Semi-quantitative method of N-isopropyl-(123I)-p-iodoamphetamine accumulation in cerebrum: cerebrum to cerebellum accumulation ratio

Semi-quantitative method of N-isopropyl-(123I)-p-iodoamphetamine (123I-IMP) accumulation in cerebrum by using 123I-IMP SPECT was tried. Ratio of counts in whole cerebrum to those in normal cerebellum was calculated, and this accumulation is defined as cerebrum to cerebellum accumulation ratio. In group of patients with dementia or patients who were remaining in bed, significantly low accumulation ratio was observed, while in "normal" group, no differences in sex and aging were observed. Calculation of cerebrum to cerebellum accumulation ratio was concluded as a useful method of evaluating the accumulation of 123I-IMP in cerebrum.     

In Vivo Optical Properties of Melanocytic Skin Lesions: Common Nevi, Dysplastic Nevi and Malignant Melanoma

We present an in vivo study of the optical properties of common nevi, dysplastic nevi and malignant melanoma skin lesions in human subjects. Reflectance spectra were measured on 1379 skin lesions, in the visible and near-infrared spectral regions, using a spectral imaging system, in a clinical setting. Analysis of the data using a reflectance model revealed differences between the optical properties of melanin present in nevi and melanoma lesions. These differences, which are in agreement with our previous observations on average reflectance spectra, may be potentially useful for the noninvasive characterization of pigmented skin lesions and the early diagnosis of melanoma.     

In vitro PHOTOSENSITIZING PROPERTIES OF RHODAMINE 123 ON DIFFERENT HUMAN TUMOR CELL LINES

Abstract— In this study, human tumor cell lines of different origin (colon carcinoma HT29, breast carcinoma MCF7 and malignant melanoma M14) were incubated for 24 h at 37*deg;C with Rhodamine 123 (Rh123) at concentrations ranging up to 4 μg/ml;. Immediately after drug removal, light irradiation was delivered at 500 W/m² for 5 min using an argon laser. After irradiation, viable cells were counted and assayed for colony formation. When only Rh123 was administered, a 50% survival was obtained at about 2.77 μg/ml and 1.48 μg/ml; for HT29 and MCF7, respectively. After light irradiation, 50% survival doses decreased to 0.47 μg/ml and 0.18 μg/ml for the two carcinoma cell lines, respectively. In the case of malignant melanoma, the decrease in survival was relatively lower than those obtained with carcinoma cells: 50% survival dose was 3.54 μg/ml with Rh123 alone and 1.32 μ/ml after irradiation. The lower sensitivity of M14 melanoma cells seems to be related to different uptake and release of drug by these cells with respect to carcinoma lines.     

Regional cerebral blood flow in cerebrovascular disease studied using N-isopropyl-p-(123I) iodoamphetamine

Twenty-one patients with cerebrovascular disease (5 with hemorrhage, 10 with infarction, 2 with TIA, 4 with motor disturbance of unknown cause) were studied using N-isopropyl-p-(123I)iodoamphetamine (123I-IMP) and single photon emission CT. In 3 of 5 cases with cerebral hemorrhage, perfusion defects were shown in and around the region of hematomas, furthermore, one of the cases with internal capsular hematoma, the perfusion defect extended to the cortical area corresponding to the neurological pathway. In one case with MCA infarction, the perfusion defect was greater and clearer than the low density area on X-ray computed tomography (CT). These results revealed the 123I-IMP study provides physiological information in contrast with X-ray CT which provides anatomical information. In 3 of 9 cases with multiple small deep hemispheric infarctions on X-ray CT, the perfusion to the basal ganglia was suspected to be decreased on 123I-IMP images. However, this visual findings was not definitive and, in fact, the diseased side was not always consistent with the clinical findings. For quantitative analysis, symmetrical regions of interest (ROIs) were constructed both basal ganglia and the ratio of average counts over the ROI to those over the whole slice was calculated. In the small infarction group, the mean +/- S.D. of the values was 0.89 +/- 0.09 in right and 0.89 +/- 0.08 in left. Although the values were not significantly different from those of normal subjects (0.99 +/- 0.02 in right, 0.97 +/- 0.03 in left), they distributed in the range less than normal in 5 of 9 cases. This method was thought to be useful and practical to evaluate the cerebral blood flow in basal ganglia of patients with deep hemispheric infarction.     

