Hyperglycaemia augments lipopolysaccharide-induced reduction in rat and human macrophage phagocytosis via the endoplasmic stress-C/EBP homologous protein pathway.

BACKGROUND: Macrophage phagocytosis constitutes an essential part of the host defence against microbes and the resolution of inflammation. Hyperglycaemia during sepsis is reported to reduce macrophage function, and thus, potentiate inflammatory deterioration. We investigated whether high-glucose concentrations augment lipopolysaccharide-induced reduction in macrophage phagocytosis via the endoplasmic stress-C/EBP homologous protein (CHOP) pathway using animal and laboratory investigations. METHODS: Peritoneal macrophages of artificially ventilated male Wistar rats, divided into four groups based on target blood glucose concentrations achieved by glucose administration with or without lipopolysaccharide, were obtained after 24 h. Human macrophages were also cultured in normal or high glucose with or without lipopolysaccharide exposure for 72 h. Changes in the phagocytic activity, intranuclear CHOP expression, and intracellular Akt phosphorylation status of macrophages were evaluated. These changes were also evaluated in human macrophages after genetic knock-down of CHOP by specific siRNA transfection or resolvin D2 treatment. RESULTS: Lipopolysaccharide impaired phagocytosis, increased intranuclear expression of CHOP, and inhibited Akt phosphorylation in both rat peritoneal and human macrophages. Hyperglycaemic glucose concentrations augmented these changes. Genetic knock-down of CHOP restored phagocytic ability and Akt phosphorylation in human macrophages. Furthermore, resolvin D2 co-incubation restored the inhibited phagocytosis and Akt phosphorylation along with the inhibition of intranuclear CHOP expression in human macrophages. CONCLUSIONS: These findings imply that controlling endoplasmic reticulum stress might provide new strategies for restoring reduced macrophage phagocytosis in sepsis-induced hyperglycaemia.

Thromboelastometry-guided intraoperative haemostatic management reduces bleeding and red cell transfusion after paediatric cardiac surgery.

BACKGROUND: Thromboelastometric evaluation of coagulation might be useful for prediction and management of bleeding after paediatric cardiac surgery. We tested the hypothesis that the use of a thromboelastometry-guided algorithm for blood product management reduces blood loss and transfusion requirements. METHODS: We studied 78 patients undergoing paediatric cardiac surgery with cardiopulmonary bypass (CPB) for the initial 12 h after operation. Stepwise multiple linear regression was used to develop an algorithm to guide blood product transfusions. Thereafter, we randomly assigned 100 patients to conventional or algorithm-guided blood product management, and assessed bleeding and red cell transfusion requirements. RESULTS: CPB time, post-bypass rotational thromboelastometry (ROTEM(®)) EXTEM amplitude at 10 min (A10), and FIBTEM-A10 were independently associated with chest tube drainage volume during the initial 12 h after operation. Discriminative analysis determined cut-off values of 30 mm for EXTEM-A10 and 5 mm for FIBTEM-A10, and estimated optimal intraoperative fresh-frozen plasma and platelet concentrate transfusion volumes. Thromboelastometry-guided post-bypass blood product management significantly reduced postoperative bleeding (9 vs 16 ml kg(-1), P<0.001) and packed red cell transfusion requirement (11 vs 23 ml kg(-1), P=0.005) at 12 h after surgery, and duration of critical care stay (60 vs 71 h, P=0.014). CONCLUSIONS: Rotational thromboelastometry-guided early haemostatic intervention by rapid intraoperative correction of EXTEM-A10 and FIBTEM-A10 reduced blood loss and red cell transfusion requirements after CPB, and reduced critical care duration in paediatric cardiac surgical patients. CLINICAL TRIAL REGISTRATION: UMIN Clinical Trials Registry UMIN000006832 (December 4, 2011).

Effects of JNK1/2 on the inflammation cytokine TNF-α-enhanced production of MMP-3 in human dental pulp fibroblast-like cells.

