IκBNS induces Muc5ac expression in epithelial cells and causes airway hyper-responsiveness in murine asthma models.

BACKGROUND: In allergic asthma, environmental allergens including house dust mite (HDM) trigger pattern recognition receptors and activate downstream signaling pathways including NF-κB pathways not only in immune cells but also in airway epithelial cells. Recent studies have shown that NF-κB activation is regulated positively or negatively depending on the cellular context by IκBNS (encoded by the gene Nfkbid), one of atypical IκB proteins, in the nucleus. Therefore, we hypothesized that IκBNS expressed in immune cells or epithelial cells is involved in the regulation of asthmatic responses. AIM: To determine the roles of IκBNS in HDM-induced asthmatic responses. METHODS: Roles of IκBNS in HDM-induced airway inflammation and airway hyper-responsiveness (AHR) were examined by using IκBNS-deficient (Nfkbid-/- ) mice. Roles of IκBNS expressed in hematopoietic cells and nonhematopoietic cells were separately evaluated by bone marrow chimeric mice. Roles of IκBNS expressed in murine tracheal epithelial cells (mTECs) were examined by air-liquid interface culture. RESULTS: House dust mite-induced airway inflammation and AHR were exacerbated in mice lacking IκBNS in hematopoietic cells. In contrast, HDM-induced airway inflammation was exacerbated, but AHR was attenuated in mice lacking IκBNS in nonhematopoietic cells. The induction of Muc5ac, a representative mucin in asthmatic airways, was reduced in Nfkbid-/- mTEC, whereas the induction of Spdef, a master regulator of goblet cell metaplasia, was not impaired in Nfkbid-/- mTEC. Moreover, IκBNS bound to and activated the MUC5AC distal promoter in epithelial cells. CONCLUSION: IκBNS is involved in inducing Muc5ac expression in lung epithelial cells and causing AHR in HDM-induced asthma models.

2-Chloroacetamidine, a novel immunomodulator, suppresses antigen-induced mouse airway inflammation.

BACKGROUND: Citrullination is a presently under-recognized posttranslational protein modification catalyzed by PAD enzymes. Immune responses to citrullinated neo-epitopes are identified in a growing number of inflammatory and autoimmune diseases. However, the involvement of hypercitrullination in the pathogenesis of bronchial asthma is still unknown. METHODS: As main experimental tool, we examined the effect of 2-chloroacetamidine (2CA), a PAD enzyme inhibitor, on OVA-immunized and airway-challenged BALB/c mice; a commonly used model of allergic airway inflammation. We also measured the effect of 2CA on ex vivo lymphocytes and cell lines. RESULTS: In vivo, 2CA dramatically suppressed lung tissue hypercitrullination, inflammatory cell recruitment, and airway-Th2 cytokine secretion. 2CA also suppressed systemic OVA-specific and total IgE production dramatically, effectively preventing de novo and diminishing established disease without measurably impacting general immunocompetence. In vitro, 2CA markedly inhibited the proliferation of mouse and human T cells with cell cycle block and apoptosis during a limited, postactivation phase. CONCLUSIONS: 2CA acts as narrow-spectrum immunosuppressant that selectively targets lymphocyte populations involved in active inflammatory tissue lesions. If hypercitrullination is generated in patients with asthma, 2CA may represent a novel disease modulator for human asthmatics/allergic diseases.

Additional strange hadrons from QCD thermodynamics and strangeness freezeout in heavy ion collisions.

We compare lattice QCD results for appropriate combinations of net strangeness fluctuations and their correlations with net baryon number fluctuations with predictions from two hadron resonance gas (HRG) models having different strange hadron content. The conventionally used HRG model based on experimentally established strange hadrons fails to describe the lattice QCD results in the hadronic phase close to the QCD crossover. Supplementing the conventional HRG with additional, experimentally uncharted strange hadrons predicted by quark model calculations and observed in lattice QCD spectrum calculations leads to good descriptions of strange hadron thermodynamics below the QCD crossover. We show that the thermodynamic presence of these additional states gets imprinted in the yields of the ground-state strange hadrons leading to a systematic 5-8 MeV decrease of the chemical freeze-out temperatures of ground-state strange baryons.

Strangeness at high temperatures: from hadrons to quarks.

