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Byline: H. Ishikawa (1), Y. Watano (2), K. Kano (2), M. Ito (1), S. Kurita (1) Keywords: Key words Codominant nuclear DNA marker; Homosporous ferns; PCR primers; PgiC; Phosphoglucose isomerase; SSCP Abstract: Polymerase chain reaction (PCR)-based nuclear DNA markers were developed for fern species. We first determined the partial nucleotide sequence of cDNA of the pgiC gene encoding cytosolic phosphoglucose isomerase from Dryopteris caudipinna, and then PCR primers for exon-primed, intron-crossing (EPIC) amplifications were designed. The EPIC primers are universally applicable to the most derived indusiate fern families such as Dryopteridaceae, Thelypteridaceae, and Woodsiaceae. The PCR products of primers 14F/16R containing two introns are moderate in size (534 bp--ca.1000 bp) and are possibly of value in phylogenetic reconstruction at specific and generic levels. Codominant nuclear DNA markers applicable to the estimation of mating systems and other population genetic studies were also developed by a combination of single-strand conformation polymorphism (SSCP) and EPIC amplification using primers 14F/15R and 15F/16R. In order to provide a case study using these markers, allelic variation of PCR products using 15F/16R was examined in populations of Arachniodes standishii (Dryopteridaceae). Author Affiliation: (1) Department of Biology, Chiba University, Inage, Chiba, Japan, JP (2) Department of Biology, Kanazawa University, Kakuma, Kanazawa, Japan, JP Article note: Received: July 4, 2001 / Accepted: September 12, 2001

Background: Microvascular decompression (MVD), the standard surgical approach for hemifacial spasm (HFS), can be divided into the interposition and transposition methods. Although the risk of HFS recurrence following interposition has been reported, there is limited data comparing long-term outcomes between both methods performed by a single surgeon. This study aimed to investigate the efficacy of MVD techniques on HFS by comparing surgical outcomes performed by a single surgeon in a single-center setting., Methods: A total of 109 patients who underwent MVD were analyzed and divided into the transposition (86 patients) and interposition (23 patients) groups. Postoperative outcomes at 1 month and 1 year were assessed and compared, including rates of spasm relief, complications, and recurrence., Results: Outcome assessment revealed higher rates of early spasm relief in the interposition group (66.3% vs. 100%, transposition vs. interposition, respectively, p = 0.0004), although spasm relief at 1-year postoperatively was comparable between the two groups (84.9% vs. 95.7%, transposition vs. interposition, respectively, p = 0.2929). No significant differences were observed in complication and recurrence rates. Kaplan-Meier analysis demonstrated no significant differences in the duration of spasm resolution by MVD method (p = 0.4347, log-rank test)., Conclusion: This study shows that both the transposition (Surgicel® and fibrin glue) and interposition (sponge) methods were excellent surgical techniques. The interposition method may achieve earlier spasm resolution compared to the transposition method., (© 2024. The Author(s).)

Background: Combined bypass, including direct and indirect procedures, has been recognized as the maximal revascularization to prevent further hemorrhagic or ischemic stroke in adult moyamoya disease (MMD). It is also important to consider cosmetic aspects when planning combined bypass for MMD. However, there are few reports that have described the cosmetic considerations in bypass surgery for MMD., Methods: We demonstrate our surgical techniques aimed at achieving extended revascularization as well as excellent cosmetic outcomes with figures and video., Conclusion: Our combined bypass procedures which focus on achieving maximal cosmetic results are effective methods that require no special instruments or techniques., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: Optimal hemostasis provides safety and reliability during neurosurgery which improves surgical outcomes. Previously, artificial cerebrospinal fluid (aCSF) and its component sodium bicarbonate were found to facilitate physiological hemostasis by amplifying platelet aggregation. This study aimed to verify whether aCSF amplifies platelet-dependent hemostasis in the presence of antiplatelet agents., Methods: We prepared platelet-rich plasma (PRP) or washed platelets using aspirin (acetylsalicylic acid, (ASA)) or normal saline (NS). We evaluated samples treated with a commercially available aCSF solution or NS for amplification of aggregation, activation of integrin αIIbβ3, phosphatidylserine (PS) exposure, P-selectin (CD62P) expression, and formation of microparticles (MPs). We assessed the effect of aCSF on in vivo hemostasis in the presence of ASA by measuring the tail bleeding time in ASA-or NS-injected C57BL/6 N mice., Results: Compared with NS, aCSF amplified ASA-inhibited platelet aggregation by recovering platelet activation including PS exposure, MP release, CD62P expression, and integrin αIIbβ3 activation. When using washed platelets, aCSF almost completely counteracted the inhibition of platelet aggregation by ASA. Prolonged bleeding time from the amputated tail of ASA-injected mice was significantly shortened by the treatment with aCSF compared to NS. Sodium bicarbonate also directly amplified ASA-inhibited platelet aggregation., Conclusions: aCSF and sodium bicarbonate facilitate physiological hemostasis through the recovery of inhibited platelet aggregation even in the presence of ASA. The utilization of aCSF in the operative field may be advantageous for facilitating hemostasis in patients with impaired platelet function and contribute to improving outcomes of neurosurgery., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: We investigated the clinical influence of the prealbumin level on the gastric cancer survival and recurrence after curative treatment., Methods: This study included 447 patients who underwent curative treatment for gastric cancer between 2013 and 2017. The risk factors for the overall survival (OS) and recurrence-free survival (RFS) were identified., Results: A prealbumin level of 20 mg/dl was regarded as the optimal point of classification, considering the 3- and 5-year survival rates. The OS rates at 3 and 5 years after surgery were 80.7% and 65.0% in the low-prealbumin group, respectively, and 93.1% and 87.9% in the high-prealbumin group, respectively, a statistically significant difference (p < 0.001). The RFS rates at 3 and 5 years after surgery were 71.7% and 68.0% in the low-prealbumin group, respectively, and 90.1% and 84.7% in the high-prealbumin group, respectively, a statistically significant difference (p = 0.031). A multivariate analysis demonstrated that the prealbumin level was a significant independent risk factor for the OS and RFS. In addition, the rate of introduction of adjuvant chemotherapy was significantly lower and the frequency of peritoneal recurrence and lymph node recurrence significantly higher in the low-prealbumin group than in the high-prealbumin group., Conclusion: Prealbumin is a risk factor for the survival in patients who undergo curative treatment for gastric cancer. It is necessary to develop an effective plan of perioperative care and surgical strategy according to the prealbumin level., (© 2021. The Author(s).)

