[Research of the Cancer-Odor].

Early detection and resection of cancer is the most effective in the treatment of solid cancer. Development of a new cancer detection method is expected to become a breakthrough to solve various problems for early detection. It has been reported that there is the specific odors of cancer by using bio olfaction such as dogs, and it has been recognized that there is the odors of cancer. Cancer cells acquire malignant traits as a result of metabolic changes originating from genetic mutation. The cancer specific odorous substances may be considered to be the end products of their metabolic changes. Omics researches such as genomics, proteomics, and metabolomics have been extensively performed to comprehensively analyze changes in DNA, RNA, protein, metabolism and its products specific to cancer for the purpose of developing a new cancer detection marker. It is thought that the research on the odor of cancer is also on the line of omics research. It is difficult to identify cancer-specific odorants buried in various environmental substances. However, it is expected that human will be able to acquire the technology, from the fact that they can be recognized by biological olfaction. We are continuing the research with the dream that identification of the odorous substances as a new cancer detection marker and sensor development for it will lead to the happiness of colleagues in the world.

Suture Granuloma Showing False-Positive Findings on FDG-PET.

We report a case of a 33-year-old male with a mixed germ-cell testicular tumor. Postoperative follow-up FDG-PET revealed concentration of FDG in the left inguinal area which is not tumor metastasis or local recurrence but suture reactivity granuloma. In this paper, we reviewed suture granulomas associated with false-positive findings on FDG-PET after surgery. If FDG-PET will be used more frequently in the future, it will be necessary to refrain from using silk thread in order to prevent any unnecessary surgery.

Colorectal cancer screening with odour material by canine scent detection.

OBJECTIVE: Early detection and early treatment are of vital importance to the successful treatment of various cancers. The development of a novel screening method that is as economical and non-invasive as the faecal occult blood test (FOBT) for early detection of colorectal cancer (CRC) is needed. A study was undertaken using canine scent detection to determine whether odour material can become an effective tool in CRC screening. DESIGN: Exhaled breath and watery stool samples were obtained from patients with CRC and from healthy controls prior to colonoscopy. Each test group consisted of one sample from a patient with CRC and four control samples from volunteers without cancer. These five samples were randomly and separately placed into five boxes. A Labrador retriever specially trained in scent detection of cancer and a handler cooperated in the tests. The dog first smelled a standard breath sample from a patient with CRC, then smelled each sample station and sat down in front of the station in which a cancer scent was detected. RESULTS: 33 and 37 groups of breath and watery stool samples, respectively, were tested. Among patients with CRC and controls, the sensitivity of canine scent detection of breath samples compared with conventional diagnosis by colonoscopy was 0.91 and the specificity was 0.99. The sensitivity of canine scent detection of stool samples was 0.97 and the specificity was 0.99. The accuracy of canine scent detection was high even for early cancer. Canine scent detection was not confounded by current smoking, benign colorectal disease or inflammatory disease. CONCLUSIONS: This study shows that a specific cancer scent does indeed exist and that cancer-specific chemical compounds may be circulating throughout the body. These odour materials may become effective tools in CRC screening. In the future, studies designed to identify cancer-specific volatile organic compounds will be important for the development of new methods for early detection of CRC.

Effects of adipocytes on the proliferation and differentiation of prostate cancer cells in a 3-D culture model.