Appropriate combinations of up to fourth order cumulants of net strangeness fluctuations and their correlations with net baryon number and electric charge fluctuations, obtained from lattice QCD calculations, have been used to probe the strangeness carrying degrees of freedom at high temperatures. For temperatures up to the chiral crossover, separate contributions of strange mesons and baryons can be well described by an uncorrelated gas of hadrons. Such a description breaks down in the chiral crossover region, suggesting that the deconfinement of strangeness takes place at the chiral crossover. On the other hand, the strangeness carrying degrees of freedom inside the quark gluon plasma can be described by a weakly interacting gas of quarks only for temperatures larger than twice the chiral crossover temperature. In the intermediate temperature window, these observables show considerably richer structures, indicative of the strongly interacting nature of the quark gluon plasma.

Antihypertensive effect of a fixed-dose combination of losartan/hydrochlorothiazide in patients with uncontrolled hypertension: a multicenter study.

BACKGROUND: Achieving adequate blood pressure (BP) control often requires more than one antihypertensive agent. The purpose of this study was to determine whether a fixed-dose formulation of losartan (LOS) plus hydrochlorothiazide (HCTZ) (LOS/HCTZ) is effective in achieving a greater BP lowering in patients with uncontrolled hypertension. METHODS: The study was a prospective, multicenter, observational trial exploring the antihypertensive effect of a single tablet of LOS 50 mg/HCTZ 12.5 mg. A total of 228 patients whose BP had previously been treated with more than one antihypertensive agents without having achieved BP goal below 130/80 mmHg enrolled in the study. RESULTS: A significant decrease in systolic and diastolic BP was observed in both clinic and home measurement after switching from the previous treatment to LOS/HCTZ. There was a significant decrease in both B-type natriuretic peptide (BNP) and urinary albumin creatinine (Cr) excretion ratio (ACR), especially in patients with elevated values. In contrast, there was a significant increase in serum Cr concentration in conjunction with a decrease in estimated glomerular filtration rate (eGFR). Overall serum uric acid (UA) concentration increased, whereas in patients with hyperuricemia there was a significant reduction in this value. CONCLUSION: Switching to LOS/HCTZ provides a greater reduction in clinic and home BP in patients with uncontrolled hypertension. This combination therapy may lead to cardio-, reno protection and improve UA metabolism.

Quantitative analysis of the spinal cord motoneuron under chronic compression: an experimental observation in the mouse.

We investigated quantitative changes in spinal cord motoneurons following chronic compression using a mouse model of cervical cord compression. Twenty-five tip-toe-walking Yoshimura (twy) mice with calcified mass lesions compressing the spinal cord posterolaterally at the C1-C2 vertebral levels were compared with five Institute of Cancer Research (ICR) mice that served as controls. Spinal cord motoneurons in the anterior grey horn between the C1 and C3 spinal cord segments were Nissl-stained and counted topographically and then analysed in relation to the extent of spinal cord compression. The number of motoneurons in C1-C3 spinal cord segments decreased significantly with a linear correlation with the transverse area of the spinal cord when the cord was compressed to 50-70% of control values. A significant reduction in the number of motoneurons occurred at the C2-C3 spinal cord segment compressed at the C1-C2 vertebral level. In contrast, at the level rostral to the C1 vertebra, the number of motoneurons increased significantly in proportion to the magnitude of compression. The current study demonstrates that a number of neurons, morphologically consistent with anterior horn cells, were observed at a rostral site absolutely free of external compression where no such cells normally exist.

Lordotic alignment and posterior migration of the spinal cord following en bloc open-door laminoplasty for cervical myelopathy: a magnetic resonance imaging study.

We investigated lordotic alignment and posterior migration of the spinal cord following en bloc open-door laminoplasty for cervical myelopathy. Fifty-five patients (32 men and 23 women) were studied, with an average follow-up of 2.4 years. Radiological examination included evaluation of lordosis of the cervical spine and spinal cord, degree of enlargement of bony spinal canal, and the magnitude of posterior cord migration. We also correlated these changes with neurological improvement. Postoperatively, there was an average of 5% loss of cervical spine lordosis (P > 0.01) on radiographs and 12% reduction in the lordotic alignment of the spinal cord (P > 0.05) on magnetic resonance imaging. Postoperatively, the size of the bony spinal canal increased by 48%. Posterior cord migration showed a significant correlation with the preoperative cervical spine and spinal cord lordosis (P < 0.05). Thirty-seven (67%) patients with neurological improvement exceeding 50% showed significant posterior cord migration following laminoplasty compared with those demonstrating less than 50% improvement (P = 0.01). Our results suggest that a significant neurological improvement is associated with posterior cord migration after cervical laminoplasty.

