Inside-the-body light delivery system using endovascular therapy-based light illumination technology.

BACKGROUND: Light-based therapies are promising for treating diseases including cancer, hereditary conditions, and protein-related disorders. However, systems, methods, and devices that deliver light deep inside the body are limited. This study aimed to develop an endovascular therapy-based light illumination technology (ET-BLIT), capable of providing deep light irradiation within the body. METHODS: The ET-BLIT system consists of a catheter with a single lumen as a guidewire and diffuser, with a transparent section at the distal end for thermocouple head attachment. The optical light diffuser alters the emission direction laterally, according to the optical fibre's nose-shape angle. If necessary, after delivering the catheter to the target position in the vessel, the diffuser is inserted into the catheter and placed in the transparent section in the direction of the target lesion. FINDINGS: ET-BLIT was tested in an animal model. The 690-nm near-infrared (NIR) light penetrated the walls of blood vessels to reach the liver and kidneys without causing temperature increase, vessel damage, or blood component alterations. NIR light transmittance from the diffuser to the detector within the organ or vessel was approximately 30% and 65% for the renal and hepatic arteries, respectively. INTERPRETATION: ET-BLIT can be potentially used in clinical photo-based medicine, as a far-out technology. ET-BLIT uses a familiar method that can access the whole body, as the basic procedure is comparable to that of endovascular therapy in terms of sequence and technique. Therefore, the use of the ET-BLIT system is promising for many light-based therapies that are currently in the research phase. FUNDING: Supported by Programme for Developing Next-generation Researchers (Japan Science and Technology Agency); JSPS KAKENHI (18K15923, 21K07217); JST-CREST (JPMJCR19H2); JST-FOREST-Souhatsu (JPMJFR2017); The Uehara Memorial Foundation; Yasuda Memorial Medical Foundation; Mochida Memorial Foundation for Medical and Pharmaceutical Research; Takeda Science Foundation; The Japan Health Foundation; Takahashi Industrial and Economic Research Foundation; AICHI Health Promotion Foundation; and Princess Takamatsu Cancer Research Fund.

cDNA array analysis for prediction of hepatic metastasis of colorectal carcinoma.

PURPOSE: Distant metastasis is a significant prognostic factor of colon carcinoma. Adjuvant chemotherapy has been shown to decrease its recurrence. However, there are no definitive methods for the diagnosis of hepatic recurrence after potentially curative surgery. The aim of this study was to evaluate the accuracy of mRNA expression profiling using samples obtained from primary tumors to predict hepatic recurrence. METHODS: Patients with stage III colorectal carcinoma without any recurrence for at least 5 years (group A: n = 9) and patients with stage IV carcinoma with hepatic metastasis (group B: n = 10) were included in this study. Tissue samples were collected from the primary tumor and adjacent normal colonic mucosa at the time of surgery in each patient. Total RNA was extracted and the mRNA expression profile was examined using a cDNA macroarray. RESULTS: A hierarchical clustering analysis revealed a dendrogram in which the patients were divided into two clusters. One cluster consisted of seven patients in group A and two in group B. The other consisted of two patients in group A and eight in group B. Therefore, the positive and negative predictive value of hierarchical clustering analysis for hepatic metastasis was 80.0% and 78.8%, respectively. Fifteen genes were revealed to be upregulated and 12 were downregulated in group B. The upregulated genes included CCNA2, TP53, and MDM2, while the downregulated genes included CDH1, GADD45A, and BCL2L2. CONCLUSIONS: mRNA expression profiling by a cDNA array analysis of specimens obtained from primary tumors was found to be useful for distinguishing patients with and without hepatic metastasis. This method is expected to contribute to the identification of patients at high risk for hepatic recurrence, while also helping in the administration of intensive adjuvant chemotherapy for such high risk patients.

Overexpression in colorectal carcinoma of two lysosomal enzymes, CLN2 and CLN1, involved in neuronal ceroid lipofuscinosis.

