Kidney fibrosis is independent of the amount of ascorbic acid in mice with unilateral ureteral obstruction.

In response to sustained damage to a kidney, fibrosis that can be characterized as the deposition of a collagenous matrix occurs and consequently causes chronic kidney failure. Because most animals used in experiments synthesize ascorbic acid (AsA) from glucose, the roles of AsA in fibrotic kidney diseases are largely unknown. Unilateral ureteric obstruction (UUO) mimics the complex pathophysiology of chronic obstructive nephropathy and is an ideal model for the investigation of the roles of AsA in kidney failure. We examined the impact of a deficiency of Akr1a, a gene that encodes aldehyde reductase and is responsible for the production of AsA, on fibrotic damage caused by UUO in mice. Oxidatively modified DNA was elevated in wild-type and Akr1a-deficient kidneys as a result of UUO to a similar extent, and was only slightly suppressed by the administration of AsA. Even though Akrla-deficient mice could produce only about 10% of the AsA produced by wild-type mice, no difference was observed in collagen I synthesis under pathological conditions. The data implied either a low demand for AsA or the presence of another electron donor for collagen I production in the mouse kidney. Next, we attempted to elucidate the potential causes for oxidative damage in kidney cells during the fibrotic change. We found decreases in mitochondrial proteins, particularly in electron transport complexes, at the initial stage of the kidney fibrosis. The data imply that a dysfunction of the mitochondria leads to an elevation of ROS, which results in kidney fibrosis by stimulating cellular transformation to myofibroblasts.

Development of a new aortoscope system for the use of endovascular intervention.

"Balloon aortoscopy" is a technique for viewing inner wall of aorta and used in clinics. By this method, endoluminal aortic surface could be clearly monitored, however, during this period, the aortic blood flow is blocked off by the inflated balloon. To solve this clinical problem, we have been developing a prototype aortoscope system without blocking off aortic flow aiming for the use of an assistive technique for endovascular interventions such as stent-graft placement for aortic aneurysm and have been evaluating through in vitro and in vivo tests. The technique introduced for this purpose was the use of intermittent and instantaneous saline jet controlled by a high-speed electromagnetic valve synchronized to heart beat (diastolic phase). In the previous study, we designed an endoscope with two channels (one for saline discharge and the other for forceps insertion), and confirmed the validity of this method by in vitro and in vivo tests. Based on these findings, in this study, we have newly designed a conventional and low price endoscope system aiming for wide clinical use. From the results of in vitro tests using a mock circulation system, it was confirmed that the newly designed system was capable of visualizing a target installed on an inner surface of the mock system suggesting an availability of the system for an aortoscope without blocking off aortic flow.

Neurological deficit after simultaneous open abdominal aortic surgery and thoracic aortic endovascular therapy.

We examined a total of 12 cases; 8 cases were identified by searching the literature on Pubmed (excluding case reports published prior to 2000) and 4 cases were ones we personally encountered. We examined age, sex, history of smoking, and preoperative risk factors as preoperative factors, the access route and coverage of the descending aorta as perioperative factors, and complications and survival time as postoperative factors. Mean coverage of the thoracic aorta was 90.8 mm. In terms of perioperative deaths, 8.3% (1 patient) were due to coagulopathy. Perioperative complications occurred in 16.7% of cases (coagulopathy in 1 patient and paralysis in another). No patients experienced complications or underwent additional treatment during a mean follow-up of 22.9 months. This study suggested that simultaneous open abdominal aortic repair and thoracic aortic endovascular therapy is feasible and also involves few postoperative complications. Paraplegia and paralysis tended to occur less frequently than with two-stage surgery, but further study is needed to explain why this is true.

Analyses of both the alkB gene transcriptional start site and alkB promoter-inducing properties of Rhodococcus sp. strain BCP1 grown on n-alkanes.