OBJECTIVE: To investigate how the mechanism of adipocyte-prostate cancer cell interaction affects the proliferation and differentiation of prostate cancer cells. METHODS: An androgen-dependent cell line (LNCaP), two androgen-independent cell lines (PC-3, DU145), and mature adipocytes harvested from male Wistar rats were used. Cancer cells were co-cultured with the isolated mature adipocytes in 3-D collagen gel matrix culture. The morphology and proliferative ability of the prostate cancer cells were examined. With regard to the activation of the phosphatidylinositol 3-kinase (PI3K) pathway, the expression of phosphatase and tensin homologue deleted on chromosome ten (PTEN), Akt and Bad were determined by immunohistochemistry. RESULTS: LNCaP cells co-cultured with adipocytes formed larger clusters than those of the control. PC-3 cells co-cultured with adipocytes did not form larger clusters, but formed spherical and spindle-shaped cells. The phosphorylation of Akt in PC-3 cells was greater in the co-cultured group compared with the controls, but there were no significant differences in the phosphorylation of Akt with regard to LNCaP and DU145 cells. CONCLUSIONS: Adipocytes could modulate the proliferation and differentiation of prostate cancer cell lines. Activation of the PI3K pathway might be involved in the prostate cancer cell-adipocyte interaction.

The effects of neoadjuvant chemotherapy and chemo-radiation therapy on MRI staging in invasive bladder cancer: comparative study based on the pathological examination of whole layer bladder wall.

OBJECTIVE: We evaluated the correlation of radiological findings obtained by MRI study with pathological diagnosis in invasive bladder cancer treated with neoadjuvant chemotherapy, with or without radiation. DESIGN, SETTING, AND PARTICIPANTS: Twenty-seven patients, who underwent total or partial cystectomy for invasive bladder tumors, were enrolled into the present study. Eight cases had received neoadjuvant chemotherapy following the staging biopsy (group A), ten cases had received chemo-radiation therapy following the staging biopsy (group B), and nine cases had received preoperative staging biopsy alone (group C). As a final treatment, 12 of the 27 patients underwent total cystectomy and the other 15 patients underwent partial cystectomy. MRI was conducted prior to total or partial cystectomy in each case. The pathological stage was assessed by histological examination of the entire layer of the bladder wall. RESULTS AND LIMITATIONS: Tumor stage assessed by MRI was consistent with pathological findings in 16 of the 27 cases (59.3%), while MRI produced over-staging in 7 cases and under-staging in 4 cases. The accuracy of staging was 75.0, 30.0, and 77.8% in groups A, B, and C, respectively. The accuracy of MRI staging in group B was lower than that in group C (P < 0.05). There was no difference in the accuracy of MRI staging between groups A and C. CONCLUSION: MRI is useful for the staging of bladder cancer. However, care needs to be taken when staging invasive bladder tumors treated with neoadjuvant chemo-radiation therapy, because inflammatory infiltrations and/or fibrous changes caused by the chemotherapy or chemo-radiation therapy make precise staging with MRI difficult.

[Metachronous testicular seminoma with an interval of 14 years from the first nonseminomatous testicular tumor: a case report].

A 32-year-old man was admitted to our hospital with a complaint of right scrotal swelling. He had a past history of left testicular nonseminomatous germ cell tumor (NSGCT), stage IIB 14 years ago. He had undergone chemotherapy with 3 courses of PEP and retroperitoneal lymphnode dissection revealing pathological CR. He was diagnosed as having right testicular tumor. He underwent right high orchiectomy. Imaging studies and pathological studies revealed pure seminoma with spermatic cord invasion, stage I, pT3N0M0. He underwent chemotherapy with 3 courses of VIP. Herein we present a case of metachronous bilateral testicular germ cell tumor with a review of literatures.

Oncologic assessment of hand-assisted retroperitoneoscopic nephroureterectomy for urothelial tumors of the upper tract: comparison with conventional open nephroureterectomy.

