Near-Infrared-Responsive Peptide that Targets Collagen Fibrils to Induce Cytotoxicity.

A novel conjugate, PHG10 dye, was synthesized using a collagen peptide and a near-infrared (NIR)-responsive dye to achieve targeted cytotoxicity. The collagen peptide motif, -(Pro-Hyp-Gly)10 - (PHG10), was incorporated for targeting collagen fibrils that are excessively produced by activated fibroblasts around tumor cells. PHG10 dye was purified by HPLC and identified by MALDI-MS. The phototoxicity and cytotoxicity of PHG10 dye were examined using human glioma cells (HGCs). Fluorescent images indicated that PHG10 dye preferably assembled to collagen-coated HGCs compared with noncoated HGCs. Under irradiation with NIR light, effective cytotoxicity was observed on collagen-coated HGCs within 20 min. Because phototoxicity and cytotoxicity are dependent on the assembled amount of PHG10 dye, the targeting of collagen fibrils by the collagen peptide motif PHG10 is assured.

Interstitial MR lymphography in mice: comparative study with gadofluorine 8, gadofluorine M, and gadofluorine P.

PURPOSE: We investigated the characteristics and capability of interstitial MR lymphography in mice using gadofluorine 8, gadofluorine M, and gadofluorine P. METHODS: We injected healthy mice with 0.5 µmol of Gd gadofluorine 8, gadofluorine M, or gadofluorine P subcutaneously into the right rear footpad and assessed the time courses of contrast enhancement in the lymph nodes. Six mice were studied for each contrast agent. We also used gadofluorine M to assess the lymphatic pathway from the right and left rear feet or tail. RESULTS: Contrast enhancement was demonstrated for the right popliteal, sacral, and iliac lymph nodes in all mice 5 minutes after injection of each of the 3 agents and decreased gradually. Enhancement in the lymph nodes was still detectable 30 minutes after injection of gadofluorine 8 or gadofluorine M. Enhancement became obscure sooner after gadofluorine P injection and was mildly stronger with the other 2 contrast agents. Clear differences were found in the hepatobiliary and urinary kinetics of the 3 agents. Gadofluorine M injected into various sites delineated the lymphatic pathway from the site of injection. CONCLUSION: Interstitial MR lymphography using gadofluorine 8, gadofluorine M, and gadofluorine P offered clear visualization of the lymphatic pathway in healthy mice during a sufficient imaging time window, and allowed repeated assessment of the pathway and clarification of the lymphatic system.

Morphological study of the human hyoid bone with three-dimensional CT images -Gender difference and age-related changes.

The human hyoid bone supports the base of the tongue and is involved in breathing, chewing, and swallowing as well as in the muscle movements associated with articulation. Accordingly it plays an important bone for a human to live. It is a very interesting organ also in multiple special area, including anatomy, mastication, swallowing, articulation, and also forensic medicine. In the morphological study of the human hyoid bone, there is a comparative anthropological research early in 1900, whereas the metrological research has been little reported later. We first used MDCT, and recorded each organic hyoid locus with a three-dimensional image for three-dimensional morphometry of gender differences, age-related changes, and the morphologic characters of the hyoid bone, and compared them with the results of our predecessors. By measuring the volume of the human hyoid bone, we identified gender difference at high rates, and estimated a certain level of ages based on the ossification at the junction area of the hyoid body and greater horns observed. Our results can be applied in the forensic medicine. By examining 600 cases, atypical horseshoes-shapes were found and the existence of the hyoid bone protrusion was demonstrated at high rates.

Integrated lymphography using fluorescence imaging and magnetic resonance imaging in intact mice.

We assessed lymph drainage in living mice by an integrated imaging method using fluorescence imaging (FLI) and magnetic resonance imaging (MRI). Mice were subcutaneously injected with quantum dots and gadofluorine 8 into the right rear footpad. They were fixed on a transparent flat plate and underwent FLI and MRI successively. Small markers were attached to the mouse surface for spatial coregistration, and image fusion of FLIs and MRIs was performed. Two-dimensional fluorescence reflectance imaging was used for FLI. FLI and MRI provided generally consistent results and demonstrated lymphatic flow to the popliteal, sacral, and iliac lymph nodes in most mice and to the renal, inguinal, and lumbar-aortic lymph nodes in some mice. On the fusion images, the locations of the lymph nodes in the mouse trunk were in good agreement between FLI and MRI, indicating successful spatial registration even for the deep structures. The popliteal node tended to be visualized a little farther caudally in FLI than in MRI, presumably because the overlying tissues were thicker in the cranial portion. Integrated FLI/MRI lymphography with image fusion appears to be a useful tool for analysis of the murine lymphatic system.

Diet and gastrointestinal signal on T1-weighted magnetic resonance imaging of mice.