Background/aim: The present study quantified the changes in the dietary and nutritional intake after gastrectomy between elderly and non-elderly patients., Patients and Methods: This prospective observational study enrolled patients who underwent curative gastrectomy for gastric cancer., Results: Twenty-three patients ≥ 75 years old were classified into the elderly group, and 127 patients < 75 years old were classified into the non-elderly group. The respective median % dietary intake losses at 1 and 3 months postoperatively were -12.4% and -5.3% in the elderly group and -8.3% and -2.8% in the non-elderly group (p = 0.075 and 0.080). On comparing the intake loss of three major nutrients, the respective median % lipid intake losses at 1 and 3 months postoperatively were -13.5% and -5.8% in the elderly group and -7.3% and 0% in the non-elderly group (p = 0.029 and 0.045)., Conclusion: Our results suggested that elderly patients experienced more serious lipid intake loss after gastrectomy than non-elderly patients., (© 2021. The Author(s).)

Purpose: Despite improvements in surgical techniques and devices and perioperative care of gastric cancer (GC), the rate of postoperative complications still has not decreased. If patients at high risk for postoperative complications could be identified early using biomarkers, these complications might be reduced. In this study, we investigated usefulness of the preoperative Glasgow Prognostic Score (GPS) as a predictive factor for complications after surgery in patients with stage II/III GC., Methods: This study retrospectively analyzed the outcomes of 424 patients who underwent curative surgery for pathological stage II/III GC from February 2007 to July 2019 at a single center. The GPS was assessed within 4 days before surgery. To identify independent risk factors for postoperative complications, univariate and multivariate analyses were performed using a Cox proportional hazards model., Results: The numbers of patients with a GPS of 0, 1, and 2 were 357, 55, and 12, respectively. The rate of complications after surgery was significantly higher among patients with a GPS of 1 or 2 than among patients with a GPS of 0 (p = 0.008). Multivariate analysis identified a GPS of 1 or 2 as an independent predictive factor for postoperative complications (p = 0.037)., Conclusion: The preoperative GPS may be a useful predictive factor for postoperative complications in patients with stage II/III GC. Being aware of the risk of complications after surgery as indicated by the GPS before surgery may promote safe and minimally invasive surgery that we expect will improve outcomes in patients with a GPS of 1 or 2., (© 2021. The Author(s).)

Purpose: To assess the utility of the Glasgow Prognostic Score (GPS) obtained before curative resection for predicting outcomes in patients with advanced gastric cancer (GC)., Methods: This study retrospectively analyzed the outcomes of 337 consecutive patients with GC who underwent curative surgery for locally advanced gastric cancer between January 2003 and June 2014. GPS was assessed within 4 days prior to surgery., Results: The number of patients with GPS scores of 0, 1, and 2 was 302, 26, and 9, respectively. There was significantly more blood loss during surgery and more postoperative complications in the GPS 1/2 group than in the GPS 0 group. Patients in the GPS 1/2 group had significantly poorer overall survival than those in the GPS 0 group (p = 0.001). On multivariate analysis, GPS 1/2 was identified as an independent factor for poor survival (p = 0.019)., Conclusion: GPS before curative resection might be a useful predictive factor for perioperative complications and survival in locally advanced GC., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Purpose: Dedifferentiated fat (DFAT) cells are mature adipocyte-derived multipotent cells that can be applicable to cell-based therapy for stress urinary incontinence (SUI). This study developed a persistence SUI model that allows long-term evaluation using a combination of vaginal distention (VD) and bilateral ovariectomy (OVX) in rats. Then, the therapeutic effects of DFAT cell transplantation in the persistence SUI model was examined., Methods: In total, 48 Sprague-Dawley rats were divided into four groups and underwent VD (VD group), bilateral OVX (OVX group), VD and bilateral OVX (VD + OVX group), or sham operation (Control group). At 2, 4, and 6 weeks after injury, leak point pressure (LPP) and histological changes of the urethral sphincter were evaluated. Next, 14 rats undergoing VD and bilateral OVX were divided into two groups and administered urethral injection of DFAT cells (DFAT group) or fibroblasts (Fibroblast group). At 6 weeks after the injection, LPP and histology of the urethral sphincter were evaluated., Results: The VD + OVX group retained a decrease in LPP with sphincter muscle atrophy at least until 6 weeks after injury. The LPP and urethral sphincter muscle atrophy in the DFAT group recovered better than those in the fibroblast group., Conclusions: The persistence SUI model was created by a combination of VD and bilateral OVX in rats. Urethral injection of DFAT cells inhibited sphincter muscle atrophy and improved LPP in the persistence SUI model. These findings suggest that the DFAT cells may be an attractive cell source for cell-based therapy to treat SUI., (© 2022. The Author(s).)

