Norihiko Shiiya

Human Breast Cancer Tissues Contain Abundant Phosphatidylcholine(36∶1) with High Stearoyl-CoA Desaturase-1 Expression

Breast cancer is the leading cause of cancer and mortality in women worldwide. Recent studies have argued that there is a close relationship between lipid synthesis and cancer progression because some enzymes related to lipid synthesis are overexpressed in breast cancer tissues. However, lipid distribution in breast cancer tissues has not been investigated. We aimed to visualize phosphatidylcholines (PCs) and lysoPCs (LPCs) in human breast cancer tissues by performing matrix assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS), which is a novel technique that enables the visualization of molecules comprehensively. Twenty-nine breast tissue samples were obtained during surgery and subjected to MALDI-IMS analysis. We evaluated the heterogeneity of the distribution of PCs and LPCs on the tissues. Three species [PC(32∶1), PC(34∶1), and PC(36∶1)] of PCs with 1 mono-unsaturated fatty acid chain and 1 saturated fatty acid chain (MUFA-PCs) and one [PC(34∶0)] of PCs with 2 saturated fatty acid chains (SFA-PC) were relatively localized in cancerous areas rather than the rest of the sections (named reference area). In addition, the LPCs did not show any biased distribution. The relative amounts of PC(36∶1) compared to PC(36∶0) and that of PC(36∶1) to LPC(18∶0) were significantly higher in the cancerous areas. The protein expression of stearoyl-CoA desaturase-1 (SCD1), which is a synthetic enzyme of MUFA, showed accumulation in the cancerous areas as observed by the results of immunohistochemical staining. The ratios were further analyzed considering the differences in expressions of the estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2), and Ki67. The ratios of the signal intensity of PC(36∶1) to that of PC(36∶0) was higher in the lesions with positive ER expression. The contribution of SCD1 and other enzymes to the formation of the observed phospholipid composition is discussed.

Lymphatic Vessel Invasion is a Significant Prognostic Indicator in Stage IA Lung Adenocarcinoma

BackgroundA radical resection is considered to be the most effective treatment for resectable non-small cell lung cancer. However, even when resected in early stages (T1aN0, T1bN0) up to 20% of patients will experience recurrence. The aim of this retrospective study was to evaluate the prognostic influence of lymphatic vessel invasion (LVI) in stage IA adenocarcinoma patients.MethodsFrom January 1983 to June 2003, a total of 229 consecutive patients with pT1a or T1b N0 M0 lung adenocarcinoma who had undergone radical resection and lymph node dissection were retrospectively reviewed. Sections stained by the hematoxylin-eosin and the Elastica van Gieson method were examined for the presence of LVI. The overall survival was estimated using the Kaplan-Meier method, log-rank test, and the Cox proportional hazards analysis.ResultsThe median follow-up was 81xa0months. A total of 143 patients (62%) were able to be diagnosed with regard to the presence of LVI, while information was not provided for 86 patients (38%), who were therefore excluded from the study. LVI was noted in 22 of the evaluable patients (15%) and was not seen in the other 121 patients (85%). The 5-year overall survival rate of the LVI-negative group and the LVI-positive group was 94.5 and 70.9%, respectively (P = .003). A multivariate analysis revealed LVI to be an independent predictive factor (hazard ratio: 0.202; P = .001).ConclusionLVI is an independent poor prognostic factor in patients with pathologic stage IA adenocarcinoma. The T1a and T1b patients with LVI both might benefit from adjuvant chemotherapy.

Array-based identification of common DNA methylation alterations in ulcerative colitis.

