VNAR development through antigen immunization of Japanese topeshark (Hemitriakis japanica).

The VNAR (Variable New Antigen Receptor) is the smallest single-domain antibody derived from the variable domain of IgNAR of cartilaginous fishes. Despite its biomedical and diagnostic potential, research on VNAR has been limited due to the difficulties in obtaining and maintaining immune animals and the lack of research tools. In this study, we investigated the Japanese topeshark as a promising immune animal for the development of VNAR. This shark is an underutilized fishery resource readily available in East Asia coastal waters and can be safely handled without sharp teeth or venomous stingers. The administration of Venus fluorescent protein to Japanese topesharks markedly increased antigen-specific IgM and IgNAR antibodies in the blood. Both the phage-display library and the yeast-display library were constructed using RNA from immunized shark splenocytes. Each library was enriched by biopanning, and multiple antigen-specific VNARs were acquired. The obtained antibodies had affinities of 1 × 10-8 M order and showed high plasticity, retaining their binding activity even after high-temperature or reducing-agent treatment. The dissociation rate of a low-affinity VNAR was significantly improved via dimerization. These results demonstrate the potential utility of the Japanese topeshark for the development of VNAR. Furthermore, we conducted deep sequencing analysis to reveal the quantitative changes in the CDR3-coding sequences, revealing distinct enrichment bias between libraries. VNARs that were primarily enriched in the phage display had CDR3 coding sequences with fewer E. coli rare codons, suggesting translation machinery on the selection and enrichment process during biopanning.

Temperature dependence of sequential chlorinated ethenes dechlorination and the dynamics of dechlorinating microorganisms.

Thermally enhanced bioremediation is a promising approach to shorten the bioremediation period of tetrachloroethene (PCE) and trichloroethene (TCE). To clarify the influence that temperature has on stepwise PCE dechlorination and associated microorganisms, this study conducted dechlorination experiments using contaminated soil and groundwater under five distinct temperature conditions (i.e., 15, 20, 25, 30, and 35 °C). PCE and TCE were dechlorinated most rapidly at 25-35 °C, whereas the preferable temperatures for the dechlorination of cis-1,2- dichloroethene (cis-1,2-DCE) and vinyl chloride (VC) were 25-30 °C and 25 °C, respectively. Microbial community analysis revealed that Sulfurospirillum and Geobacter may have a dominant contribution to the dechlorination of PCE to cis-1,2-DCE, whereas Dehalococcoides harboring VC reductase genes are likely major contributors to the dechlorination of cis-1,2-DCE and VC. These results suggest that temperature influences various microbial groups, including major dechlorinating microorganisms, resulting in the different extent of PCE dechlorination. In addition, the microbial community structure greatly changed after the onset of the experiment, whereas the temperature influence of 15-30 °C on the microbial community structure was minor; however, the microbial community was significantly impacted at 35 °C. Collectively, these results suggest that thermally enhanced anaerobic dechlorination at 25 °C is useful for successful dechlorination of chlorinated ethenes in a short period.

Prevention of end-stage renal disease with percutaneous endoscopic gastrojejunostomy tubes in a patient with recurrent episodes of binge eating and purging.

Background: Bulimia nervosa (BN) and the binge-eating/purging subtype of anorexia nervosa (b/p AN) are characterized by binge eating and unsafe compensatory behaviors, such as laxatives or diuretic abuse, self-induced vomiting, and excessive exercise. BN often causes miscellaneous physical complications that can lead to death. However, there have been very few prior reports on the physical complications of chronic BN. Case Presentation: We report a case of chronic BN of over 10 years. Her compensatory behaviors caused dehydration more easily than before. Repeated dehydration may result in end-stage renal disease (ESRD). Therefore, we had to prevent dehydration by placing a percutaneous endoscopic gastrojejunostomy (PEG-J) tube. After PEG-J placement, dehydration did not reoccur, thereby protecting her renal involvement from progressing to ESRD. Conclusion: Chronic BN exposure may cause repeated dehydration with time. Therefore, the PEG-J tube would be effective in preventing dehydration, which leads to ESRD.