PURPOSE: To evaluate the oncologic results of our operative technique, hand-assisted retroperitoneoscopic nephroureterectomy (HRNU), for the treatment of upper-tract urothelial cancer, various perioperative parameters and oncologic outcomes were compared for HRNU and conventional open nephroureterectomy (CONU). PATIENTS AND METHODS: Thirty-six patients with clinical stage T(1,2)N(0)M(0) renal-pelvic and ureteral tumors underwent HRNU. A retroperitoneoscopic nephrectomy was carried out with hand assistance via a lower-abdominal midline incision. The lower ureter was resected by open surgery through the same incision, and the operative specimen was extracted via the same incision. Thirty-seven cases of CONU were reviewed as historical controls. Various perioperative and parameters and oncologic results were compared for the two procedures. RESULTS: The HRNU was completed in all but one case, which was converted to CONU. The mean operating time (395 minutes) was longer than that for CONU (289 minutes), and the mean estimated blood loss with HRNU (497 g) was greater than that with CONU (296 g). The mean time to oral intake (1.4 days) was shorter than that after CONU (2.3 days), and the mean time to walking was shorter (2.1 days v 2.6 days). There were no statistical differences in the cause-specific survival rate, the disease-free survival rate, or the bladder recurrence-free survival rate between HRNU (median follow-up 23 months) and CONU (median follow-up 56 months). CONCLUSION: The HRNU, a combination of endoscopic and conventional open surgery, seems to be a reasonable surgical procedure, because the lower-abdominal incision can be utilized, not only as a route for hand assistance, but also as a window for open surgery when resecting the distal ureter as well as for extraction of surgical specimens. The procedure is a safe alternative to conventional open surgery for upper urinary-tract tumors from an oncologic viewpoint.

PACAP- and PHI-mediated sustained relaxation in circular muscle of gastric fundus: findings obtained in PACAP knockout mice.

Mediators of neurogenic responses of the gastric fundus were studied in wild type and pituitary adenylate cyclase activating peptide (PACAP) knockout mice. Electrical field stimulation (EFS) to the circular muscle strips of the wild type mouse fundus induced a tri-phasic response, rapid transient contraction and relaxation, and sustained relaxation that was prolonged for an extended period after the end of EFS. The transient relaxation and contraction were completely inhibited by N(G)-nitro-L-arginine and atropine, respectively. The sustained relaxation was completely inhibited by a PACAP receptors antagonist, PACAP(6-38). The strips prepared from PACAP knockout mice exhibited a large contraction without rapid relaxation and unexpectedly, a sustained relaxation. However, the sustained relaxation was decreased to about a half of that observed in wild type mice. Anti-peptide histidine isoleucine (PHI) serum abolished the sustained relaxation in the knockout mice. The serum partially inhibited the sustained relaxation in wild type mice and PACAP(6-38) abolished the relaxation that remained after the antiserum-treatment. PHI relaxed the strips prepared from wild type mice. The relaxation was completely inhibited by PACAP(6-38). It was concluded that PACAP and PHI separately mediate the sustained relaxation in the mouse gastric fundus, and that nitric oxide and ACh mediate transient relaxation and contraction, respectively.

Renal-tissue damage induced by laparoscopic partial nephrectomy using microwave tissue coagulator.

BACKGROUND AND PURPOSE: The microwave coagulator is a useful instrument that enables surgeons to perform partial nephrectomy without vascular clamping. The extent of postoperative thermal damage in surgically spared renal tissue has not been well examined. The present study was conducted to evaluate the tissue damage caused by microwave coagulation in laparoscopic partial nephrectomy (LPN) for small renal tumors. MATERIALS AND METHODS: Seven cases of LPN with a microwave tissue coagulator were entered in the present study. The median tumor diameter was 1.5 cm, and the median size of the resected specimen was 2.2 cm. Postoperative tissue damage was evaluated by contrast-enhanced CT 1 month after surgery. Surgically spared renal-tissue volume and functioning renal-tissue volume were estimated from the images by NIH Image 1.62 software. RESULTS: Postoperative CT revealed unenhanced renal tissue adjacent to the surgical margin. The median estimated volumes of surgically spared and functioning renal tissue were 96.1% (range 74.3%-99.8%) and 88.4% (range 68.0%-92.7%) of preoperative normal renal tissue, respectively. The percentile volume of functioning to surgically spared renal tissue ranged from 89.9% to 96.0% (median 92.8%). CONCLUSIONS: The microwave coagulator enables us to carry out partial nephrectomy without vascular clamping. Although 96% of normal renal tissue was surgically spared, 4% to 10% of this tissue was nonfunctioning as a result of microwave-induced thermal damage.