In magnetic resonance (MR) imaging of small animals, the gastrointestinal contents may give rise to intense signals on T1-weighted images. The aim of this study was to determine the optimal dietary preparation to reduce gastrointestinal signals in mice and to evaluate the usefulness of this approach. Images of the mouse trunk were obtained using a T1-weighted, three-dimensional fast low-angle shot sequence under various dietary conditions and were compared with respect to the gastrointestinal signals and image quality. The dietary preparation studied included giving alternative diets for 24 h, intestinal cleansing, and 6-h fasting. Mice with and without dietary preparation underwent MR lymphography using gadofluorine 8, and the visualization of abdominal lymph nodes was compared. In the absence of dietary preparation, hyperintense areas were conspicuous in the gastrointestinal system, whereas on the images taken from mice fed potato or sweet potato for 24 h before imaging, gastrointestinal hyperintensity was less prominent. This preparation also reduced artifactual signals and resulted in higher-quality images of the kidneys. Intestinal cleansing, which consisted of 24-h fasting and laxative intake, did not reduce the gastrointestinal signals and caused signal changes that were indicative of fatty liver development. Some of the abdominal lymph nodes of the mice that did not receive dietary preparation were visualized on MR lymphography source images but not on maximum intensity projection (MIP) images. In contrast, on the MIP images of mice fed potato, all the lymph nodes delineated on the source images were successfully visualized. In conclusion, feeding mice potato or sweet potato for 24 h before MR imaging reduces the gastrointestinal signals and image degradation due to artifacts. Appropriate dietary preparations facilitate the display of target structures on MIP images and are expected to enhance the capabilities of small animal MR imaging.

Anti-hypertensive effect of gamma-aminobutyric acid (GABA)-rich Chlorella on high-normal blood pressure and borderline hypertension in placebo-controlled double blind study.

The anti-hypertensive effect of GABA-rich Chlorella was studied after oral administration for 12 weeks in the subjects with high-normal blood pressure and borderline hypertension in the placebo-controlled, double-blind manner in order to investigate if GABA-rich Chlorella, a dietary supplement, is useful in control of blood pressure. Eighty subjects with Systolic blood pressure (SBP) 130-159 mmHg or diastolic blood pressure (DBP) 85-99 mmHg (40 subjects/group) took the blinded substance of GABA-rich Chlorella (20 mg as gamma-aminobutyric acid) or placebo twice daily for 12 weeks, and had follow-up observation for an additional 4 weeks. Systolic blood pressure in the subjects given GABA-rich Chlorella significantly decreased compared with placebo (p < 0.01). Diastolic blood pressure had the tendency to decrease after intake of GABA-rich Chlorella. Neither adverse events nor abnormal laboratory findings were reported throughout the study period. Reduction of SBP in the subjects with borderline hypertension was higher than those in the subjects with high-normal blood pressure. These results suggest that GABA-rich Chlorella significantly decreased high-normal blood pressure and borderline hypertension, and is a beneficial dietary supplement for prevention of the development of hypertension.

Evaluation of gadoxetate disodium as a contrast agent for mouse liver imaging: comparison with gadobenate dimeglumine.

We investigated the characteristics of gadoxetate disodium (Gd-EOB-DTPA) as a contrast agent for magnetic resonance imaging of the mouse liver. Mice were imaged sequentially under isoflurane anesthesia using a T1-weighted, three-dimensional fast low-angle shot (3D FLASH) sequence after an intravenous injection of Gd-EOB-DTPA or gadobenate dimeglumine (Gd-BOPTA), and the time course of the contrast effect was examined. The time course of the contrast effect of Gd-EOB-DTPA was also assessed after intravenous injection under pentobarbital anesthesia and after subcutaneous injection while awake or under isoflurane or pentobarbital anesthesia. Moreover, different doses of Gd-EOB-DTPA or Gd-BOPTA were injected subcutaneously into conscious mice, and the clarity of the liver border was evaluated visually. Intravenous injection under isoflurane anesthesia caused rapid contrast enhancement in the liver with both Gd-EOB-DTPA and Gd-BOPTA, and the contrast effect was 41% stronger with Gd-EOB-DTPA. Subcutaneous injection of Gd-EOB-DTPA caused delayed but favorable contrast enhancement in the liver. Washout of Gd-EOB-DTPA was faster in mice injected while awake than in those injected under anesthesia. After intravenous injection, washout was faster under pentobarbital anesthesia than under isoflurane anesthesia. The peak liver contrast was 11% and 18% stronger under pentobarbital anesthesia than under isoflurane anesthesia, after intravenous and subcutaneous injections, respectively. Subcutaneous injection of Gd-EOB-DTPA or Gd-BOPTA caused dose-dependent contrast effects in the liver. At a given dose, the contrast effect tended to be stronger and liver demarcation tended to be clearer with Gd-EOB-DTPA than with Gd-BOPTA. In conclusion, intravenous or subcutaneous injection of Gd-EOB-DTPA produces a favorable contrast effects in the mouse liver, indicating its potential in investigating mouse models of liver diseases. The contrast effects vary between conscious mice and anesthetized mice and among anesthetic agents used.

