Association of lysophosphatidic acid molecules with liver fibrosis: different roles indicated.

Lysophosphatidic acid is composed of lysophosphatidic acid (LPA) molecules with varied chemical forms. The present cross-sectional study was conducted to investigate the associations of various LPA molecules with liver fibrosis. Forty-six patients affected by various types of liver disease who underwent an ultrasound-guided liver biopsy were recruited for this study. Liver fibrosis was evaluated using histological grading, as well as shear wave velocity (Vs) and serum level of type IV collagen 7S (T4c7s). Serum levels of LPA molecules were determined using liquid-chromatography tandem mass-spectrometry (LC-MSMS). Total LPA showed a significant positive association with fibrosis severity evaluated based on histological grading, Vs, and T4c7s used as parameters, following adjustment for other confounding factors, including disease type, age, gender, body mass index, and high-sensitivity C-reactive protein. This association was replicated when 16:0-LPA was substituted for total LPA. In contrast, when 20:4-LPA was substituted for total LPA, no significant association with liver fibrosis was observed. In conclusion, the degree of association varied among the different LPA molecule chemical forms, suggesting different pathophysiological roles of individual LPA molecules, although total LPA concentration was shown to be associated with liver fibrosis.

Hypothermic effects on gas exchange performance of membrane oxygenator and blood coagulation during cardiopulmonary bypass in pigs.

INTRODUCTION: Whether hypothermic cardiopulmonary bypass could attenuate both blood coagulation and platelet activation compared to normothermic cardiopulmonary bypass remains elusive. METHODS: Biocompatibility of a polymer-coated cardiopulmonary bypass circuit was comparatively assessed by plasma proteomics between juvenile pigs undergoing hypothermic (23°C) cardiopulmonary bypass and those undergoing normothermic (37°C) cardiopulmonary bypass (n = 6, respectively). Plasma samples were taken three times: 5 minutes after initiation of cardiopulmonary bypass (T5, before cooling), just before declamping and rewarming (Tc), and just before termination of cardiopulmonary bypass (Trw, 120 minutes). Proteomic analysis was quantitively performed by isobaric tags for relative and absolute quantification labeling. Thrombin-antithrombin complexes (TAT III) were measured by enzyme immunoassay, and vitamin K-dependent protein C (PROC), ß-thromboglobulin (TG), and P-selectin were measured by enzyme-linked immunosorbent assay. Blood gas analyses evaluated oxygenator performance. RESULTS: Hypothermic cardiopulmonary bypass had a significantly higher PaO2 at Tc and lower PaCO2 at Trw than normothermic cardiopulmonary bypass. Two hundred twenty-four proteins were identified with statistical criteria of both protein confidence (>95%) and false discovery rate (<5%). Six of these proteins significantly decreased at Tc than at T5 in hypothermic cardiopulmonary bypass (p = 0.02-0.04), with three related to platelet degranulation. Protein C decreased at Trw compared with T5 in normothermic cardiopulmonary bypass (p = 0.04). Thrombin-antithrombin complex had a slightly larger increase with normothermic cardiopulmonary bypass at Trw than with hypothermic cardiopulmonary bypass. ß-thromboglobulin and P-selectin levels were significantly lower at Trw with hypothermic cardiopulmonary bypass than with normothermic cardiopulmonary bypass (p = 0.04). CONCLUSION: Hypothermic cardiopulmonary bypass attenuated platelet degranulation/blood coagulation and maintained better oxygenator performance compared to normothermic cardiopulmonary bypass in juvenile pigs.

[A Case of Retroperitoneal Bronchogenic Cyst Successfully Resected with Retroperitoneoscopic Surgery].

A 32-year-old woman was referred to our hospital for consultation with a suspected left adrenal tumor detected by ultrasonography at a health check. Computed tomography and magnetic resonance imaging revealed a 3×1×4 cm multilocular cystic mass arising from the diaphragmatic crura, suggesting a retroperitoneal bronchogenic cyst. The patient underwent excision of the cyst and adjacent diaphragm using a retroperitoneoscopic approach. Retroperitoneal bronchogenic cysts are rare. We review the cases of 26 patients who underwent laparoscopic resection of a retroperitoneal bronchogenic cyst.

Polymer-coated cardiopulmonary bypass circuit attenuates upregulation of both proteases/protease inhibitors and platelet degranulation in pigs.