This study sought to clarify whether suspended particles containing high Cu concentrations are present in the sea-surface microlayer (S-SML). For this reason, suspended particles (10-2000 μm) in the S-SML were collected periodically from a ship mooring pond during 2018-2020, and the acid-soluble Cu concentration in the suspended particles was measured as particulate Cu (P-Cu). The highest concentration of P-Cu in the S-SML of the pond was 75 μg L -1 with a 90th percentile value of 2.5 μg L -1 . This is below P-Cu values reported for the S-SML in North American ports, but 140 times higher than this found in bulk seawater in the Atlantic Ocean. The highest P-Cu concentration in the S-SML of non-organism (abiotic) origin was 17 μg L -1 , and the abiotic P-Cu to P-Cu ratio varied from 0.2 to 100%, likely depending on the quality and quantity of biogenic material in the S-SML samples. It is assumed that the S-SML particles examined here contain high Cu concentrations originating from ship antifouling paints., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: The lymph node (LN) ratio (LNR) has been proposed as a sensitive prognosticator in patients with esophageal squamous cell carcinoma (ESCC), especially when the number of LNs harvested is insufficient. We investigated the association between the LNR and survival in patients with locally advanced ESCC who received neoadjuvant chemotherapy (NAC) and explored whether the LNR is a prognosticator in these patients when stratified by their response to NAC., Methods: We retrospectively reviewed 199 locally advanced ESCC patients who received curative resection after NAC between January 2011 and December 2019. The predictive accuracy of the adjusted X-tile cut-off values for LNR of 0 and 0.13 was compared with that in the Union for International Cancer Control pathological N (UICC pN) categories. The association between survival rate and clinicopathological features was examined., Results: Multivariate analysis identified that the LNR was an independent risk factor for recurrence-free survival [RFS; hazard ratio (HR) 6.917, p < 0.001] and overall survival (OS) (HR 4.998, p < 0.001). Moreover, even when stratified by response to NAC, the LNR was a significant independent risk factor for RFS and OS (p < 0.001). The receiver operating characteristic curves identified that the prognostic accuracy of the LNR tended to be better than that of the UICC pN factor in all cases and responders., Conclusion: The LNR had a significant prognostic value in patients with locally advanced ESCC, including in those who received NAC., (© 2021. Society of Surgical Oncology.)

Background: Naples prognostic score (NPS) is a scoring system based on albumin, cholesterol concentration, lymphocyte-to-monocyte ratio, and neutrophil-to-lymphocyte ratio reflecting host systemic inflammation, malnutrition, and survival for several malignancies. This study was designed to assess the prognostic significance of NPS in patients with locally advanced esophageal squamous cell carcinoma (ESCC) and to compare its prognostic accuracy with that of other systemic inflammatory and nutritional index., Methods: We retrospectively examined 165 patients with locally advanced ESCC who underwent neoadjuvant therapy followed by curative resection between January 2011 and September 2019. Patients were divided into three groups based on their NPS before neoadjuvant therapy (Group 0: NPS = 0; Group 1: NPS = 1-2; Group 2: NPS = 3-4). We compared the clinicopathological characteristics and survival rates among the groups., Results: The 5-year recurrence-free survival (RFS) and overall survival (OS) rates were significantly different between the groups (P < 0.001). The NPS was superior to other systemic inflammatory and nutritional index for predicting prognoses, as determined using area under the curves (P < 0.05). Multivariate analysis demonstrated that the NPS was a significant predictor of poor RFS (Group 1: hazard ratio [HR] 1.897, P = 0.049; Group 2: HR 3.979, P < 0.001) and OS (Group 1: HR 2.152, P = 0.033; Group 2: HR 3.239, P = 0.006)., Conclusions: The present study demonstrated that NPS was an independent prognostic factor in patients with locally advanced ESCC and more reliable and accurate than the other systemic inflammatory and nutritional index.

A liquid-membrane type nitrate-selective electrode was improved to lower the influence of contaminants by modifying its inner electrode system from Ag | AgCl | Cl - to Ag | Ag + . The NO 3 - -selective electrode displayed a linear response to the concentration of NO 3 - with a Nernstian slope of -53 ± 1 mV decade -1 , in the concentration region between 10 -5 and 2 mol dm -3 (M). The NO 3 - detection limit was about 10 -5 M. The electrochemical response of this electrode was stable for more than 30 days. The deterioration in responding characteristics due to the coexistence of Cl - was suppressed by use of the Ag | Ag + redox couple in the absence of Cl - inside the NO 3 - -selective electrode.

