Chitosanase-immobilized magnetite-agar gel particles as a highly stable and reusable biocatalyst for enhanced production of physiologically active chitosan oligosaccharides.

A novel immobilized chitosanase was developed and utilized to produce chitosan oligosaccharides (COSs) via chitosan hydrolysis. Magnetite-agar gel particles (average particle diameter: 338 µm) were prepared by emulsifying an aqueous agar solution dispersing 200-nm magnetite particles with isooctane containing an emulsifier at 80 °C, followed by cooling the emulsified mixture. The chitosanase from Bacillus pumilus was immobilized on the magnetite-agar gel particles chemically activated by introducing glyoxyl groups with high immobilization yields (>80%), and the observed specific activity of the immobilized chitosanase was 16% of that of the free enzyme. This immobilized chitosanase could be rapidly recovered from aqueous solutions by applying magnetic force. The thermal stability of the immobilized chitosanase improved remarkably compared with that of free chitosanase: the deactivation rate constants at 35 °C of the free and immobilized enzymes were 8.1 × 10-5 and 3.9 × 10-8 s-1, respectively. This immobilized chitosanase could be reused for chitosan hydrolysis at 75 °C and pH 5.6, and 80% of its initial activity was maintained even after 10 cycles of use. COSs with a degree of polymerization (DP) of 2-7 were obtained using this immobilized chitosanase, and the product content of physiologically active COSs (DP ≥ 5) reached approximately 50%.

Facilitated encapsulation of a nonionic contrast agent for X-ray computed tomography into lipid vesicles by the multiple emulsification-solvent evaporation method.

We studied the encapsulation of iohexol (Ihex), a nonionic contrast agent used for X-ray computational tomography, into lipid vesicles using the multiple emulsification-solvent evaporation method to formulate a nanosized contrast agent. This lipid vesicle preparation method consists of three steps: (1) primary emulsification for producing water-in-oil (W/O) emulsions containing fine water droplets that will be converted to the internal water phase of the lipid vesicles, (2) secondary emulsification for formulating multiple water-in-oil-in-water (W/O/W) emulsions encapsulating the fine water droplets containing Ihex, and (3) solvent evaporation to remove the oil phase solvent (n-hexane) and to form lipid bilayers surrounding the fine inner droplets, resulting in the formation of lipid vesicles encapsulating Ihex. As the diameter and Ihex concentration of the primary W/O emulsion droplets decreased, a higher Ihex encapsulation yield was obtained for the final lipid vesicles. The entrapment yield of Ihex in the final lipid vesicles varied significantly with the emulsifier (Pluronic® F-68) concentration in the external water phase of W/O/W emulsion, and the highest yield (65%) was obtained when the emulsifier concentration was 0.1 wt%. We also investigated the powderization of lipid vesicles encapsulating Ihex via lyophilization. The powderized vesicles were dispersed in water after rehydration and maintained their controlled diameters. The entrapment yield of Ihex in powderized lipid vesicles was maintained for over 1 month at 25 ˚C, while significant leakage of Ihex was observed in the lipid vesicles suspended in the aqueous phase.

One-Step Encapsulation of Capsaicin into Chitosan-Oleic Acid Complex Particles: Evaluation of Encapsulation Ability and Stability.

Capsaicin (CAP) demonstrates a potential for application in the food and pharmaceutical industries owing to its various attractive health benefits, including anti-cancer, anti-inflammatory, and antioxidant activities. However, the application of CAP is often limited by its low solubility in water, low bioavailability, and strong pungency. In this study, a simple one-step method for the stable encapsulation and dispersion of CAP in aqueous media was developed using polyelectrolyte complex particles formed by chitosan (CHI) and oleic acid (OA). Homogeneous particles with mean diameters below 1 µm were successfully prepared via spontaneous molecular complexation by mixing an aqueous solution of CHI with an ethanolic solution of OA and CAP. CAP was incorporated into the hydrophobic domains of the CHI-OA complex particles through hydrophobic interactions between the alkyl chains of OA and CAP. The factors affecting CAP encapsulation were investigated, and a maximum encapsulation yield of approximately 100% was obtained. The CHI-OA-CAP complex particles could be stored for more than 3 months at room temperature (22-26 °C) without resulting in macroscopic phase separation or degradation of CAP. We believe that our findings provide a useful alternative encapsulation technique for CAP and contribute to expanding its practical application.

