control system of the cell cycle in malignant tumors during or after radiotherapy, highlighting the differences between normal tissue and tumor characteristics.


Subject(s)
Cell Cycle , Fluorescent Dyes/chemistry , Synchrotrons/instrumentation , Ubiquitination , Uterine Cervical Neoplasms/pathology , Cell Survival , Female , HeLa Cells , Humans , Microscopy, Fluorescence , X-Rays
4.
Radiat Res ; 195(6): 561-567, 2021 06 01.
Article in English | MEDLINE | ID: mdl-33826740

ABSTRACT

The genomic landscape neighboring large deletions including the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus on human X chromosome in 6-thioguanine-resistant mutants originating from immortalized human fibroblast cells exposed to X rays was characterized by real-time quantitative PCR (qPCR)-based analyses. Among the 13 mutant clones with large deletions extending over several Mb, including the HPRT locus, revealed by 10 conventional sequence-tagged site (STS) markers, three clones bearing the largest deletions were selected for further qPCR analysis using another 21 STS markers and 15 newly designed PCR primer pairs. The results indicated that the major deletions were in very specific regions between the 130-Mb and 140-Mb positions containing the HPRT locus on the X chromosome and, contrary to our initial expectations, additional minor deletions were distributed in a patchwork pattern. These findings strongly indicate that the complex deletion patterns in the affected chromosome are related to the radiation track structure with spatially heterogeneous energy deposition and the specific structure of the chromatin-nuclear membrane complex. The uncovered complex deletion patterns are in agreement with the idea of complex chromatin damage, which is frequently associated with carcinogenesis.


Subject(s)
Chromosome Deletion , Genome, Human/genetics , Carcinogenesis/genetics , Carcinogenesis/radiation effects , Genetic Loci/genetics , Humans , Polymerase Chain Reaction , X-Rays/adverse effects
5.
Online J Issues Nurs ; 20(3): 7, 2015 Aug 25.
Article in English | MEDLINE | ID: mdl-26882516

ABSTRACT

Sixty nurses from five countries (Canada, India, Ireland, Japan, and Korea) took part in 11 focus groups that discussed the question: Do you consider your work meaningful? Fostering meaning and mentorship as part of the institutional culture was a central theme that emerged from the discussions. In this article, we begin with a background discussion of meaning and meaningful work as presented in the literature related to existentialism and hardiness. Next, we describe the method and analysis processes we used in our qualitative study asking how nurses find meaning in their very challenging work and report our findings of four themes that emerged from the comments shared by nurses, specifically relationships, compassionate caring, identity, and a mentoring culture. After offering a discussion of our findings and noting the limitations of this qualitative study, we conclude that nursing leaders and a culture of mentorship play an important role in fostering meaningful work and developing hardy employees.


Subject(s)
Attitude of Health Personnel , Job Satisfaction , Nursing , Empathy , Focus Groups , Humans , Leadership , Mentoring , Qualitative Research
6.
Seishin Shinkeigaku Zasshi ; 105(9): 1181-5, 2003.
Article in Japanese | MEDLINE | ID: mdl-14639942

ABSTRACT

Japan had a long history of long-term hospital care and is now trying to change it into community-based psychiatry. Implementing comprehensive approach that has already been proven effective through evidence based data in one particular community, one has to overcome several local barriers, if the same approach is to be implemented in different situations, such as differences in socio-cultural background and health care system. This is a report of the activities of Minato Net 21 pointing out the "local barriers" on implementing comprehensive approach in an urban district of Tokyo.


Subject(s)
Community Mental Health Services/organization & administration , Delivery of Health Care, Integrated/organization & administration , Urban Health Services/organization & administration , Community Mental Health Services/trends , Comprehensive Health Care/organization & administration , Comprehensive Health Care/trends , Culture , Delivery of Health Care, Integrated/trends , Humans , Tokyo , Urban Health Services/trends
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