Pz peptidase A from the thermophilic bacterium Geobacillus collagenovorans MO-1, hydrolyzes a synthetic peptide substrate, 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-PLGPR), which contains a collagen-specific tripeptide sequence, -Gly-Pro-X, but does not act on collagen proteins themselves. The mammarian enzyme, thimet oligopeptidase (TOP), which has comparable functions to bacterial Pz peptidases but limited identity at the primary sequence level, has recently been subjected to X-ray crystallographic analysis; however, no crystal structure has yet been reported for complexes of TOP with substrate analogues. Here, we report crystallization of recombinant Pz peptidase A in complex with two phosphinic peptide inhibitors (PPIs) that also function as inhibitors of TOP, and determination of the crystal structure these complexes at 1.80-2.00 A resolution. The most striking difference between Pz peptidase A and TOP is that there is no channel running the length of bacterial protein. Whereas the structure of TOP resembles an open bivalve, that of Pz peptidase A is closed and globular. This suggests that collagenous peptide substrates enter the tunnel at the top gateway of the closed Pz peptidase A molecule, and reactant peptides are released from the bottom gateway after cleavage at the active site located in the center of the tunnel. One of the two PPIs, PPI-2, which contains the collagen-specific sequence, and helped to clarify the exquisite structure and reaction mechanism of Pz peptidase A toward collagenous peptides. This study describes the mode of substrate binding and its implication for the mammalian enzymes.

Abstract The aim was to evaluate the role of the combination of olmesartan, an angiotensin II (Ang II) receptor blocker (ARB), with daunorubicin (DNR) in reducing cardiac toxicity in rats. DNR was administered at a dose of 3 mg/kg/day every other day for 12 days. Olmesartan was administered orally every day for 12 days. Rats treated with DNR alone showed cardiac toxicity as evidenced by worsening cardiac function, elevation of malondialdehyde level in heart tissue and decreased in the level of total glutathione peroxidase activity; treatment with ARB reversed these changes. Furthermore, ARB treatment down-regulated matrix metalloproteinase-2 expression, myocardial expression of Ang II, attenuated the increased protein expressions of p67(phox) and Nox4 and reduced oxidative stress-induced DNA damage evaluated by expression of 8-hydroxydeoxyguanosine. In conclusion, the result demonstrated that Ang II and oxidative stress play a key role in anthracycline-induced cardiotoxicity and that treatment with ARB will be beneficial against DNR-induced cardiotoxicity.

Abstract Experimental autoimmune myocarditis (EAM) is mediated by myocardial infiltration by myosin-specific T-cells secreting inflammatory cytokines. In this study, rat models of EAM were prepared by injection with porcine cardiac myosin. One week after immunization, edaravone was administered intraperitoneally at 3 or 10 mg/kg/day to rats for 2 weeks. Cardiac function was measured by haemodynamic and echocardiographic studies and TUNEL assay was performed. Left ventricular (LV) expression of NADPH oxidase sub-units (p47(phox) and p67(phox)), pro-inflammatory cytokines (TNF-alpha), endoplasmic reticulum (ER) stress signalling proteins (GRP78, caspase-12 and GADD153) and mitogen-activated protein kinase (MAPK) family proteins (phospho-p38 MAPK and phospho-JNK) were measured by western blotting. Edaravone improved LV function in a dose-dependent manner. Central venous pressure was significantly low and LV ejection fraction and fractional shortening was significantly high in edaravone groups compared with those in the vehicle group. In addition, edaravone treatment down-regulated LV expressions of p47(phox), TNF-alpha, GADD153, phospho-p38 MAPK and phospho-JNK. Furthermore, the LV expressions of p67(phox), GRP78, caspase-12 and TUNEL-positive cells of rats with EAM treated with edaravone were significantly low compared with those of the vehicle group. These findings suggest that edaravone ameliorated the progression of EAM by inhibiting oxidative and ER stress and, subsequently, cardiac apoptosis.