Plasticity of the spinal cord contributes to neurological improvement after treatment by cervical decompression. A magnetic resonance imaging study.

To investigate the relationship between morphological plasticity of the spinal cord and neurological outcome after surgery for compressive lesions, we correlated the transverse area of the cervical spinal cord measured by transaxial magnetic resonance imaging (MRI) obtained during the early postoperative period (1-6 months) with neurological function assessed at a median postoperative follow-up period of 2.5 years. Measurements on MRI in 56 patients (35 men and 21 women) included evaluation of the cross-sectional area of the cervical cord and the subarachnoidal space at the level of decompression. The transverse area of the cervical cord increased by 30 to 62% postoperatively and that of the subarachnoidal space by 57 to 95%. Neurological improvement was noted in all patients and averaged 63% in our assessment scale. Expansion of the cervical cord during the early postoperative period correlated significantly with the late postoperative neurological status (P = 0.009). Our results suggest that an increase in the cross-sectional area of the cervical spinal cord, representing spinal cord morphological plasticity, is a significant factor in determining the late neurological improvement following decompressive surgery.

Histological investigation of spinal cord lesions in the spinal hyperostotic mouse (twy/twy): morphological changes in anterior horn cells and immunoreactivity to neurotropic factors.

We examined the morphology of spinal accessory motoneurons and immunoreactivity to neurotrophins, brain-derived neurotropic factor (BDNF) and neurotrophin (NT)-3, as well as the presence of reactive astrocytosis in 70 tiptoe walking Yoshimura (twy) mice that develop calcification at C1-C2 vertebral level compressing the spinal cord. At the level of compression, the area of neuronal soma and total length of dendrites of wheat germ agglutinin-horseradish peroxidase (WGA-HRP)-labelled accessory motoneurons in the medial cell pool decreased significantly with decrement in motoneuron population, relative to the control. In contrast, at sites rostral to the compressive lesion, a significant enlargement of the neuron soma and dendritic elongation were noted, associated with increased motoneuron population and decreased transverse area of the cord at the level of compression. At this site, enhanced BDNF and NT-3 immunoreactivities were evident in the anterior horn cells. In mice with a more severe degree of compression, astrocyte-like cells showing BDNF immunoreactivity became abundant and axons in the anterior column demonstrated a marked NT-3 immunoreactivity. Our results suggest increased functional activity of anterior horn cells at levels rostra] to the site of compression. We speculate that the presence of BDNF and NT-3 in neurons and astrocyte-like cells is proportionate to the severity of chronic mechanical compression and may contribute to the heterotropic neuronal reserve and survival.

Three-dimensional topographic analysis of spinal accessory motoneurons under chronic mechanical compression: an experimental study in the mouse.

We investigated the effect of chronic mechanical compression of the cervical spinal cord on the number of spinal accessory motoneurons in 25 tiptoe-walking Yoshimura mice. The animals had calcified deposits in the atlantoaxial membrane at the C1-C2 vertebral level, compressing the spinal cord posterolaterally. Motoneurons of the spinal accessory nerve between C1 and C5 segments were labelled using wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injected into the sternocleidomastoid muscles. The counted cells were processed into a three-dimensional computer display to analyse the cytoarchitectonic changes caused by external cord compression. The number of WGA-HRP-labelled spinal accessory motoneurons was significantly reduced on the affected side. The number of motoneurons in compromised C2 and C3 cord segments correlated linearly with the extent of mechanical compression, but no such relationship was present on the contralateral side. There was an increase in the number of WGA-HRP-labelled spinal accessory motoneurons in the medial cell pools of the anterior grey horn at a level most rostral to the compression, and in the ventrolateral cell pools at levels immediately rostral to the compression. Our findings suggest that the spinal accessory motoneurons translocate rostral to the area of external compression in order to avoid mechanical injury.