Rhodococcus sp. strain BCP1, known for its capacity to grow on short-chain n-alkanes (C(2) to C(7)) and to cometabolize chlorinated solvents, was found to also utilize medium- and long-chain n-alkanes (C(12) to C(24)) as energy and carbon sources. To examine this feature in detail, a chromosomal region which includes the alkB gene cluster encoding a non-heme di-iron monooxygenase (alkB), two rubredoxins, and one rubredoxin reductase was cloned from the BCP1 genome. Furthermore, the activity of the alkB gene promoter (P(alkB)) was examined in the presence of gaseous, liquid, and solid n-alkanes along with intermediates of the putative n-alkane degradation pathway. A recombinant plasmid, pTP(alkB)LacZ, was constructed by inserting the lacZ gene downstream of P(alkB), and it was used to transform Rhodococcus sp. strain BCP1. Measurements of ß-galactosidase activity showed that P(alkB) is induced by C(6) to C(22) n-alkanes. Conversely, C(2) to C(5) and >C(22) n-alkanes and alkenes, such as hexene, were not inducers of alkB expression. The effects on P(alkB) expression induced by alternative carbon sources along with putative products of n-hexane metabolism were also evaluated. This report highlights the great versatility of Rhodococcus sp. strain BCP1 and defines for the first time the alkB gene transcriptional start site and the alkB promoter-inducing capacities for substrates different from n-alkanes in a Rhodococcus strain.

Evaluation of a newly designed endoscope for observing inner wall of large arteries for the use of endovascular intervention.

A prototype endoscope for observing inner wall of large arteries was specially designed and evaluated through in vitro and in vivo tests. The purpose of this endoscope is to visualize the inner wall of large arteries, e.g., an aorta, without blocking off the blood stream aiming for the use of an assistive technique for endovascular interventions such as stent-graft placement for aortic aneurysm. The technique newly introduced for this purpose was the use of intermittent high-pressure saline jet synchronized to heart beat (diastolic phase). In the previous studies using commercially available bronchoscopes, we confirmed the validity of the system utilizing this technique [1, 2]. Based on these findings, in this study, we have specially designed a new endoscope with two channels, one for saline discharge and the other for forceps, and evaluated its performance through in vitro and in vivo tests. From the results of in vitro tests using a mock circulation system, it was confirmed that the newly designed endoscope was capable of visualizing a target installed on an inner surface of the mock system. Also confirmed through in vivo tests using swine was that we could observe bifurcation in descending aorta, e.g., left renal artery, without stopping off the blood stream.

Elective Matsui-Kitamura stent graft repair for descending thoracic aortic aneurysm and chronic type-B aortic dissection.

BACKGROUND: The position of thoracic endovascular aortic repair (TEVAR) compared to open surgery of the thoracic aorta has changed. This study evaluates outcomes after TEVAR performed electively using our original Matsui-Kitamura stent graft (MKSG) to treat descending thoracic aortic aneurysms (dTAA) and chronic type-B aortic dissection (type-B AD), and elucidates the risk factors for postoperative spinal cord ischemia (SCI). METHODS: TEVAR was performed using an MKSG in 66 patients (age: 70.8+/-9.2 years). The underlying etiology was atherosclerotic change in 39 patients, chronic type-B aortic dissection in 23 patients, and other in 4 patients. RESULTS: No perioperative deaths occurred. Three patients showed temporary paralysis due to postoperative SCI. Abdominal aortic aneurysm (AAA) surgery was a risk factor for postoperative SCI (P=0.04). The 5-year survival rate was 81.2%. CONCLUSION: The present study demonstrated that TEVAR of patients with dTAA and chronic type-B AD using an MKSG can be performed with high technical success rates and low rates of severe acute complications. AAA surgery was a risk factor for postoperative SCI.

[Video-assisted thoracic surgery for clinical stage I lung cancer].