BACKGROUND: Lysosomal proteases are implicated in cancer progression and metastasis. In the current study, using subtraction cloning for genes that are differentially expressed in metastasis, the authors isolated a clone encoding ceroid lipofuscinosis, neuronal 2 (CLN2), which is a lysosomal serine protease defective in neuronal ceroid lipofuscinosis (NCL). Increased CLN2 activity has been reported in breast carcinoma and the antiapoptotic effect of another causative gene of NCL, ceroid lipofuscinosis, neuronal 1 (CLN1), is known. METHODS: The mRNA levels of CLN2, CLN1, and cathepsins B, D, H, and L were investigated in colorectal carcinoma patients with different clinical stages using real-time quantitative reverse transcriptase polymerase chain reaction. A polyclonal antibody was raised against a recombinant CLN2 protein for immunoblotting and immunohistochemistry. RESULTS: The mRNA levels of CLN1 and cathepsins B, D, and L were significantly higher in metastatic lesions than in primary tumors. In the primary tumors, mRNA expressions of CLN2 and cathepsin D were associated with advanced clinical stages (P < .015 and P < .031, respectively). Among the lysosomal enzymes examined, only the mRNA expression of CLN2 in both the primary tumors of all patients and the pT3 tumors was correlated with the presence of liver metastases (P < .0049 and P < .029, respectively). The polyclonal antibody prepared in the current study demonstrated CLN2 overexpression by immunoblotting and immunohistochemistry. CONCLUSIONS: The results indicate that there is a close correlation between CLN2 and CLN1 expression and colorectal carcinoma progression and metastasis and suggest that they may be potential molecular targets.

Comparison of larvicidal, adulticidal and acaricidal activity of two geometrical butylidenephthalide isomers.

Insecticidal and acaricidal activities of two geometrical isomers, (E)- and (Z)-butylidenephthalide isolated from Angelica acutiloba, against larvae and adults of fruit fly (Drosophila melanogaster), cat fleas (Ctenocephalides felis) and house dust mites (Dermatophagoides farinae and Tyrophagus putrescentiae) were investigated and compared with that of positive controls. (E)- and (Z)-Butylidenephthalide exhibited 50% lethal concentration (LC50) values of 2.07 and 0.94 micromol/ml of diet concentration against larvae of D. melanogaster, respectively. This indicated that two isomers of butylidenephthalide have geometrical stereoselectivity for larvicidal effect. Even though both (E)- and (Z)-butylidenephthalide also showed potent adulticidal and acaricidal activity against adults of D. melanogaster and two mites, there was no significant difference between two isomers. Insecticidal activity of both (E)- and (Z)-butylidenephthalide toward adults of C. felis was not detected even at the maximum concentration of 200 microg/cm2.

Clinicopathological characteristics of colorectal cancers with loss of imprinting of insulin-like growth factor 2.

Loss of imprinting (LOI), the biallelic expression of an imprinting gene, of insulin-like growth factor 2 (IGF2) has been reported to be associated with colorectal carcinogenesis because of its high prevalence in normal colorectal mucosa as well as cancerous tissue in patients with colorectal cancer. However, the characteristics of colorectal cancer associated with IGF2 LOI have not been clearly demonstrated. In this study, we investigated the IGF2 LOI status of tumor and normal mucosa in 255 consecutive patients with colorectal cancer. Of these, 95 were informative for IGF2 LOI, by direct sequencing of mRNA of IGF2. Regarding the LOI status in each patient, the prevalence of LOI in nontumorus normal mucosa was significantly higher in cases with LOI-positive cancer than in those with LOI-negative cancer (p < 0.001). Concerning the clinicopathological characteristics of LOI-positive cancer, the prevalence of poorly differentiated or mucinous carcinoma (p = 0.016) and of right-sided locations (p = 0.009) were significantly higher than those of LOI-negative cancer. Contrary to past reports that revealed a significant correlation between microsatellite instability (MSI) and IGF2 LOI in a relatively small series of noncohort patients, our study did not find a statistically significant difference in LOI-positive rate between MSI-positive and -negative cases. Our results suggested the presence of a particular type of colorectal cancer associated with the proximal colon and poor differentiation, but independent of MSI. These results may contribute to clarification of the mechanism of colorectal tumorigenesis and to determining an appropriate screening strategy for colorectal carcinoma.

