Construction of a novel bioanode for amino acid powered fuel cells through an artificial enzyme cascade pathway.

OBJECTIVE: The construction of a novel bioanode based on L-proline oxidation using a cascade reaction pathway comprised of thermostable dehydrogenases. RESULTS: A novel multi-enzymatic cascade pathway, containing four kinds of dehydrogenases from thermophiles (dye-linked L-proline dehydrogenase, nicotinamide adenine dinucleotide (NAD)-dependent Δ1-pyrroline-5-carboxylate dehydrogenase, NAD-dependent L-glutamate dehydrogenase and dye-linked NADH dehydrogenase), was designed for the generation of six-electrons from one molecule of L-proline. The current density of the four-dehydrogenase-immobilized electrode, with a voltage of + 450 mV (relative to that of Ag/AgCl), was 226.8 µA/cm2 in the presence of 10 mM L-proline and 0.5 mM ferrocene carboxylate at 50 °C. This value was 4.2-fold higher than that of a similar electrode containing a single dehydrogenase. In addition, about 54% of the initial current in the multi-enzyme cascade bioanode was maintained even after 15 days. CONCLUSIONS: Efficient deep oxidation of L-proline by multiple-enzyme cascade reactions was achieved in our designed electrode. The multi-enzyme cascade bioanode, which was built using thermophilic dehydrogenases, showed high durability at room temperature. The long-term stability of the bioanode indicates that it shows great potential for applications as a long-lived enzymatic fuel cell.

A L-proline/O2 biofuel cell using L-proline dehydrogenase (LPDH) from Aeropyrum pernix.

To utilize amino acids from food waste as an energy source, L-proline/O2 biofuel cell was constructed using a stable enzyme from hyperthermophilic archaeon for long-term operation. On the anode, the electrocatalytic oxidation of L-proline by L-proline dehydrogenase from Aeropyrum pernix was observed in the presence of ferrocenecarboxylic acid as mediator. On the cathode, electrocatalytic oxygen reduction was detected. Ketjenblack modification of carbon cloth substrate increased the current density due to increased laccase loading and enhanced electron transfer reaction. The biofuel cell using these electrodes achieved a current density of 6.00 µA/cm2. We successfully constructed the first biofuel cell that generates power from L-proline.

Vector synthesis high-resolution electrocardiography, atrial natriuretic peptide and N-terminal prohormone brain natriuretic peptide for estimation of cardiac load in pregnancy.

AIM: We analyzed atrial natriuretic peptide (ANP), N-terminal pro-brain natriuretic natriuretic peptide (NT-proBNP) and vector synthesis high-resolution electrocardiography (ECG), to estimate cardiac load with circulatory dynamic change from pregnancy through the post-partum period. METHODS: The subjects were singleton pregnant women (n = 19), who were divided into three stages: stage 1, 34-36 weeks of gestation; stage 2, 2-6 post-partum days; and stage 3, 1-3 months after delivery. Vector synthesis high-resolution ECG, ANP and NT-proBNP were analyzed for all subjects. RESULTS: A pregnant woman with massive uterin liomyoma expressed largest the corrected recover time (RTc) dispersion in I + II of tow Dimensional (2D) color distribution map ANP and NT-proBNP were significantly higher in stage 2 than in stages 1 and 3. CONCLUSIONS: ANP, NT-proBNP and vector synthesis high-resolution ECG there might be able to evaluate cardiac load of normal pregnancy.

Suitable biopsy site for detection of esophageal eosinophilia in eosinophilic esophagitis suspected cases.