The magnitude of rater differences, instead of interrater reliability, in the assessment scales of schizophrenia has rarely been investigated and was therefore addressed in this study.Thirty-six patients with schizophrenia were independently assessed by 4 expert physicians, using clinical rating scales including the Positive and Negative Syndrome Scale (PANSS). The scores obtained by the physician in charge (PIC), who had a long close contact with the patients, served as the referent answer for the purpose of this study. The scores rated by the other 3 non-PIC psychiatrists, who had a first formal examination with them, were evaluated for percentage deviance from the referent answer.The results showed that the PIC raters endorsed the numerically highest score in 20 (56%) of the 36 patients, whereas they rated the lowest in only 2 (6%) in the PANSS total score. The non-PIC assessors on the average underrated the PANSS total score by 10%, and such a tendency of underestimating the severity was noted across other clinical scales. Furthermore, the PANSS total score by one of the non-PIC physicians was deviant from the referent answer by at least 20% in 15 (42%) of 36 instances. Importantly, this magnitude of deviance was noted in the context of an intraclass correlation coefficient of 0.92.This unique investigation disclosed clinically pertinent differences among raters, even under an excellent interrater reliability. The magnitude of differences described herein seems to be an underestimation, and the baseline scores by the independent new raters might need to be corrected for those by the PICs.

BACKGROUND AND OBJECTIVES: The aim of this study was to evaluate the feasibility of en bloc colorectal resection combined with radical prostatectomy as an alternative to total pelvic exenteration (TPE) for patients with locally advanced rectal cancer involving the lower urinary tract organs. METHODS: Twenty men with primary rectal cancer clinically involving the lower urinary tract organs underwent extended colorectal resection combined with radical prostatectomy. Data were entered prospectively into a database. Oncological and functional outcomes were analyzed. RESULTS: Anal sphincter-preserving operation (SPO) with radical prostatectomy was performed in 12 patients, abdominoperineal resection with radical prostatectomy in 8, and urinary reconstruction in 16. Morbidity and mortality rates were 35.0% and 0%, respectively. Five-year overall and disease-free survival rates were 83.6% and 42%, respectively. The cumulative 5-year local recurrence rate was 20.0%. All patients with urinary reconstruction achieved good voiding function, and patients with SPO showed acceptable anal function. CONCLUSIONS: For lower rectal cancers involving lower urinary tract, en bloc rectal resection combined with radical prostatectomy appears oncologically acceptable and can reduce the number of TPEs. J. Surg. Oncol. (c) 2010 Wiley-Liss, Inc.

STUDY DESIGN.: Prospective clinical series. OBJECTIVE.: To determine how many thoracic scoliotic pedicles have cancellous versus cortical versus absent channels. SUMMARY OF BACKGROUND DATA.: Although morphologic evaluations of thoracic pedicles have been well reported, the results do not practically reflect clinical findings during actual pedicle screw placement. We propose a novel pedicle channel classification describing the osseous anatomy encountered during pedicle probe insertion. METHODS.: We noted 4 pedicle types in 53 consecutive scoliosis patients. Type A: pedicle probe smoothly inserted without difficulty; the morphology is described as a "Large Cancellous Channel." Type B: pedicle probe inserted snugly with increased force; described as a "Small Cancellous Channel." Type C: pedicle probe cannot be manually pushed but must be tapped with a mallet down the pedicle into the body; described as a "Cortical Channel." Type D: pedicle probe cannot locate a channel thus necessitating a "juxtapedicular" screw position; described as a "Slit/Absent Channel." The average age at time of surgery was 23.4 +/- 16.7 years. Diagnoses included idiopathic scoliosis (n = 38) and syndromic scoliosis (n = 15). The average main thoracic Cobb angle was 73 degrees +/- 26 degrees . Evaluation of pedicle morphology of the 4 types was also performed in 21 consecutive cases of adolescent idiopathic scoliosis using preoperative computed tomography images. RESULTS.: A total of 1021 pedicles with screws placed were evaluated. The average percent per type was as follows: 61.0% type A; 29.2% type B, 6.8% type C, and 3.0% type D. On the convexity, 98.2% of pedicles were type A or B versus 81.5% on the concavity (P < 0.05). There were significant differences between adolescent versus adult idiopathic scoliosis (P = 0.007), and syndromic scoliosis versus adult idiopathic scoliosis (P = 0.017) regarding pedicle morphologic proportions. There was a significant tendency toward a decrease in the proportion of type A pedicles, an increase in the proportion of type B pedicles as the Cobb angle increased (P < 0.0001). Evaluation based on 312 thoracic pedicles in 21 consecutive adolescent idiopathic scoliosis patients using preoperative computed tomography axial images confirmed assumptions of the 4 pedicle types. CONCLUSION.: We propose a classification for pedicle channels describing the osseous anatomy encountered during pedicle probe insertion. Based on the classification, surprisingly, we found during surgery that 90% of thoracic pedicles had a cancellous channel, whereas 7% had a cortical channel and only 3% had an absent channel.