[A modified retroperitoneal approach to the kidney in patients with a highly deformed thorax: obtaining a wide operative field through subperiosteal resection of the 10th, 11th and 12th ribs].

We herein report a technique which facilitates a retroperitoneal approach to the kidney in cases of highly deformed thorax due to kyphoscoliosis. The operation consists of a lumbar oblique incision with removal of the 11th rib, combined with the additional removal of the 12th and 10th ribs. Resection of the upper two ribs was performed subperiosteally, leaving the periosteum of the deep side untouched. However, the deep side periosteum of the 12th rib was incised caudal from the pleural margin in order to facilitate exposure of the diaphragm. The retroperitoneal space was entered through the tip of the 11th rib bed. The diaphragm was incised dorso-medially at a level 1 cm caudal from the lower margin of the pleura, to an extent necessary to enable the pleura together with the cranial diaphragm to be manoeuvred in an upward direction. Two cases with renal tuberculosis associated with high-grade kyphosis and one case with staghorn calculi accompanied with lordosis were operated on utilizing this technique. In the former two cases, the thoracic cage was in direct contact with the iliac bone and there was practically no space between the rib border and the iliac crest. This was also true of the third case, but the grade of deformity was not as extensive as in the former two cases. Removal of the 10th, 11th and 12th ribs could be achieved without injuring the pleura and a satisfactorily large operating field could thus be developed which enabled a simple nephrectomy to be performed without difficulty. The characteristic feature of the described approach is that resection of the 10th and 11th ribs is simply to facilitate manoevrability of the wound margin, without going through the rib bed. The technique could be advantageous in selected cases where there is a highly deformed thorax.

Silencing effect of CpG island hypermethylation and histone modifications on O6-methylguanine-DNA methyltransferase (MGMT) gene expression in human cancer.

O6-methylguanine-DNA methyltransferase (MGMT) repairs the cytotoxic and mutagenic O6-alkylguanine produced by alkylating agents such as chemotherapeutic agents and mutagens. Recent studies have shown that in a subset of tumors, MGMT expression is inversely linked to hypermethylation of the CpG island in the promoter region; however, how the epigenetic silencing mechanism works, as it relates to hypermethylation, was still unclear. To understand the mechanism, we examined the detailed methylation status of the whole island with bisulfite-sequencing in 19 MGMT non-expressed cancer cell lines. We found two highly methylated regions in the island. One was upstream of exon 1, including minimal promoter, and the other was downstream, including enhancer. Reporter gene assay showed that methylation of both the upstream and downstream regions suppressed luciferase activity drastically. Chromatin immunoprecipitation assay revealed that histone H3 lysine 9 was hypermethylated throughout the island in the MGMT negative line, whereas acetylation on H3 and H4 and methylation on H3 lysine 4 were at significantly high levels outside the minimal promoter in the MGMT-expressed line. Furthermore, MeCP2 preferentially bound to the CpG-methylated island in the MGMT negative line. Given these results, we propose a model for gene silencing of MGMT that is dependent on the epigenetic state in cancer.

Significant reduction of WT1 gene expression, possibly due to epigenetic alteration in Wilms' tumor.

WT1 at 11p13 is a tumor suppressor gene, an aberration of which causes Wilms' tumor (WT). Since WT1 expression is reduced in a certain proportion of WTs and its mutation is found only in 10-20% of WTs, we examined WT1 gene silencing due to epigenetic alteration in a total of 22 WTs. WT1 expression was significantly reduced in half of WTs without any mutation in the WT1 gene itself, suggesting that the reduction of expression was possibly epigenetic. We found promoter hypermethylation in one WT with loss of heterozygosity (LOH) and showed that promoter methylation reduced reporter gene activity by a reporter assay. These data suggested that methylation was an epigenetic mechanism leading to WT1 silencing and that the expression-reduced allele by hypermethylation combined with LOH was consistent with the revised two-hit model. In addition, as the beta-catenin mutation is frequently associated with the WT1 mutation, the association of WT1 silencing with the beta-catenin mutation was also investigated. beta-catenin mutated in only one WT without WT1 silencing, suggesting that the beta-catenin mutation was not associated with the reduction of WT1 expression.