BACKGROUND AND AIM: Esophageal eosinophilia (EE) is the most important finding for the diagnosis of eosinophilic esophagitis. We conducted the present retrospective study to clarify the most suitable site of the esophagus to examine for EE detection. METHODS: We enrolled 4999 subjects who underwent upper gastrointestinal endoscopy screening examinations as part of a detailed health check-up. When endoscopic esophageal abnormalities characteristic of eosinophilic esophagitis were observed, two or three biopsy specimens were obtained from the middle or lower esophagus, and endoscopic findings of fixed rings, exudates, furrows, edema, stricture, and crepe paper esophagus in biopsied sites were analyzed. RESULTS: Thirty-five subjects underwent histological examination for EE, of whom 20 showed positive findings in biopsied specimens. Higher grade endoscopic findings of exudates, furrows, and edema were observed in cases with EE in comparison to the 15 without EE, whereas statistically significant higher grade endoscopic findings of exudates, furrows, and edema were also observed in biopsied sites with EE in comparison to those without EE. In addition, a positive finding of EE in biopsied specimens from the lower esophagus was significantly more frequently noted as compared to those from the middle esophagus. Multiple logistic regression analysis showed that a lower esophagus biopsy site and severe exudates were significant factors related to a positive EE finding in biopsied specimens. CONCLUSION: The most suitable conditions for detection of EE are a lower esophagus biopsy site and the presence of exudates in cases suspicious of eosinophilic esophagitis shown by endoscopy.

Analysis of negative result in serum anti-H. pylori IgG antibody test in cases with gastric mucosal atrophy.

The purpose is to elucidate factors related to negative results of anti-H. pylori antibody test in cases with gastric mucosal atrophy. A total of 859 individuals without past history of eradication therapy for H. pylori (545 males, 314 females; mean age 52.4 years) who underwent an upper GI endoscopy examination and serological test were enrolled as subjects. Serological testing was performed using SphereLight H. pylori antibody J®, and endoscopic findings of gastric mucosal atrophy by the classification of Kimura and Takemoto and post-eradication findings were analyzed. The positive rates for the anti-H. pylori antibody test in subjects with and without gastric mucosal atrophy were 85.6% and 0.9%, respectively. In analysis of subjects with gastric mucosal atrophy, a low positive rate and serum titer was observed in subjects with C1, C2 and O3 atrophy. When the analysis was performed separately in male and female subjects, low positive rate was observed in males with O3 atrophy and females with C2 atrophy. Suspected post-eradication endoscopic findings were more frequently observed in cases with C2 atrophy. In conclusion, negative result of anti-H. pylori antibody test was frequently observed in middle-aged subjects with C1, C2 and O3 gastric mucosal atrophy.

Pd/NHC-catalyzed enantiospecific and regioselective Suzuki-Miyaura arylation of 2-arylaziridines: synthesis of enantioenriched 2-arylphenethylamine derivatives.

A palladium-catalyzed stereospecific and regioselective cross-coupling of enantiopure 2-arylaziridines with arylboronic acids under mild conditions to construct a tertiary stereogenic center has been developed. N-heterocyclic carbene (NHC) ligands efficiently promote the coupling, suppressing ß-hydride elimination. The enantiospecific cross-coupling allowed us for preparation of a series of biologically important 2-arylphenethylamine derivatives in an enantiopure form.

Usefulness of Practitioner-Led Pancreatic Cancer Screening.

The 5-year survival rate for pancreatic cancer has improved (10%) but remains worse than that for other cancers. Early pancreatic cancer diagnosis is challenging, and delayed diagnosis can delay treatment, which impairs survival. Practitioners do not promptly refer cases to a general hospital, causing delayed discovery. Herein, we aimed to examine the usefulness of the Pancreatic Cancer Project in Matsue, whose objective is to detect pancreatic cancer in patients presenting at any medical institution in Matsue City. Clinical data were extracted from medical records, and abdominal ultrasonography and tumor marker blood level assessments were performed (n = 234; median age, 71 [range, 41-94] years; 51% male). Cases with abnormal abdominal ultrasonography or blood test findings were referred for specialist imaging and followed up. The pancreatic cancer detection rate was 6.0% (n = 14); all cases were referred to a general hospital by practitioners within 1 month. Patients had stage IA (n = 1), IIA (n = 6), IIB (n = 2), III (n = 1), and IV (n = 4) disease. Overall, pancreatic cancer could be detected at an earlier stage (I-II), but referral to a general hospital by visiting practitioners should be prompt. The Pancreatic Cancer Project in Matsue may help improve the detection and prognosis of pancreatic cancer.

Different sex-related influences of eating habits on the prevalence of reflux esophagitis in Japanese.