INTRODUCTION: Interaction of blood with a cardiopulmonary bypass (CPB) circuit activates the coagulation-fibrinolysis, complement and kinin-kallikrein systems that are mainly supported by proteases and their inhibitors. METHODS: Biocompatibility of a new polymer-coated (SEC-coated) CPB circuit was globally evaluated and compared with that of a non-coated CPB circuit by quantitative proteomics, using isobaric tags for relative and absolute quantification labeling tandem mass spectrometry. Plasma samples were taken three times (5 min after initiation of CPB, just before declamping and just before termination of CPB) in 12 pigs undergoing 120 min of CPB with the SEC-coated CPB circuit or a non-coated CPB circuit (n = 6, respectively). RESULTS: Identified were 224 proteins having high protein confidence (>99%) and false discovery rate (FDR) <5%. Among these proteins, there were 25 significantly upregulated proteins in the non-coated CPB group compared to those in the SEC-coated CPB group. Dominant protein functions were platelet degranulation, serine-type (cysteine-type) endopeptidase inhibitor activity and serine-type endopeptidase activity in the 25 proteins. Bioinformatics analysis similarly revealed upregulation of proteins belonging to platelet degranulation and negative regulation of endopeptidase activity in the non-coated CPB group; these upregulations were effectively attenuated in the SEC-coated CPB group. CONCLUSION: The new polymer (SEC)-coated CPB circuit effectively attenuated upregulation of proteins compared to the non-coated CPB circuit. These proteins were associated with both proteases/protease inhibitors and platelet degranulation.

Does the Rewarmed Heart Restore the Myocardial Proteome to That of the Pre-Cooled State?--A Proteomic Analysis of Surgical Samples.

BACKGROUND: Hypothermia is utilized in cardiac and aortic surgery to protect organs from ischemic reperfusion injury. Although the cooled body is invariably rewarmed after the procedure, it is still unknown whether the rewarmed body regains its former biological state. This study determined the modulatory effects of hypothermia on the human myocardial proteome and whether subsequent rewarming restores the proteome to the state prior to cooling. METHODS AND RESULTS: A quantitative proteomic analysis was performed using isobaric tags for relative and absolute quantification labeling tandem mass spectrometry. Right atrial samples were taken 3 times (pre, during and post cooling) during deep hypothermic cardiopulmonary bypass (CPB) from 8 patients with aortic arch aneurysms and 3 corresponding time points during normothermic CPB from 8 patients with ascending aortic or valsalva aneurysms. In total, 697 proteins were identified, with 222 proteins having high protein confidence. Bioinformatic analyses revealed significant downregulation of 19 proteins associated with energy production at hypothermic cardioplegic arrest. On rewarmed beating, 10 proteins remained downregulated, including those regulating cardiac contraction and adaptor proteins, although levels of the aforementioned 19 downregulated proteins returned to their initial values. Additional echocardiographic evaluation demonstrated that hypothermia preserved the variables of diastolic function to a greater extent than normothermic surgery. CONCLUSIONS: Rewarming restores the human myocardial proteome to the pre-cooled state, except for proteins regulating cardiac contraction and adaptor proteins.

Effectiveness of onabotulinumtoxin A for refractory overactive bladder with cough-associated detrusor overactivity.

Introduction: We report a case of refractory overactive bladder with cough-associated detrusor overactivity treated by onabotulinumtoxin A. Case presentation: A 79-year-old woman who underwent mid-urethral sling surgery 8 years ago complained mainly of urinary incontinence following abdominal pressure. Various medicines to treat overactive bladder symptoms were ineffective. Cystometry revealed cough-associated detrusor overactivity. Onabotulinumtoxin A injections in her bladder improved subjective symptoms, and cough-associated detrusor overactivity disappeared on cystometry. Conclusions: Onabotulinumtoxin A injection effectively resolved refractory overactive bladder with urgency urinary incontinence due to cough-associated detrusor overactivity.

Hypothermia produces rat liver proteomic changes as in hibernating mammals but decreases endoplasmic reticulum chaperones.

Hypothermia is used in the clinic for protection of organs such as the brain against ischemic injury during aortic/complex congenital cardiac surgery or post-resuscitation encephalopathy. The principal mechanism of hypothermic protection is suppression of metabolism, however, the pleiotropic effects of cooling are incompletely understood. Here, we used a rat model system to evaluate metabolic changes induced by deep hypothermia. The hypothermia-induced changes were identified using fluorescence-based two-dimensional (2-D) difference gel electrophoresis (DIGE) and matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF/TOF) tandem mass spectrometry. Rats were randomly assigned to a normothermic control group (37°C, n=6) or hypothermia group (23°C, n=6) that received surface cooling for 3h. Liver tissue was excised for assessment. Functional profiling of differently expressed proteins was performed as an enrichment analysis of Gene Ontology (GO) terms and pathways. We found that the livers of anesthetized rats with deep hypothermia showed significant downregulation of proteins in the endoplasmic reticulum and mitochondria, and of those involved in ATP binding, amino acid metabolism and urea cycle, response to oxidative stress, anti-apoptosis, negative regulation of apoptosis. The changes in the proteome of the hypothermic rats showed similarities, except with regard to endoplasmic reticulum chaperones, to those identified elsewhere in mammals undergoing hibernation.

