Tissue engineering of corneal stroma with rabbit fibroblast precursors and gelatin hydrogels

PURPOSE: To isolate fibroblast precursors from rabbit corneal stroma using a sphere-forming assay, to engineer corneal stroma with the precursors and gelatin, and to establish the therapeutic application of precursors in a rabbit corneal stroma. METHODS: In the in vitro study, a sphere-forming assay was performed to produce precursors from rabbit corneal stroma. Corneal stroma was engineered by cultivating precursors in porous gelatin for one week. In the in vivo study, the engineered corneal stromal sheet with precursors (precursor/gelatin group) or with fibroblasts (fibroblast /gelatin group) or without cells (gelatin group) was transplanted to a pocket of rabbit corneal stroma. Gene expression and extracellular matrix production were examined immunohistochemically in each group one week and four weeks after surgery. RESULTS: In the in vitro study, cells in the spheres were BrdU-positive, and their progeny were keratocan-positive. The study also showed that the corneas transplanted with a porous gelatin sheet did not show any opacity four weeks after transplantation in any group. In the gelatin sheet of the precursor/gelatin group, a more intense expression of type I collagen was observed relative to the other two groups four weeks after the surgery. CONCLUSIONS: Our findings demonstrate that the transplantation of fibroblast precursors combined with gelatin hydrogel into the corneal stroma is a possible treatment strategy for corneal stromal regeneration

Report of a study using phantom materials, and clinical experience with simultaneous radio-hyperthermotherapy.

Simultaneous radiohyperthermotherapy (SRH) is a combined hyperthermia-radiation therapy in which irradiation is given during heating. Mutual interference between the high energy radiotherapy system (Toshiba LMR-15A) and the 13.56 MHz capacitive heating system (Omron HEH-500C) was tested with phantom materials prior to a clinical trial with SRH. The energy and flatness of irradiation were not affected by the heating system within the range of clinical use. The high energy radiotherapy system did not affect the increase or distribution of temperature during simultaneous treatment. The results of this phantom study indicated that these apparatuses would not produce clinically significant mutual interference during SRH. A clinical trial was performed on a 57-year-old woman with postoperative recurrence of rectal cancer. This is the first reported clinical case treated with true SRH in which external irradiation was administered during mid capacitive heating. Twelve SRH treatments were performed on the recurrent lesion at a frequency of twice a week for six weeks using the apparatuses described above. There was a significant reduction in pain after treatment. The tumor marker carcinoembryonic antigen (CEA) level decreased after treatment. On CT images taken after treatment, the tumor site became a low density area which indicated necrosis. There were no side effects. These results suggest that further clinical study of SRH should be performed to clarify its advantages.</p

Concomitant Nrf2- and ATF4-Activation by Carnosic Acid Cooperatively Induces Expression of Cytoprotective Genes

Carnosic acid (CA) is a phytochemical found in some dietary herbs, such as Rosmarinus officinalis L., and possesses antioxidative and anti-microbial properties. We previously demonstrated that CA functions as an activator of nuclear factor, erythroid 2 (NF-E2)-related factor 2 (Nrf2), an oxidative stress-responsive transcription factor in human and rodent cells. CA enhances the expression of nerve growth factor (NGF) and antioxidant genes, such as HO-1 in an Nrf2-dependent manner in U373MG human astrocytoma cells. However, CA also induces NGF gene expression in an Nrf2-independent manner, since 50 μM of CA administration showed striking NGF gene induction compared with the classical Nrf2 inducer tert-butylhydroquinone (tBHQ) in U373MG cells. By comparative transcriptome analysis, we found that CA activates activating transcription factor 4 (ATF4) in addition to Nrf2 at high doses. CA activated ATF4 in phospho-eIF2α- and heme-regulated inhibitor kinase (HRI)-dependent manners, indicating that CA activates ATF4 through the integrated stress response (ISR) pathway. Furthermore, CA activated Nrf2 and ATF4 cooperatively enhanced the expression of NGF and many antioxidant genes while acting independently to certain client genes. Taken together, these results represent a novel mechanism of CA-mediated gene regulation evoked by Nrf2 and ATF4 cooperation