BACKGROUND AND AIM: Many types of food have been shown to affect lower esophageal sphincter pressure and esophageal motor function, and thus, the prevalence of reflux esophagitis. The present study was performed to clarify the different eating habits that predominantly affect the prevalence of reflux esophagitis in Japanese. METHODS: The study included 2303 individuals (males: 1599, females: 704, mean age: 49.9 years) who underwent upper gastrointestinal endoscopy for gastric cancer screening. The daily dietary contents of the patients were analyzed using a self-administered questionnaire. RESULTS: A total of 201 patients had endoscopically-proven reflux esophagitis, and the percentage of males with reflux esophagitis was significantly higher than their female counterparts (11.3% vs 2.8%). The body mass indexes of individuals with reflux esophagitis were significantly higher than those without, both for males and females. Total energy intake was the most important risk factor for the occurrence of reflux esophagitis in males, but the food content was not a significant risk factor. Dietary habit did not affect the prevalence of reflux esophagitis in the female patients. The age and height of females with reflux esophagitis significantly exceeded those of females without reflux esophagitis, and were independent risk factors for the occurrence of reflux esophagitis only in the female patients. CONCLUSION: There is a sex-related difference in the influence of eating habits on the prevalence of reflux esophagitis in Japanese.

Splicing modulator FR901464 is a potential agent for colorectal cancer in combination therapy.

FR901464 (FR) was first described as an anticancer drug and later identified as a modulator of splicing factor 3B subunit 1 (SF3B1). Although the effectiveness of splicing modulators has been investigated in colorectal cancer (CRC) cells, their usefulness in animal experiments has not been confirmed. The association of SF3B1 with CRC progression and the influence of FR on transcriptional activity in CRC has not been fully elucidated. FR showed strong cytotoxicity against CRC cell lines, SF3B1-mutated cancer cell lines, and human fibroblasts with IC50 values less than 1 ng/ml. FR-resistant clones derived from HCT116, DLD1, Lovo, and CT26 cells showed IC50 values greater than 100 ng/ml. SF3B1 sequencing demonstrated low frequencies of SF3B1 mutations in CRC and mutations in codon 1074 of exon 22 in all FR-resistant clones. Unlike hematological malignancies, SF3B1 expression was not associated with CRC progression. Although FR showed significant growth inhibition in a xenograft model of RKO cells, severe toxicity was also induced. These data indicated CRC might be a suitable target of FR unless toxicity occurs. Microarray analysis and real-time quantitative PCR demonstrated downregulation of genes associated with Fanconi anemia (BRCA1 and BRCA2) and 28 driver oncogenes. These data suggested combination treatment of FR with other anticancer drugs whose sensitivity is associated with genes affected by FR treatment. Combination treatment with PARP1 inhibitor olaparib, whose sensitivity was enhanced by BRCA 1/2 deficiency, showed synergistic effects in CRC cells. Our data indicates the potential of FR in combination therapy rather than monotherapy for CRC treatment.

Helicobacter pylori infection does not accelerate the age-related progression of arteriosclerosis: a 4-year follow-up study.

BACKGROUND AND AIM: Helicobacter pylori (H. pylori) infection has been reported to correlate with the onset of cardiovascular diseases. However, the relationship between H. pylori infection and the progression of arteriosclerosis has not been fully investigated. The aim of this study was to clarify the possible role of H. pylori infection in the development of arteriosclerosis. METHODS: Study subjects were 258 cases who attended their annual medical check-ups and were twice investigated by the parameters of arteriosclerosis at a 4-year interval. Arteriosclerotic parameters (systolic blood pressure [SBP]), ankle brachial index [ABI], and pulse wave velocity [PWV]) were measured non-invasively using an automatic device. H. pylori-infection status was determined by assaying serum anti-H. pylori IgG antibodies. The changes in the arteriosclerotic parameters during the 4 years were compared between H. pylori-seropositive and seronegative individuals. RESULTS: A total of 166 subjects (64.3%) were H. pylori-seropositive. After adjustment for gender, age, body mass index, and smoking and drinking habits, there were no differences in any of the arteriosclerotic parameters between the groups, not only at enrollment but after the 4 years, although the high-density lipoprotein cholesterol levels of the seropositive groups were significantly lower than those of seronegative groups. SBP and PWVs were significantly increased during 4 years in both the seropositive and seronegative groups. The percentage of changes in SBP and PWVs, which were calculated by ([values after 4 years--values at enrollment/values at enrollment] x 100), did not differ between the groups. CONCLUSION: H. pylori infection does not accelerate the age-related progression of arteriosclerosis in Japanese individuals.