Clinical significance of 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) in the diagnosis of intrahepatic masses

Hepatobiliary scintigraphy with 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) was carried out on 48 patients with intrahepatic masses, 44 with hepatocellular carcinoma and one each of hepatocellular adenoma, focal nodular hyperplasia, cholangiocellular carcinoma, and adenoid cystic carcinoma. Scans were performed twice, early scan (30 min post i.v.) and delayed scan (2.5 h post i.v.), and the delayed scan was used for assessing the accumulation of 99mTc-PMT in the intrahepatic masses. In the hepatocellular carcinoma group, based on individual patients, 17 out of 44 (38.6%) showed accumulation of 99mTc-PMT in various degrees; and based on individual masses, accumulation was noted in 21 out of 55 masses (38.2%). However, only the cases which had not received transarterial infusion of anti-cancer drugs (TAI) and/or blocking agents (TAE) were taken into consideration, 9 out of 18 patients (50%) and 12 out of 25 masses (48.0%) were found capable of picking up 99mTc-PMT. A case of hepatocellular adenoma showed a strong accumulation of 99mTc-PMT in the mass which was depicted as a defect on the 99mTc-colloid scan and did not show a significant accumulation of 67Ga. In a case of focal nodular hyperplasia, there were two space-occupying lesions (SOLs), one of which showed a clear-cut defect on the 99mTc-colloid scan and the other which showed only a distorted uptake pattern. However, both masses were strongly positive with 99mTc-PMT. 99mTc-PMT scintigraphy is useful in connection with 99mTc-colloid scan and sometimes with 67Ga-citrate in the diagnosis of intrahepatic masses originating from hepatocytes.     

Selection of malignant melanoma variant cell lines for ovary colonization

Murine melanoma line B16-F1, which shows some specificity for metastatic organ colonization of lung but rarely metastasizes to ovary, was used to select variant cell lines with increased preference for experimental ovary metastasis. Ovary-colonizing melanoma cell lines were sequentially selected in syngeneic C57BL/6 mice by repeated intravenous administration and surgical recovery of ovarian melanoma tumors for tissue culture. After ten selections for experimental ovary metastasis, line B16-010 was established which formed experimental metastatic ovary tumors in almost every test animal. In tissue culture B16-010 cells grew in circular colonies with rounded, smooth cell peripheries compared to B16-F1 cells which were flatter, grew in irregular patterns, and exhibited long cellular projections. Ovary-selected B16 lines contained less melanin pigment (B16-010 less than B16-05 less than B16-01 approximately equal to B16-F1) compared to the parental melanoma line. Together with previous cloning and selection data, these results are consistent with the preexistence of highly malignant cells in the parental tumor population that possess the ability to metastasize to specific organs.     

Electron spin resonance imaging of mouse B16 melanoma

An X-band electron spin resonance (ESR) imaging apparatus with a pin-hole TE102 mode cavity and a rapid scan coil was constructed. Using this apparatus, ESR imaging of melanin in mouse B16 melanoma was observed for the first time. The ESR spectrum of B16 melanoma is similar to that of natural melanin extracted from sepia officinalis in microwave power dependence.     

Optical properties of human melanocytic nevi in vivo

We present an in vivo study of the optical properties of melanin present in melanocytic nevi of human subjects with Fitzpatrick skin type III (Caucasian descent) using optical spectroscopy. We show that the melanin absorption spectrum exhibits an exponential dependence on wavelength with a decay constant which follows a normal distribution characteristic of a random biological variable. Moreover, we demonstrate lack of correlation among melanin optical properties, melanin concentration and skin light scattering properties, which indicates that the true optical absorption of melanin can be measured free from confounding scattering effects. We also show that the average melanin absorption spectrum in vivo is in very good agreement with a previously reported oxygen photoconsumption action spectrum of melanin. Finally, we provide an overview of the emerging picture of the melanin absorption properties in vivo among various skin types and also among various skin lesions such as melanocytic nevi and melanoma.     

