Near-Infrared Fluorescent Probes for Imaging of Intracellular Mg2+ and Application to Multi-Color Imaging of Mg2+, ATP, and Mitochondrial Membrane Potential.

The magnesium ion (Mg2+) is an essential cation to maintain proper cellular activities. To visualize the dynamics and functions of Mg2+, there is a great need for the development of Mg2+-selective fluorescent probes. However, conventional Mg2+ fluorescent probes are falling behind in low selectivity and poor fluorescence color variation. In this report, to make available a distinct color window for multi-color imaging, we designed and synthesized highly Mg2+-selective and near-infrared (NIR) fluorescent probes, the KMG-500 series consisting of a charged ß-diketone as a selective binding site for Mg2+ and a Si-rhodamine residue as the NIR fluorophore, which showed photoinduced electron transfer (PeT)-type OFF-ON response to the concentration of Mg2+. Two types of KMG-500 series probes, tetramethyl substituted Si-rhodamine KMG-501 and tetraethyl substituted Si-rhodamine KMG-502, were synthesized for the evaluation of cell permeability. For intracellular application, the membrane-permeable acetoxymethyl derivative KMG-501 (KMG-501AM) was synthesized and allowed to stably stain cultured rat hippocampal neurons during imaging of intracellular Mg2+. On the other hand, KMG-502 was cell membrane permeable without AM modification, preventing the probe from staying inside cells during imaging. KMG-501 distributed mainly in the cytoplasm and partially localized in lysosomes and mitochondria in cultured rat hippocampal neurons. Mg2+ increase in response to the FCCP uncoupler inducing depolarization of the mitochondrial inner membrane potential was detected in the KMG-501 stained neurons. For the first time, KMG-501 succeeded in imaging intracellular Mg2+ dynamics with NIR fluorescence. Moreover, it allows one to simultaneously visualize changes in Mg2+ and ATP concentration and also mitochondrial inner membrane potential and their interactions. This probe is expected to be a strong tool for multi-color imaging of intracellular Mg2+.

[A Study of Patients Who Underwent Breast-Conserving Surgery and Developed Cryptogenic Organizing Pneumonia(COP)].

Of the 210 patients who underwent breast-conserving surgery for breast cancer and received radiation therapy for 3 years from April 2012 to March 2015 at the Department of Therapeutic Radiology of our hospital, 6 were diagnosed with cryptogenic organizing pneumonia(COP)-like pneumonia and treated as reported. The mean age of the patients was 51years(40- 65 years), and the pathological subtypes were the luminal type(5 cases)and HER2 type(1case ), all of which were treated with radiation therapy for breast conservation. Postoperative systemic therapy included hormonal therapy with anastrozole in 2 cases, tamoxifen plus LH-RH agonist in 3 cases, and chemotherapy in 1case. The mean onset time of COP was 4.2 months after the completion of irradiation therapy, and all of the 5 patients who received endocrine therapy received it concurrently with radiation therapy. The major symptoms were fever(4 cases)and cough(6 cases). Chest radiography showed an infiltrative shadow consistent with pneumonia. Steroid therapy was effective in all cases while antibiotics were not. It was inferred that COP should be regarded as one of the complications of radiation therapy after breast-conserving surgery.

Morphology and distribution of blood fluke eggs and associated pathology in the gills of cultured Pacific bluefin tuna, Thunnus orientalis.

Infestations of blood flukes of the genus Cardicola have been observed in juvenile Pacific bluefin tuna (PBT) cultured in Japan. Infected fish harbor large numbers of parasite eggs in their gills. Although the link between blood fluke infection and juvenile mortality is not clear, accumulation of parasite eggs appears to be pathogenic to the fish. We investigated the origins, general morphology/distribution, and histopathology of these eggs in artificially produced 0 yr old PBT. Dead and live fish were sampled on several occasions from two culture facilities in Wakayama prefecture, Japan. The number of eggs in each gill filament was enumerated under a microscope. In addition, we estimated the total number of eggs by dissolving the gills in a weak NaOH solution. We observed two morphologically distinct egg types in the gill filaments, smaller, oval shaped eggs in the gill lamellae and larger, crescent shaped eggs that occurred primarily in the filamentary arteries. Based on the ITS2 sequence, the ovoid and crescent shaped eggs were identified as C. orientalis and C. opisthorchis, respectively. Eggs of the former species were more abundant (maximum: 6400 per filament) than the latter (maximum: 1400), but the number was highly variable among filaments. The eggs of the latter species were relatively evenly distributed among the filaments. In a heavily infected individual, we estimated a total of >4.5 million eggs were present in the gills on one side of the fish. The number of eggs from the two species was positively correlated to each other and the dead fish tended to harbor more eggs than the live fish. Histological observation revealed host responses around the eggs, including encapsulation by fibroblasts and nodule formation, as seen in response to other aporocotylid eggs. In addition, we observed widespread fusion of gill lamellae and blockage of the filamentary arteries in some instances. Our results provide information that can be used for routine diagnosis of Cardicola blood flukes in cultured tuna and suggest they represent a risk to juvenile PBT.

Spectral sensitivity of juvenile chub mackerel (Scomber japonicus) in visible and ultraviolet light.

Although chub mackerel (Scomber japonicus) is widely distributed all over the world, the relevance of its visual sensitivity to its ecology is not yet fully understood. We investigated spectral sensitivity in juvenile chub mackerel in the range of ultraviolet (UV) to visible light (369-652 nm) by electroretinogram (ERG) using light-emitting diodes (LEDs). Sensitivity peaked at a wavelength of approximately 482 nm in dark-adapted fish and 525 nm in light-adapted fish. A secondary sensitivity peak in the UV range at approximately 382 nm was found in both dark- and light-adapted fish. The UV sensitivity of chub mackerel may be attributable to UV transmissibility of the optical media and to the presence of a beta-band of visible light-sensitive visual pigments, and not to an alpha-band of UV visual pigments. This UV sensitivity may be useful for feeding or communication with other fishes.