Visualization of the morphological changes in the median nerve after carpal tunnel release using three-dimensional magnetic resonance imaging.

OBJECTIVES: This study aimed to compare the pre- and postoperative morphology of the median nerve using three-dimensional (3-D) MRI in patients with carpal tunnel syndrome (CTS). METHODS: We assessed 31 patients with CTS who underwent open carpal tunnel release and T2*-weighted MRI of the wrist preoperatively and at 6 months postoperatively. The median nerve morphology was evaluated on the basis of the cross-sectional areas (CSAs) and cross-sectional volumes (CSVs). The association between these MRI findings and nerve conduction studies was also evaluated. RESULTS: The mean preoperative CSA and CSV values at the proximal carpal tunnel level significantly decreased from 22.2 mm2 and 24.4 mm3 to 16.5 mm2 and 18.1 mm3, respectively, postoperatively. Median nerve swelling at the proximal carpal tunnel level was observed in 29 (94%) and 23 (74%) patients before and after surgery, respectively. The mean preoperative CSA and CSV values at the hamate level significantly increased from 9.9 to 12.3 mm2 and from 10.9 to 13.5 mm3 after surgery, respectively. Nerve narrowing at the hamate bone level was preoperatively observed in 28 (90%) patients and postoperatively in 21 (68%) patients. Preoperative CSA and CSV values at the proximal carpal tunnel were significantly associated with preoperative distal motor and sensory latency. CONCLUSIONS: Visual confirmation of the median nerve morphology using 3-D MRI is useful when considering postoperative recovery and explaining the nerve condition to the patients. KEY POINTS: • The 3-D morphology of the median nerve after carpal tunnel release can be delineated using 3-D MRI. • Preoperative swelling of the median nerve in the 2-D and 3-D planes reflects the severity of carpal tunnel syndrome. • Visual confirmation of the median nerve morphology is useful when considering median nerve recovery after carpal tunnel release and for explaining the condition of the nerve to patients.

T2 mapping of the median nerve in patients with carpal tunnel syndrome and healthy volunteers.

INTRODUCTION: We investigated the changes in MRI T2 mapping values in subjects with carpal tunnel syndrome (CTS) compared to healthy controls. METHODS: We enrolled 71 patients with CTS and 26 healthy controls. Median nerve T2 values were measured at the distal carpal tunnel, hamate bone, proximal carpal tunnel, and forearm levels. These were compared between patients and controls and correlated with median nerve cross-sectional area (CSA) and nerve conduction measurements. RESULTS: The mean T2 values at the proximal carpal tunnel levels were higher in the CTS group (56.7 ms) than in the control group (51.2 ms, P = .02) and also were higher than at the distal carpal tunnel (51.0 ms, P < .001) and forearm levels (47.6 ms, P < .001). T2 values were not significantly associated with CSA or nerve conduction measurements. DISCUSSION: T2 mapping of the carpal tunnel provides qualitative information on median nerve pathology but does not reflect CTS severity.

Protein-Stabilized Palm-Oil-in-Water Emulsification Using Microchannel Array Devices under Controlled Temperature.

Microchannel (MC) emulsification for the preparation of monodisperse oil-in-water (O/W) and water-in-oil-in-water (W/O/W) emulsions containing palm oil as the oil phase was investigated for application as basic material solid/semi-solid lipid microspheres for delivery carriers of nutrients and drugs. Emulsification was characterized by direct observation of droplet generation under various operation conditions, as such, the effects of type and concentration of emulsifiers, emulsification temperature, MC structure, and flow rate of to-be-dispersed phase on droplet generation via MC were investigated. Sodium caseinate (SC) was confirmed as the most suitable emulsifier among the examined emulsifiers, and monodisperse O/W and W/O/W emulsions stabilized by it were successfully obtained with 20 to 40 µm mean diameter (dm) using different types of MCs.

Effect of dehydrocholic acid conjugated with a hydrocarbon on a lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine.