STUDY DESIGN.: Case report. OBJECTIVE.: To describe an iatrogenic aortic injury by pedicle screw instrumentation during posterior reconstructive surgery of spinal deformity. SUMMARY OF BACKGROUND DATA.: Iatrogenic major vascular injuries during anterior instrumentation procedures have been reported by several authors, but there have been few reports regarding iatrogenic major vascular injuries during posterior instrumentation procedures. METHODS.: A 57-year-old woman with thoracolumbar kyphosis due to osteoporotic T12 vertebral fracture underwent posterior correction and fusion (T10-L2), using segmental pedicle screw construct concomitant with T12 pedicle subtraction osteotomy. Postoperative routine plain radiographs and computed tomography myelography demonstrated a misplaced left T10 pedicle screw, which was in contact with the posteromedial aspect of the thoracic aorta, and suspected penetration of the aortic wall. The patient underwent removal of the pedicle screw, and repair of the penetrated aortic wall through a simultaneous anterior-posterior approach. RESULT.: The patient tolerated the procedure well without neurologic sequelae, and was discharged several days after removal of a left tube thoracostomy. Plain radiographs demonstrated solid fusion at the osteotomy site and no loosening of hardware. Preoperative neurologic symptoms improved completely at 18-months follow-up. CONCLUSION.: Use of pedicle screw instrumentation has the potential to cause major vascular injury during posterior spinal surgery, and measures to prevent this complication must be taken. Timely diagnosis and treatment are essential to prevent both early and delayed complications and death.

A facultatively anaerobic, prosthecate bacterium, strain Mfc52T, was isolated from a microbial fuel cell (MFC) inoculated with soil and fed with cellulose as the sole fuel. Cells were Gram-negative, non-spore-forming, straight or slightly curved rods, and some of them had one or two polar prosthecae (stalks). Cells reproduced by binary fission or by budding from mother cells having prosthecae. Strain Mfc52T fermented various sugars and produced lactate, acetate and fumarate. Ferric iron, nitrate, oxygen and fumarate served as electron acceptors, while sulfate and malate did not. Nitrate was reduced to nitrite. The DNA G + C content was 64.7 mol%. On the basis of 16S rRNA gene sequence phylogeny, strain Mfc52T was affiliated with the genus Rhizomicrobium in the class Alphaproteobacteria and most closely related to R. palustre with a sequence similarity of 97 %. Based on these physiological and phylogenetic characteristics, the name Rhizomicrobium electricum sp. nov. is proposed; the type strain is strain Mfc52T (= JCM 15089T = KTCC 5806T).

