Chitosan Hydrogels Crosslinked with Oxidized Sucrose for Antimicrobial Applications.

Oxidized sucrose (OS) reacts with amino-group-containing polysaccharides, including chitosan, without catalyst, resulting in hydrogels entirely composed of carbohydrates. The presence of imine bonds with low structural stabilities and unreacted aldehydes in the structures of these hydrogels hinder their application as biomaterials. Therefore, herein, the chitosan hydrogels (CTSGs) obtained after the crosslinking of chitosan with OS were reduced using sodium borohydride to convert imine bonds to secondary amines and aldehydes to alcohols. The structures of CTSGs were comprehensively characterized using Fourier transform infrared and 13C nuclear magnetic resonance spectroscopies, and the results implied that the degree of crosslinking (CR) depended on the OS feed amount used during CTSG preparation. The properties of CTSGs were significantly dependent on CR; with an increase in CR, the thermal stabilities and dynamic moduli of CTSGs increased, whereas their swelling properties decreased. CTSGs exhibited antimicrobial properties against the gram-negative bacterium Escherichia coli, and their performances were also dependent on CR. The results indicated the potentials of CTSGs completely based on carbohydrates as antimicrobial hydrogels for various medical and pharmaceutical applications. We believe that this study will contribute to the development of hydrogels for application in the food, medical, and pharmaceutical fields.

Preparation and characterization of carboxymethylated Aureobasidium pullulans ß-(1 → 3, 1 → 6)-glucan and its in vitro antioxidant activity.

Aureobasidium pullulans ß-(1 â†’ 3, 1 â†’ 6)-glucan (APG) has a high degree of ß-(1 â†’ 6)-glucosyl branching and a regular triple helical structure similar to that of schizophyllan. In this study, APG was carboxymethylated to different degrees of substitution (DS = 0.51, 1.0, and 2.0, denoted CMAPG 1-3, respectively) using a heterogeneous reaction. With increasing DS, the triple-helix structure drastically decreased and converted to a random coil structure in CMAPG 3. Further, aqueous solutions of CMAPG changed from pseudoplastic fluids to perfect Newtonian liquids with increasing DS, indicating that the intra- and intermolecular hydrogen bonds had been cleaved by the substituents to form a random coil structure. In addition, APG and CMAPG solutions exhibited scavenging ability against hydroxyl, organic, and sulfate radicals. It was also found that the carboxymethylation of APG drastically enhanced the organic radical scavenging ability. On the basis of the relationship between the DS and radical scavenging ability of the CMAPG samples, we believe hydroxyl and organic radicals were preferably scavenged by the donation of hydrogen atoms from the glucose rings and the methylene moieties of the carboxymethyl groups, respectively. Considering the obtained results, CMAPG and APG are expected to have applications in pharmaceuticals, functional foods, and cosmetics as antioxidant polysaccharides.

Orai2 channel regulates prostaglandin E2 production in TNFα/IL1α-stimulated astrocytes.

Astrocytes are glial cells that serve homeostatic functions in the central nervous system (CNS). Recent research, however, suggests that under pathological conditions, astrocytes are stimulated by various factors and actively participate in CNS inflammation. In the present study, we found that astrocytes upregulate various inflammatory factors including prostaglandin E2 (PGE2 ) by co-stimulation with tumor necrosis factor-alpha (TNFα) and interleukin-1alpha (IL1α). These TNFα/IL1α-stimulated astrocytes also showed increased Ca2+ release from the endoplasmic reticulum (ER) and increased expression of Orai2, a member of the store-operated calcium channel (SOCC) family. To reveal the role of Orai2, we used astrocytes in which Orai2 was knocked-down (KD) or knocked-out (KO). The expression of the prostaglandin E synthase Ptges and the production of PGE2 were higher in Orai2-KD astrocytes than in WT astrocytes when stimulated with TNFα and IL1α. Orai2-KO astrocytes also showed increased expression of Ptges and increased PGE2 production. The expression of Ptgs2, another PGE2 synthetic enzyme, was also upregulated in Orai2-KO astrocytes. Moreover, Orai2-KO astrocytes showed increased store-operated calcium entry (SOCE) and increased Orai1 expression. These results suggest that Orai2 is upregulated in TNFα/IL1α-stimulated astrocytes and reduces PGE2 production to some extent, modulating CNS inflammation. Our findings may aid in understanding how astrocytes are associated with inflammatory responses, and the identification of new targets that modulate astrocytic reactivity.

Preparation and Enzyme Degradability of Spherical and Water-Absorbent Gels from Sodium Carboxymethyl Cellulose.