[Study on the recognition of upper abdominal symptoms by Japanese adults ].

Four hundred and sixty-three cases, who visited medical center for annual check-up, were enrolled in this study. All subjects were required to answer three symptoms in the ten upper abdominal symptoms and the frequency of these symptoms by which they might visit hospital to take medical care. Upper abdominal pain, nausea and chest pain were best three symptoms, which might cause hospital visit and only a few subjects thought that they make a hospital visit by heartburn, acid regurgitation and early satiety. More than 30% of subjects think that they might visit medical center for medical care when these symptoms occur over three times per week. In contrast, upper abdominal pain, nausea, vomiting, heartburn and early satiety were frequently observed in 100 patients, who visited medical center to take medical care. In addition, about half of patients visited medical center when these symptoms occurred every day.

S100A12 facilitates osteoclast differentiation from human monocytes.

Osteoclasts play a critical role not only in bone homeostasis but also in inflammatory osteolysis, such as that occurring in inflammatory arthritis and systemic inflammation. In both inflammation conditions, inflammatory cytokines like Interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF)-α induce RANKL expression in osteoblasts, but the roles of these cytokines in osteoclast activation remain unclear. S100A12, an S100 family member, is a low-molecular-weight calcium-binding protein. Although it has a pro-inflammatory role, its effects on osteoclast differentiation have been unclear. Here we examined the direct effects of S100A12 on human osteoclasts in vitro. S100A12 facilitated osteoclast formation in the presence of RANKL, as judged by the cells' morphology and elevated expression of osteoclast-related molecules, including NFATc1, ACP5, CALCR, and ITGß3. In addition, S100A12 administration markedly enhanced the osteoclasts' bone resorption ability, consistent with their increased expression levels of CTSK and CA2. Blocking RAGE and TLR4 cancelled the effects of S100A12. Our results indicate that S100A12 is a potential therapeutic target for inflammatory osteolysis.

Evaluation of appropriate follow-up after curative surgery for patients with colorectal cancer using time to recurrence and survival after recurrence: a retrospective multicenter study.

INTRODUCTION: The follow-up schedule for colorectal cancer patients after curative surgery is inconsistent among the guidelines. Evaluation of time to recurrence (TTR) and survival after recurrence (SAR) may provide evidence for appropriate follow-up. METHODS: We assessed 3039 colon cancer (CC) and 1953 rectal cancer (RC) patients who underwent curative surgery between 2007 and 2008. We evaluated the pre- and post-recurrent clinicopathological factors associated with TTR and SAR in each stage of CC and RC. RESULTS: The recurrence rates of stages I, II, and III were 1.2%, 13.1%, and 26.3%, respectively, for CC, and 8.4%, 20.0%, and 30.4%, respectively, for RC. In CC patients, high carcinoembryonic antigen (CEA) level and lymphovascular invasion were independent predictors of short TTR. In RC patients, metastatic factors (liver metastasis in stage III) and venous invasion (stage III) were independent predictors of short TTR. The prognostic factors of SAR were age (stage II CC and stage III RC), female gender (stage III RC), high CEA level (stage II RC), histological type (stage III CRC), nodal status (stage III CC), recurrence within 1 year (stage III RC), M1b recurrence (stage II CRC), local recurrence (stage II CC), and no surgical resection after recurrence (stage II and III CRC). CONCLUSIONS: The follow-up schedule for stage I should be different from that for the other stages. We recommend that intensive follow-up is appropriate in stage III CC patients with undifferentiated adenocarcinoma or N2 nodal status, stage II RC patients with high preoperative CEA level, and stage III RC patients.

Circumferential Locations of Different Shapes of Short-segment Barrett's Esophagus.