AIM: To investigate the effects of the c-Jun N-terminal kinase (JNK1/2) on the inflammation cytokine tumour necrosis factor-alpha (TNF-α)-enhanced production of matrix metalloproteinase-3 (MMP-3) in human dental pulp fibroblast-like cells (HPFs). METHODOLOGY: HPFs were grown from pulp explants from healthy donors. Primary cultures were established by culturing the cells for 20 to 30 days. The experiments with HPFs were performed between passages 3 and 10. The HPFs were incubated in serum-free medium containing TNF-α for 24 h. The medium in each well was prepared in SDS sample buffer and was analysed for MMP-3 by Western blotting. RESULTS: JNK inhibitor SP601245 markedly inhibited the production of MMP-3 in TNF-α-stimulated human dental pulp fibroblasts. MMP-3 production was enhanced by TNF-α in HPFs; silencing JNK1 and JNK2 expression inhibited this activation. cAMP response element-binding protein (CREB) was activated by TNF-α in HPFs; silencing JNK1 and JNK2 expression inhibited this activation. CONCLUSION: The activation of CREB via JNK pathways in the presence of TNF-α occurred with enhancement of MMP-3 production in dental pulp fibroblasts.

Assessment of speech production with dentures by electromagnetic articulography.

This report investigates the use of electromagnetic articulography (EMA) to compare basic speech patterns between a patient with traditional dentures to those of a normally dentate person. The goal is to assess the efficacy of traditional dentures in order to generate clinical data and works towards the improvement of denture design. Kinematic and acoustic data were acquired for these two subjects using a variety of repetitive vowel-consonant-vowel tasks. Spatiotemporal parameters indicating dynamic properties of the tongue blade and jaw movements, and timing coordination of the movements between them and with the output acoustic signal, were measured and compared within and between the participants. The results show significant differences in both spatial and temporal patterns and variation between individual tasks within each subject's data, as well as a difference in the two subjects' performance of the same task (cross-subject) for select calculated kinematic and latency parameters. It is concluded that there is more variation in spatiotemporal parameters in speech patterns for patients with dentures than without; in particular, latencies of the tongue blade and jaw movements and acoustic landmarks of the consonants, show strategies of movements timing coordination, typical of the speaker with denture.

Postnatal treatment factors affecting craniofacial morphology of unilateral cleft lip and palate (UCLP) patients in a Japanese population.

We have evaluated the craniofacial morphology of Japanese patients with unilateral cleft lip and palate (UCLP) and assessed the various postnatal factors that affect it. Lateral cephalograms of 140 subjects (mean (SD) aged 7 (2) years) with UCLP were taken before orthodontic treatment. Surgeons from Hokkaido University Hospital had done the primary operations. The craniofacial morphology was assessed by angular and linear cephalometric measurements. Cheiloplasty, palatoplasty, and preoperative orthopaedic treatment were chosen as postnatal factors. To compare the assessments of the postnatal factors, we made angular and linear cephalometric measurements for each subject and converted them into Z scores in relation to the mean (SD) of the two variables. Subjects treated by the modified Millard cheiloplasty had larger sella-nasion-point A (SNA) and nasion-point A-pogonion (NA-POG) measurements than subjects treated by the modified Millard with a vomer flap cheiloplasty. Two-stage palatoplasty showed consistently better craniofacial morphology than the other palatoplasty. Subjects who had preoperative orthopaedic treatment with a Hotz plate had significantly larger upper incisor/sella-nasion (U1-SN) measurements than who had no preoperative orthopaedic treatment or an active plate. We conclude that in subjects treated by a modified Millard type of cheiloplasty, a two-stage palatoplasty, and a Hotz plate there were fewer adverse effects on craniofacial morphology.

Histological study of the nasal septal cartilage in BALB/c-bm/bm mouse which spontaneously induces malocclusion.