We evaluated our results of video-assisted thoracic surgery (VATS) performed for lung cancer over 8 years. Between April 2000 and October 2008, a total of 409 (60.9%) underwent VATS for lung cancer. Operative procedures as a radical operation were partial resection in 58 patients, segmentectomy in 64 patients, and lobectomy in 229 patients. There was 1 patient with operative death including hospital death due to pulmonary thromboembolism. In a median follow-up period of 21 months, the 5-year cause specific survival rate was 93.7%. According to operative procedures, the 5-year survival rate was 100% in patients underwent partial resection and segmentectomy, and 91.1% in patients underwent lobectomy. According to pathological stages, the 5-year survival rate was 98.8% in 289 patients with stage IA, 69.1% in 34 patients with stage IB, and 68.2% in 14 patients with stage IIIA. In conclusion, VATS lobectomy and VATS intentional limited resection can be performed with low mortality and good prognosis for clinical stage IA lung cancer patients.

Twisted ovarian tumor causing progressive hemothorax: a case report of porous diaphragm syndrome.

BACKGROUND: Porous diaphragm syndrome is caused by a defect in the diaphragm. The defect may induce pleural effusion in a patient with an ovarian tumor. CASE REPORT: A 59-year-old Japanese woman with an ovarian tumor and right hemothorax underwent thoracotomy and presented with a fenestra in the right diaphragm through which bloody fluids were flowing from the peritoneal cavity into the pleural space. Following suturing of the fenestra, laparotomy revealed intraabdominal bleeding due to torsion of an ovarian tumor. CONCLUSION: This is the first report in which the diaphragmatic defect was identified in a patient with an ovarian tumor and hemothorax. Porous diaphragm syndrome may be involved in the pathophysiology of right pleural effusion observed in other medical conditions such as Meigs' syndrome, ovarian hyperstimulated syndrome, and ovarian cancer.

[Quality of life of long-term survivors of surgically treated lung cancer].

Quality of life (QOL) of long-term survivors (more than 3 years after surgery) of primary non-small cell lung cancer was studied. QOL was analyzed using European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, 30-Item version 3.0 (QLQ-C30) and Hospital Anxiety and Depression Scale (HADS). Sixty of 91 patients (66%) participated in this study 87 +/- 5 (38-172) months postoperatively. In QLQ-C30, calculated scores of physical (84.0 +/- 2.4), role (81.3 +/- 3.6), cognitive (79.7 +/- 2.6), emotional (86.8 +/- 1.9), and social (91.0 +/- 1.9) functioning, and global QOL (72.6 +/- 2.9) were obtained. Calculated HADS A (anxiety) was 3.3 +/- 0.3 and HADS D (depression) was 4.0 +/- 0.4. Postoperative follow-up duration was correlated with financial impact only. QOL of long-term survivors was influenced by gender histology, marital status, employment status, and academic carrier.

Experimental study of aortic anastomosis using a circular stapling device in the porcine model.

OBJECTIVES: The aim of this study was to assess the strength (pressure resistance) and histological findings of aortic anastomoses performed using a circular stapling device. MATERIALS AND METHODS: A circular stapling device was used for anastomosing a porcine aorta and a Dacron graft. The maximum pressure resistance of the anastomotic site of a porcine aortic specimen and a Dacron graft was examined (n=10). A porcine aorta with Dacron graft was anastomosed to a beating heart, and pressure overload was induced by adrenaline (n=5). Specimens of the anastomotic sites were harvested after 14 days and examined histologically. RESULTS: The maximum pressure resistance of the anastomotic site was 427.3+/-34.4 (375-511) mmHg. No anastomotic sites leaked as a result of pressure overloading at 227.6+/-21.1 (201-260) mmHg. Histologically, good incorporation and cell coverage were observed, and the inner surfaces of the anastomotic sites were smooth and without stenoses. CONCLUSIONS: Aortic anastomosis using a circular stapling device is feasible and worthy of further investigation.

Inorganic polyphosphate stimulates lon-mediated proteolysis of nucleoid proteins in Escherichia coli.