The effects of bile acids, dehydrocholic acid (DHA) and DHA conjugated with a hydrocarbon (6-aminohexanoate; 6A-DHA) were evaluated using a lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). DOPC formed a homogenous thin membrane in presence or absence of the DHA, while 20 mol% 6A-DHA induced phase separation on the DOPC thin membrane. It was observed formation of a stomatocyte-like liposomes when these membranes were suspended in a basic solvent. Generally, liposome formation can be prevented by some bile acids. It was found that DHA and 6A-DHA did not disrupt liposome formation, while DHA and 6A-DHA perturbed the liposomal membrane, resulting in increased local-fluidity due to the bent structure of DHA and 6A-DHA. DHA and 6A-DHA showed completely different effects on the hydrophobicity of the boundary surface of DOPC liposome membranes. The steroidal backbone of DHA was found to prevent the insertion of water molecules into the liposomal membrane, whereas 6A-DHA did not show the same behavior which was attributed to its conjugated hydrocarbon.

Reliability of radiographic measurement of lateral capitellohumeral angle in healthy children.

This retrospective cohort study was designed to validate the reliability of measurement of the lateral capitellohumeral angle (LCHA), an index of sagittal angulation of the elbow, in healthy children. The results were compared to the Baumann angle (BA), which is a similar concept to LCHA.Sixty-two radiographs of the elbow in healthy children (range, 2-11 years) were reviewed by 6 examiners at 2 sessions. The mean value and reliability of the measurement of LCHA and BA were assessed. Intraobserver reliability and interobserver reliability were calculated using intraclass correlation coefficients (ICCs).The mean LCHA value was 45° (range, 22° to 70°) and the mean BA was 71° (range, 56° to 86°). The ICCs for intraobserver reliability of the LCHA measurements were almost perfect for 2 examiners, substantial for 3 examiners, and moderate for 1 examiner with a mean value of 0.77 (range, 0.57-0.95). For BA measurements, the ICCs were almost perfect for 1 examiner and substantial for 5 examiners with a mean value of 0.74 (range, 0.66-0.83). The ICCs for interobserver reliability between the first and second measurements were both moderate for LCHA (0.56 and 0.51) and for BA (0.52 and 0.50).LCHA showed almost the same reliability in measurement as BA, which is the gold standard assessment for coronal alignment of the elbow. LCHA showed moderate-to-good reliability in the evaluation of sagittal plane elbow alignment.

Sauvé-Kapandji procedure with headless compression screw in patients with rheumatoid arthritis.

OBJECTIVE: We examined the surgical outcomes of the Sauvé-Kapandji (S-K) procedure using a headless compression screw and a metal cancellous screw in patients with rheumatoid arthritis (RA). METHODS: This retrospective study included 41 RA patients who underwent the S-K procedure for distal radioulnar joint disorders with two screws: headless compression screws (HCS group, n = 20) and cannulated cancellous screws (CCS group, n = 21). Clinical and radiographic outcomes were assessed 1 year after surgery. Radiographic outcomes included bony union of the distal radioulnar joint (DRUJ), bone resorption around the screw, a screw back-out, and use of additional K-wire. We investigated any complications related to the screw head. RESULTS: All 20 patients in the HCS group showed bone fusion of the DRUJ. In the CCS group, an asymptomatic non-union was observed in one patient and additional K-wire was needed to stabilize the DRUJ in three patients. No patients complained of any complications related to the screw head in the HCS group, while the CCS group demonstrated the hardware protrusion in two patients who complained of tenderness or discomfort at the screw head. CONCLUSIONS: The use of a headless compression screw in the S-K procedure is useful in patients with RA.

Hydration-aggregation pretreatment for drastically improving esterification activity of commercial lipases in non-aqueous media.

We investigated a novel, simple method for activating lipases in non-aqueous reaction media. Lipase powders were suspended in n-fatty alcohols and were then hydrated by adding a small amount of water. A paste-like aggregate was recovered from the mixture followed by lyophilization for obtaining activated lipases as dry powders. Lipase activity was evaluated for esterification between myristic acid and methanol in n-hexane. The activated lipases exhibited high esterification activity depending on the experiment conditions during hydration-aggregation pretreatment such as the amount of added water, the temperature, the pH of added buffer solutions, and the carbon chain length of the n-fatty alcohols used as pretreatment solvents. Various commercial lipases from different origins could be activated by this method. Changes in lipase conformation induced by the hydration-aggregation pretreatment were studied based on fluorescence and Fourier-transform infrared spectroscopy.

