[Effects of 14-, 15-, and 16-membered ring macrolides on IL-8 gene expression in BET-1A cells].
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Anti-Bacterial Agents↗
Biomedical subjects
K Otake
Publications and source records attributed to K Otake.
Extracellular Nef of HIV-1 can target CD4 memory T population.
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CD4-Positive T-Lymphocytes↗
Nucleic acid amplification testing of hepatitis B virus.
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Blood Donors↗
Immunohistochemical study of the distribution of Ca(2+)/calmodulin-dependent protein kinase phosphatase in the rat central nervous system.
Distribution of Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaM-K Pase) which dephosphorylate multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaM-kinases) in the rat brain and spinal cord were examined immunohistochemically by using an antibody against this enzyme. CaM-K Pase was localized only in the cytoplasm as has been investigated in PC 12 cells, and was never observed in the nucleus. Immunostainability varied from cell group to cell group. Mitral cells in the olfactory bulb, pyramidal neurons in the fifth layer of the cerebral cortex, hippocampal and striatal interneurons, dorsal and ventral pallidal, entopeduncular, and the reticular part of the substantia nigra neurons were intensely immunolabeled. Motoneurons in all the cranial nerve nuclei and the anterior horn of the spinal cord also revealed intense immunolabeling. On the contrary, pyramidal neurons in the Ammon's horn of the hippocampal formation, granule cells in the olfactory bulb, dentate gyrus and cerebellar cortex, Purkinje cells, neurons in the medial habenular nucleus and the inferior olivary nucleus have not shown immunoreactivity. Axons in the white matter or nerve root of the cranial nerve nuclei were immunolabeled. Glial cells in the white matter also showed immunostaining. Because the substrate of CaM-K Pase is multifunctional CaM-kinase II, I and IV, localization of each CaM-kinase was compared with that of CaM-K Pase. The distribution of CaM-K Pase and these CaM-kinases was found to overlap in various regions in the brain and spinal cord. It was concluded, therefore, that CaM-K Pase could regulate the activity of these CaM-kinases by dephosphorylation, when they existed together in neurons.
Animals↗
Sites of action of thyrotropin-releasing hormone on central nervous system neurons revealed by expression of the immediate-early gene c-fos in the rat.
Centrally administered thyrotropin-releasing hormone produces a number of physiological and behavioral changes, e.g., a general antidepressant effect, increasing body temperature, and elevated blood pressure. However, the specific brain sites of action responsible for the centrally activating property of thyrotropin-releasing hormone have not been precisely determined. Using chloral hydrate-anesthetized adult Sprague-Dawley rats, we compared the distribution of Fos-like immunoreactivity after intracerebroventricular administration of thyrotropin-releasing hormone with the results after intracerebroventricular injection of vehicle alone. Some rats were paralysed and artificially ventilated to avoid possible Fos expression secondarily induced by autonomic (e.g., respiratory) disturbances. In thyrotropin-releasing hormone administered rats, selective Fos-like immunoreactivity was observed in V/VI layers of the pre- and infralimbic areas of the medial prefrontal cortex, the ventral midline thalamus, and the nucleus of the solitary tract as well as in the adjacent reticular formation. Fos-like immunoreactivity was significantly reduced in most areas of the cerebral cortex (II/III layers), the shell of the nucleus accumbens, the medial amygdaloid nucleus, parts of the hypothalamus, and the periaqueductal gray. These data suggest that various behavioral and autonomic responses induced by centrally administered thyrotropin-releasing hormone might be produced through the complex neural circuitry comprising the above structures, which are presumed to be implicated in limbic and/or autonomic functions.
Animals↗
Possible pathways through which neurons of the shell of the nucleus accumbens influence the outflow of the core of the nucleus accumbens.
The nucleus accumbens (Acb), a major sector of the ventral striatum, is considered to be an integral part of the striatal complex. The Acb has been shown to be composed of two subdivisions, core and shell, which are distinguishable in several aspects, suggesting that these two subdivisions play different functional roles. The aim of this study was to identify pathways of the efferents of the shell of the Acb to influence the outflow of the core of the Acb. Potential disynaptic projections of the shell to the core of the Acb were investigated in chloral hydrate-anesthetized male Sprague-Dawley rats. Following ipsilateral injections of biotinylated dextran amine (BDA) into the shell of the Acb and cholera toxin B subunit (CT-B) into the core, strong overlapping distributions of BDA-labeled terminals and CT-B-labeled neuronal cell somata were found in the medial part of the ventral tegmental area, medial part of the lateral hypothalamic area, and dorsolateral part of the basolateral amygdaloid nucleus. The significance of multiple sites of relay between the efferents of the shell and the afferents of the core of the Acb at different levels of the neuraxis may be related to the functional specificity of each relay site.
Animals↗
Sleep apnea syndrome in patients with pulmonary thromboembolism.
Seven patients (one man and six women) with a diagnosis of pulmonary thromboembolism (PTE) were examined by polysomnography in order to clarify the relationship between sleep-related breathing disorders and PTE. In the chronic stage of PTE, sleep apnea syndrome (SAS) was recognized in two patients (a man and a woman) among the subjects. Four of the five patients without SAS showed nocturnal hypoxemia. The female predominance was different from ordinary SAS without the disease background of PTE even though PTE is accompanied by hypoxemia and pulmonary hypertension like SAS. A pathophysiological relationship between PTE and SAS was not found in this study.
Aged↗
Internalization of adenovirus by alveolar macrophages initiates early proinflammatory signaling during acute respiratory tract infection.