Background/Aims: Excessive endoplasmic reticulum stress (ERS) triggers apoptosis in various conditions including diabetic cardiomyopathy and pressure overload-induced cardiac hypertrophy and heart failure. The primary function of 14-3-3 protein is to inhibit apoptosis, but the roles of this protein in protecting against cardiac ERS and apoptosis are largely unknown. Methods: We investigated the roles of 14-3-3 protein in vivo during cardiac ERS and apoptosis induced by pressure overload or thapsigargin injection using transgenic (TG) mice that showed cardiac-specific expression of dominant negative (DN) 14-3-3eta. Results: Cardiac positive apoptotic cells and the expression of glucose-regulated protein (GRP)78, inositol-requiring enzyme (Ire)1alpha, tumor necrosis factor receptor (TNFR)-associated factor (TRAF)2, CCAAT/enhancer binding protein homology protein (CHOP), caspase-12, and cleaved caspase-12 protein were significantly increased in the pressure-overload induced DN 14-3-3eta mice compared with that in the WT mice. Furthermore, thapsigargin injection significantly increased the expression of GRP78 and TRAF2 expression in DN 14-3-3eta mice compared with that in the WT mice. Conclusion: The enhancement of 14-3-3 protein may provide a novel protective therapy against cardiac ERS and ERS-initiated apoptosis, at least in part, through the regulation of CHOP and caspase-12 via the Ire1alpha/TRAF2 pathway.

Although previous studies have claimed that Pungtungia herzi (Cyprinidae) is an obligate brood parasite, in this study a significant proportion of P. herzi egg masses was found in crevices formed by stones and rocks, which was not guarded by hosts. Egg masses were also found in almost all nests of a goby Odontobutis obscura, suggesting that P. herzi has two alternative reproductive tactics: brood parasitic spawning and non-parasitic crevice spawning, the latter of which may occur when a shortage of reproductive resources (i.e. host nests) exists. The hatching rates of parasitic egg masses were significantly higher than those of non-parasitic egg masses, and the hatching rate of P. herzi eggs exhibited a strong positive correlation with continuous egg guarding by a reproductive O. obscura male. These results indicate that brood parasitism effectively improves reproductive success in P. herzi. The alternative reproductive tactic of this species suggests that brood parasitism has evolved directly from crevice spawning via the sharing of spawning habitat with host species.

The functionality of sense-antisense transcripts (SATs), although widespread throughout the mammalian genome, is largely unknown. Here, we analyzed the SATs expression and its associated promoter DNA methylation status by surveying 12 tissues of mice to gain insights into the relationship between expression and DNA methylation of SATs. We have found that sense and antisense expression positively correlate in most tissues. However, in some SATs with tissue-specific expression, the expression level of a transcript from a CpG island-bearing promoter is low when the promoter DNA methylation is present. In these circumstances, the expression level of its opposite strand transcript, especially when it is poly(A)-negative was coincidentally higher. These observations suggest that, albeit the general tendency of sense-antisense simultaneous expression, some antisense transcripts have coordinated expression with its counterpart sense gene promoter methylation. This cross-strand relationship is not a privilege of imprinted genes but seems to occur widely in SATs.

Graphene devices on standard SiO(2) substrates are highly disordered, exhibiting characteristics that are far inferior to the expected intrinsic properties of graphene. Although suspending the graphene above the substrate leads to a substantial improvement in device quality, this geometry imposes severe limitations on device architecture and functionality. There is a growing need, therefore, to identify dielectrics that allow a substrate-supported geometry while retaining the quality achieved with a suspended sample. Hexagonal boron nitride (h-BN) is an appealing substrate, because it has an atomically smooth surface that is relatively free of dangling bonds and charge traps. It also has a lattice constant similar to that of graphite, and has large optical phonon modes and a large electrical bandgap. Here we report the fabrication and characterization of high-quality exfoliated mono- and bilayer graphene devices on single-crystal h-BN substrates, by using a mechanical transfer process. Graphene devices on h-BN substrates have mobilities and carrier inhomogeneities that are almost an order of magnitude better than devices on SiO(2). These devices also show reduced roughness, intrinsic doping and chemical reactivity. The ability to assemble crystalline layered materials in a controlled way permits the fabrication of graphene devices on other promising dielectrics and allows for the realization of more complex graphene heterostructures.