Patients with long-standing ulcerative colitis (UC) have higher risk of developing colorectal cancer. Albeit the causes remain to be understood, epigenetic alterations have been suggested to play a role in the long-term cancer risk of these patients. In this work, we developed a novel microarray platform based on methylation-sensitive amplified fragment length polymorphism (MS-AFLP) DNA fingerprinting. The over 10,000 NotI sites of the human genome were used to generate synthetic primers covering these loci that are equally distributed into CpG rich regions (promoters and CpG islands) and outside the CpG islands, providing a panoramic view of the methylation alterations in the genome. The arrays were first tested using the colon cancer cell line CW-2 showing the reproducibility and sensitivity of the approach. We next investigated DNA methylation alterations in the colonic mucosa of 14 UC patients. We identified epigenetic alterations affecting genes putatively involved in UC disease, and in susceptibility to develop colorectal cancer. There was a strong concordance of methylation alterations (both hypermethylation and hypomethylation) shared by the cancer cells of the CW-2 cell line and the non-cancer UC samples. To the best of our knowledge, this work defines the first high-throughput aberrant DNA methylation profiles of the colonic mucosa of UC patients. These epigenetic profiles provide novel and relevant knowledge on the molecular alterations associated to the UC pathology. Some of the detected alterations could be exploited as cancer risk predictors underlying a field defect for cancerization in UC-associated carcinogenesis.

Single-cell time-of-flight secondary ion mass spectrometry reveals that human breast cancer stem cells have significantly lower content of palmitoleic acid compared to their counterpart non-stem cancer cells

Lipids comprise the primary component of cell membranes. Imaging mass spectrometry is increasingly being used to visualize membranous lipids in clinical specimens, and it has revealed that abnormal lipid metabolism is related to the development of diseases. To characterize cell populations which are rare and sparsely localized in tissues, we conducted time-of-flight secondary ion mass spectrometry (TOF-SIMS) analyses of individual cells sorted by fluorescence activated cell sorting (FACS) and applied the method to analyze breast cancer stem cells (CSCs). TOF-SIMS analyses visualized phosphoric acids and four fatty acid (FA) species in the sorted CD45(-)/CD44(+)/CD24(-) CSCs, and these ions are suspected to have originated from membranous phospholipids as they were uniformly detected from the locus where the cells attached. Integrated ion intensity of palmitoleic acids [FA(16:1)] normalized by phosphoric acid signals were decreased significantly in CSCs as compared to that of CD45(-)/CD44(-)/CD24(+) non-stem cancer cells (NSCCs). This finding was supported by liquid chromatography coupled electrospray ionization-tandem mass spectrometry analysis, which revealed phosphatidylcholine (PC)(16:0/16:1) to be less abundant and PC(16:0/16:0) to be more abundant in CSCs as compared to NSCCs. Therefore, our novel method successfully provided lipid composition analysis of individual cells classified by the expression of a complex combination of cell-surface markers. The lipid compositions of CSCs originating from the heterogeneous cellular populations of clinical specimens were successfully characterized by this method.

Laparoscopic transabdominal preperitoneal inguinal hernia repair using needlescopic instruments: a 15-year, single-center experience in 317 patients

BackgroundLaparoscopic inguinal hernia repair is associated with a decrease in postoperative pain, shortened hospital stay, earlier return to normal activity, and decrease in chronic pain. Moreover, laparoscopic surgery performed with needlescopic instruments has more advantages than conventional laparoscopic surgery. However, there are few reports of large-scale laparoscopic transabdominal preperitoneal inguinal hernia repair using needlescopic instruments (nTAPP). This report reviews our experiences with 352 nTAPP in 317 patients during the 15-year period from April 1996 to April 2011.MethodsWe performed nTAPP as the method of choice in 88.5% of all patients presenting with inguinal hernia. To perform the nTAPP, 3-mm instruments were used. A 5-mm laparoscope was inserted from the umbilicus, and surgical instruments were inserted through 5- and 3-mm trocars. After reduction of the hernia sac and dissection of the preperitoneal space, we placed polyester mesh or polypropylene soft mesh with staple fixation. The peritoneum was closed with 3–0 silk interrupted sutures.ResultsThe mean operative time was 102.9xa0min for unilateral hernias and 155.8xa0min for bilateral hernias. There was no conversion to open repair. Forty-three patients (13.6%) used postoperative analgesics, and the mean frequency of use was 0.5 times. Regarding intraoperative complications, we observed one bladder injury, but no bowel injuries or major vessel injuries. Postoperative complications occurred in 32 patients (10.1%). One patient with a retained lipoma required reoperation. There was no incidence of chronic pain or mesh infection. The operative time for experienced surgeons (≥20 repairs) was significantly shorter than that of inexperienced surgeons (<20 repairs; Pxa0<xa00.05).ConclusionsThe nTAPP was a safe and useful technique for inguinal hernia repair. Large prospective, randomized controlled trials will be required to establish the benefit of nTAPP.