Background: The lymph node (LN) ratio (LNR) and the log odds of positive LNs (LODDS) have been proposed as sensitive prognosticators in patients with primary gastric cancer, especially in patients with an insufficient number of harvested LNs. We investigated the association of LNR and LODDS with survival in patients with remnant gastric cancer (RGC) and explored whether these staging methods are prognostic factors in patients with an insufficient number of harvested LNs., Methods: The present study retrospectively examined 95 patients with RGC who received gastrectomy between January 2000 and December 2018. The patients were classified according to the adjusted X-tile cutoff for LNR and LODDS. The association between survival rates and clinicopathological features was investigated. The predictive accuracy of the LNR and LODDS was compared with that of the Union for International Cancer Control pathological N factor., Results: Multivariate analysis revealed that the LNR and LODDS were independent risk factors for recurrence-free survival (RFS) [hazard ratio (HR) 2.623, p = 0.020; HR 3.404, p = 0.004, respectively] and overall survival (OS) (HR 3.694, p = 0.003; HR 2.895, p = 0.022, respectively) in patients with RGC. Moreover, even in patients with 15 or fewer harvested LNs, only the LNR was a significant independent risk factor for RFS (HR 21.890, p < 0.001) and OS (HR 6.597, p = 0.002). The receiver operating characteristic curves revealed that the prognostic accuracy of the three methods was comparable (p > 0.05)., Conclusion: LNR has significant prognostic value for patients with RGC, including those with an insufficient number of harvested LNs.

Background: Lymph node ratio (LNR), defined as the ratio of metastatic nodes to the total number of examined lymph nodes, has been proposed as a sensitive prognostic factor in patients with gastric cancer (GC). We investigate its association with survival in pathological stage (pStage) II/III GC and explore whether this is a prognostic factor in each Union for International Cancer Control pStage (7th edition)., Patients and Methods: We retrospectively examined 838 patients with pStage II/III GC who underwent curative gastrectomy between June 2000 and December 2018. Patients were classified into low-LNR (L-LNR), middle-LNR (M-LNR), and high-LNR (H-LNR) groups according to adjusted X-tile cutoff values of 0.1 and 0.25 for LNR, and their clinicopathological characteristics and survival rates were compared., Results: The 5-year recurrence-free survival (RFS) and overall survival (OS) rates postsurgery showed significant differences among the groups (P < 0.001). Multivariate analysis demonstrated that LNR was a significant predictor of poor RFS [M-LNR: hazard ratio (HR) 3.128, 95% confidence interval (CI) 2.254-4.342, P < 0.001; H-LNR: HR 5.148, 95% CI 3.546-7.474, P < 0.001] and OS (M-LNR: HR 2.749, 95% CI 2.038-3.708, P < 0.001; H-LNR: HR 4.654, 95% CI 3.288-6.588, P < 0.001). On subset analysis stratified by pStage, significant differences were observed between the groups in terms of the RFS curves of pStage II and III GC (P < 0.001 and < 0.001, respectively) and OS curves of pStage II and III GC (P = 0.001 and < 0.001, respectively)., Conclusions: High LNR is a predictor of worse prognosis in pStage II/III GC, including each substage.

In order to improve plant factories, an appropriate control system on fertilization is urgently required. An automatic management system to control nutrient concentration was constructed using a programmable logic controller (PLC) and ion selective electrodes (ISEs) of nitrate, phosphate, and potassium ion. The concentration of nutrient components in a culture solution was monitored using these ISEs. When the concentration of the nutrient components diminished to the threshold set as an optimum condition (0.1 - 2.0 mM), an appropriate amount of a concentrated solution of each nutrient component was added to the culture solution using solenoid valves connected with the PLC. The present cultivation system was simply constructed without any computers and pumps. Three kinds of automatic control systems simultaneously worked and did not influence each other.

A phosphate ion-selective electrode using molybdenum metal was constructed. The modified molybdenum electrode responded to HPO 4 2- in the presence of molybdenum dioxide and molybdophosphate (PMo 12 O 40 3- ) on the surface. The electrode exhibited a linear response to HPO 4 2- in the concentration range between 1.0 × 10 -5 and 1.0 × 10 -1 M (mol dm -3 ) in the pH range from 8.0 to 9.5 with a detection limit of 1.0 × 10 -6 M. The sensor showed near Nernstian characteristics (27.8 ± 0.5 mV dec -1 ) at pH 9.0. Since the responding potential was attributed to the activity of HPO 4 2- , the potential at a given concentration of phosphate depended on the pH. The electrode indicated a good selectivity with respect to other common anions such as NO 3 - , SO 4 2- , Cl - , HCO 3 - and CH 3 COO - . The modified molybdenum electrode can be continuously used for over a 1 month with good reproducibility. The feasibility of the electrochemical sensor was proved by successful for the detection of phosphate in real samples.