OBJECTIVES: The BALB/c-bm/bm mouse is characterized by short limbs and short tail attributed to undersulfated glycosaminoglycans. Anterior transverse crossbite sometimes spontaneously appears in BALB/c-bm/bm mice. The BALB/c-bm/bm mouse shows a short nose and cranium. The reason for hypo-growth of anterior craniofacial structures has not been clarified, although the nasal septal cartilage might be related to the growth of anterior craniofacial structures. Therefore, the purpose of this study was to evaluate histological findings of the nasal septal cartilage at the border region of the ethmoid and sphenoid bone in BALB/c-bm/bm mice. MATERIALS AND METHODS: BALB/c mice (wild type) and BALB/c-bm/bm mice with normal occlusion (bm/bm) were used. Sagittal sections of female mice aged 2, 4, and 8 weeks were stained with hematoxylin and eosin for histological analysis. RESULTS: At the border region between the nasal septal cartilage and the ethmoid bone in bm/bm, the area of proliferative zone was significantly smaller than that in wild type. At the border regions between the nasal septal cartilage and both the ethmoid and sphenoid bones, the number of proliferative chondrocytes was significantly smaller. Normal endochondral ossification was not observed at the border region between the nasal septal cartilage and the sphenoid bone in bm/bm. CONCLUSION: The findings suggest that disorder of endochondral ossification in the nasal septal cartilage contributes to the hypo-growth of anterior craniofacial structures in bm/bm.

Morphological evaluation of cranial and maxillary shape differences of the brachymorphic mouse with spontaneous malocclusion using three-dimensional micro-computed tomography.

OBJECTIVES: The aim of this study was to determine whether significant cranial and maxillary deformity exists in BALB/c-bm/bm (brachymorphism) mouse with spontaneous malocclusion using three-dimensional (3D) images. MATERIALS AND METHODS: Thirty female mice were divided into the following three groups: control group (BALB/c mice, n = 10), Norm group (BALB/c-bm/bm mice with normal occlusion, n = 10), and Mal group (BALB/c-bm/bm mice with malocclusion, n = 10). Nine points in the skull were selected, and transverse and antero-posterior distances were measured using three-dimensional images of micro-computed tomography (CT). Moreover, 3D images were superimposed at the median plane to visualize the skull shape asymmetry. RESULTS: The transverse distances at the posterior cranial and maxillary region and the antero-posterior distances in the Norm and Mal groups were significantly shorter than those in the control group. The nasal septum of the Mal group was significantly shorter than that of the Norm group. Morphological measurements and superimposed 3D images showed that lateral deviation occurred at the anterior cranial and maxillary region in the Mal group. CONCLUSION: The 3D micro-CT images revealed that the antero-posterior length and posterior transverse width at the cranium and maxilla in BALB/c-bm/bm mice were significantly smaller than those in BALB/c mice. It was quantitatively and morphologically clear that BALB/c-bm/bm mice show a spontaneous transverse crossbite owing to lateral deviation of the maxilla and nasal bone.

Emdogain stimulates matrix degradation by osteoblasts.

Emdogain has been used clinically for periodontal regeneration, although the underlying molecular mechanisms are not clear at present. In this study, we hypothesized that Emdogain stimulated degradation of type I collagen via osteoblasts. We showed that Emdogain enhanced cell-mediated degradation of type I collagen in an MMP-dependent manner. Although MG-63 cells spontaneously produced a zymogen form of MMP-1, treatment with Emdogain significantly induced the generation of the active form of this enzyme. We demonstrated that MMP-3 was produced from MG63 cells in response to Emdogain in a MEK1/2-dependent manner. Concomitantly, blocking of MEK1/2 activation by U0126 significantly inhibited the generation of the active form of MMP-1 without affecting the total production of this collagenase. These results suggest that Emdogain facilitates tissue regeneration through the activation of the collagenase, MMP-1, that degrades matrix proteins in bone tissue microenvironments.

Platelets treated with ticlopidine are less reactive to unusually large von Willebrand factor multimers than are those treated with aspirin under high shear stress.