Calcium crystal deposition in the ligamentum flavum of the lumbar spine.

OBJECTIVE: To investigate the histological and immunohistochemical properties of the lumbar ligamentum flavum with calcium crystal deposition. METHODS: We examined the histological characteristics of the ligamentum flavum of the lumbar spine containing calcium deposits, obtained from 16 surgical cases with symptomatic lumbar spinal stenosis. Sections of the ligaments were also immunostained for elastase, chymotrypsin and S-100 protein, and examined by energy dispensive X-ray microanalysis and scanning electron microscopy. The results were compared with those of ligaments without calcium deposits. RESULTS: The elastic fibres of ligaments with calcium deposits showed marked degeneration (irregular arrangement and fragmentation of the fibre bundles) and nodular granulomatous lesions. Calcium crystal deposits were present in this area and in the nodular granulomatous areas. The calcified areas were surrounded by neutrophils and new small blood vessels. Immunostaining for elastase and chymotrypsin was positive in these areas, and S-100 protein-containing chondrocytes were detected around and within the calcified areas. Calcium pyrophosphate dihydrate crystals were identified in the calcified areas in all patients. CONCLUSION: Nodular degeneration of the elastic fibres of the ligament is probably caused by the action of proteolytic enzymes. Degeneration results in small granulomatous areas that form the nidus for calcium crystal deposition. Accumulated neutrophils, and S-100 protein-containing chondrocytes appear to precipitate the deposition of calcium crystals in the granulomatous lesions of the degenerated lumbar ligamentum flavum.

Serum levels of integrins in chronic liver diseases.

The serum levels of beta 1 integrin (microgram/ml) were significantly higher in the patients with chronic persistent hepatitis (2.59 +/- 0.04), chronic active hepatitis (3.45 +/- 0.13), cirrhosis (4.77 +/- 0.30) and hepatocellular carcinoma (4.71 +/- 0.49) than in normal subjects (2.11 +/- 0.08). Serum levels of beta 3 integrin (microgram/ml) were significantly higher in the patients with chronic active hepatitis (10.48 +/- 1.22), liver cirrhosis (13.55 +/- 1.54) and hepatocellular carcinoma (14.1 +/- 1.77) when compared with normal subjects (5.51 +/- 0.52). A positive correlation was found between serum levels of beta 1 and beta 3 integrins (p < 0.001). A strong positive correlation was observed between serum levels of beta 1 integrin and histologic features, particularly in the degree of hepatic fibrosis, while no correlation was found between serum levels of beta 3 integrin and hepatic fibrosis. Immunohistochemical studies revealed that the beta 1 integrin was present on the plasma membranes of hepatocytes and sinusoidal lining cells in the normal liver, and was increased in fibrotic areas, and on the plasma membranes of hepatocytes and sinusoidal lining cells of the chronic liver disease. However, no positive staining for beta 3 integrin was observed in fibrotic area. The serum level of beta 1 integrin in patients with chronic liver diseases may therefore be a useful marker of hepatic fibrosis.

Calcium crystal deposition in the ligamentum flavum of the cervical spine.

This article reports eight patients who underwent posterior decompressive surgery for myeloradiculopathy caused by calcium crystal deposition in the ligamentum flavum of the cervical spine. All were women with an average age of 72 years and showed neurologic improvement postoperatively. Four patients had diabetes mellitus, and knee meniscus calcification on radiographs were noted in four patients. The ligamentum flavum of C5-6 and C6-7 were sites frequently involved. Crystallographic examination confirmed resected deposits as calcium pyrophosphate dihydrate crystal in all patients and additional hydroxyapatite crystal in two. Histology confirmed marked degeneration in elastic fibers about the calcium deposits. This pathologic condition is possibly spinal manifestation of systemic calcium crystal deposition disease.

Potential usefulness of 18F-2-fluoro-deoxy-D-glucose positron emission tomography in cervical compressive myelopathy.