Vertebral artery variations in thoracic aortic patients.

OBJECTIVESnArterial variation is common in the vertebral artery, and simple occlusion of the left subclavian artery may result in brain infarction, especially when it terminates in the posterior inferior cerebellar artery (PICA). We report the results of preoperative vertebral artery evaluation by magnetic resonance angiography (MRA) and its impact upon operative strategy.nnnMETHODSnAmong the 214 patients who underwent thoracic aortic surgery from 2009 through 2012, 159 patients with preoperative MRA were retrospectively analysed. Patients age ranged from 35 to 88 (median 72), 122 were male and 115 had degenerative aneurysms. Prevalence rates of vertebral artery variations and occlusive lesions were reported, together with operative strategies and outcomes.nnnRESULTSnThere were 19 hypoplasia (12%), 10 PICA termination (6%) and 12 occlusive lesion (8%) on the right vertebral artery and 10 hypoplasia (6%), 5 PICA termination (3%), 7 direct arch origin (4%) and 3 occlusive lesion (2%) on the left. Two of the seven arch-originated arteries terminated in the PICA. In aortic arch replacement, these were reconstructed together with the left subclavian artery while hypothermia was maintained. During thoracic endovascular aortic repair with Zone-2 proximal landing, debranching bypass was employed to preserve left subclavian perfusion when there was PICA termination, hypoplasia or occlusive lesion. In 1 patient with hypoplasia between the basilar artery and the left PICA, bypass was added immediately after deployment because radial pressure dropped critically. No brain infarction occurred with this strategy.nnnCONCLUSIONSnPICA termination and right side hypoplasia/occlusive lesion, where left subclavian perfusion is important for brain protection, is present in ∼ 30%. Left vertebral artery that originated from the arch should be managed with care, because PICA termination is highly prevalent.

UV damage-induced phosphorylation of HBO1 triggers CRL4DDB2-mediated degradation to regulate cell proliferation

ABSTRACT Histone acetyltransferase binding to ORC-1 (HBO1) is a critically important histone acetyltransferase for forming the prereplicative complex (pre-RC) at the replication origin. Pre-RC formation is completed by loading of the MCM2-7 heterohexameric complex, which functions as a helicase in DNA replication. HBO1 recruited to the replication origin by CDT1 acetylates histone H4 to relax the chromatin conformation and facilitates loading of the MCM complex onto replication origins. However, the acetylation status and mechanism of regulation of histone H3 at replication origins remain elusive. HBO1 positively regulates cell proliferation under normal cell growth conditions. Whether HBO1 regulates proliferation in response to DNA damage is poorly understood. In this study, we demonstrated that HBO1 was degraded after DNA damage to suppress cell proliferation. Ser50 and Ser53 of HBO1 were phosphorylated in an ATM/ATR DNA damage sensor-dependent manner after UV treatment. ATM/ATR-dependently phosphorylated HBO1 preferentially interacted with DDB2 and was ubiquitylated by CRL4DDB2. Replacement of endogenous HBO1 in Ser50/53Ala mutants maintained acetylation of histone H3K14 and impaired cell cycle regulation in response to UV irradiation. Our findings demonstrate that HBO1 is one of the targets in the DNA damage checkpoint. These results show that ubiquitin-dependent control of the HBO1 protein contributes to cell survival during UV irradiation.

Aortic arch replacement for degenerative aneurysms: advances during the last decade