Indigo enhances wound healing activity of Caco-2 cells via activation of the aryl hydrocarbon receptor.

Indigo Naturalis, also known as Qing Dai (QD) is a compound obtained from Indigofera tinctoria, Isatis tinctoria, and Polygonum tinctoria and is known to ameliorate refractory ulcerative colitis (UC) by an unknown mechanism. QD maintains both homeostasis and the integrity of colon epithelia in mice that have experimentally induced colitis. The primary component of QD, indigo, comprises 42.4% of the compound. Indigo efficiently suppresses rectal bleeding and reduces the erosion of the colon epithelium, whereas it does not reduce weight loss or increase survival in a certain condition. Indigo is a ligand of the aryl hydrocarbon receptor (AhR), which is involved in the anti-colitis activity of QD. Here we investigate the effects of indigo on wound (erosion) closure in colon epithelial cells. Oral administration of indigo induced expression of Cytochrome P450 1A1 (Cyp1a1) in the colon but not in the liver, suggesting that indigo stimulates AhR from the luminal side of the colon. The erosion-closure activity tested in the scratch assays using Caco-2 cells was accelerated by addition of QD and indigo to the culture medium. QD and indigo also induced nuclear localization of AhR and expression of CYP1A1 in the Caco-2 cells. Acceleration of scratch wound closure was abolished by addition of the AhR-antagonist CH223191. Cell proliferation and actin polymerization were also shown to contribute to erosion closure. The results suggest that indigo exerts its erosion-healing effects by increasing proliferation and migration of colon epithelial cells via activation of AhR in intestinal epithelia.

Severe necrosis of the glans penis associated with calciphylaxis treated by partial penectomy.

INTRODUCTION: Calciphylaxis is characterized by marked vascular calcification and painful skin ulcers, and it has a poor prognosis. CASE PRESENTATION: The patient was a 72-year-old male. He was referred for penile pain. He had a 4-year history of dialysis therapy under a diagnosis of diabetic nephropathy. Black and yellow necrosis was observed involving the entire glans, accompanying severe pain. Computed tomography revealed marked calcification involving the thoracoabdominal aorta to iliac arteries, the dorsal artery of the penis and the corpus cavernosum, leading to a diagnosis of calciphylaxis. Penile pain gradually exacerbated and partial penectomy was performed. After surgery, penile pain promptly subsided. Pathological examination confirmed marked calcification of the microvascular wall and narrowing of the lumen. CONCLUSION: We reviewed 15 Japanese patients with calciphylaxis who had undergone penile surgery. Surgical treatment was considered to be effective at relieving penile pain, but the prognosis remained poor.

Insomnia in patients on hemodialysis for a short versus long duration.

BACKGROUND: Many studies have investigated insomnia and the factors associated with this condition in hemodialysis (HD) patients, although the influence of HD duration has not been thoroughly investigated. In the present study, we investigated the factors, especially the duration of HD, associated with insomnia in HD patients. PATIENTS AND METHODS: A total of 138 patients undergoing HD were recruited, and the Japanese version of the Pittsburgh Sleep Quality Index (PSQI) was used to assess the quality of sleep. Subjects with a total PSQI score up to 4 and those with a score of at least 5 were identified as normal subjects and subjects with insomnia, respectively. Additionally, we assessed restless legs syndrome, depression using the Center for Epidemiologic Studies Depression Scale, and health-related quality of life (QOL) using the Short Form 8 Health Survey. We divided the subjects into two groups according to the median HD duration. RESULTS: The prevalence rate of insomnia was 54.3% among all the subjects. Twenty-one subjects (15.2%) had depression, 26 (18.8%) had restless legs syndrome, and 75 (54.3%) had insomnia. The median HD duration was 4 years. The scores of components 1 and 4 of the PSQI, subjective sleep quality and habitual sleep efficiency, did not show a significant difference between the normal and insomnia groups. The score of component 7, daytime dysfunction, showed a significant difference between the short and long HD duration groups. In multiple regression analysis, the score of the Short Form 8 Health Survey showed a significant association with the PSQI score in the long HD duration group, but no variable showed a significant association in the short HD duration group. CONCLUSION: Patients with a longer duration of HD indicated that insomnia has an influence on their daily activities, with a significant association between insomnia and QOL. Greater attention should be paid to poor QOL and troubles in daily activities caused by insomnia in patients with a longer HD duration.