STUDY DESIGN: This case study describes the usefulness of high-resolution 18F-2-fluoro-deoxy-D-glucose (18FDG)-positron emission tomography (PET) for metabolic neuroimaging of the cervical spinal cord in patients with compressive myelopathy. OBJECTIVE: To examine whether 18FDG-PET imaging could visualize deterioration of cervical spinal cord function associated with a variable degree of compression and to determine its potential usefulness during assessment of myelopathy. SUMMARY OF BACKGROUND DATA: A few studies have described the use of 18FDG-PET imaging in cervical cord diseases, but visualization of the cervical spinal cord before and after surgical decompression for compressive myelopathy has not been reported. The potential usefulness of 18FDG-PET imaging for assessment of the function of compressed cervical cord has not been discussed previously. METHODS: An 18FDG-PET scan was performed before and after surgery in seven patients with cervical compressive myelopathy. The correlation between the metabolic rate of glucose of the cervical spinal cord and neurologic scores was evaluated. The metabolic rate of glucose in different vertebral levels was also measured. RESULTS: Preoperative metabolic rate of glucose was high in two patients but low in the other five. At the time of the second postoperative examination, metabolic rate of glucose was higher in six of the seven patients, and the increase was associated with neurologic improvement. Use of 18FDG was not related to changes in signal intensities on T2-weighted magnetic resonance images. The metabolic rate of glucose decreased at the affected vertebral level in four patients, increased in two, and did not change in one, relative to the unaffected levels. CONCLUSIONS: High-resolution 18FDG-PET neuroimaging may provide clinically useful qualitative and quantitative estimation of impaired metabolic activity of the compromised cervical spinal cord in compressive myelopathy. 18FDG-PET images may also offer additional information related to neuronal dysfunction induced by mechanical compression.

Herniation of cervical intervertebral disc: immunohistochemical examination and measurement of nitric oxide production.

STUDY DESIGN: Surgically obtained cervical herniated intervertebral discs were examined histologically and immunohistochemically. The production of nitric oxide (NO) in the local tissue was examined using the electron spin resonance (ESR) method. OBJECTIVES: To investigate the local histologic and immunohistochemical changes in cervical disc herniation, including NO production, and to compare such changes with those in autopsy cases. SUMMARY OF BACKGROUND DATA: Very little is known about the histopathologic processes of cervical disc herniation. In addition, no information is available on the level of in vivo NO production in cervical disc herniation. METHODS: Thirty-six herniated cervical discs obtained from 31 patients were immunohistochemically examined for localization of blood vessels, matrix metalloproteinase (MMP)-3, and inducible NO synthetase (iNOS). We also compared the production of NO, measured by the ESR method, in eight specimens with that of five control discs obtained from fresh cadavers. RESULTS: The presence of herniated discs correlated with the degeneration of cartilaginous endplate and torn anulus fibrosus. Formation of new blood vessels around the herniated discs was detected, using von Willebrand factor antibody, in seven uncontained hernias and 20 contained hernias. Immunohistochemical studies showed the presence of cells positive for MMP-3 (chondrocytes), iNOS (chondrocytes and granulation tissue) in cervical disc hernias. ESR analysis showed a significantly higher NO production in herniated cervical discs than in disc samples of fresh cadavers. CONCLUSIONS: Herniated cervical intervertebral disc is characterized by the presence of an inflammatory process associated with neovascularization and increased expression of MMP-3. Production of NO was markedly high in both contained- and uncontained-type hernias.

Expression of various growth factors for cell proliferation and cytodifferentiation during fracture repair of bone.