Freeze-dryable lipid vesicles with size tunability and high encapsulation efficiency prepared by the multiple emulsification-solvent evaporation method.

We investigated the extent of potential applicability of our recently developed method for preparing lipid vesicles [T. Kuroiwa et al., J. Am. Oil Chem. Soc., 93 (2016) 421], designated as the multiple emulsification-solvent evaporation method, with the intention of controlling the vesicle diameter and achieving high entrapment efficiency for water-soluble compounds. Using this method, the diameter of lipid vesicles could be varied by selecting the methods for preparing the primary water-in-oil emulsion, which contained water droplets as templates for the internal water phases of lipid vesicles. We obtained lipid vesicles with mean diameters of 0.2-4.4µm from water-in-oil-in-water multiple emulsions after solvent evaporation. A high entrapment yield of calcein, a water-soluble fluorescent dye, into the lipid vesicles was obtained for each vesicle sample, depending on their diameter and the type of emulsifier added to the external water phase. The use of polymeric emulsifier was more effective in achieving a high entrapment yield. The obtained lipid vesicles were powderized via freeze-drying. Vesicles could be powderized while maintaining their original diameter, as confirmed by scanning electron microscopy. Furthermore, the powderized vesicles could be rehydrated and resuspended without significant change in their diameter. However, the entrapment yield of calcein decreased after freeze-drying and rehydration. The calcein leakage during the freeze-drying followed by rehydration could be suppressed by adding an appropriate amount of trehalose as a lyoprotectant.

Reliability and Validity of Radiographic Measurement of the Humerus-Elbow-Wrist Angle in Healthy Children.

This article was updated on May 4, 2017, because of a previous error. The proximal line drawn in Figure 1 was different from the line described in the Materials and Methods section, which reads "The proximal line was drawn at the level of the radial tuberosity, and the distal line was made at the level of the top of the radial bowing (Fig. 1)." The correct figure is presented in this version of the article. An erratum has been published: JBJS Open Access. 2017 May 26;2(2):e0012ER. BACKGROUND: We conducted a retrospective cohort study to evaluate the normal value, range, reliability, and validity of measurement of the humerus-elbow-wrist angle, an index of valgus-varus angulation of the elbow, in healthy children. This measurement has been used to assess postoperative radiographic results. METHODS: Radiographs of the elbow in 62 healthy children ranging from 2 to 11 years of age were reviewed by 6 examiners at 2 sessions. The mean value and the reliability of measurement of the humerus-elbow-wrist angle, the carrying angle, and the Baumann angle were assessed. Intraobserver and interobserver reliability were calculated with use of intraclass correlation coefficients (ICCs). To determine concurrent validity, the association between the humerus-elbow-wrist angle and carrying angle measurements was examined with use of Pearson correlation coefficients. RESULTS: The mean humerus-elbow-wrist angle value was 12.0° (range, 1° to 24°), and the mean carrying angle was 14.6° (range, 4° to 28°). The ICCs for intraobserver measurements of the humerus-elbow-wrist angle were almost perfect for 4 examiners and were substantial for 2 examiners, with a mean value of 0.85 (range, 0.73 to 0.94). The ICCs for interobserver reliability with regard to the first and second measurements of the humerus-elbow-wrist angle were both substantial (0.76 and 0.78). A significant association between the humerus-elbow-wrist angle and the carrying angle was observed, with the Pearson correlation coefficients ranging from 0.74 to 0.90 (p < 0.001). CONCLUSIONS: Measurement of the humerus-elbow-wrist angle demonstrated good reliability and validity. The humerus-elbow-wrist angle is a reliable radiographic measure of coronal alignment of the humerus and forearm.

Glomerular cytokine expression in murine lupus nephritis.