Total Skin Electron Beam Therapy

The peripherally T-cell lymphoma; Mycosis fungoides etc, has the good radiation sensitivity, and has been adapted for total skin electron beam therapy. In this study the pendular irradiation method was used for the purpose of total skin electron beam therapy in Mycosis fungoides, and physical data on the radiation field and the electron beam energy were useful clinically.皮膚に限局した一連の末梢型T細胞リンパ腫は放射線に対する感受性が高く、電子線治療の適応となる疾患である。こららの疾患は一般的に全身の皮膚に浸潤するため、治療に際してはTarget Volumeの深さに合わせた最小限のエネルギーで全身隈なく照射する必要がある。筆者等は最近臨床で遭遇した菌状息肉症の患者を治療するため、その患者に合った物理的なデータを測定した。照射野の拡大には振子照射法を用い、エネルギー低減方法は装置に装備されている鉛のスキャタラーを低原子番号で、しかも加工のしやすい塩化Vinyl板に交換する方法を工夫した。データとして治療効果、副作用に関係する線量率、エネルギー、及び照射野内平坦度について測定した結果、距離が長くなる関係から線量率が低下する全身照射法の欠点は解消できなかったが、エネルギー及び平坦度については使用可能なデータを得ることができた

Granularity of Asymmetric Screen-Film System

非対称増感紙フィルム系について、両面乳剤フィルムのウィナースペクトル測定に及ぼす影響を検討した。濃度0.5～2.5のノイズ試料のフロント乳剤、バック乳剤、支持体層それぞれで測定したスペクトルの和と、両面同時に測定した全スペクトルとを比較した。その結果、全ウィナースペクトルの値は、フロント乳剤、バック乳剤の和より高くなった。また、試料濃度が高いほど、その差は大きくなった。これらは、マイクロデンシトメータの光学系の配置と両面乳剤フィルムの構造に起因している。したがって、非対称システムのようにフィルムの前面と後面で特性を分けて考える場合、注意を要する。We have examined a factor affecting the Wiener spectrum measurement of double-emulsion film. An asymmetric screen-film system (Kodak HC/INSIGHT system) was used in this study. The results indicated that the total Wiener spectra of double-emulsion films are higher than the sum of spectra obtained with the front emulsion and back emulsion separately. These differences are attributed to the arrangement of the optical system of the microdensitometer used to measure and the structure of double-emulsion film

In Vitro Assessment of Factors Affecting the Apparent Diffusion Coefficient of Jurkat Cells Using Bio-phantoms

It is well known that many tumor tissues show lower apparent diffusion coefficient (ADC) values, and that several factors are involved in the reduction of ADC values. The aim of this study was to clarify how much each factor contributes to decreases in ADC values. We investigate the roles of cell density, extracellular space, intracellular factors, apoptosis and necrosis in ADC values using bio-phantoms. The ADC values of bio-phantoms, in which Jurkat cells were encapsulated by gellan gum, were measured by a 1.5-Tesla magnetic resonance imaging device with constant diffusion time of 30sec. Heating at 42℃ was used to induce apoptosis while heating at 48℃ was used to induce necrosis. Cell death after heating was evaluated by flow cytometric analysis and electron microscopy. The ADC values of bio-phantoms including non-heated cells decreased linearly with increases in cell density, and showed a steep decline when the distance between cells became less than 3μm. The analysis of ADC values of cells after destruction of cellular structures by sonication suggested that approximately two-thirds of the ADC values of cells originate from their cellular structures. The ADC values of bio-phantoms including necrotic cells increased while those including apoptotic cells decreased. This study quantitatively clarified the role of the cellular factors and the extracellular space in determining the ADC values produced by tumor cells. The intermediate diffusion time of 30msec might be optimal to distinguish between apoptosis and necrosis