Epidemiological Support for an Hypothesis for Melanoma Induction Indicating a Role for UVA Radiation

An hypothesis for melanoma induction is presented: UV radiation absorbed by melanin in melanocytes generates products that may activate the carcinogenic process. Products formed by UV absorption in the upper layers of the epidermis cannot diffuse down as far as to the melanocytes. Thus, melanin in the upper layer of the skin may be protective, while that in melanocytes may be photocarcinogenic. Observations that support this hypothesis include: (1) Africans with dark skin have a reduced risk of getting all types of skin cancer as compared with Caucasians, but the ratio of their incidence rates of cutaneous malignant melanoma to that of squamous cell carcinoma is larger than the corresponding ratio for Caucasians. (2) Albino Africans, as compared with normally pigmented Africans, seem to have a relatively small risk of getting cutaneous malignant melanomas compared to nonmelanomas. This is probably also true for albino and normally pigmented Caucasians. (3) Among sun-sensitive, poorly tanning persons, frequent UV exposures are associated with increased risk of melanoma, whereas among sun-resistant, well-tanning persons, increased frequency of exposure is associated with decreased melanoma risk. (4) It is likely that UVA, being absorbed by melanin, might have a melanoma-inducing effect. This is in agreement with some epidemiological investigations which indicate that sun-screen lotions may not protect sufficiently against melanoma induction. The relative latitude gradient for UVA is much smaller than that for UVB. The same is true for the relative latitude gradient of cutaneous malignant melanoma as compared with squamous cell carcinoma and basal cell carcinoma. Under the assumption that the average slopes of the curves relating incidence rates with fluences of carcinogenic UV radiation are similar for melanomas and nonmelanomas, these facts are in agreement with the assumption that UVA plays a significant role in the induction of melanomas in humans. This is in agreement with the experimental results with Xiphophorus.     

Artocarpus plants as a potential source of skin whitening agents

Artocarpus plants have been a focus of constant attention due to the potential for skin whitening agents. In the in vitro experiment, compounds from the Artocarpus plants, such as artocarpanone, norartocarpetin, artocarpesin, artogomezianol, andalasin, artocarbene, and chlorophorin showed tyrosinase inhibitory activity. Structure-activity investigations revealed that the 4-substituted resorcinol moiety in these compounds was responsible for their potent inhibitory activities on tyrosinase. In the in vitro assay, using B16 melanoma cells, the prenylated polyphenols isolated from Artocarpus plants, such as artocarpin, cudraflavone C, 6-prenylapigenin, kuwanon C, norartocarpin, albanin A, cudraflavone B, and brosimone I showed potent inhibitory activity on melanin formation. Structure-activity investigations revealed that the introduction of an isoprenoid moiety to a non-isoprenoid-substituted polyphenol enhanced the inhibitory activity of melanin production in B16 melanoma cells. In the in vivo investigation, the extract of the wood of Artocarpus incisus and a representative isolated compound from it, artocarpin had a lightening effect on the skin of guinea pigs' backs. Other in vivo experiments using human volunteers have shown that water extract of Artocarpus lakoocha reduced the melanin formation in the skin of volunteers. These results indicate that the extracts of Artocarpus plants are potential sources for skin whitening agents.     

Inhibitory effect of capsaicin on B16-F10 melanoma cell migration via the phosphatidylinositol 3-kinase/Akt/Rac1 signal pathway

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), the major pungent ingredient of red pepper, has been reported to possess anti-carcinogenic and anti-mutagenic activities. In this study, the anti-migration activity of capsaicin on highly metastatic B16-F10 melanoma cells was investigated. Capsaicin significantly inhibited the migration of melanoma cells without showing obvious cellular cytotoxicity at low doses. This effect correlated with the down-regulation of phosphatidylinositol 3-kinase (PI3-K) and its downstream target, Akt. Although B16-F10 cell migration was increased by the PI3-K activator through the activation of Akt, these PI3-K activator-induced phenomena were attenuated by capsaicin. Moreover, capsaicin was found to significantly inhibit Rac1 activity in a pull-down assay. These results demonstrate that capsaicin inhibits the migration of B16-F10 cells through the inhibition of the PI3-K/Akt/Rac1 signal pathway. The present investigation suggests that capsaicin targets PI3-K/Akt/ Rac1-mediated cellular events in B16-F10 melanoma cells. Consequently, capsaicin administration should be considered an effective approach for the suppression of invasion and metastasis in malignant melanoma chemotherapy.     