To synthesize a biodegradable alternative to spherical polyacrylic acid absorbent resin, spherical hydrogel particles were prepared from carboxymethyl cellulose (CMC) dissolved in an aqueous solution, using ethylene glycol diglycidyl ether (EGDE) as a crosslinking agent. The effect of varying the initial CMC concentration and feed amount of EGDE on the shape, water absorbency, water-holding capacity, and enzyme degradability of the resultant CMC hydrogels was determined. The reaction solution was poured into fluid paraffin, and spherical hydrogel particles were obtained via the shear force from stirring. The shape and diameter of the spherical hydrogel particles in the swollen state depended on the CMC concentration. The spherical hydrogel particles obtained by increasing the amount of EGDE resulted in a decrease in absorbency. Additionally, all the spherical hydrogel particles were degraded by cellulase. Thus, spherical biodegradable hydrogel particles were prepared from CMC, and the particle size and water absorption of the hydrogel could be controlled in the range of 5-18 mm and 30-90 g·g-1 in the swollen state, respectively. As an alternative to conventional superabsorbent polymers, the spherical CMC hydrogels are likely to be useful in industrial and agricultural applications.

NMR characterization of cellulose acetate: Mole fraction of monomers in cellulose acetate determined from carbonyl carbon resonances.

Cellulose acetate (CA) samples with varying degrees of substitution were prepared via homogeneous acetylation in 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) and by the acid-hydrolysis of cellulose triacetate in acetic acid. Quantitative analysis of the 13C NMR spectra facilitated the assignment of the carbonyl carbon shifts of the 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted anhydroglucose units (AGUs), and the determination of the mole fraction of 7 AGUs and unsubstituted AGU in the CA chains. This shed some light on the mechanism of CA production in homogeneous reaction systems. In addition, comparison of the mole fractions of the 8 AGUs suggested that the acetone solubility of CA strongly related to the AGU composition.

NMR characterization of methylcellulose: Chemical shift assignment and mole fraction of monomers in the polymer chains.

Methylcellulose samples with different degrees of substitution were prepared by a heterogeneous reaction of cellulose. Two-dimensional NMR spectroscopy provided complete assignment of the 1H and 13C chemical shifts of the un-, 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted anhydroglucose units (AGUs). Comparative analysis of the chemical shift data revealed the relationship between the methyl substituents at the 2-, 3-, and 6-positions and the 1H and 13C chemical shifts of the AGUs; additivity could be applied to the 1H and 13C chemical shifts of methylcellulose. Quantitative 13C NMR spectra of the samples determined the composition of the eight AGUs and the substituent distribution at the 2-, 3-, and 6-positions of cellulose. This provided estimations of the hydroxyl group reactivity toward methylation and the interactions between the substituent groups within the same AGU.

Exercise pressor reflex function in female rats fluctuates with the estrous cycle.

In women, sympathoexcitation during static handgrip exercise is reduced during the follicular phase of the ovarian cycle compared with the menstrual phase. Previous animal studies have demonstrated that estrogen modulates the exercise pressor reflex, a sympathoexcitatory mechanism originating in contracting skeletal muscle. The present study was conducted in female rats to determine whether skeletal muscle contraction-evoked reflex sympathoexcitation fluctuates with the estrous cycle. The estrous cycle was judged by vaginal smear. Plasma concentrations of estrogen were significantly (P < 0.05) higher in rats during the proestrus phase of the estrus cycle than those during the diestrus phase. In decerebrate rats, either electrically induced 30-s continuous static contraction of the hindlimb muscle or 30-s passive stretch of Achilles tendon (a maneuver that selectively stimulates mechanically sensitive muscle afferents) evoked less renal sympathoexcitatory and pressor responses in the proestrus animals than in the diestrus animals. Renal sympathoexcitatory response to 1-min intermittent (1- to 4-s stimulation to relaxation) bouts of static contraction was also significantly less in the proestrus rats than that in the diestrus rats. In ovariectomized female rats, 17ß-estradiol applied into a well covering the dorsal surface of the lumbar spinal cord significantly reduced skeletal muscle contraction-evoked responses. These observations demonstrate that the exercise pressor reflex function and its mechanical component fluctuate with the estrous cycle in rats. Estrogen may cause these fluctuations through its attenuating effects on the spinal component of the reflex arc.

Numerous FUS-positive inclusions in an elderly woman with motor neuron disease.

We report an autopsy case of a 75-year-old Japanese woman with motor neuron disease (MND) showing numerous neuronal and glial inclusions immunostained with anti-fused in sarcoma (FUS) antibody. At 73 years, she received a diagnosis of MND and died of respiratory insufficiency 2 years later. No mutation was found in all exons of the FUS gene. Neuropathological examination revealed a reduced number of anterior horn cells and degeneration of the pyramidal tracts. Neither Bunina bodies nor inclusions positive for ubiquitin/phosphorylated TAR DNA binding protein of 43 kD (pTDP-43), such as skein-like or round inclusions, were observed. However, basophilic inclusions (BIs) were frequently observed in the remaining neurons of the anterior horns, facial nuclei, hypoglossal nuclei, vestibular nuclei, dentate nuclei and inferior olivary nuclei. In an immunohistochemical analysis, the BIs showed strong immunoreactivity with anti-FUS and anti-ubiquitin-binding protein p62 (p62) antibodies. The nuclear staining of FUS was preserved in some neurons with FUS-positive inclusions, and a few FUS-positive glial inclusions were found. FUS-positive inclusions were more common than p62-positive inclusions in some anatomical regions, and in some neurons, p62 immunoreactivity was observed in only parts of the BIs. These results suggest that BI formation and TDP-43 aggregation have different pathogenic mechanisms, and FUS may play an important role in the pathogenesis of MND with BIs. This patient has the oldest reported age of onset for MND with BIs, and clinical features observed in this patient were indistinguishable from those of classic sporadic MND. Therefore, we consider that the age of onset and clinical features of FUS-related disorders may be variable.