BACKGROUND: Aberrant expression of T helper cell (Th) cytokines is believed to play a central role in the pathogenesis of systemic lupus erythematosus (SLE). While the glomerulus is one of the major targets of lupus inflammation, little is known about the cytokine expression in glomeruli. The current study aimed to explore the profiles of Th cytokine gene expressions in isolated glomeruli of lupus-prone mice. METHODS: Glomeruli were purified from lupus-prone MRL/lpr mice using the magnetic microbead method. Expressions of cytokine genes representing the Th subset and FoxP3 were examined using real-time polymerase chain reaction. Serum levels of these cytokines were also measured by enzyme-linked immunosorbent assay. MRL/n mice were used as controls. Histologic glomerular damages were scored semiquantitatively. To examine the role of TNF-α in glomerular damage, we administered etanercept, a TNF-α antagonist, into the subjects. RESULTS: Glomerular gene expressions of TNF-α in lpr mice increased with week postpartum and reached statistically significant levels at 16 weeks compared with those of the glomeruli from control mice. Expressions of IFN-γ, IL-4 and FoxP3 also increased, but the difference was not significant. There was a significant increase in serum levels of TNF-α, IFN-γ, and IL-17 and decrease in those of IL-4. Among the genes examined, TNF-α significantly correlated with glomerular damage score. Administration of etanercept did not affect glomerular cytokine expressions or proteinuria and failed to ameliorate histologic glomerular damages. CONCLUSION: Our data suggest that Th1 cytokines, especially TNF-α, are dominantly expressed in the glomeruli of lupus-prone mice, but its pathophysiological role remains unclear.

Formulation and stabilization of nano-/microdispersion systems using naturally occurring edible polyelectrolytes by electrostatic deposition and complexation.

This review paper presents an overview of the formulation and functionalization of nano-/microdispersion systems composed of edible materials. We first summarized general aspects on the stability of colloidal systems and the roles of natural polyelectrolytes such as proteins and ionic polysaccharides for the formation and stabilization of colloidal systems. Then we introduced our research topics on (1) stabilization of emulsions by the electrostatic deposition using natural polyelectrolytes and (2) formulation of stable nanodispersion systems by complexation of natural polyelectrolytes. In both cases, the preparation procedures were relatively simple, without high energy input or harmful chemical addition. The properties of the nano-/microdispersion systems, such as particle size, surface charge and dispersion stability were significantly affected by the concerned materials and preparation conditions, including the type and concentration of used natural polyelectrolytes. These dispersion systems would be useful for developing novel foods having high functionality and good stability.

Elevated interleukin-10: a new cause of dyslipidemia leading to severe HDL deficiency.

BACKGROUND: Low high-density lipoprotein cholesterol (HDL-C) is a risk factor for coronary artery disease. Investigating mechanisms underlying acquired severe HDL deficiency in noncritically ill patients ("disappearing HDL syndrome") could provide new insights into HDL metabolism. OBJECTIVE: To determine the cause of low HDL-C in patients with severe acquired HDL deficiency. METHODS AND RESULTS: Patients with intravascular large B-cell lymphoma (n = 2), diffuse large B-cell lymphoma (n = 1), and autoimmune lymphoproliferative syndrome (n = 1) presenting with markedly decreased HDL-C, low low-density lipoprotein cholesterol (LDL-C), and elevated triglycerides were identified. The abnormal lipoprotein profile returned to normal after therapy in all 4 patients. All patients were found to have markedly elevated serum interleukin-10 (IL-10) levels that also normalized after therapy. In a cohort of autoimmune lymphoproliferative syndrome patients (n = 93), IL-10 showed a strong inverse correlation with HDL-C (R(2) = 0.3720, P < .0001). A direct causal role for increased serum IL-10 in inducing the observed changes in lipoproteins was established in a randomized, placebo-controlled clinical trial of recombinant human IL-10 in psoriatic arthritis patients (n = 18). Within a week of initiating subcutaneous recombinant human IL-10 injections, HDL-C precipitously decreased to near-undetectable levels. LDL-C also decreased by more than 50% (P < .0001) and triglycerides increased by approximately 2-fold (P < .005). All values returned to baseline after discontinuing IL-10 therapy. CONCLUSION: Increased IL-10 causes severe HDL-C deficiency, low LDL-C, and elevated triglycerides. IL-10 is thus a potent modulator of lipoprotein levels, a potential new biomarker for B-cell disorders, and a novel cause of disappearing HDL syndrome.