Domain regulation of imprinting cluster in Kip2/Lit1 subdomain on mouse chromosome 7F4/F5: large-scale DNA methylation analysis reveals that DMR-Lit1 is a putative imprinting control region.

Mouse chromosome 7F4/F5, where the imprinting domain is located, is syntenic to human 11p15.5, the locus for Beckwith-Wiedemann syndrome. The domain is thought to consist of the two subdomains Kip2 (p57(kip2))/Lit1 and Igf2/H19. Because DNA methylation is believed to be a key factor in genomic imprinting, we performed large-scale DNA methylation analysis to identify the cis-element crucial for the regulation of the Kip2/Lit1 subdomain. Ten CpG islands (CGIs) were found, and these were located at the promoter sites, upstream of genes, and within intergenic regions. Bisulphite sequencing revealed that CGIs 4, 5, 8, and 10 were differentially methylated regions (DMRs). CGIs 4, 5, and 10 were methylated paternally in somatic tissues but not in germ cells. CGI8 was methylated in oocyte and maternally in somatic tissues during development. Parental-specific DNase I hypersensitive sites (HSSs) were found near CGI8. These data indicate that CGI8, called DMR-Lit1, is not only the region for gametic methylation but might also be the imprinting control region (ICR) of the subdomain.

Local administration of IL-12-transfected dendritic cells induces antitumor immune responses to colon adenocarcinoma in the liver in mice.

Colorectal cancer is one of the most common fatal malignancies in the United States, with an incidence second only to lung cancer. The liver is the most common site of colorectal metastases and frequently the only affected organ once the primary tumor has been surgically removed. The only potentially curative treatment for metastatic colorectal cancer in the liver is surgery, although most patients are not eligible for resection. We have therefore, evaluated the therapeutic efficacy of dendritic cells (DCs) engineered to express IL-12 in a liver metastasis model. Direct administration of DCs into the portal vein significantly inhibited the growth of established MC38 colon carcinoma in the liver in C57BL/6 mice. This effect was accompanied by an intratumoral accumulation of CD4+, CD8+, and NLDC-145+ immune effector cells, and also resulted in a systemic immune response as determined by enhanced production of IFN-gamma by T lymphocytes isolated from both spleen and draining lymph nodes. Evaluation of homing of Cy3-labeled DCs following the portal vein injection confirmed their distribution in the liver and lymphoid tissue. Thus, a local delivery of DCs transduced with the IL-12 gene can not only inhibit colorectal tumor growth in vivo but also mount systemic antitumor immune responses. This approach is likely to improve the outcome of immunotherapy for metastatic colorectal cancer since high numbers of tumor-associated DCs positively correlate with a more favorable prognosis. Simultaneous local gene therapy with IL-12 will further improve clinical efficacy without placing the patient at risk for systemic toxicity.

PAC1 receptor-mediated relaxation of longitudinal muscle of the mouse proximal colon.

Since pituitary adenylate cyclase-activating polypeptide (PACAP) was shown to partially mediate nonadrenergic, noncholinergic (NANC) relaxation of longitudinal muscle of the proximal colon of ICR mice, we further studied the receptor subtype activated by PACAP by using a mutant mouse whose PAC1 receptors are markedly reduced. In wild-type mice, the PACAP-mediated component of NANC relaxation was 33%, but it was absent in the mutant mice. The potency of exogenous PACAP in inducing relaxation in the mutant mice was one hundredth of that in wild-type mice. VPAC1 and VPAC2 receptors were not suggested to have any role in the relaxation. These results suggest that PACAP mediates NANC relaxation of longitudinal muscle of mouse proximal colon via PAC1 receptors.