Gadobenate dimeglumine as a contrast agent for MRI of the mouse liver.

We investigated the characteristics and utility of gadobenate dimeglumine (Gd-BOPTA) for MRI of the mouse liver. Mice were imaged under isoflurane anesthesia using a T(1)-weighted, three-dimensional fast low-angle shot (3D FLASH) sequence before and after intravenous or subcutaneous injection of Gd-BOPTA, and the time course of the contrast effect was examined. The appropriate dose for subcutaneous injection was determined visually, and the inter- and intra-observer reproducibilities in liver volumetry were evaluated with and without contrast injection. When mice were imaged sequentially before and after Gd-BOPTA injection and isoflurane anesthesia was maintained throughout the experiment, a long-lasting contrast effect was noted in the liver. Subcutaneous injection caused delayed, but favorable, enhancement. Washout from the liver was definitely accelerated in conscious mice in comparison with anesthetized mice. Visual evaluation indicated that a dose of 0.1 mmol/kg was appropriate for clear delineation of the entire liver margin, and the application of Gd-BOPTA significantly improved the inter- and intra-observer reproducibilities of liver volumetry. In conclusion, the intravenous or subcutaneous injection of Gd-BOPTA has a favorable contrast effect for the mouse liver, resulting in clear visualization of the liver border and improved reproducibility of liver volumetry. The possible influence of anesthesia on the pharmacokinetics of a contrast agent should be considered in determining the optimal scan timing.

A preliminary study of the draining lymph node basin in advanced lower rectal cancer using a radioactive tracer.

PURPOSE: This study was designed to examine the draining lymph node basin at highest risk of metastasis in lower rectal cancer using 99 mTc-tin colloid. METHODS: In 43 patients, the area with highest hot nodes density was defined as the draining lymph node basin using a gamma probe. Metastatic states of all removed lymph nodes were examined histologically. RESULTS: A total of 203 hot nodes were identified in 39 patients (91 percent) with a mean of 5.2 nodes. The number of removed lymph nodes was 808 nodes: 670 nodes in the mesorectum, and 138 nodes in the pelvis. In 21 patients, the metastatic states of 119 nodes were investigated histologically. From the distribution of the identified hot nodes, the draining lymph node basins were classified into two patterns: lateral type (n = 17), and mesorectal type (n = 22). Only 20 (17 percent) of 119 positive nodes were identified as hot node. Tumor cells that occupied the lymph node diffusely or massively probably interfered with the exact diagnosis of metastasis. Lymph node metastasis in the pelvis was observed in 5 patients (13 percent). One false negative was of the mesorectal type with diffuse metastases in the para-aortic lymph nodes. The remaining four patients were of the lateral type and all positive lymph nodes, including positive nonhot nodes, were located within the draining lymph node basin. Consequently, in 20 (95.2 percent) of 21 patients with lymph node metastasis, all positive lymph nodes were located within the draining lymph node basin of the tumor. CONCLUSIONS: Two types of the draining lymph node basin of advanced lower rectal cancer were identified using this method. The concordance between lymph node metastases and the draining lymph node basin is good.

[Clinical significance and problems of sentinel node identification for individualization of rectal cancer therapy].

In the surgical treatment of advanced rectal cancer, wide lymphadenectomy has been replaced by autonomic nerve preservation because of serious problems such as postoperative urinary and sexual dysfunction. This may offer a considerable clue to as whether metastasis of lateral lymph node can be diagnosed by a new technique in order to provide more benefits to many patients with advanced rectal cancer. Moreover the sentinel lymph node (SLN) concept has lately attracted attention as a new technique. We studied SLN in 40 patients with advanced lower rectal cancer using 99mTc-Sn colloid as a tracer. SLN was successfully identified in 35 (87.5%) of 40 patients, and the mean number of SLNs identified per patient was 5.6 lymph nodes (range 1-14). In this study overall accuracy was 86%, and it was suggested that lateral lymphadenectomy was excluded in 91%, thanks to application of the SLN technique. Although the role this technique plays in the individualization of the therapy for rectal cancer will be large in the near future, many problems remain to be solved in the exact identification of SLN.

Contrast-enhanced MR angiography in rats with hepatobiliary contrast agents.