We report the case of a 51-year-old woman with pulmonary adenocarcinoma with enteric differentiation (PAED) that is indistinguishable from metastatic colorectal carcinoma by immunohistochemistry as well as histology. A chest computed tomography scan revealed a 1cm nodule in the right upper lobe and a 3cm mass in the left lower lobe. Initial examination showed no evidence of any other tumor. She underwent partial resection of the right upper lobe and left lower lobectomy. Histopathological examination revealed that both tumors were composed of medium to large complex glands with central necrosis. The tumor cells were cuboidal to tall columnar with eosinophilc cytoplasm, oval nuclei, and brush-border. Immunohistochemical study yielded the following results: tumor cells were diffusely positive for cytokeratin (CK) 20 and CDX-2, and negative for CK7, thyroid transcription factor-1, and Napsin A. MUC2 was partially observed, while MUC5AC was not detected. These findings were strongly indicative of metastatic colorectal carcinoma. However, no primary colorectal cancer was detected in any clinical examination, including fluorine 18-labeled fluorodeoxyglucose-positron emission tomography scan and video capsule endoscopy, and she has not presented with any characteristic symptoms at any follow-up to date, approximately 4 years after operation. From all features, the final diagnosis was primary PAED, suggestive of multifocal primary lung cancer. So far, only 1 case of CK7-negative PAED has been reported. This is the second case of primary PAED resembling metastatic colorectal cancer morphologically and immunohistologically.

Summary Background: A few studies have explored the prevalence of dyslexia among children who speak Japanese as their native language by evaluating them individually by means of reading-based tasks. The present study was designed to clarify the frequency of suspected dyslexia among second-graders attending ordinary classes. Methods: The subjects were 40 children (22 male, 18 female; 7 years 4 months - 8 years 4 months; mean age, 7 years 11 months) out of 182 second-graders at a public elementary school situated in a local city. Each subject underwent a monomoraic syllable reading task, a word reading task, a non-word reading task, and a short sentence reading task. Results: The scores on the four tests were not normally distributed; rather, they were strongly skewed to shorter reading time or fewer reading errors. Additionally, they were significantly extended toward either longer reading time or more reading errors. Except in the non-word reading task, most subjects only made a few reading errors. Seven subjects (17.5%) showed at least one score that was more than 1.5 IQR (interquartile range) higher than the third quartile of that subject's 8 scores on the 4 tasks. Assuming that those 7 children are potentially dyslexic, at least 3.8 % of second-graders (7 out of 182) are suspected to be suffering from dyslexia. Conclusion: It is likely that the prevalence of dyslexia in Japan is comparable to that in Europe and the US. To confirm this, a more comprehensive study on a larger scale should be implemented in the future.

The judgment standards of R-K method include ambiguities and are thus compensated by subjective interpretations of sleep stage scorers. This paper presents a novel method to compensate uncertainties in judgments by the subjective interpretations by the sleep model estimation approach and by describing the judgments in probabilistic terms. Kalman filter based on the two sleep models with no body movement and with body movement was designed. Sleep stages judged by three different scorers were re-judged by the filter. The two sleep models were stochastically estimated from bio-signals from 15 nights data and the re-judged scores by the filter were evaluated by the data from 5 nights. The average values of kappa statistics, which show the degree of agreement, were 0.85, 0.89 and 0.81, respectively, for the original sleep stages. Because the new method provides probabilities on how surely the sleep belongs to each sleep stage, we were able to determine the most, second most and third most probable sleep stage. The kappa statistics between the most probable sleep stages were improved to 0.90, 0.93 and 0.84, respectively. Those of sleep stages determined from the most and second most probable were 0.92, 0.94 and 0.89 and those from the most, second most and third most probable were 0.95, 0.97 and 0.92. The sleep stages estimated by the filter are expressed by probabilistic manner which are more reasonable in expression than those given by deterministic manner. The expression could compensate the uncertainties in each judgments and thus were more accurate than the direct judgments.