Much attention has recently been focused on the interaction between unusually large von Willebrand factor multimers (UL-VWFM) and platelets under high shear stress in pathological thrombus formation. The antiplatelet drugs acetylsalicylic acid (aspirin) and a thienopyridine derivative (ticlopidine) are commonly used to treat cerebral ischemia but exert different effects on high-shear-stress-induced platelet aggregation (H-SIPA) in the plasma. To examine the effects of these drugs in the absence of plasma factors, we studied H-SIPA using washed platelets (WPs) and purified UL-VWFM. WPs were prepared from the blood of 9 aspirin-treated and 11 ticlopidine-treated patients with cerebral ischemia, and H-SIPA in the presence of UL-VWFM was measured using a cone plate aggregometer. Plasma levels of VWF antigen with its multimer analysis, ristocetin cofactor and VWF-cleaving protease (ADAMTS13) activity were also measured. Forty-six healthy volunteers from 2 age groups, 20-40 years (n=20) and 41-60 years old (n=26), were also tested as controls. H-SIPA was significantly inhibited for ticlopidine-treated platelets, but it was observed to a lesser extent for aspirin-treated platelets. For both groups, no difference in the plasma levels of VWF antigen, ristocetin cofactor and ADAMTS13 activity was noted. All patients possessed UL-VWFM, and it was detected in healthy volunteers with increasing frequency with increasing age. Under plasma-free conditions, platelets from aspirin-treated patients exhibit marginal but significant inhibition of H-SIPA. Furthermore, the presence of UL-VWFM in the plasma of patients and normal volunteers is directly related to their age rather than being a consequence of underlying disease.

Clinical features and management of brain arteriovenous malformations in elderly patients.

BACKGROUND: Brain arteriovenous malformations (AVMs) of the elderly have not received sufficient attention, given the increase in age of individuals in recent years. We therefore designed a retrospective study to clarify features of brain AVMs in this age group in comparison with their counterparts in the general population. METHODS: A retrospective study was performed, based on data for AVMs treated in Nara Medical University Hospital and affiliated hospitals over the past 13 years. The series included all cases of brain AVMs, except for pure dural AVMs, diagnosed from June 1989 to June 2003. A total of 175 patients were diagnosed as having an AVM during this period, including 32 patients more than 60 years old. Clinical features and effective treatment of brain AVMs in those over and under 60 were explored and outcome at 3 to 6 months after surgery was evaluated according to a modified neurological scale. FINDINGS: The most common mode of presentation was intracranial hemorrhage in both groups, and this was remarkable in the elderly. Epilepsy at presentation was less frequent in the elderly (P< 0.05). In the elderly group infratentorial lesions were encountered more frequently (P< 0.05). Good or excellent outcomes of surgery were accomplished in 82.6% of the non-elderly group, and in 69.6% of the elderly group. When restricted to the grades I or II of Spetzler and Martin (S & M) grading, postoperative neurological scores of both groups were significantly better than preoperative values (P < 0.01). In the grade III cases, the non-elderly demonstrated significant improvement after surgery (P <0.01), but the elderly did not. INTERPRETATION: Elderly patients with a brain AVM had clinical features of less frequent epileptic presentation and more frequent infratentorial lesions. It was suggested that surgery was acceptable in elderly patients with pallial AVMs of grade I and II. Surgery for grade III AVMs of the elderly remains to be clarified.

Static-dynamic friction transition of FRP esthetic orthodontic wires on various brackets by suspension-type friction test.

A new testing apparatus for the measurement of frictional properties was designed and the frictional coefficients were obtained and compared with each other in various combinations of brackets and orthodontic wires, including esthetic fiber-reinforced plastic (FRP) wire that was especially designed and manufactured. Three kinds of wires (stainless steel, nickel-titanium, and FRP) and four brackets (single-crystal alumina, polycrystalline alumina, polycarbonate, and stainless steel) were used. The testing was done under dry and wet conditions. The friction testing equipment was designed to attach the bracket to a C-shaped bar suspended with a variable mass, and sliding along a fixed wire. The transition between static and dynamic friction was measured as a breakaway force, with the use of a universal test machine. In addition to material properties, this testing fixture eliminates geometrical factors, such as the rotational moment at the edge of the bracket slot, deflection of the orthodontic wire, and tension of the ligature wire. Nearly ideal frictional properties between materials are obtained. The frictional properties of FRP wire were similar to those of metal wires on all brackets, except the polycrystalline alumina bracket. The frictional coefficient between the polycrystalline ceramic bracket and FRP wire was larger than that of other combinations. There was little difference in frictional coefficients between dry and wet conditions.

Association between minor physical anomalies and lateral ventricular enlargement in childhood and adolescent onset schizophrenia.