Objects: There is growing evidence that sphingosine 1-phosphate (S1P) is involved in inflammatory diseases. As endometriosis is known as an inflammatory disease, we investigated the role of S1P system in the development of endometriosis., Methods: The expression of sphingosine kinase (SphK) 1 in endometriosis lesions was examined by immunohistochemistry. The cystic fluid of ovarian cysts/tumors were obtained to measure S1P concentrations. Endometriotic stromal cells (ESC) derived from endometrioma were used for in vitro experiments., Results: Sphingosine kinase 1 was detected in epithelium and stromal cells of endometriotic lesions. The mean S1P concentration in the cystic fluid of endometriomas was higher than that in nonendometriomas significantly (98.2 nM vs less than 1.5 nM, P < .01). Interleukin-1β (IL-1β) or transforming growth factor-β exhibited 2.7-fold and 11.5-fold increase in SphK1 messenger RNA (mRNA) expression in ESC, respectively ( P < .01). Higher dose of S1P (125nM) increased the cell number of ESC by 20%, and low dose of S1P (1.25 nM and 12.5 nM) induced IL-6 mRNA production and IL-6 secretion by ESC dose-dependently. JTE013, an antagonist for S1PR2, partially suppressed IL-6 induction by S1P ( P < .05). JTE013 and VPC23019, an antagonist for S1PR1 and S1PR3, suppressed the ESC proliferation induced by S1P., Conclusion: The present study for the first time proved that the SphK-S1P-S1PR axis play a role of accelerating inflammation and growth of endometriotic cells.

An automatic management system for nutrient solutions was constructed using a programmable logic controller (PLC) and a K + -ion selective electrode (K + -ISE). The concentration of K + was monitored by the K + -ISE. When the concentration of K + fell to the threshold limit, an appropriate amount of a concentrated K + solution was added to the hydroponic solution. The volume was also maintained at a constant level by addition of water. This system can be constructed simply and inexpensively without any computers and pumps.

A liquid-membrane type nitrate-selective electrode was constructed, in which the responding membrane contained polyvinylchloride, o-nitrophenyloctylether and tetraheptylammonium nitrate. The NO 3 - -selective electrode displayed a linear response to the concentration of NO 3 - with a Nernstian slope of -53.3 ± 1.0 mV decade -1 , in the 10 -5 - 10 -1 mol dm -3 (M) NO 3 - concentration range. The NO 3 - detection limit was about 10 -6 M. The electrochemical response of this electrode was stable for more than 30 days. Measurements performed using the NO 3 - -sensor indicated that in the presence of green plants, the concentration of NO 3 - in a hydroponic solution decreased from 0.20 to 0.05 mM over a three-day period.

In this study, nanostructured porous gold electrodes were prepared by the anodization of gold in the presence of oxalic acid or glucose as a reductant, and applied as scaffolds for direct electron transfer (DET)-type bioelectrocatalysis. Gold cations generated in the anodization seem to be reduced by the reductant to construct a porous gold structure. The DET-type performance of the electrode was examined using two DET-type model enzymes, bilirubin oxidase (BOD) and peroxidase (POD), for the four-electron reduction of dioxygen and the two-electron reduction of peroxide, respectively. BOD and POD on the anodized porous gold electrodes exhibited well-defined sigmoidal steady-state waves corresponding to DET-type bioelectrocatalysis. Scanning electron microscopy images revealed sponge-like pores on the electrodes. The anodized porous gold electrodes demonstrate promise as scaffolds for DET-type bioelectrocatalysis.

Amperometric biosensors were constructed for the simultaneous detection of lactate enantiomers. The enantioselectivity of the sensor is based on NAD-dependent l- and d-lactate dehydrogenases that, respectively, oxidize l- and d-lactates into pyruvate. The NADH formed during the enzymatic reduction was catalytically oxidized at Meldola's blue-adsorbed mesoporous electrodes. Stable amperometric measurements were performed in a two-electrode system using Ag|AgCl|sat. KCl as a counter electrode via a salt bridge. The response of the sensor reached a pseudo-steady state within 60 s. The agreement of the sensitivities for l- and d-lactates and the pseudo-steady-state characteristics of the sensors demonstrate that the current is strongly influenced by the diffusion of lactates at the edge of the electrode, enabling reproducible measurements. The pseudo steady-state characteristics are also realized at the chip-type electrode. The sensor was successfully applied for the detection of d- and l-lactates in horse serum.

Shewanella can transfer electrons to various extracellular electron acceptors. We electrochemically investigated the pathway of extracellular electron transfer from Shewanella strain Hac319 to electrodes. A resting cell suspension of Shewanella strain Hac319 containing lactate produced a steady-state sigmoidal wave in the presence of flavin mononucleotide (FMN) in cyclic voltammetry, but not in the absence of FMN. A harvested cell suspension without cell-washing also produced a similar catalytic wave without any external addition of free FMN. The midpoint potentials of the two sigmoidal waves were identical to the redox potential of free FMN. The data indicate that FMN secreted from the Shewanella strain Hac319 works as an electron-transfer mediator from the cell to electrodes. An addition of cyanide to a resting cell suspension of Shewanella strain Hac319 increased the rate of the FMN reduction in the presence of lactate, while it decreased the respiration rate. By considering the fact that cyanide is coordinated to the heme moiety of hemoproteins and shifts the redox potential to the negative potential side, the data indicate that the electron derived from lactate is predominantly transferred in a down-hill mode from an electron donor with a redox potential more negative than that of FMN without going through outer membrane cytochromes c molecules.