"Green odor" inhalation by stressed rat dams reduces behavioral and neuroendocrine signs of prenatal stress in the offspring.

Chronic maternal stress during pregnancy results in the "prenatally stressed" offspring displaying behavioral and neuroendocrine alterations that persist into adulthood. We investigated how inhalation of green odor (a mixture of equal amounts of trans-2-hexenal and cis-3-hexenol) by stressed dams might alter certain indices of prenatal stress in their offspring. These indices were depression-like behavior (increased immobility time in the forced-swim test) and acute restraint stress-induced changes in hypothalamo-pituitary-adrenocortical (HPA) axis activity [plasma corticosterone (CORT) and ACTH levels and the number of Fos-immunoreactive cells in the hypothalamic paraventricular nucleus (an index of neuronal activity)]. Pregnant rats were exposed to restraint stress for 60 min/day for 10 days (gestational days 10-19). The prenatally stressed offspring exhibited significant increases in depression-like behavior and in restraint stress-induced ACTH, CORT, and Fos responses, unless their dam had been exposed to green odor. The behavioral effect of the odor was also seen in offspring that were fostered by unstressed dams. The results obtained in the dams themselves were as follows. In vehicle-exposed stressed dams, but not in green odor-exposed ones, total body and adrenal weights were significantly decreased or increased, respectively. Depression-like behavior was not observed in the vehicle-exposed stressed dams themselves. Green odor inhalation prevented the impairment of maternal behavior induced by restraint stress. Thus, exposure of dams to stress may affect both the fetal brain and fetal HPA axis, and also maternal behavior, leading to altered behavioral and neuroendocrine responses in the offspring. Such effects may be prevented by the stressed dams inhaling green odor.

Retrospective evaluation of craniofacial growth of Japanese children with isolated cleft palate: from palatoplasty to adolescence.

OBJECTIVES: The purposes of this study were to analyze the craniofacial growth in women with an isolated cleft palate, to compare their matured craniofacial form with that of women with normal occlusion, and to survey the factors that influenced the matured craniofacial morphology of the adults with cleft palate during their growth process. MATERIALS AND METHODS: Eighteen women with nonsyndromic isolated cleft palate were chosen from patients who received a palatoplasty at the Kyushu University Hospital, Fukuoka, Japan. Their lateral cephalometric radiographs were taken longitudinally from palatoplasty to adolescence. Fifty women with normal occlusion were chosen as controls. From their lateral cephalographs, linear and angular variables were calculated using the x,y coordinates of 20 skeletal landmarks. RESULTS: Maxillary length was shorter and the nasomaxillary complex was positioned more posteriorly in relation to the anterior cranial base of the adults with isolated cleft palate, compared with the controls. The mandible was shorter and was rotated inferiorly and posteriorly. However, remarkable deviation from the average craniofacial growth pattern was not recognized from palatoplasty to adolescence. The factors that influenced the craniofacial growth in the subjects with cleft palate were the forward growth of the A point from 2 to 5 years of age, the downward growth of the Ba point, the anterior upper facial height N-Ans in puberty, and the vertical position of the point Ba at the time of palatoplasty. CONCLUSION: This study provided evidence of the growth tendency and the factors influencing the intermaxillary relationship in subjects with isolated cleft palate. These are significant for orthodontic treatment planning.

Detection of increased icb-1 transcript levels in maturing HL-60 cells: a novel marker for granulocytic and monocytic in vitro differentiation.

Human gene icb-1 was initially described as a gene with increased expression in endometrial tumor cells differentiated in vitro by culture on a reconstituted basement membrane. We provide evidence for a more general involvement of icb-1 gene function in cellular differentiation processes. We report the up-regulation of icb-1 transcript levels in HL-60 promyelocytic leukemia cells during their in vitro differentiation induced by all-trans retinoic acid, vitamin D(3) or DMSO. Increased icb-1 mRNA levels could be observed both in monocytic and granulocytic differentiation. We also report the identification of the novel icb-1 splice variants icb-1beta and gamma, and the spleen-specific isoform icb-1delta. Expression of icb-1 can be used as a novel marker for in vitro differentiation processes of HL-60 cells.