Cell growth and P(3HB) accumulation from CO2 of a carbon monoxide-tolerant hydrogen-oxidizing bacterium, Ideonella sp. O-1.

Cell growth and accumulation of polyhydroxybutyric acid, P(3HB), from CO(2) in autotrophic condition of a newly isolated hydrogen-oxidizing bacterium, the strain O-1, was investigated. The bacterium, which was deposited in the Japan Collection of Microorganisms as JCM17105, autotrophically grows by assimilating H(2), O(2), and CO(2) as substrate. 16S rRNA gene sequence of the bacterium was the closest to Ideonella dechloratans (99%). Specific growth rate of the strain O-1 was faster than a hydrogen-oxidizing bacterium, Ralstonia eutropha, which is well-known P(3HB)-producing microorganism. The strain O-1 is tolerant to high O(2) concentration and it can grow above 30% (v/v) O(2), while the growth of R. eutropha and Alcaligenes latus was seriously inhibited. In culture medium containing 1 g/L (NH(4))(2)SO(4), cell concentration of the strain O-1 and P(3HB) increased to 6.75 and 5.26 g/L, respectively. The content of P(3HB) in the cells was 77.9% (w/w). The strain O-1 was very tolerant to carbon monoxide (CO) and it grew even at 70% (v/v) CO, while the growth of R. eutropha and A. latus were seriously inhibited at 5% (v/v) CO. From these results, it is expected that the strain O-1 will be useful in the manufacture of P(3HB) because the industrial exhaust gas containing CO(2), H(2), and CO can be directly used as the substrate in the fermentation process.

Retroperitoneal lymph node dissection for testicular cancer in a patient with a double inferior vena cava.

INTRODUCTION: A double inferior vena cava is a rare anomaly with an incidence ranging from 0.3% to 3.0%. In patients with a double inferior vena cava, it is important to understand the precise anatomy and possible irregular lymph node flow when performing surgery for malignancies. CASE PRESENTATION: A 60-year-old man with a non-seminoma was referred to our hospital after left high orchiectomy. Computed tomography revealed a double inferior vena cava and swollen masses in the para-aortic region. After four cycles of chemotherapy with etoposide and cisplatin, retroperitoneal lymph node dissection was safely performed with a modified template extended to the right side of the paracaval region by referring to three-dimensional images created by SYNAPSE VINCENT® software. CONCLUSION: Preoperative three-dimensional images were useful to understand this patient's unusual and complicated anatomical positions.

Adsorption and Release of Rose Bengal on Layer-by-Layer Films of Poly(Vinyl Alcohol) and Poly(Amidoamine) Dendrimers Bearing 4-Carboxyphenylboronic Acid.

Phenylboronic acid-bearing polyamidoamine dendrimer (PBA-PAMAM)/poly(vinyl alcohol) (PVA) multilayer films were prepared through the layer-by-layer (LbL) deposition of PBA-PAMAM solution and PVA solution. PBA-PAMAM/PVA films were constructed successfully through the formation of boronate ester bonds between the boronic acid moiety in PBA and 1,3-diol units in PVA. When the (PBA-PAMAM/PVA)5 films were immersed in rose bengal (RB) solution, RB was adsorbed onto the LbL films. The amount of RB adsorbed was higher in the LbL films immersed in acidic solution than in basic solution. The release of RB from the LbL films was also promoted in the basic solution, while it was suppressed in the acidic solution. The boronic acid ester is oxidized to phenol by hydrogen peroxide (H2O2) and the carbon-boron bond is cleaved, so that the (PBA-PAMAM/PVA)5 films can be decomposed by immersion in H2O2 solution. Therefore, when RB-adsorbed (PBA-PAMAM/PVA)5 films were immersed in H2O2 solution, the release of RB was moderately promoted when the solution was weakly acidic.

Plasma proteomic changes during therapeutic hypothermia in resuscitated patients after cardiac arrest.