Background: This study was designed to investigate whether postoperative infectious complications (ICs) are a risk factor for the prognosis in esophageal cancer patients who receive neoadjuvant chemotherapy by stratifying the response to neoadjuvant chemotherapy., Methods: The present study retrospectively examined patients who received neoadjuvant chemotherapy followed by esophagectomy between January 2011 and September 2015. Risk factors for overall survival (OS) were examined by Cox proportional hazard analyses. Pathological responders to neoadjuvant chemotherapy were defined as those with a tumor disappearance of more than one-third of the initial tumor. Postoperative ICs were defined using the Clavien-Dindo classification., Results: Of the 111 patients examined, 45 (40.5%) developed postoperative ICs. A pathological response to neoadjuvant chemotherapy was observed in 54 (48.6%) patients. The multivariate analysis demonstrated that postoperative ICs were a significant independent risk factor for the OS (hazard ratio [HR] 2.359; 95% confidence interval [CI] 1.057-5.263, p = 0.036). In the subset analysis, postoperative ICs were a marginally significant independent risk factor for OS in the nonresponders (HR 2.862; 95% CI 0.942-8.696, p = 0.063) but not in the responders (HR 0.867; 95% CI 0.122-6.153, p = 0.886)., Conclusions: These results suggested that the negative survival impact of postoperative ICs can be canceled out in esophageal cancer patients who respond to neoadjuvant chemotherapy.

Rotary ATPases are unique rotary molecular motors that function as energy conversion machines. Among all known rotary ATPases, F 1 -ATPase is the best characterized rotary molecular motor. There are many high-resolution crystal structures and the rotation dynamics have been investigated in detail by extensive single-molecule studies. In contrast, knowledge on the structure and rotation dynamics of V 1 -ATPase, another rotary ATPase, has been limited. However, recent high-resolution structural studies and single-molecule studies on V 1 -ATPase have provided new insights on how the catalytic sites in this molecular motor change its conformation during rotation driven by ATP hydrolysis. In this review, we summarize recent information on the structural features and rotary dynamics of V 1 -ATPase revealed from structural and single-molecule approaches and discuss the possible chemomechanical coupling scheme of V 1 -ATPase with a focus on differences between rotary molecular motors.

We performed numerical simulations on an extremely fast, mediated, electron transfer-type bioelectrocatalytic reaction using a microband electrode. The simulations under fast-enzyme-kinetics conditions predicted that the decrement of the current density by increaseing the microband thickness would effectively improve the upper limit of detection. These predictions were accurate for an ultrathin-ring with thickness of 100 nm and gold leaf with thickness of 10 μm electrodes, acting as novel amperometric glucose sensors with FAD-dependent glucose dehydrogenase. The gold leaf electrode provided pseudo-steady-state currents which were proportional to the glucose concentration up to a concentration of 20 times higher than the mediator concentration.

Non-catalytic direct electron transfer (DET) signal of Compound I of horseradish peroxidase (POD) was first detected at 0.7 V on POD/carbon nanotube mixture-modified electrodes. Excellent performance of DET-type bioelectrocatalysis was achieved with POD immobilized with glutaraldehyde on Ketjen Black (KB)-modified electrodes for H 2 O 2 reduction with an onset potential of 0.65 V (vs. Ag | AgCl | sat. KCl) without any electrode surface modification. The POD-immobilized KB electrode was found to be suitable for detecting H 2 O 2 with a low detection limit (0.1 μM at S/N = 3) at -0.1 V. By co-immobilizing glucose oxidase (GOD) and POD on the KB-modified electrode, a bienzyme electrode was constructed to couple the oxidase reaction of GOD with the DET-type bioelectrocatalytic reduction of H 2 O 2 by POD. The amperometric detection of glucose was performed with a high sensitivity (0.33 ± 0.01 μA cm -2 μM -1 ) and a low detection limit (2 μM at S/N = 3).

Purpose: Autologous cells potentially provide an ideal injectable substance for management in vesicoureteral reflux (VUR). The aim of this study is to examine the effects of mature adipocyte-derived dedifferentiated fat (DFAT) cell transplantation on VUR in a rat bladder pressurization-induced VUR model., Methods: To create VUR, Sprague-Dawley rats underwent urethral clamping and placement of cystostomy followed by intravesical pressurization. Rat DFAT cells (1 × 10 6 cells, DFAT group, n = 5) or saline (control group, n = 5) was then injected into the bilateral vesicoureteral junctions. Two weeks later, VUR grade was evaluated on cystography. The number of apoptotic cells in the renal pelvic urothelium, the ureteral inner/outer diameter ratio and the area of connective tissue in the posterior bladder wall were measured., Results: The reflux grade in the DFAT group was significantly lower than that in the control group. The number of apoptotic cells in the renal pelvic urothelium, ureteral inner/outer diameter ratio and connective tissue area in DFAT group were significantly lower in comparison with the control group., Conclusions: DFAT cell transplantation improved VUR and exerted nephroprotective effects in a rat VUR model.