We experienced anaphylactic shock after introduction of the general anesthesia twice in the same patient. After the first incidence of anaphylactic shock, we judged that the allergen was a latex. For the second time we planned the latex free environment, but the anaphylactic shock occurred again. As a result of the investigation, it turned out that the allergens were due to latex and sevoflurane. After the first incidence, it was necessary to retrieve the antigen from all the agents used. When we encounter the anaphylactic shock, it is necessary to examine all the medicines, and should have two or more suspectible medicines in mind.

AIM OF THE STUDY: Acanthopanax senticosus Harms extract (ASE) is used as an ingredient of over-the-counter drugs and functional foods, such as health supplements, in Japan. ASE exhibits a hypoglycemic effect; however, the mechanism of the hypoglycemic effect is not clear. In the present study, we investigated whether ASE has a glucose absorption inhibitory action. MATERIALS AND METHODS: We examined the effects of ASE on alpha-amylase and alpha-glucosidase activities, and on glucose uptake in Caco-2 cells. We also examined the effects of ASE oral administration on glucose tolerance in type 2 diabetes mellitus model db/db mice. RESULTS: The addition of ASE inhibited alpha-glucosidase activity but not alpha-amylase activity. The alpha-glucosidase inhibitory activity of ASE was approximately 1/13 of that of acarbose. The addition of ASE inhibited 2'-deoxy-D-glucose (DG) uptake in human intestinal Caco-2 cells, and the inhibitory activity of ASE was approximately 1/40 of that of phloretin. Kinetic analysis of glucose uptake indicated that ASE has no effects on DG uptake through passive diffusion, but that ASE inhibits intracellular DG uptake chiefly by inhibiting transport via a glucose transporter. In the glucose tolerance study, db/db mice orally administered ASE for 3 days showed significantly lower plasma glucose level than the control group 30min after sucrose loading, without affecting plasma insulin levels. In addition, ASE oral administration significantly inhibited alpha-glucosidase activity in the small intestine mucosa extirpated from the mice. CONCLUSION: These findings indicate that ASE may be useful as an ingredient of functional foods to improve postprandial hyperglycemia and prevent type II diabetes mellitus.

Arginine vasopressin attracted attention as a potentially important neurohormonal mediator of the heart failure syndrome and hyponatremic states in humans because arginine vasopressin influences renal handling of free water, vasoconstriction and myocyte biology through activation of V(2) and V(1a) receptors. Current research is exploring V(2)- and dual V(1a)/V(2)-receptor antagonism for the treatment of hyponatremia, as well as for the congestion and edema associated with chronic heart failure, because vasopressin receptor antagonists might offer benefits in comparison with conventional loop diuretics. The purpose of this review is to update the current status of experimental and clinical studies with available vasopressin receptor antagonists (conivaptan and tolvaptan) and their potential role in the treatment of heart failure and hyponatremia of multiple causes.

Brain-derived neurotrophic factor (BDNF) belongs to the neurotrophin family, and enhances the growth and maintenance of several neuronal systems. In addition, BDNF may promote neurogenesis and protect against hippocampal volume loss in depressive disorders. Although first detected in brain, BDNF also exists in peripheral tissues and is mainly stored in platelets and circulates in blood. Recent reports indicate that serum BDNF levels in depressive patients are lower than in control subjects, and antidepressant treatment increases serum BDNF levels in responders. A single report suggests that decreased serum BDNF in major depression is related to mechanisms of platelet BDNF release; however, the mechanisms of changes in BDNF blood levels are still poorly understood. In the present study, we investigated the direct influence of antidepressants on BDNF release from platelets and their effects on serum levels. We used samples of washed platelets prepared from rat blood, and investigated the platelet BDNF release and serum BDNF concentration changes in response to adding antidepressants. We found that BDNF was dose-dependently released from platelets by direct treatment with various kinds of antidepressants in vitro, and serum BDNF concentration was also increased by intravenous antidepressant treatment. These results confirm that BDNF release from platelets is affected by antidepressants, which may relate to the circulating BDNF level change in peripheral blood. The response of BDNF release differs depending on the type and amount of antidepressants, making BDNF a serious candidate as a predictor of antidepressant treatment response.