Evaluation by using radionuclide uptake of bone in Paget's disease of bone: special reference to treatment with calcitonin

Bone scintigraphy using 99mTc-MDP was performed on 2 patients with Paget's disease of bone before and after the treatment with a synthetic eel calcitonin analogue [Asu1,7)-eel calcitonin, ECT] at a dose of 40 U per day. All pagetic lesions showed markedly the increased accumulation of the radionuclide. The uptake ratio, defined as the count rate of 99mTc-MDP over each bone lesion to that over the control bone, was calculated. The response to the calcitonin therapy was evaluated with the uptake ratio of the radionuclide. The uptake ratio decreased markedly within the first 3 months of the treatment, in association with a palliation of bone pain, while the serum alkaline phosphatase activities which had been within the normal range or slightly high before the treatment did not show any significant change or did not reflect a clinical feature (e.g. bone pain) with the treatment. Thus, the uptake ratio on the bone scintigram seemed to offer the most sensitive and most reliable information for the evaluation of calcitonin treatment of Paget's disease of bone.     

Scintigraphic evaluation of brain death with 99mTc-d,l-hexamethyl-propyleneamine oxime (HMPAO)

Lately, the criteria of brain death is being discussed. Cerebral scintigram, especially scintigraphic evaluation of brain death by dynamic study, has been previously reported. Cerebral imaging using radiolabeled amines such as 123I-IMP N-isopropyl-p-iodoamphetamin (IMP) or 99mTc d,l-hexamethyl-propyleneamine oxime (HMPAO) offers a significant information of brain death by the finding of "non visualized brain". However, the dynamic scintigram acquired during the bolus injection of 99mTc-HMPAO provides an additional information of brain death by classical "hot nose sign". 99mTc-HMPAO is able to be administered relatively in a large amount of dose. This cerebral perfusion tracer is lipophilic and remains in the central nervous system, which characterize its role as an reliable indicator of cerebral blood flow. As a result, this compound became suitable for the non-invasive study of brain circulation when the diagnosis of brain death is uncertain. We report a case of brain death in which diagnosis was made by the classical "hot nose sign" in dynamic scintigraphy performed when 99Tc-HMPAO was injected as well as the SPECT which showed a lack of cerebral visualization at the equilibrium state. As far as we are informed, this additional procedure used in the diagnosis of brain death has not reported before. The importance of performing a dynamic scintigram at the administration of 99mTc-HMPAO is also discussed in this report.     

FTIR microspectroscopy of melanocytic skin lesions: a preliminary study

Fourier transform infrared (FTIR) microspectroscopy has been employed to investigate benign (ordinary dermal and Reed nevi), dysplastic and malignant (invasive melanoma) skin lesions through the analysis of spectral changes of melanocytes as well as in the evaluation of the presence of melanin. Hierarchical cluster analysis and principal component analysis led to a satisfactory separation of malignant from dysplastic and normal melanocytes. Also, on enlarging the clustering with spectra from Reed and dermal nevi, the multivariate analysis segregated well the spectral data into discrete clusters, allowing the obtaining of reliable average spectra for analysis at the molecular level of the main groups or components responsible for the biological and biochemical changes. The most significant spectral characteristics appear to be related to differences in secondary protein structures, in nucleic acid conformation, in intra- and intermolecular bonding. In all cases, supervised and unsupervised spectral analyses resulted in satisfactory agreement with histopathological findings.     

A simple technique to measure regional cerebral blood flow during intravascular balloon clamping

A case of giant internal carotid ophthalmic aneurysm was presented. In order to clarify whether the patient could tolerate carotid occlusion, a balloon clamping test was performed. before surgery. The cerebral blood flow was measured using early imaging by single photon emission computed tomography (SPECT) with N-isopropyl-(iodine-123)-p-iodoamphetamine (123I-IMP). When the balloon clamping test was performed the tracer was injected, and scanning was performed 35 minutes after removing the catheter. This tracer enabled a "memory of blood flow" during temporary ischemia to determine the character of quick diffusion and slow wash out, that could not be performed by other methods of cerebral blood flow measurement. SPECT with 123I-IMP can simplify the measurement of cerebral blood flow during the balloon clamping test.     