Inorganic polyphosphate (polyP) accumulates in response to amino acid starvation in Escherichia coli. Previously, we found that the complex formation of Lon with polyP stimulates proteolysis of free ribosomal proteins. In the current studies, we examined the effects of polyP on the degradation of major nucleoid proteins. Fusions of green fluorescent protein with HimA, Fis, HupA, and HupB were clearly associated with polyP in vivo. Lon degraded His-tagged HimA protein only in the presence of polyP in vitro as well as in vivo. Whereas, when HimA and HimD formed a heterodimer, Lon could not degrade it even in the presence of polyP. In addition, Lon degraded His-tagged Fis protein in the presence of polyP. However, in vivo, Lon did not efficiently degrade the Fis protein even when cells accumulated polyP in response to amino acid starvation. It appears that this is due to tighter binding of Fis to DNA than to polyP and resistance of the DNA-Fis to Lon-mediated proteolysis. Indeed, we found that at least a five-fold excess of polyP was necessary to displace DNA from the DNA-Fis complex. Furthermore, Lon degraded His-tagged HupA protein efficiently in the presence of polyP. We also showed that degradation of the translational initiation factor InfC depends on polyP.

Establishment of an immortalised human ovarian surface epithelial cell line without chromosomal instability.

Epithelial ovarian carcinoma is thought to derive from ovarian surface epithelium (OSE). The black box of the early molecular changes in ovarian carcinogenesis is being interpreted by the development of experimental systems employing immortalised human OSE cells. However, the existing cell lines of the OSE cells have limited utility due to chromosomal instability. Our goal was to establish new immortalised human OSE cells that retain the original characteristics of the primary cells without chromosomal alterations. Using primary human OSE cells obtained from a postmenopausal patient with endometrial cancer, five cell lines ('HOSE1' lines) were newly established by infection with retroviral expression vectors containing type 16 human papillomavirus (HPV-16) E6, E7, a variant E6 (E6delta151), and Bmi1 polycomb gene, in combination with telomerase reverse transcriptase (hTERT). Consequently, five HOSE1s cell lines, HOSE1s-E6/hTERT, -E7/hTERT, -E6/E7/hTERT, -E6delta151/E7/hTERT, and -E6delta151/Bmi1/hTERT, grew beyond the population doubling number of 200. These cell lines, except for HOSE1-E6/hTERT, essentially showed the original features of the primary human OSE cells. Of them, HOSE1-E7/hTERT preserved diploidy in a kariotype analysis, and did not show transformed phenotypes in anchorage-independent growth and tumour formation. Thus, HOSE1-E7/hTERT may provide a novel model system with which to investigate the mechanisms of early molecular changes.

Beta-synuclein gene alterations in dementia with Lewy bodies.

OBJECTIVE: To determine whether mutations in the genes for alpha-synuclein or beta-synuclein are responsible for dementia with Lewy bodies (DLB), a disorder closely related to Parkinson disease (PD). METHODS: The authors ascertained 33 sporadic cases of DLB and 10 kindreds segregating DLB. DNA samples from the 43 index cases were screened for alterations in the genes for alpha-synuclein and beta-synuclein, as alpha-synuclein alterations cause PD and beta-synuclein may modulate alpha-synuclein aggregation and neurotoxicity. RESULTS: Two amino acid alterations were identified in unrelated DLB index cases: a valine to methionine substitution at codon 70 (V70M) and a proline to histidine substitution at codon 123 (P123H), both in the beta-synuclein gene. These amino acid substitutions occur at conserved residues in highly conserved regions of the beta-synuclein protein. Screening of at least 660 chromosomes from control subjects matched to the patients' population groups failed to identify another V70M or P123H allele. Cosegregation analysis of an extended pedigree segregating the P123H beta-synuclein alteration suggested that it is a dominant trait with reduced penetrance or a risk factor polymorphism. Histopathology and immunohistochemistry analysis of index case brain sections revealed widespread Lewy body pathology and alpha-synuclein aggregation without evidence of beta-synuclein aggregation. CONCLUSION: Mutations in the beta-synuclein gene may predispose to DLB.

Adult-onset leukoencephalopathy with vanishing white matter with a missense mutation in EIF2B5.