Objective To clarify the relationship between the shape and circumferential location of non-circumferential short-segment Barrett's esophagus (SSBE). Methods We examined 3,788 subjects (2,497 males, 1,291 females; mean age 52.4 years) who underwent upper GI endoscopy as part of a detailed medical checkup. The presence of columnar-appearing mucosa ≥10 mm long in the distal esophagus was diagnosed as BE and then divided into circumferential and non-circumferential localized types. Localized SSBE was further divided into round and sharp types based on the shape of the proximal margin. Results SSBE was endoscopically observed in 197 subjects (5.2%). The numbers of patients with circumferential SSBE, round localized SSBE, and sharp localized SSBE were 38, 114 and 69, respectively. Round and sharp types of localized SSBE were simultaneously observed in 25 patients. Reflux esophagitis was more frequently observed in subjects with BE, regardless of type, in comparison to those without BE. Round localized SSBE was found mainly in the left posterior wall of the esophagus in a location similar to the main area of the esophageal cardiac glands. In contrast, sharp localized SSBE was observed mainly in the right anterior wall of the esophagus in a location similar to that of esophageal mucosal injury caused by mild type reflux esophagitis. Conclusion The location differs between round and sharp localized SSBE, possibly due to differences in the process of BE development.

A Study on the Relationship between Reflux Esophagitis and Periodontitis.

Objective Metabolic syndrome and dental erosion have been demonstrated to correlate with gastroesophageal acid reflux disease (GERD), while periodontitis has been reported to have a positive relationship with metabolic syndrome. However, no correlation between periodontitis and GERD has yet been reported. We therefore investigated the relationship between periodontitis and GERD. Methods The subjects consisted of 280 individuals who visited the Health Center for a detailed medical checkup examination. Each underwent upper endoscopy and periodontitis examinations, with the latter performed by measuring the concentrations of lactate dehydrogenase and hemoglobin in saliva. The subjects were divided into those with positive and negative periodontitis findings, and the prevalence rates of endoscopically proven reflux esophagitis, dyslipidemia, hypertension, and hyperglycemia were compared. Results The number of subjects positive for periodontitis was 93, while 187 had negative findings. The prevalence of reflux esophagitis was not different between the positive and negative groups (8.6% vs. 8.0%). In addition, a multiple logistic regression analysis did not identify a positive relationship between the presence of periodontitis and reflux esophagitis. On the other hand, dyslipidemia and hypertension were more frequently observed in the subjects that were positive for periodontitis. Conclusion We did not find an association between periodontitis and reflux esophagitis in the present study. On the other hand, the presence of periodontitis was found to correlate with hypertension and dyslipidemia.

Expression and Functions of Immediate Early Response Gene X-1 (IEX-1) in Rheumatoid Arthritis Synovial Fibroblasts.

In rheumatoid arthritis (RA), synovial fibroblasts (RA-SFs) accumulate in affected joints, where they play roles in inflammation and joint destruction. RA-SFs exhibit tumor-like proliferation and are resistant to apoptosis. Although RA-SF activation is well described, negative regulators of RA-SF activation are unknown. We previously reported that histone deacetylase (HDAC) inhibitors facilitate apoptosis in RA-SFs. Here we found that RA-SFs treated with the HDAC inhibitor Trichostatin A (TSA) exhibited an upregulation of the immediate early response gene X-1 (IEX-1). IEX-1 has roles in apoptosis sensitivity, cell-cycle progression, and proliferation, and is reported to be involved in immune responses, inflammation, and tumorigenesis, and to have anti-arthritic properties. To investigate IEX-1's role in RA-SFs, we used in vitro-cultured synovial fibroblasts from RA and osteoarthritis (OA) patients. We confirmed that TSA upregulated the IEX-1 protein and mRNA expressions in RA-SFs by western blotting and quantitative RT-PCR. Inhibiting HDAC1, 2, and 3 (but not 6 or 8) also upregulated IEX-1. The IEX-1 mRNA levels were higher in RA-SFs than in OA-SFs, and were further upregulated in RA-SFs by the pro-inflammatory cytokines TNFα and IL-1ß. The staining of surgical specimens showed that IEX-1 was present in the pannus from affected RA joints. Si-RNA-mediated IEX-1 knockdown upregulated the lipopolysaccharide (LPS)-induced expression of TNFα and various chemokine mRNAs, indicating that IEX-1 downregulates TNFα and chemokines. Furthermore, apoptosis analysis showed that IEX-1 knockdown protected RA-SFs from apoptosis induced by TSA or by an anti-Fas mAb, indicating that IEX-1 is pro-apoptotic in RA-SFs. Collectively, our results showed that IEX-1 is induced by TNFα and IL-1ß in RA-SFs, in which it suppresses TNFα and chemokine production and induces apoptosis; thus, IEX-1 negatively regulates RA-SF activation. Further investigation of IEX1's functions in RA-SFs may lead to new therapeutic approaches for RA.