Hypothermia is used for several h during cardiac and aortic surgery to protect ischemic organs. Therapeutic hypothermia (TH) is used for ≤24 h as a treatment for comatose patients after the return of spontaneous circulation (ROSC) following cardiac arrest. The proteomic approach may provide unbiased data on alterations in the abundance of proteins during TH. The objective of this study was to assess the effects of cooling/rewarming on the plasma proteome during TH after ROSC and to identify the mechanism underlying its therapeutic effects. A total of nine comatose adult patients, resuscitated shortly after cardiac arrest, were cooled to 34°C for 24 h and slowly rewarmed to 36°C. A quantitative gel-free proteomic analysis was performed using the isobaric tag for relative and absolute quantification labeling tandem mass spectrometry. Plasma samples were obtained prior to cooling and rewarming, and immediately after rewarming, from all patients during TH after ROSC. A total of 92 high-confidence proteins were identified. Statistically significant alterations were observed (>1.2-fold increase or <0.833-fold decrease) in the levels of 15 of those proteins (P=0.003-0.047), mainly proteins belonging to the acute-phase response or platelet degranulation. Unexpectedly, the levels of free hemoglobin (hemoglobin subunits α and ß) were significantly downregulated during TH (P<0.05). The level of the terminal complement complex (SC5b-9) showed significant reduction after cooling (P=0.023). Although the acute-phase response proteins were upregulated, the abundance of complement proteins did not change, and the levels of SC5b-9 and free hemoglobin decreased during TH in patients after ROSC.

Rapidly Neutralizable and Highly Anticoagulant Thrombin-Binding DNA Aptamer Discovered by MACE SELEX.

We present a rapidly neutralizable and highly anticoagulant thrombin-binding aptamer with a short toehold sequence, originally discovered by systematic evolution of ligands by exponential enrichment (SELEX) with microbead-assisted capillary electrophoresis (MACE). MACE is a novel CE-partitioning method for SELEX and able to separate aptamers from a library of unbound nucleic acids, where the aptamer and target complexes can be detected reliably and partitioned with high purity even in the first selection cycle. Three selection rounds of MACE-SELEX discovered several TBAs with a nanomolar affinity (Kd = 4.5-8.2 nM) that surpasses previously reported TBAs such as HD1, HD22, and NU172 (Kd = 118, 13, and 12 nM, respectively). One of the obtained aptamers, M08, showed a 10- to 20-fold longer prolonged clotting time than other anticoagulant TBAs, such as HD1, NU172, RE31, and RA36. Analyses of the aptamer and thrombin complexes using both bare and coated capillaries suggested that a large number of efficient aptamers are missed in conventional CE-SELEX because of increased interaction between the complex and the capillary. In addition, the toehold-mediated rapid antidote was designed for safe administration. The efficient aptamer and antidote system developed in the present study could serve as a new candidate for anticoagulant therapy.

Bone-targeting poly(ethylene sodium phosphate).

Poly(ethylene sodium phosphate) (PEP·Na) showed excellent cytocompatibility and in vivo bone affinity. Moreover, PEP·Na did not interact with thrombin, which is a coagulation-related protein. Because immobilization of therapeutic agents and imaging probes on PEP·Na is easily performed, PEP·Na is a promising polymer for bone-targeted therapies.

Fabrication of two-dimensional layered perovskite [NH3(CH2)12NH3]PbX4 thin films using a self-assembly method.

Ultra-thin films of the two-dimensional layered perovskites [NH3(CH2)12NH3]PbX4 (X = Br, I) with the quantum confinement effect have been fabricated by a convenient self-assembly method.

Plasma proteomic changes during hypothermic and normothermic cardiopulmonary bypass in aortic surgeries.

Deep hypothermic circulatory arrest (DHCA) is a protective method against brain ischemia in aortic surgery. However, the possible effects of DHCA on the plasma proteins remain to be determined. In the present study, we used novel high­throughput technology to compare the plasma proteomes during DHCA (22˚C) with selective cerebral perfusion (SCP, n=7) to those during normothermic cardiopulmonary bypass (CPB, n=7). Three plasma samples per patient were obtained during CPB: T1, prior to cooling; T2, during hypothermia; T3, after rewarming for the DHCA group and three corresponding points for the normothermic group. A proteomic analysis was performed using isobaric tag for relative and absolute quantification (iTRAQ) labeling tandem mass spectrometry to assess quantitative protein changes. In total, the analysis identified 262 proteins. The bioinformatics analysis revealed a significant upregulation of complement activation at T2 in normothermic CPB, which was suppressed in DHCA. These findings were confirmed by the changes of the terminal complement complex (SC5b­9) levels. At T3, however, the level of SC5b­9 showed a greater increase in DHCA compared to normothermic CPB, while 48 proteins were significantly downregulated in DHCA. The results demonstrated that DHCA and rewarming potentially exert a significant effect on the plasma proteome in patients undergoing aortic surgery.