Amelioration of ultraviolet-induced photokeratitis in mice treated with astaxanthin eye drops

Purpose: Ultraviolet (UV) acts as low-dose ionizing radiation. Acute UVB exposure causes photokeratitis and induces apoptosis in corneal cells. Astaxanthin (AST) is a carotenoid, present in seafood, that has potential clinical applications due to its high antioxidant activity. In the present study, we examined whether topical administration of AST has preventive and therapeutic effects on UV-photokeratitis in mice. Methods: C57BL/6 mice were administered with AST diluted in polyethylene glycol (PEG) in instillation form (15 μl) to the right eye. Left eyes were given vehicle alone as controls. Immediately after the instillation, the mice, under anesthesia, were irradiated with UVB at a dose of 400 mJ/cm2. Eyeballs were collected 24 h after irradiation and stained with H&E and TUNEL. In an in vitro study, mouse corneal epithelial (TKE2) cells were cultured with AST before UV exposure to quantify the UV-derived cytotoxicity. Results: UVB exposure induced cell death and thinning of the corneal epithelium. However, the epithelium was morphologically well preserved after irradiation in AST-treated corneas. Irradiated corneal epithelium was significantly thicker in eyes treated with AST eye drops, compared to those treated with vehicles (p<0.01), in a doses dependent manner. Significantly fewer apoptotic cells were observed in AST-treated eyes than controls after irradiation (p<0.01). AST also reduced oxidative stress in irradiated corneas. The in vitro study showed less cytotoxicity of TKE2 cells in AST-treated cultures after UVB-irradiation (p<0.01). The cytoprotective effect increased with the dose of AST. Conclusions: Topical AST administration may be a candidate treatment to limit the damages by UV irradiation with wide clinical applications

Polymer Relaxation Time Enhancement at Temperatures above Glass Transition Temperatures Predicted by Idealized Mode-Coupling Theory

The mode-coupling theory of glass transition predicts the relaxation time divergence of glass-forming materials at the crossover temperature, which is approximately 1.2 times the calorimetric glass transition temperature. However, this divergence has not been experimentally observed. This is known as the most serious drawback of the mode-coupling theory. The use of viscosity-sensitive single molecule fluorescence probes enables the detection of the poly(vinyl acetate) and poly(ethyl methacrylate) relaxation time enhancement around the crossover temperature, thereby supporting the prediction by the mode-coupling theory

Evaluation of Changes in Activities of Daily Living and Quality of Life of Patients with Bone Metastasis Who Underwent Conservative Therapy through Bone Metastasis Cancer Boards

Background and Objectives: Changes in activities of daily living (ADL) and quality of life (QOL) of patients with bone metastasis who underwent surgical treatment through Bone Metastasis Cancer Boards (BMCBs), a recent multidisciplinary approach for managing bone metastases, have been reported; however, no reports exist on patients who undergo conservative treatment. In this study, we aimed to evaluate these patients’ ADL and QOL and examine the factors influencing changes in these parameters. Materials and Methods: We retrospectively reviewed 200 patients with bone metastases who underwent conservative therapy through BMCBs between 2013 and 2021. A reassessment was conducted within 2–8 weeks after the initial assessment. Patients’ background and changes in performance status (PS), Barthel Index (BI), EuroQol five-dimension (EQ-5D) scores, and Numerical Rating Scale (NRS) scores were initially assessed. Furthermore, we categorized patients into two groups based on improvements or deteriorations in ADL and QOL and performed comparative analyses. Results: Significant improvements in EQ-5D (0.57 ± 0.02 versus [vs.] 0.64 ± 0.02), NRS max (5.21 ± 0.24 vs. 3.56 ± 0.21), and NRS average (2.98 ± 0.18 vs. 1.85 ± 0.13) scores were observed between the initial assessment and reassessment (all p p = 0.06, and 0.054, respectively). In addition, spinal cord paralysis (odds ratio [OR]: 3.69, p = 0.049; OR: 8.42, p p = 0.02; OR: 0.25, p = 0.007), and NRS average scores (OR: 0.38, p = 0.02; OR: 0.14, p Conclusions: Patients with bone metastases who underwent conservative treatment through BMCBs exhibited an increase in QOL without a decline in ADL. The presence of spinal cord paralysis, absence of chemotherapy, and poor pain control were associated with a higher risk of deterioration in ADL and QOL

Transcutaneous Application of Carbon Dioxide (CO₂) Induces Mitochondrial Apoptosis in Human Malignant Fibrous Histiocytoma <em>In Vivo</em>