The histological analysis, colorimetric evaluation, and chemical quantification of melanin content in 'suntanned' fish.

Human melanocytes respond to UV irradiation by increasing the synthesis of melanin. While much is now understood of the pathways governing this process and the nature of the melanin synthesized, little is known of melanins produced by lower vertebrates and their capacity to respond to UV. Here we report that a fish, red seabream, can undergo 'suntanning'. Histological, colorimetric and chemical assays were performed for suntanned red seabream fish bred in net cages to analyse the melanins and compared with shaded or wild red seabream fish. For color evaluation, the L* values of suntanned fish were dramatically lower than those in the other two groups. Pyrrole-2,3,5-tricarboxylic acid (PTCA), an indicator of eumelanin, was detected in suntanned fish at five times higher levels than in shaded or wild fish while 4-amino-3-hydroxyphenyl-alanine (4-AHP), a marker for pheomelanin, could not be detected in any of the samples. Histological analysis showed that melanocytes in the suntanned skin enlarged and increased in number to form a monolayer at the surface of the skin. Analysis of L* values and PTCA levels showed quite a high correlation coefficient (r = -0.843). When comparing shaded and wild red seabream fish, the scores were closer but some significant differences were still found in some body areas. These results indicate that eumelanin accumulates in suntanned fish during the increase in skin color, which is induced by sunlight, presumably by ultraviolet radiation.

Extraosseous accumulation of 99mTc-MDP in lymph node metastases of small cell carcinoma of the esophagus.

We report a case of esophageal carcinoma that showed extraosseous accumulation of 99mTc-MDP in lymph node metastases to the cervical and paracardial lymph nodes. There are few cases showing abnormal extraosseous accumulation of 99mTc-MDP in esophageal cancer lesion. The patient was a 53-year-old man with advanced esophageal cancer. Bone scintigraphy demonstrated extraosseous accumulations in left supraclavicular and paracardial lymph node metastases. The histopathological diagnosis was small cell carcinoma of the esophagus, which is a rare disease with aggressive behavior and poor prognosis. Our patient underwent 2 courses of systemic chemotherapy (CDDP + VP16), but died of rapidly growing systemic metastases 5 months after the initial treatment.

Adult embryonal rhabdomyosarcoma in axilla.

A case of embryonal rhabdomyosarcoma (RMS) arising from an axillary space is described. RMS is one of the most common soft-tissue neoplasms in children, but adult embryonal RMS is rare. The patient was a 55-year-old man with a tumor of the left axillary area. RMS arising from an axillary area is also uncommon. The histological diagnosis was embryonal RMS. He received irradiation combined with chemotherapy (VAC: VCR, act-D, CPA) for the primary and metastatic lesions, obtained complete remission, and has remained disease-free for over four years as of this report.

Intense Ga-67 accumulation in pure primary mediastinal seminomas.

The authors examined three patients with pure primary mediastinal seminomas, all of whom had marked Ga-67 uptake on scintigraphy. The unusual presentation of sclerotic bone metastases with remarkable extraosseous soft tissue involvement was observed in one patient. Another had a seminoma associated with a multiloculated thymic cyst, which is a rare lesion.

Scintigraphic findings of MALT lymphoma of the thyroid.

Mucosa-associated lymphoid tissue (MALT) lymphoma has been established as a distinct entity among non-Hodgkin's lymphomas, and the most common primary site is the stomach. We describe scintigraphic findings in a patient with MALT lymphoma of the thyroid. A 71-year-old woman with Hashimoto's thyroiditis suffered from rapid cervical swelling, and ultrasonography and CT revealed a thyroid nodule. The nodule showed accumulation of 99mTc pertechnetate comparable to the surrounding thyroid tissue, mimicking a benign nodule. Both 67Ga and 201Tl imaging visualized the lesion as an increased uptake area. After radiotherapy, abnormally increased uptake disappeared on 67Ga images, which predicted a favorable outcome. MALT lymphoma of the thyroid may be visualized as a warm nodule on 99mTc pertechnetate scintigraphy.

Quantitative lung perfusion scintigraphy and detection of intrapulmonary shunt in liver cirrhosis.

OBJECTIVE: Frequent association between liver cirrhosis and hypoxemia has been well documented. It is mostly attributable to intrapulmonary shunt due to dilation of pulmonary vasculature. We performed quantitative lung perfusion scintigraphy to detect an intrapulmonary shunt in cirrhosis patients. METHODS: Prior to injection, Tc-99m MAA was applied to thin layer chromatography for quality control. Three cirrhosis patients who had hypoxemia were examined as well as 11 control subjects. After i.v. injection of Tc-99m MAA, whole body anterior and posterior images were taken at 5 min in patients with cirrhosis and at 8 time points up to 60 min in control subjects. Regions of interest were placed at the bilateral lungs and the whole body, and pulmonary accumulation was calculated. RESULTS: All the control subjects demonstrated more than 90% of radioactivity in the lungs until 20 min. In contrast, all the patients showed values less than 80% at 5 min. In the cirrhosis patients with hypoxemia, the presence of intrapulmonary shunt was confirmed on quantitative lung perfusion scan. In control subjects, pulmonary accumulation of Tc-99m MAA dropped as a function of time and became less than 90% after 30 min. CONCLUSION: The timing of measurements is essential in evaluating intrapulmonary shunt.