We examined immunohistochemically the fracture repair process in rat tibial bone using antibodies to PCNA, BMP2, TGF-beta 1,-2,-3, TGF-beta R1,-R2, bFGF, bFGFR, PDGF, VEGF, and S-100. The peak level of cell proliferation as revealed by PCNA labelling appeared first in primitive mesenchymal cells and inflammatory cells at the fracture edges and neighboring periosteum at 2-days after fracture, followed by the peaks of periosteal primitive fibroblasts and chondroblasts, which appeared at fracture edges at 3- and 4-days after fracture, respectively. BMP2 was weakly positive in primitive mesenchymal cells, osteoblasts and chondroblasts. At 3-days post-fracture, periosteal osteoblasts produced osteoid tissue and callus with marrow spaces lined by osteoblasts and osteoclasts, and all primitive mesenchymal cells and osteoblasts were positive for TGF-beta 1,-2,-3, and TGF-beta R1,-R2. They were also positive for vascular growth factors bFGF, FGFR and PDGF, but negative for VEGF, and the peak of PCNA labelling of vascular endothelial cells in the marrow space was delayed to 4-days after fracture. Chondroblasts at fracture edges produced hypertrophic chondrocytes at 5-days after fracture and they were positive for TGF-beta 1,-2,-3, and TGF-beta R1,-R2. Primitive chondroblasts were positive for vascular growth factors VEGF as well as bFGF, FGFR, and the peak of PCNA labelling of vascular endothelial cells in the cartilage was at 5-days after fracture. Hypertrophic chondrocytes were also positive for these growth factors but negative for bFGF and bFGFR. S-100 protein-induced calcification was only positive on chondroblasts and hypertrophic chondrocytes. At 7-days after fracture, bone began to be formed from the cartilage at fracture edges, by a process similar to bone formation in the growth plate. Enchondral ossification established a bridge between both fracture edges and periosteal membranous ossification encompassed the fracture site like a sheath at 14 day after fracture. Our study of fracture repair of bone indicates that this process is complex and occurs through various steps involving various growth factors.

Potential role of streptozotocin in enhancing ossification of the posterior longitudinal ligament of the cervical spine in the hereditary spinal hyperostotic mouse (twy/twy).

We investigated the effects of streptozotocin-induced diabetes mellitus on ossification of the posterior longitudinal ligament (OPLL) in the murine cervical spine. A genetically-bound spinal hyperostotic mouse, the tip-toe walking Yoshimura (twy) mouse, was used in these experiments. Histological examination showed that streptozotocin enhanced membranous and enchondral ossification of the posterior longitudinal ligament of the cervical spine, particularly in the area of the ligamentous enthesis. It also increased the number of alkaline phosphatase-positive osteoblast-like mesenchymal cells particularly around the enthesis, while the number of such cells was less in control twy mice and ICR mice treated with streptozotocin. The area of OPLL subsequently increased in size in streptozotocin-treated twy mice. We suggest that streptozotocin-induced diabetes enhances OPLL in the genetically-bound spinal hyperostotic mouse (twy/twy).

Herniated cervical intervertebral discs: histological and immunohistochemical characteristics.

We examined the histological and immunohistochemical changes within and around herniated cervical intervertebral discs. A total of 28 herniated discs were harvested en bloc during anterior decompressive surgeries and examined together with the surrounding tissues. The presence of herniated discs correlated with the degeneration of cartilaginous endplate and torn annulus fibrosus. Formation of new blood vessels around the herniated discs was detected, using von Willebrand factor antibody, in seven (25% of all) uncontained hernias and eight (38%) contained hernias. Immunohistochemical studies using specific antibodies showed the presence of cells positive for matrix metalloproteinase-3 (chondrocytes), CD68 (macrophages and monocytes), and interleukin-1 beta (endothelial cells), in cervical disc hernias. Our results suggested that the magnitude and degree of immunohistochemical tissue reaction in cervical disc herniation correlate with the extent as well as location of herniated disc material.

A diabetic case of Hb Riyadh with a low HbA1c value.

A 74-year-old male patient with diabetes mellitus and hypertension who had been treated for a long period was admitted to our hospital. Laboratory data on admission revealed high values for fasting blood sugar and fructosamine, 219 mg/dl and 389 mumol/l respectively, while the concentration of glycated hemoglobin (HbA1c) was low (3.0%). High performance liquid chromatography and isoelectric focusing analysis of the patient's Hb disclosed abnormal Hb with the content being 41.3%. The structural analysis indicated that this abnormal Hb was Hb Riyadh [beta 120 (GH3) Lys-->Asn]. The low value of HbA1c despite the high blood glucose level may be attributed to this abnormal hemoglobin.

Ligamentum flavum hematoma in the thoracic spine.

We report a case of a hematoma of ligamentum flavum at T11-12 in a 66-year-old man who presented with progressive weakness of the right foot and numbness of both legs. Past history was negative and no precipitating episode of lower back sprain or trauma. The resected T11 and T12 laminas showed old hematoma with degenerative changes in the ligamentum flavum. Hematoma occurring in the thoracic spine has never been reported previously.