<div><p>Mitochondria play an essential role in cellular energy metabolism and apoptosis. Previous studies have demonstrated that decreased mitochondrial biogenesis is associated with cancer progression. In mitochondrial biogenesis, peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) regulates the activities of multiple nuclear receptors and transcription factors involved in mitochondrial proliferation. Previously, we showed that overexpression of PGC-1α leads to mitochondrial proliferation and induces apoptosis in human malignant fibrous histiocytoma (MFH) cells <em>in vitro</em>. We also demonstrated that transcutaneous application of carbon dioxide (CO₂) to rat skeletal muscle induces PGC-1α expression and causes an increase in mitochondrial proliferation. In this study, we utilized a murine model of human MFH to determine the effect of transcutaneous CO₂ exposure on PGC-1α expression, mitochondrial proliferation and cellular apoptosis. PGC-1α expression was evaluated by quantitative real-time PCR, while mitochondrial proliferation was assessed by immunofluorescence staining and the relative copy number of mitochondrial DNA (mtDNA) was assessed by real-time PCR. Immunofluorescence staining and DNA fragmentation assays were used to examine mitochondrial apoptosis. We also evaluated the expression of mitochondrial apoptosis related proteins, such as caspases, cytochorome c and Bax, by immunoblot analysis. We show that transcutaneous application of CO₂ induces PGC-1α expression, and increases mitochondrial proliferation and apoptosis of tumor cells, significantly reducing tumor volume. Proteins involved in the mitochondrial apoptotic cascade, including caspase 3 and caspase 9, were elevated in CO₂ treated tumors compared to control. We also observed an enrichment of cytochrome c in the cytoplasmic fraction and Bax protein in the mitochondrial fraction of CO₂ treated tumors, highlighting the involvement of mitochondria in apoptosis. These data indicate that transcutaneous application of CO₂ may represent a novel therapeutic tool in the treatment of human MFH.</p> </div

Effect of transcutaneous application of CO₂ on MFH cell growth <i>in vivo</i>.

<p>Mice were treated with CO₂ or control air three days after MFH cell implantation. Treatment was administered twice weekly for two weeks. (A) MFH tumors in CO₂ treated and control mice, two weeks post-implantation. (B) Tumor volume (mm³) in CO₂ treated or control mice was monitored for two weeks post-implantation. (C) Body weight (g) of CO₂ treated or control mice was monitored for two weeks post-implantation. Data represent the mean ± S.E of at least three independent experiments (*<i>p</i><0.05, **<i>p</i><0.01).</p

Effect of transcutaneous application of CO₂ treatment on mitochondrial proliferation in tumors.

<p>qRT-PCR for <i>PGC-1α</i> (A) and <i>TFAM</i> (B) in CO₂ treated or control tumor specimens collected two weeks post-treatment. Expression was normalized to <i>β-actin</i> control. Data represent the mean ± S.E of at least three independent experiments (*<i>p</i><0.05). (C) mtDNA was measured in CO₂ treated or control tumor samples by PCR and the relative copy number was determined by normalizing to nDNA. Data represent the mean ± S.E. of at least three independent experiments (*<i>p</i><0.05). (D) Immunofluorescence staining of mitochondria in CO₂ treated or control tumors after two weeks (<i>Blue</i>, nuclear; <i>Red</i>, mitochondria).</p

Effect of transcutaneous CO₂ application on intracellular Ca²⁺ concentration in a mouse model of human MFH.

<p>Implanted tumors were isolated from mice at 0 (n = 12), 6 (n = 6) and 24 hours (n = 12) after transcutaneous CO₂ exposure, and the intracellular Ca²⁺ concentration was assessed using the Calcium Assay Kit. Data represent the mean ± S.E. of at least three independent experiments (*<i>p</i><0.05, **<i>p</i><0.01).</p

Evaluation of mitochondrial induced apoptosis in CO₂ or control treated tumors.

<p>(A) DNA fragmentation analysis of tumor samples from CO₂ treated and control mice two weeks post-treatment by immunofluorescence. (<i>Blue</i>, nuclear; <i>Green</i>, apoptosis nuclear) (B) DNA fragmentation was assessed by flow cytometry in CO₂ treated tumors (<i>Blue dots</i>) and control tumors (<i>Red</i>) two weeks post-treatment. (C) Immunoblot analyses determined that increased expression of the cleavage products of caspase 3 and 9, and PARP occurred in the CO₂ treated tumors compared to the control tumors. Tubulin was used as an endogenous loading control. (D) Immunoblot analysis of cytochrome c and Bax in mitochondrial and cytoplasmic fractions of CO₂ treated and control tumors. Tubulin was used as an endogenous loading control. (C, D) Positive bands in immunoblot analyses were semiquantified using densitometrical analyses using the Image J program (NIH, USA, <a href="http://rsb.info.nih.gov/ij/" target="_blank">http://rsb.info.nih.gov/ij/</a>).</p