Gender differences in the time-course changes of reflux esophagitis in Japanese patients.

OBJECTIVE: Obesity is related to an increased prevalence of reflux esophagitis and gastroesophageal reflux disease symptoms. This study was performed to clarify the influence of gender on time-course changes in the rate of a high BMI and incidence of reflux esophagitis in Japanese subjects. METHODS: The subjects included individuals who visited a medical center for medical checkups between April 2000 and March 2001, April 2005 and March 2006 and April 2010 and March 2011. At each examination, the subjects underwent upper gastrointestinal endoscopy to determine the presence of reflux esophagitis, size of the diaphragmatic hiatus and degree of gastric mucosal atrophy. A body mass index (BMI) of ≥ 25 kg/m(2) was defined as a high BMI. RESULTS: A multiple logistic regression analysis showed that a high BMI, milder degree of gastric mucosal atrophy and larger size of diaphragmatic hiatus were significant predictive factors for the presence of reflux esophagitis in both men and women. The number of male subjects with reflux esophagitis and a high BMI increased during the 10-year examination period. In contrast, the number of individuals with reflux esophagitis and a high BMI was not increased among women. For both men and women, the proportions of patients with a large diaphragmatic hiatus and mild gastric mucosal atrophy increased during the 10-year period. CONCLUSION: The prevalence of reflux esophagitis in the female subjects remained constant over 10 years, different from that observed in the men. A lack of change in BMI may be an important factor accounting for the constant prevalence of reflux esophagitis in women.

Relationship between esophageal cardiac glands and gastroesophageal reflux disease.

OBJECTIVE: The role of esophageal cardiac glands has not been fully determined. This study was performed to clarify the protective role of esophageal cardiac glands against the development of gastroesophageal reflux disease (GERD). METHODS: The subjects included 2,656 Japanese individuals who visited our institution for a detailed medical checkup. GERD symptoms were assessed using the Japanese version of the Carlsson-Dent self-administered questionnaire (QUEST) and an upper gastrointestinal endoscopy examination in each subject. The presence of reflux esophagitis, size of diaphragmatic hiatus, degree of gastric mucosal atrophy and existence of visible esophageal cardiac glands in the distal esophagus, based on the detection of yellowish elevated areas, were determined using endoscopy. RESULTS: Esophageal cardiac glands were observed in 355 cases (13.4%). Reflux esophagitis was significantly less frequent in the cases with esophageal cardiac glands than in those without. The esophageal cardiac glands were mainly located on the left-posterior side of the esophageal wall of the distal esophagus. A multiple regression analysis showed that the presence of esophageal cardiac glands was an independent factor for preventing reflux esophagitis. On the other hand, the existence of these glands did not correlate with the presence of GERD symptoms (QUEST score of 6 or more). CONCLUSION: The presence of visible esophageal cardiac glands may have a protective role against the development of reflux esophagitis.

A case of fallopian tube adenofibroma: difficulties associated with differentiation from ectopic pregnancy.

Benign solid tumors of the fallopian tubes are extremely rare and often difficult to differentiate from tumors associated with adjacent organs or from various inflammatory diseases. Here, we present a patient who was diagnosed with ectopic pregnancy, based on preoperative tests and intraoperative macroscopic findings, but was later diagnosed with a fallopian tube adenofibroma, based on histopathological evidence, and intrauterine pregnancy. Although initial pregnancy test results were positive, no gestational sac (GS) was seen in the uterus and the patient was diagnosed with an ectopic pregnancy and underwent emergency laparoscopic surgery. A 20-mm, fetus-like solid mass was noted inside the GS-like cystic tumor of the left fallopian tube. From histopathological findings, the lesion was identified as a serous fallopian tube adenofibroma. The baby was born healthy with no problems. This case report suggests that fallopian tube adenofibroma should be considered in the differential diagnosis of suspected ectopic pregnancies.