Study of penetration behavior of PCB-DNAPL in a sand layer by a column experiment.

To better understand the infiltration performances of high concentration PCB oils (KC-300 and KC-1000 polychlorinated biphenyl (PCB) mixtures), representative dense non-aqueous phase liquid (DNAPL), under both saturated and unsaturated conditions, we conducted experiments on a sand column filled with Toyoura Standard Sand. When PCB oil with the volume comparable to the total porosity in the column was supplied, the residual PCB concentrations under PCB-water conditions were 4.9×10(4)mgkg(-1) in KC-300 and 3.9×10(4)mgkg(-1) in KC-1000. Under PCB-air conditions, residual PCB concentrations were 6.0×10(4)mgkg(-1) and 2.4×10(5)mgkg(-1) in the upper and lower parts for KC-300 and 3.6×10(4)mgkg(-1) and 1.5×10(5)mgkg(-1) in those for KC-1000, respectively, while the rest of the PCBs were infiltrated. On the other hand, when a small amount of PCB oil with the volume far smaller than the total porosity in the column was supplied, the original PCBs were not transported via water permeation. However, lower-chlorinated PCB congeners-e.g., di- or tri-chlorinated biphenyls-preferentially dissolved and were infiltrated from the bottom of the column. These propensities on PCB oil infiltration can be explained in conjunction with the degree of PCB saturation in the sand column.

Preparation of antibodies against a novel immunosuppressant, FTY720, and development of an enzyme immunoassay for FTY720.

FTY720 (1) is a novel immunosuppressant (immunomodulator), derived from ISP-I (2: myriocin and thermozymocidin). To clarify the pharmacokinetic properties of 1, antibodies against 1 were prepared and a competitive enzyme immunoassay (EIA) was developed. Two kinds of haptens, 3 and 4, for 1 were synthesized and coupled to ovalbumin (OVA). Rabbits were immunized with 3-OVA or 4-OVA, and corresponding antibodies were obtained. Both antibodies recognized the 2-amino-2-(2-phenylethyl)propane-1,3-diol moiety in 1. Using the anti-3-OVA antibody, a competitive EIA for 1 was developed and evaluated. The range of quantification by the EIA was 0.06-10 ng/mL. The application of the EIA has enabled us to measure the FTY720 concentration in serum after oral administration of 1 (1mg/kg) to rats.

Geographic variation of the Japanese spinous loach, Cobitis takatsuensis, inferred from allozyme analysis.

Genetic variation among river populations of the Japanese spinous loach, Cobitis takatsuensis, on Shikoku Island, was demonstrated by allozyme electrophoresis based on 40 presumed loci representing 20 enzymes and one non-enzymatic protein. Complete allelic replacement was not evident between populations in any one river system, but existed between populations in different river systems. The maximum genetic distance among populations reached 0.13 and the populations were divided genetically into four or five groups by river system on UPGMA and Neighbor-joining trees. In view of the genetic diversity of C. takatsuensis on Shikoku Island, the populations in three small river systems are thought to have originated from populations in one or the other of two large river systems with a bottle-neck effect. C. takatsuensis is currently designated as an endangered species in Japan. This study suggested that the populations of C. takatsuensis in each river system should be regarded as independent conservation units.