The aim of this animal study was to evaluate the feasibility of contrast-enhanced magnetic resonance (MR) angiography with two hepatobiliary contrast agents, Gd-DTPA-DeA and Gd-EOB-DTPA. Coronal images of the rat abdomen were acquired using a three-dimensional spoiled gradient recalled sequence before and after the administration of Gd-DTPA-DeA, Gd-EOB-DTPA, or Gd-DTPA. Four sets of postcontrast images were collected every 90 s. Contrast ratios were calculated for the abdominal aorta on the source images, and the retention index was defined as the ratio of the contrast ratio on the last imaging to that on the first postcontrast imaging. Partial minimum intensity projection (MIP) images covering the abdominal aorta were generated from the first and second postcontrast imagings, and the image quality was visually evaluated. The contrast ratio on the first postcontrast imaging was the highest for Gd-DTPA-DeA, followed by Gd-EOB-DTPA and Gd-DTPA. Retention indices were higher with Gd-DTPA-DeA than with Gd-EOB-DTPA and Gd-DTPA, implying a prolonged contrast effect with Gd-DTPA-DeA. On the MIP image from the first postcontrast imaging, delineation of the abdominal aorta tended to be better with Gd-DTPA-DeA and Gd-EOB-DTPA than with Gd-DTPA, and the difference was evident at low injection doses. Image quality for the second postcontrast imaging was higher with Gd-DTPA-DeA than with the other two agents, suggesting a longer imaging window for Gd-DTPA-DeA. In conclusion, Gd-DTPA-DeA and Gd-EOB-DTPA showed stronger contrast enhancement for the rat abdominal aorta and provided MR angiograms of higher image quality when compared with Gd-DTPA at the same injection dose. These hepatobiliary agents may make it possible to perform contrast-enhanced MR angiography even at a low injection dose.

Phantom and animal studies of a new hepatobiliary agent for MR imaging: comparison of Gd-DTPA-DeA with Gd-EOB-DTPA.

PURPOSE: To investigate the characteristics of Gd-DTPA-DeA as a hepatobiliary contrast agent for MR imaging in comparison with those of Gd-EOB-DTPA. MATERIALS AND METHODS: We undertook phantom experiments to assess T1 relaxivity for Gd-DTPA-DeA, Gd-EOB-DTPA, and Gd-DTPA in human plasma. For Gd-DTPA-DeA and Gd-EOB-DTPA, we evaluated the contrast effect in rats using an SPGR sequence. The contrast ratios of liver and abdominal aorta were measured up to 21 minutes after intravenous administration of the agents. Visualization of the bile duct and renal pelvis was also assessed. RESULTS: In human plasma, T1 relaxivity was similar for Gd-DTPA-DeA and Gd-EOB-DTPA, and higher than those for Gd-DTPA. Whereas the contrast ratio of liver peaked about five minutes after the injection of Gd-EOB-DTPA and was followed by a subsequent decline, a continuous rise was shown for Gd-DTPA-DeA, resulting in a larger maximal contrast effect. Contrast ratios of the abdominal aorta were larger for Gd-DTPA-DeA. Biliary excretion was observed for both agents but occurred earlier with Gd-EOB-DTPA. While renal excretion was shown for all rats three minutes after the injection of Gd-EOB-DTPA, it was not observed for Gd-DTPA-DeA. CONCLUSION: Gd-DTPA-DeA may be used as a hepatobiliary contrast agent and shows different pharmacokinetics from Gd-EOB-DTPA.

Interstitial magnetic resonance lymphography: comparative animal study of gadofluorine 8 and gadolinium diethylenetriamine-pentaacetic acid.

PURPOSE: We evaluated the usefulness of gadofluorine 8 (G8) and gadolinium diethylenetriamine-pentaacetic acid (Gd-DTPA) for interstitial magnetic resonance lymphography (MRL). METHOD: Phantom imaging studies were conducted with G8 and Gd-DTPA corresponding to 0.1-0.005 micromol Gd/mL, and signal intensity was measured. Popliteal lymph node (PLN) accumulation was investigated in rabbits. Imaging was performed before and after subcutaneous administration of G8 (50 micromol Gd/mL) and Gd-DTPA (50 micromol Gd/mL). Contrast enhancement ratio measurements of PLN were determined both prior to and following the administration. RESULTS: Signal intensity of phantom for G8 are higher than those for Gd-DTPA from 0.1 to 0.002 micromol Gd/mL (P < 0.0001). Imaging after 5 minutes can clearly detect PLN accumulation on two contrast agents. Nevertheless, PLN accumulation of Gd-DTPA disappeared after about 30 minutes. A statistically significant difference between G8 and Gd-DTPA can be seen from 5 to 90 minutes in CER (P < 0.0001). PLN became clearly visible at 5 minutes after the injection of G8 and Gd-DTPA. Nevertheless, PLN accumulation of Gd-DTPA disappeared at 30 minutes. Otherwise, PLN accumulation of G8 continued up to 90 minutes. CONCLUSION: These studies indicated that G8 was more suitable than Gd-DTPA as a contrast agent for interstitial MRL up to 90 minutes.