Experimental study on the collimator for increasing the resolution of N-isopropyl-p-(123I)iodoamphetamine (IMP) SPECT imaging of the brain

This study was carried out to design a new collimator for the present 123I-IMP SPECT imaging of the brain, which is hindered by the contamination of 124I and 126I. In this study we intended to increase spacial resolution along the transaxial direction and, at the same time, to compensate for the decrease of sensitivity by sacrificing the resolution along the axial direction to some extent. For this purpose, we developed 4 kinds of slat type units; ultrahigh resolution (UHR), high resolution (HR), high sensitivity (HS), and ultrahigh sensitivity (UHS). In practice, either UHR or HR is set to the detector together with either HS or UHS. After testing 4 kinds of combinations, we found that the combination of UHR-HS gave us far better images than those obtained with the conventional medium energy parallel hole collimator and was best suited for 123I-IMP SPECT imaging of the brain at present. We are now thinking of fusing these two units together into one collimator.     

Silkworm-pheophorbide alpha mediated photodynamic therapy against B16F10 pigmented melanoma

In order to apply photodynamic therapy (PDT) to pigmented melanoma, the efficacy of PDT mediated by pheophorbide alpha from silkworm excreta (SPbalpha) and commercial Photofrin against B16F10 melanoma was comparatively studied from the in vivo assay using C57BL/6J mice. From in vitro PDT assay, the proliferation of B16F10 cells treated with SPbalpha (more than 0.5 microg/ml) and light illumination (1.2 J/cm2) were significantly inhibited with the necrotic response. This indicated that the photocytotoxicity of SPbalpha (665 nm) was not influenced by melanin from melanoma. From the assessment of the in vivo photosensitizing activity, the tumor growth was further delayed in groups treated with SPbalpha/PDT compared to that treated with Photofrin /PDT. The survival rate of tumor bearing mice treated with SPbalpha/PDT was closely associated with its photosensitizing effect. In addition, the photosensitizing effect of SPbalpha/PDT showed a dose dependent tendency in light illumination. These results demonstrated that B16F10 melanoma cells were significantly photosensitized by SPbalpha/PDT, regardless of the influence of melanin from melanoma, and SPbalpha/PDT at very low drug dose (1 mg/kg) and light dose (1.2 J/cm2) showed the photosensitizing efficacy surpassing Photofrin against B16F10 melanoma in mice system.     

Brown Macroalgae Sargassum cristaefolium Extract Inhibits Melanin Production and Cellular Oxygen Stress in B16F10 Melanoma Cells

The brown macroalgae Sargassum has been reported for its anti-UV and photoprotective potential for industrial applications. This study evaluated the melanin inhibition activity of Sargassum cristaefolium (SCE) ethanol extract. Melanogenesis inhibition by SCE was assessed in vitro with B16-F10 melanoma cell models and in silico against melanin regulatory proteins Tyrosinase (TYR) and Melanocortin 1 Receptor (MC1R). The regulatory properties evaluated were the melanin content, intracellular tyrosinase activity and cellular antioxidant activities. In addition, the bioactive compounds detected in SCE were subjected to molecular docking against TYR and MC1R. Based on the results, 150 µg/mL SCE effectively inhibited the production of melanin content and intracellular tyrosinase activity. Cellular tyrosinase activity was reduced by SCE-treated cells in a concentration-dependent manner. The results were comparable to the standard tyrosinase inhibitor kojic acid. In addition, SCE effectively decreased the intracellular reactive oxygen species (ROS) levels in B16-F10 cells. The antioxidant properties may also contribute to the inhibition of melanogenesis. In addition, LCMS UHPLC-HR-ESI-MS profiling detected 33 major compounds. The results based on in silico study revealed that the bioactive compound putative kaurenoic acid showed a strong binding affinity against TYR (−6.5 kcal/mol) and MC1R (−8.6 kcal/mol). However, further molecular analyses are needed to confirm the mechanism of SCE on melanin inhibition. Nevertheless, SCE is proposed as an anti-melanogenic and antioxidant agent, which could be further developed into cosmetic skin care products.