Larvicidal and adulticidal activity of alkylphthalide derivatives from rhizome of Cnidium officinale against Drosophila melanogaster.

The insecticidal activity of the chloroform extract of Cnidium officinale rhizomes and its constituents was investigated against larvae and adults of Drosophila melanogaster and compared with that of rotenone. Bioassay-guided isolation of the chloroform extract of C. officinale resulted in the isolation and characterization of four alkylphthalides, cnidilide (1), (Z)-ligustilide (2), (3S)-butylphthalide (3), and neocnidilide (4). The structures of these compounds were established by spectroscopic analysis. The isolated compounds 2, 3, and 4 exhibited LC50 values of 2.54, 4.99, and 9.90 micromol/mL of diet concentration against larvae of D. melanogaster, respectively. Against both sexes (males/females, 1:1) of adults (5-7 days old), compound 3 showed the most potent activity of the compounds isolated with the LD50 value of 5.93 microg/adult, comparable to that of rotenone (LD50 = 3.68 microg/adult). Structure-activity relationships of phthalides isolated suggest that the presence of conjugation with the carbonyl group in the lactone ring appeared to play an important role in the larvicidal activity. Acetylcholinesterase (prepared from the adult heads of D. melanogaster) inhibitory activity was also investigated in vitro to determine the insecticide mode of action for the acute adulticidal activity.

Insecticidal effect of phthalides and furanocoumarins from Angelica acutiloba against Drosophila melanogaster.

Insecticidal activity of Angelica acutiloba extract and its constituents was investigated and compared with that of rotenone. Bioassay-guided isolation of the chloroform extract of A. acutiloba against larvae of Drosophila melanogaster afforded two phthalides, (Z)-butylidenephthalide (1) and (Z)-ligustilide (2), and two furanocoumarins, xanthotoxin (3) and isopimpinellin (4). The structures of these compounds were established by spectroscopic analysis. The isolated compounds 1, 2, 3, and 4 exhibited LC(50) values of 0.94, 2.54, 3.35, and 0.82 micromol/mL of diet concentration against larvae of D. melanogaster, respectively. Against both sexes (males/females, 1:1) of adults (5-7 days old), compound 1 showed the most potent activity with a LD(50) value of 0.84 microg/adult. Compound 1 is a more active insecticide than rotenone (LD(50) = 3.68 microg/adult) and has potential as a novel insect control agent. However, compound 2 was inactive against adults. The structure-activity relationship of phthalides isolated indicated that the aromaticity appeared to play an important role in the activity of both larvae and adults. To determine the insecticide mode of action for acute adulticidal activity, acetylcholinesterase (AChE) inhibitory activity was also investigated in vitro, and the result indicated that the acute adulticidal activity of compounds 3 and 4 was due to the inhibition of AChE.

An early stage small bowel adenocarcinoma with microsatellite instability phenotype in a case of hereditary nonpolyposis colorectal cancer.

BACKGROUND: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominantly inherited disease characterized by onset at a relatively early age, excess of synchronous and metachronous tumors, and a variety of extracolorectal malignancies. Small bowel carcinoma is included in the tumor spectrum of HNPCC, but the frequency of occurrence of this tumor in HNPCC patients is comparatively rare. CASE PRESENTATION: We report the case of a 55-year-old woman who had a history of multiple colon cancers at 33 years of age, sigmoid colon cancer at 47, and endometrial carcinoma at 51. This case fulfills the Amsterdam criteria for HNPCC and followed from the patient's age of 47 at our institute. Surveillance colonoscopy showed a sessile polyp in the ileum that was 9 mm in diameter and located about 10 cm proximal to the ileorectal anastomosis, and that was resected by endoscopic mucosal resection. Histopathological studies showed an adenoma with well-differentiated adenocarcinoma in the mucosa. A molecular analysis of the adenoma component of this polyp was performed, and microsatellite instability was found in four of the nine analyzed loci. The patient was a mutation carrier in hMSH2, one of the mismatch repair genes responsible for HNPCC. CONCLUSION: Reports of early-stage carcinoma of the small bowel in HNPCC are very rare, and an adenoma-carcinoma sequence was present in the small bowel tumor of this patient. The molecular findings of this tumor are discussed.