Background: Lean body mass loss after surgery, which decreases the compliance of adjuvant chemotherapy, is frequently observed in gastric cancer patients who undergo gastrectomy for gastric cancer. However, the risk factors for loss of lean body mass remain unclear., Methods: The current study retrospectively examined the patients who underwent curative gastrectomy for gastric cancer between June 2010 and March 2014 at Kanagawa Cancer Center. All the patients received perioperative care for enhanced recovery after surgery. The percentage of lean body mass loss was calculated by the percentile of lean body mass 1 month after surgery to preoperative lean body mass. Severe lean body mass loss was defined as a lean body mass loss greater than 5 %. Risk factors for severe lean body mass loss were determined by both uni- and multivariate logistic regression analyses., Results: This study examined 485 patients. The median loss of lean body mass was 4.7 %. A lean body mass loss of 5 % or more occurred for 225 patients (46.4 %). Both uni- and multivariate logistic analyses demonstrated that the significant independent risk factors for severe lean body mass loss were surgical complications with infection or fasting (odds ratio [OR] 3.576; p = 0.001), total gastrectomy (OR 2.522; p = 0.0001), and gender (OR 1.928; p = 0.001)., Conclusions: Nutritional intervention or control of surgical invasion should be tested in future clinical trials for gastric cancer patients with these risk factors to maintain lean body mass after gastrectomy.

Ion transport through a single channel of gramicidin A (GA) within the bilayer lipid membrane (BLM) between two aqueous phases (W1 and W2) has been analyzed based on the electroneutrality principle. The single-channel current increases in proportion to the magnitude of the applied membrane potential and is also dependent on the permeability coefficients of electrolyte ions (K(+) and Cl(-)). By varying the ratio of the concentration of KCl in W1 to that in W2, the ratio of the diffusion coefficient of K(+) in the BLM to that of Cl(-) in the BLM can be evaluated.

The fabrication of ultrathin-ring electrodes with a diameter of 2 mm and a thickness of 100 nm is established. The ultrathin-ring electrodes provide a large density of pseudo-steady-state currents, and realize pseudo-steady-state amperometry under quiescent conditions without a Faraday cage. Under the limiting current conditions, the current response at the ultrathin-ring electrode can be well explained by the theory of the microband electrode response. Cyclic voltammograms at the ultrathin-ring electrode show sigmoidal characteristics with some hysteresis. Numerical simulation reveals that the hysteresis can be ascribed to the time-dependence of pseudo-steady-state current. The performance of amperometry with the ultrathin-ring electrode has been verified in its application to redox enzyme kinetic measurements.

The aggregation formation of monoclonal antibodies as biopharmaceuticals induced by heat stress was evaluated by size-exclusion chromatography, and the formation rate was correlated with several physicochemical parameters of the antibodies to clarify the factors to govern the aggregate formation. The parameters we studied were: the melting temperature (Tm) and the standard enthalpy of the melting point (ΔmH°) evaluated by differential scanning calorimetry under given and common conditions; the wavelength (λmax) and the intensity (Fint) of the maximum fluorescence peak of 1-anilinonaphthalene-8-sulfonate as a probe dye; the z-average diameter (D) evaluated by dynamic light scattering; and the isoelectric point (pI) and the hydrophobic index (Hpho) of the complementarity determining region calculated from the amino acid sequence. Multivariate statistical analysis with these explanatory variables based on Akaike's information criterion indicates that the soluble aggregate formation is negatively correlated with Tm and pI, while the insoluble aggregate formation is positively correlated with Fint and pI. Based on these results, the mechanisms of the aggregate formation and methods to prevent the formation are discussed.

The propagation of the change in potential differences across liquid membranes from the potential-sending cell to the potential-receiving cell was investigated by use of a system combined with three liquid membrane cells, which were composed of two aqueous phases and a 1,2-dichloroethane solution phase. The ionic composition of one potential-sending cell (S) was identical to that of the receiving cell (Rec), and that of another potential-sending cell (Ap) was different from that of the Rec. When the connection of cell Rec was switched from cell S to cell Ap, the change in the membrane potential was caused by the circulating current. The greater the ratio of the interfacial area of the membrane of cell Ap to that of cell Rec, the faster the change in the membrane potential propagated from cell Ap to cell Rec.

We studied vascular structure of the rabbit placenta, especially on three-dimensional morphological patterns and developmental process. Basic structure of maternal arterial system was re-constructed during day 13-18 of pregnancy, forming main routes for blood supply through the arterial sinuses and radial arteries. Intra-villous spaces were drastically developed showing as branches from the terminal radial arteries. Fetal arterial system was generated accompanied with maternal vascular development, showing characteristic features such as the perforating linear artery, hairpin flexion, and circular anastomoses in the capillaries. From the correlation of maternal and fetal blood currents, gas-exchange style in the rabbit placenta was considered as counter-current and pool mixed patterns. These data demonstrated an original feature for the placental arterial systems in rabbits, which differed from other animals having a property for discoid placenta.

This paper describes a diffusion-controlled electrolysis of glucose with mediated bioelectrocatalysis at microdisk-electrodes. Under conditions of an extremely fast enzyme reaction, compared with the diffusion of glucose, the current in potential-step chronoamperometry reaches an almost steady state within 10 s, and is proportional to the glucose concentration. The current can be detected at +0.1 V (vs. Ag|AgCl) with 9,10-phenanthrenequinone as a mediator. The addition of carboxymethylcellulose increased the linear range up to 10 mM.