We report of a woman aged 52 years born to consanguineous parents and seeking treatment for progressive dementia and delusion. Neurologic examination revealed dementia and emotional instability, indifference, and confabulation. There was also mild spasticity of the bilateral lower limbs. MRI revealed diffuse white matter hyperintensity on T2-weighted images accompanied by hypointense areas on fluid-attenuated inversion recovery images. A homozygous missense mutation was identified in EIF2B5.

A patient with dementia with Lewy bodies and codon 232 mutation of PRNP.

The authors describe a patient who had a point mutation at codon 232 of the prion protein gene, resulting in the substitution of methionine for arginine (M232R). The patient developed dementia and died 6 years after its onset. Autopsy revealed dementia with Lewy bodies, not Creutzfeldt-Jakob disease. Although the M232R mutation has been reported to cause Creutzfeldt-Jakob disease, findings in our patient suggest that not all patients presenting progressive dementia with M232R mutation have Creutzfeldt-Jakob disease.

Clear cell sarcoma arising in the retroperitoneum.

Clear cell sarcoma is a rare soft-tissue neoplasm, arising most commonly in the tendons and aponeuroses of young adults. We report here the first female case of clear cell sarcoma arising in the retroperitoneum with clinical features similar to those of malignant ovarian tumors. Aspects of clinical presentation, histopathologic evaluation, and treatment are described.

Analysis of prognostic factors in newly diagnosed patients with acute promyelocytic leukemia: the APL92 study of the Japan Adult Leukemia Study Group (JALSG).

All-trans-retinoic acid (ATRA) has been incorporated in front-line therapy for newly diagnosed acute promyelocytic leukemia (APL). We conducted a multicenter study of differentiation therapy with ATRA alone or in combination with chemotherapy followed by intensive postremission chemotherapy in patients with APL (the JALSG APL92 study), and analyzed prognostic factors to increase the cure rate in our subsequent trial. From 1992 to 1997, adult patients with newly diagnosed APL received oral ATRA 45 mg/m2 daily alone until complete remission (CR) if initial leukocyte counts were < 3.0x10(9)/l, and ATRA daily plus daunorubicin (DNR) 40 mg/m2x3 days plus enocitabine (BHAC) 200 mg/m2x5 days if leukocyte counts were > or =3.0 x 10(9)/l. If peripheral blasts exceeded 1.0x10(9)/l during therapy, DNRx3 days plus BHACx5 days was added. After CR was achieved, three courses of consolidation and six courses of maintenance/intensification chemotherapy were administered. Of 376 patients enrolled, 369 were evaluable (median age 46 years, range 15-86 years; median leukocyte counts 2.0x10(9)/l), and 333 (90%) achieved CR (94% of patients treated with ATRA alone, 88% with ATRA plus later chemotherapy, 89% with ATRA plus initial chemotherapy, and 86% with ATRA plus initial and later chemotherapy). At a median follow-up of 45 months, the predicted 6-year overall and event-free survival (EFS) rates for all patients were 65% and 52%, respectively. Favorable prognostic factors for CR were younger age, no or mild purpura, high serum total protein level, low lactate dehydrogenase level, and no or mild disseminated intravascular coagulation (DIC). Favorable prognostic factors for EFS were leukocyte counts < 10.0x10(9)/l, mild DIC, and no sepsis during induction therapy. In the JALSG APL97 study, we intensified chemotherapy for patients with leukocyte counts > or =3.0x10(9)/l, and are randomly testing whether further chemotherapy is required for APL patients with negative PCR for PML/retinoic acid receptor alpha in the maintenance phase.

Role of inorganic polyphosphate in promoting ribosomal protein degradation by the Lon protease in E. coli.

Inorganic polyphosphate (polyP), a polymer of hundreds of phosphate (Pi) residues, accumulates in Escherichia coli in response to stresses, including amino acid starvation. Here we show that the adenosine 5'-triphosphate-dependent protease Lon formed a complex with polyP and degraded most of the ribosomal proteins, including S2, L9, and L13. Purified S2 also bound to polyP and formed a complex with Lon in the presence of polyP. Thus, polyP may promote ribosomal protein degradation by the Lon protease, thereby supplying the amino acids needed to respond to starvation.