During the last decade, treatment paradigm for degenerative aortic arch aneurysms has been changed by a better understanding of the pathophysiology of brain complication and introduction of endovascular technologies. To avoid neurocognitive dysfunction, safe duration of deep hypothermic circulatory arrest is now considered <25xa0min, and retrograde cerebral perfusion became less frequently used. Selective cerebral perfusion (SCP) is not associated with neurocognitive decline unless profound hypothermia (<20xa0°C) is used, which may suggest profound hypothermic SCP is not advantageous but may be detrimental. Attempts have been made to use mild to moderate hypothermia during SCP, and safe duration of distal circulatory arrest seems <60xa0min at 28xa0°C to avoid ischemic spinal cord injury. Three-vessel perfusion seems advantageous to provide adequate brain and spinal cord protection. To avoid aortogenic brain atheroembolism in the high risk patients, we previously proposed the “isolation” technique, where SCP is established before systemic perfusion. This technique has subsequently been modified to use both axillary and left carotid arteries for systemic arterial return, so that aortogenic emboli may not enter the brain circulation. In the TEVAR (thoracic endovascular aortic repair) era, hybrid operations such as the frozen elephant trunk or TEVAR completion after the elephant trunk are increasingly performed for extensive or distal arch aneurysms. It should be noted, however, that the frozen elephant trunk operation for extensive aneurysms carries an increased risk of paraplegia, and for distal arch aneurysms its outcome is not better than that after the standard open repair in Japan.

Characterizing saccular aortic arch aneurysms from the geometry-flow dynamics relationship

Objective: Low wall shear stress (WSS) has been reported to be associated with accelerated atherosclerosis, aneurysm growth, or rupture. We evaluated the geometry of aortic arch aneurysms and their relationship with WSS by using the 4‐dimensional flow magnetic resonance imaging to better characterize the saccular aneurysms. Methods: We analyzed the geometry in 100 patients using multiplanar reconstruction of computed tomography. We evaluated WSS and vortex flow using 4‐dimensional flow magnetic resonance imaging in 16 of them, which were compared with 8 age‐matched control subjects and eight healthy young volunteers. Results: Eighty‐two patients had a saccular aneurysm, and 18 had a fusiform aneurysm. External diameter/aneurysm length ratio and sac depth/neck width ratio of the fusiform aneurysms were constant at 0.76 ± 0.18 and 0.23 ± 0.09, whereas those of saccular aneurysms, especially those involving the outer curvature, were higher and more variable. Vortex flow was always present in the aneurysms, resulting in low WSS. When the sac depth/neck width ratio was less than 0.8, peak WSS correlated inversely with luminal diameter even in the saccular aneurysms. When this ratio exceeded 0.8, which was the case only with the saccular aneurysms, such correlation no longer existed and WSS was invariably low. Conclusions: Fusiform aneurysms elongate as they dilate, and WSS is lower as the diameter is larger. Saccular aneurysms dilate without proportionate elongation, and they, especially those occupying the inner curvature, have higher and variable sac depth/neck width ratio. When this ratio exceeds 0.8, WSS is low regardless of diameter, which may explain their malignant clinical behavior.

A nationwide survey of aortic valve surgery in Japan: current status of valve preservation in cases with aortic regurgitation

ObjectiveAlthough aortic valve-sparing operations are performed throughout Japan, the indications, specific repair techniques, and outcomes have not been reported in full. Thus, we conducted the first nationwide survey of aortic valve-sparing surgery.MethodsWe mailed a questionnaire to 508 institutions across Japan to obtain information on elective aortic valve and aortic root surgeries performed in 2014. Included in the mailing was a secondary questionnaire that sought further information from institutions reporting aortic valve-sparing surgeries.ResultsTwo hundred and fifty (49%) institutions responded and reported a total of 7859 aortic valve operations and 771 aortic root operations. Aortic valve operations performed strictly for aortic regurgitation totaled 2080, 156 (8%) of which were aortic valve repairs. Of the 699 aortic root surgeries performed for aortic regurgitation, 236 (34%) were valve-sparing root replacement surgeries. The valve-sparing root replacement surgeries comprised aortic valve reimplantation (nxa0=xa0173, 73%) and aortic root remodeling (nxa0=xa063, 27%). Five of 57 (9%) institutions were responsible for 42% (99/233) of the total aortic valve-sparing surgeries performed. Detailed information that was obtained for 233 patients who underwent aortic valve repair or valve-sparing root replacement showed 30-day mortality and reoperation for regurgitation after aortic valve repair (nxa0=xa097), aortic root remodeling (nxa0=xa037), and aortic valve reimplantation (nxa0=xa099) to be 1, 0, and 1% and 3, 3, and 1%, respectively.ConclusionTo date, aortic valve-sparing operations have been performed for limited patients at limited institution in Japan, but the early outcomes have been excellent.