Ion transports from one aqueous phase (W1) to another (W2) across bilayer lipid membranes (BLM) containing KAT1 potassium channel were investigated by recording current fluctuations. KAT1 channel is a voltage-gated K(+) channel from Arabidopsis thaliana and has been suggested to have a key role in stomatal opening by osmoregulation in guard cells. Although KAT1 channel is a K(+)-specific transporter, the species of the counter anions significantly affected the level of the single channel current, suggesting that KAT1 also transported these anions. When various potassium salts were used as electrolytes, the single channel conductance decreased with an increase of the ionic radius of the coexisting anions except for F(-). This indicates the existence of selective permeation for anions, but the anion-selectivity is less noticeable than the cation-selectivity.

Ion transport from one aqueous phase (W1) to another (W2) across a planar bilayer lipid membrane (BLM) in the presence of inhalation anesthetics was electrochemically investigated. In the absence of inhalation anesthetics in the BLM system, no ion transport current flowed between W1 and W2 across the BLM. When inhalation anesthetics such as halothane, chloroform, diethyl ether and trichloroethylene were added to the two aqueous phases or the BLM, the ion transport current quite clearly appeared. When the ratio of the concentration of KCl or NaCl in W1 to that in W2 was varied, the zero current potential across the BLM was shifted. By considering the magnitude of the potential shift, we concluded that the ion transport current can be predominantly ascribed to the transport of Cl(-) across the BLM. Since the dielectric constants of these anesthetics are larger than that of the inner hydrophobic domain of the BLM, the concentration of hydrophilic electrolyte ions in the BLM increases with the increase in the dielectric constant of the inner hydrophobic domain caused by addition of these anesthetics. These situations lead to an increase in the ion permeability coefficient.

Ion transport from one aqueous (W1) to another (W2) across bilayer lipid membranes (BLM) containing gramicidin A (GA) was investigated by recording current fluctuations, when various alkali metal chlorides and potassium salts were used as supporting electrolytes. The magnitude of the single-channel current at a given membrane potential depended on not only the cationic species, but also on the anionic species, and then it decreased with an increase in the diameter of the anion when the diameter of the anion was larger than the pore size of the GA channel. The baseline of the recording current, however, increased with an increase in the diameter of the anion, and its height depended on the concentration of GA in the BLM. The results indicate that GA serves as not only a channel-forming compound, but also as a carrier compound in the BLM.

This is the first report of a case of hypocalcemia in a female infant with holoprosencephaly. Hypocalcemia developed 60 days after birth, secondary to decreased serum 1,25-dihydroxyvitamin D, as a result of renal tubular dysfunction which may have been induced by prerenal acute renal failure, the administration of anticonvulsants, and hypothyroidism. However, there was no evidence of rickets, and her serum 25-hydroxyvitamin D value was normal. She was treated with high-dose (0.5 microg/kg) 1alpha-hydroxyvitamin D3 and calcium lactate, and her calcium and 1,25-dihydroxyvitamin D values were consequently, normalized. However, she died at 268 days after birth.

We present a girl with type I membranoproliferative glomerulonephritis (MPGN) diagnosed by the third renal biopsy. The first renal biopsy was performed at age 11.2 years after microscopic hematuria (which was revealed by school urinary screening) had persisted for 3 months, along with a low level of serum C3. Pathological examination of the biopsied specimen revealed endocapillary proliferative glomerulonephritis with multiple humps. The serum C3 level increased to within the normal range 2 months after the first renal biopsy, and the microscopic hematuria disappeared at age 12.3. However, microscopic hematuria, proteinuria, and the low serum complement level reappeared at age 12.8. Pathological examination of a further renal biopsy that was performed at age 13.2 revealed focal MPGN with humps. Prednisolone therapy was subsequently initiated. Fluvastatin was added to her treatment regime when she developed hypercholesterolemia at age 13.6 and was continued even after normal cholesterol levels were reestablished. Pathological examination of the third renal biopsy, which was performed at age 15.2, revealed type I MPGN with humps. Serum C3 normalized 6 months after the cessation of prednisolone at age 15.9. It is clinically important that patients with nontypical acute glomerulonephritis should be observed over a long period and repeated renal biopsies should be performed.

The 1H NMR signals of the heme methyl, propionate and related chemical groups of cytochrome c3 from Desulfovibrio vulgaris Miyazaki F (D.v. MF) were site-specifically assigned by means of 1D NOE, 2D DQFCOSY and 2D TOCSY spectra. They were consistent with the site-specific assignments of the hemes with the highest and second-lowest redox potentials reported by Fan et al. (Biochemistry, 29 (1990) 2257-2263). The site-specific heme assignments were also supported by NOE between the methyl groups of these hemes and the side chain of Val18. All the results contradicted the heme assignments for D.v. MF cytochrome c3 made on the basis of electron spin resonance (Gayda et al. (1987) FEBS Lett., 217 57-61). Based on these assignments, the interaction of cytochrome c3 with D.v. MF ferredoxin I was investigated by NMR. The major interaction site of cytochrome c3 was identified as the heme with the highest redox potential, which is surrounded by the highest density of positive charges. The stoichiometry and association constant were two cytochrome c3 molecules per monomer of ferredoxin I and 10(8) M-2 (at 53 mM ionic strength and 25 degrees C), respectively.