Production of single-chain variable fragment antibody (scFv) in fed-batch and continuous culture of Pichia pastoris by two different methanol feeding methods

This study examined the effects of two methods of methanol feeding, DO-stat and methanol concentration control, in fed-batch and continuous cultures of Pichia pastoris on cell growth and single-chain variable fragment antibody (scFv) expression. By maintaining the methanol concentration at 3.9 g l(-1) in fed-batch culture, a scFv concentration of 198 mg l(-1) was obtained. In continuous culture using both methanol feeding methods, the scFv concentration in the fermentation broth increased with a decreasing dilution rate. A maximum scFv concentration of 810 mg l(-1) at a dilution rate of 0.0094 h(-1) was obtained by maintaining the methanol concentration at 3.9 g l(-1). Although the specific methanol consumption rate was the same for both methods, the specific productivity of scFv was higher in methanol concentration control from 0.0094 to 0.049 h(-1) than it was in DO-stat control. Therefore, continuous culture with methanol feeding by the concentration control method shows promise for the industrial scale production of recombinant proteins by Pichia pastoris.     

Phase unwrapping method for three-dimensional stress analysis by scattered-light photoelasticity with unpolarized light. 2. Experiment

A phase unwrapping method that employs scattered-light photoelasticity with unpolarized light was proposed for automated three-dimensional stress analysis [Appl. Opt. 45, 8848 (2006)]. I now demonstrate the validity of this method by performing nondestructive measurements at three different wavelengths of the secondary principal stress direction psi(j) and the total relative phase retardation rho(jtot) in the plane that contains the rotated principal stress directions in a spherical frozen stress model and compare the results obtained with mechanically sliced models. The parameters psi(j) and rho(jtot) were measured nondestructively over the entire field of view for the first time, to the best of my knowledge.     

Stimulation of porphyrin production by application of an external magnetic field to a photosynthetic bacterium, Rhodobacter sphaeroides

The effects of an external magnetic field on the production of porphyrin were investigated using Rhodobacter sphaeroides IF012203 under anaerobic-light conditions. Upon application of a 0.13-0.3-T magnetic field, the growth was slightly suppressed and porphyrin extracellular production was activated at both the N and S poles, particularly at the N pole up to about 5.3 times that in the control experiment (6.7 mg/1) (without magnetic filed application). The maximum production was 35.8 mg/1 at the N pole with a magnetic field of 0.3 T. At the same time, the 5-aminolevulinic acid dehydratase (ALAD) concentration was enhanced in the cells at the N and S poles, but particularly at the the N pole. This suggested the possibility that the magnetic field might activate ALAD gene expression in R. sphaeroides IF012203 and result in enhanced porphyrin production.     

Omeprazole Suppresses Oxaliplatin-Induced Peripheral Neuropathy in a Rodent Model and Clinical Database

(1) Background: Oxaliplatin is used as first-line chemotherapy not only for colorectal cancer but also for gastric and pancreatic cancers. However, it induces peripheral neuropathy with high frequency as an adverse event, and there is no effective preventive or therapeutic method. (2) Methods: The effects of omeprazole, a proton pump inhibitor (PPI), on oxaliplatin-induced peripheral neuropathy (OIPN) was investigated using an in vivo model and a real-world database. (3) Results: In a rat model, oxaliplatin (4 mg/kg, i.p., twice a week for 4 weeks) caused mechanical hypersensitivity accompanied by sciatic nerve axonal degeneration and myelin sheath disorder. Repeated injection of omeprazole (5–20 mg/kg, i.p., five times per week for 4 weeks) ameliorated these behavioral and pathological abnormalities. Moreover, omeprazole did not affect the tumor growth inhibition of oxaliplatin in tumor bearing mice. Furthermore, clinical database analysis of the Food and Drug Administration Adverse Event Reporting System (FAERS) suggests that the group using omeprazole has a lower reporting rate of peripheral neuropathy of oxaliplatin-treated patients than the group not using (3.06% vs. 6.48%, p < 0.001, reporting odds ratio 0.44, 95% confidence interval 0.32–0.61). (4) Conclusions: These results show the preventing effect of omeprazole on OIPN.     

Metabolomic Profiling of Plasma,Urine, and Saliva of Kidney Transplantation Recipients

Kidney biopsy is commonly used to diagnose kidney transplant dysfunction after transplantation. Therefore, the development of minimally invasive and quantitative methods to evaluate kidney function in transplant recipients is necessary. Here, we used capillary electrophoresis-mass spectrometry to analyze the biofluids collected from transplant recipients with impaired (Group I, n = 31) and stable (Group S, n = 19) kidney function and from donors (Group D, n = 9). Metabolomics analyses identified and quantified 97 metabolites in plasma, 133 metabolites in urine, and 108 metabolites in saliva. Multivariate analyses revealed apparent differences in the metabolomic profiles of the three groups. In plasma samples, arginine biosynthesis and purine metabolism between the I and S Groups differed. In addition, considerable differences in metabolomic profiles were observed between samples collected from participants with T cell-mediated rejection (TCR), antibody-mediated rejection, and other kidney disorders (KD). The metabolomic profiles in the three types of biofluids showed different patterns between TCR and KD, wherein 3-indoxyl sulfate showed a significant increase in TCR consistently in both plasma and urine samples. These results suggest that each biofluid has different metabolite features to evaluate kidney function after transplantation and that 3-indoxyl sulfate could predict acute rejection.     

Dissolution behavior of core structure materials by molten corium in boiling water reactor plants during severe accidents

For the decommissioning of the Fukushima Daiichi Nuclear Power Plant, it is necessary to consider the access route to the fuel debris for its removal, which can be determined by knowing the corruption situation of the core support structure. To predict the damage condition of reactor vessel, dissolution behavior of the core structure material should be understood. In this study, the dissolution behavior of core structure materials (stainless steel) by molten metallic corium (stainless steel + B₄C) originated from control rod and its cladding was investigated. As a result of immersion experiment, it was found that there were two types of dissolution mode in this system: (1) chemical dissolution by eutectic reaction between Fe and B and (2) physical dissolution caused by the grains falling off from solid steel due to infiltration of molten metal. Moreover, on the basis of kinetic analysis, it was considered that the chemical dissolution in this system was slow. Therefore, the dissolution is considered to mainly occur through the mechanism that physical dissolution precedes chemical dissolution.