OBJECTIVE: The primary purpose of this study was to investigate the association between morphological abnormalities of brain and minor physical anomalies (MPAs) in childhood and adolescent onset schizophrenia. METHOD: Twenty-seven patients who had been diagnosed with schizophrenia according to DSM-IV criteria before 18 years of age were included in the study. MPAs were evaluated with the modified version of Waldrop scale (WS) by Green et al. Morphological abnormalities of brain was evaluated with ventricular-brain ratio (VBR) by using cerebral magnetic resonance imaging (MRI) examination. RESULTS: A significant positive correlation was observed between WS scores and VBRs. CONCLUSION: This result indicates a relationship between MPAs and lateral ventricular enlargement, and supports neurodevelopmental etiology in childhood or adolescent onset schizophrenia.

Melanoma chondroitin sulfate proteoglycan regulates matrix metalloproteinase-dependent human melanoma invasion into type I collagen.

Tumor cell adhesion and proteolysis of the extracellular matrix proteins surrounding the cells are tightly linked processes in tumor invasion. In this study, we sought to identify components of the cell surface of a vertical growth phase melanoma cell line, WM1341D, that mediate invasive cellular behavior. We determined by antisense inhibition that melanoma chondroitin sulfate proteoglycan (MCSP) and membrane-type 3 matrix metalloproteinase (MT3-MMP) expressed on WM1341D are required for invasion of type I collagen and degradation of type I gelatin. MT3-MMP co-immunoprecipitated with MCSP in WM1341D melanoma cells cultured on type I collagen or laminin. The association between MT3-MMP and MCSP was largely disrupted by removing chondroitin sulfate glycosaminoglycan (CS) from the cell surface, suggesting CS could mediate the association between the two cell surface core proteins. Recombinant MT3-MMP and MT3-MMP from whole cell lysates of WM1341D cells were specifically eluted from CS- conjugated affinity columns. The results indicate that MT3-MMP possesses the potential to promote melanoma invasion and proteolysis and that the formation of a complex between MT3-MMP and MCSP may be a crucial step in activating these processes.

Presence of third molar germs in orthodontic patients in Japan.

The purpose of this investigation was to evaluate the existence of third molar germs in orthodontic patients in Japan and to examine the relationship between the existence of third molars and sagittal maxillomandibular jaw relationships. The subjects comprised 306 patients from the orthodontic clinic of Hokkaido University Dental Hospital who were younger than 15 years. The subjects were divided into 2 groups: 1 group included 144 patients who were born between 1966 and 1969 (60s group), and the other group included 162 patients who were born between 1980 and 1987 (80s group). Assessments were made from panoramic radiographs and lateral cephalograms. The following results were obtained: (1) all 4 third molar germs were present in 77% of subjects, (2) mandibular third molars were present significantly more often than maxillary third molars, and (3) the percentage of skeletal Class III subjects who had all 4 third molars was lower than that of skeletal Class II subjects. The chi(2) test was used to determine statistical significance in differences.

Isolation and characterization of a new immortal rat astrocyte with a high expression of NGF mRNA.

We have established a new line of immortalized rat astrocytes through transfection of plasmid pSV3-neo encoding the large T antigen of simian virus 40 into normal astrocytes. One of these immortalized astrocytes (ACT-57) with a flat and polygonal cell shape, exhibited stable growth in a chemically defined medium (modified N-2 medium) as well as in medium containing ordinary serum. ACT-57, retained a detectable level of expression of glial fibrillary acidic protein (GFAP) and its mRNA, and exhibited a stronger expression of nerve growth factor (NGF) mRNA than that of normal rat astrocytes or C6 glioma cells. NGF mRNA was significantly up-regulated by phorbol ester (12-O-tetradecanoylphorbol 13-acetate, TPA) and gamma-amino-n-butyric acid (GABA) but not by hydrocortisone. None of stimulants (TPA, dibutyryl cyclic AMP (db-cAMP), hydrocortisone, L-glutamate, carbacol, GABA, dopamine, or isoproterenol) changed the expression level of either brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3). There was a discrete difference between ACT-57 and normal astrocytes in the response to GABA and isoproterenol. These findings imply that normal cortical astrocytes possess a functional heterogeneity whereas the clonal astrocyte, ACT-57, does not, indicating that ACT-57 cells may be useful for in vitro studies of neuron-astrocyte interactions involving the induction of neurotrophic factors such as NGF.