Densely methylated MLH1 promoter correlates with decreased mRNA expression in sporadic colorectal cancers.

It has been reported that MLH1 is silenced by promoter methylation, and that this phenomenon is associated with microsatellite instability (MSI) in sporadic colorectal cancer (CRC). To clarify the significance of MLH1 promoter methylation in sporadic CRC, we examined the correlation between methylation status over the entire promoter region and mRNA expression in cases showing high-frequency MSI (MSI-H). MLH1 promoter methylation was analyzed using the bisulfite modification sequencing in 48 MSI-H cases. We also screened for somatic mutation, loss of heterozygosity, and immunohistochemical staining of MLH1. The results showed that methylation patterns could be subdivided into three types: methylation of more than 80% of the CpG sites analyzed (type 1 methylation), methylation of less than 20% (type 2 methylation), and methylation mainly in the region 500 to 921 bases upstream from the translation start site (type 3 methylation). Of the three types, only type 1 methylation correlated with decreased mRNA expression. The frequency of type 1 methylation was significantly higher in cases involving the proximal colon (66.7%, 18/27) compared to that of the distal colon and rectum (23.8%, 5/21, P = 0.004). Immunohistochemical staining of MSI-H cases showed that decreased MLH1 was found in 77.1% (37/48). Of the cases with decreased MLH1, type 1 methylation was present in 59.5% (22/37). Overall, our data suggested that the type 1 methylation pattern may affect MLH1 mRNA expression, such that the majority of MSI-H cases in sporadic CRC, especially proximal colon cancer, exhibited type 1 methylation.

Evaluation of screening strategy for detecting hereditary nonpolyposis colorectal carcinoma.

BACKGROUND: The Amsterdam criteria are used worldwide for the clinical diagnosis of hereditary nonpolyposis colorectal carcinoma (HNPCC). In Japan, clinical criteria (JCC) have been proposed to identify as many HNPCC cases as possible, but the suitability of the JCC remains uncertain. In this article, the authors evaluate retrospectively whether the JCC are adequate to diagnose HNPCC compared with the Bethesda guidelines (BG) and also investigated useful screening methods for HNPCC. METHODS: The authors studied 452 colorectal carcinoma cases, of which 69 cases fulfilled the JCC (A, 12; B, 57) and 106 fulfilled the BG. Microsatellite instability (MSI) was examined for 452 cases. TGF beta RII, immunohistochemical staining, and germline mutations of hMLH1 and hMSH2 were analyzed in high-frequency MSI cases. RESULTS: High-frequency MSI was found in 21.7% (98 of 452). Germline mutations were detected in eight cases (hMLH1, three, hMSH2; five). Six cases fulfilled the JCC (A, four; B, two), and six fulfilled the BG. The germline mutation rate was significantly higher in the JCCA than in non-JCCA cases (33.3% vs. 0.91%; P < 0.001) and in cases with an age at onset younger than 50 years than older than 50 years (9.3% vs. 0.27%, P < 0.001). All germline mutation carriers had the TGF beta RII mutation. Immunohistochemically, a decreased nuclear staining was found in 57.3% (47 of 82) for hMLH1 and in 18.3% (15 of 82) for hMSH2. The frequency of predicted germline mutations was higher in cases with decreased hMSH2 than hMLH1 (33.3% vs. 6.4%; P = 0.016). CONCLUSIONS: The JCCA are suitable for selecting cases to analyze for gene mutations, but the JCCB are not useful for the clinical setting. The authors suggest that an age at onset younger than 50 years is also important for screening. Analyzing TGF beta RII mutations and immunohistochemical staining of hMLH1 or hMSH2 for cases with MSI phenotype are useful for selecting cases who should be tested for germline mutations.