Crystal structure and charge distribution of YbFeMnO4.

The structure of synthetic YbFeMnO(4) has been refined by single-crystal X-ray diffraction. Space group R3m, a = 3.4580 (1), c = 25.647 (3) A, V = 265.59 (3) A(3), Z = 3. Yb is in octahedral coordination, whereas Fe and Mn are disordered on a single crystallographic type of trigonal bipyramid, in which the cation is off-centred from the basal plane. Assuming perfect stoichiometry, R(1) = 0.0195, but the charge distribution (CD) analysis suggests incomplete occupation of the Yb site. Refinement of the occupancy lowers R(1) to 0.0175, resulting in s.o.f.(Yb) = 0. 963 (3), with a significant improvement of the Fourier difference. The electroneutrality is likely preserved through incomplete occupancy of one of the two oxygen sites: the compound is thus non-stoichiometric, with the formula Yb(0.963)FeMnO(3.945). Another mechanism for preserving the electroneutrality is the oxidation of a small amount of Mn(2+) to Mn(3+), which is, however, less probable because of the reduction conditions in which the sample was synthesized. Both models give a satisfactorily CD result, but they cannot be definitively distinguished by X-ray data.

Brain imaging in childhood- and adolescence-onset schizophrenia associated with obsessive-compulsive symptoms.

OBJECTIVE: Childhood- and adolescence-onset schizophrenic patients with obsessive-compulsive symptoms (OCS) constitute a specific subgroup of schizophrenia. We performed magnetic resonance imaging in this group seeking evidence of neurodevelopmental insults. METHOD: Thirty-two schizophrenic patients were compared with 19 controls. Schizophrenic subjects were divided into 15 patients with OCS (SOCS+ group; onset at 15.5 +/- 1.6 years) and 17 without OCS (SOCS- group; onset at 15.3 +/- 1.3 years). Areas of the hippocampus, frontal lobe, corpus callosum and putamen were analysed morphometrically. RESULTS: The left hippocampus was significantly smaller in the SOCS+ group than in the SOCS- and control groups. CONCLUSION: Reduced size of the left hippocampus in the SOCS+ group supports a neurodevelopmental etiology in this subgroup.

Spinal dural arteriovenous fistula with perimesencephalic subarachnoid haemorrhage.

A case is reported of a 66 year old woman presenting with perimesencephalic subarachnoid haemorrhage (SAH) which was caused by a spinal dural arteriovenous fistula at the C1 level. The fistula drained into the venous system of the posterior cranial fossa through a perimedullary vein. The bleeding was thought to result from venous hypertension induced by the fistula. This case may support the hypothesis that perimesencephalic non-aneurysmal SAH can be ascribed to venous bleeding and that venous hypertension is the key to its pathology.

Ser787 in the proline-rich region of human MAP4 is a critical phosphorylation site that reduces its activity to promote tubulin polymerization.

p34cdc2 kinase-phosphorylation sites in the microtubule (MT)-binding region of MAP4 were determined by peptide sequence of phosphorylated MTB3, a fragment containing the carboxy-terminal half of human MAP4. In addition to two phosphopeptides containing Ser696 and Ser787 which were previously indicated to be in vivo phosphorylation sites, two novel phosphopeptides, containing Thr892 or Thr901 and Thr917 as possible phosphorylation sites, were isolated, though only in in vitro phosphorylation. The role of phosphorylation at Ser696 and Ser787, which were differently phosphorylated during the cell cycle (Ookata et al., (1997). Biochemistry, 36: 15873-15883), was investigated in MT-polymerization, using MAP4 Ser to Glu mutants, which mimic phosphorylation at each site. Mutation of Ser787 to Glu strikingly reduced the MAP4's MT-polymerization activity, while Glu-mutation at Ser696 did not. These results suggest that Ser787 could be the critical phosphorylation site causing MTs to be dynamic at mitosis.