Adenovirus is a common respiratory pathogen which causes a broad range of distinct clinical syndromes and has recently received attention for its potential for in vivo gene delivery. Although adenovirus respiratory tract infection (ARTI) results in dose-dependent, local inflammation, the pathogenesis of this remains unclear. We hypothesized that alveolar macrophages (AMphi) rapidly internalize adenovirus following in vivo pulmonary administration and then initiate inflammatory signaling within the lung. To evaluate the role of AMphi in the induction of lung inflammation during ARTI in vivo, we directly assessed adenovirus uptake by murine AMphi and correlated uptake with the initiation of proinflammatory gene expression. Stimulation of cytokine (tumor necrosis factor alpha [TNF-alpha], interleukin-6 [IL-6], macrophage inflammatory protein-2 [MIP-2], and MIP-1alpha) expression in the lung was evaluated at the level of mRNA (by reverse transcription-PCR [RT-PCR]) and protein (by enzyme-linked immunosorbent assay) and by identification of cells expressing TNF-alpha and IL-6 mRNA in lung tissues (by in situ hybridization) and isolated lung lavage cells (by RT-PCR). Adenovirus, labeled with the fluorescent dye (Cy3), was rapidly and widely distributed on epithelial surfaces of airways and alveoli and was very rapidly ( approximately 1 min) localized within AMphi. At 30 min after infection AMphi but not airway epithelial or vascular endothelial cells expressed mRNA for TNF-alpha and IL-6, thus identifying AMphi as the cell source of initial cytokine signaling. IL-6, TNF-alpha, MIP-2, and MIP-1alpha levels progressively increased in bronchoalveolar lavage fluid after pulmonary adenovirus infection, and all were significantly elevated at 6 h (P < 0.05). To begin to define the molecular mechanism(s) by which adenovirus initiates the inflammatory signaling in macrophages, TNF-alpha expression from adenovirus-infected RAW264.7 macrophages was evaluated in vitro. TNF-alpha expression was readily detected in adenovirus-infected RAW cell supernatant with kinetics similar to AMphi during in vivo infection. Blockage of virus uptake at specific cellular sites, including internalization (by wortmannin), endosome acidification and/or lysis (by chloroquine) or by Ca(2+) chelation (by BAPTA) completely blocked TNF-alpha expression. In conclusion, results showed that during ARTI, (i) AMphi rapidly internalized adenovirus, (ii) expression of inflammatory mediators was initiated within AMphi and not airway epithelial or other cells, and (iii) the initiation of inflammatory signaling was linked to virion uptake by macrophages occurring at a point after vesicle acidification. These results have implications for our understanding of the role of the AMphi in the initiation of inflammation following adenovirus infection and adenovirus-mediated gene transfer to the lung.
Acute Disease↗
Primary pulmonary hypertension with central sleep apnea: sudden death after bilevel positive airway pressure therapy.
An obese 23-year-old man with sleep-disordered breathing and primary pulmonary hypertension (PPH) had been administered oral beraprost sodium, anticoagulant warfarin, and home oxygen therapy, at another hospital as treatment for the PPH, but he had not experienced any symptomatic improvement. The patient had a body mass index of 32.4kg/m2, and complained of fatigue, shortness of breath on exertion, excessive daytime sleepiness, and snoring. Arterial blood gas analysis showed a PaO2 and a PaCO2 of 70.9 and 31.2mmHg, respectively. A polysomnographic study revealed central sleep apnea with an apnea-hypopnea index (AHI) of 29.7episodes/h. The patient showed improvement of daytime sleepiness after starting nocturnal nasal bilevel positive airway pressure (BiPAP) therapy for the central sleep apnea, but his pulmonary hypertension, measured in the daytime, worsened. The patient died suddenly while walking to the bathroom in the morning 1 month after initiation of BiPAP therapy. It is necessary to consider the possibility of sudden death when nasal BiPAP therapy is given to a PPH patient with central sleep apnea.
Adult↗
[The examinations for diffuse lung diseases].
Diffuse lung diseases show an abnormal shadow that is widely scattered on the bilateral lung fields in the chest X ray view and includes many respiratory diseases such as the infectious or the non-infectious disease; neoplasms. Among these, idiopathic pulmonary fibrosis(IPF) has been studied extensively because of its high frequency and difficulty of treatment. IPF is defined by the respiratory functions, the radiological findings, which depend on HRCT, and the histopathological evaluation by surgical lung biopsy. In particular, the histopathological appearance of usual interstitial pneumonia(UIP) is essential for the diagnosis of IPF. Most serum examinations such as angiotensin-converting enzyme, anti-nuclear antibodies are applied to rule out other diffuse lung diseases. SP-D or KL-6, which is the marker of the type II epithelial cells, is thought to be very useful for revealing the disease activity, but since it is not increased in the early stages of IPF, it is not applied in the diagnosis of IPF. The definitive serum examinations for the diagnosis or determinations of the therapeutic effect or prognosis of IPF have not been established. Easier, more useful and critical examinations including genetic diagnosis are required to manage patients with IPF.
Aged↗
Activation process of dipteran-specific insecticidal protein produced by Bacillus thuringiensis subsp. israelensis.
Dipteran-specific insecticidal protein Cry4A is produced as a protoxin of 130 kDa in Bacillus thuringiensis subsp. israelensis. Here we performed the in vitro processing of Cry4A and showed that the 130-kDa protoxin of Cry4A was processed into the two protease-resistant fragments of 20 and 45 kDa through the intramolecular cleavage of a 60-kDa intermediate. The processing into these two fragments was also observed in vivo. To investigate functional properties of the two fragments, GST (glutathione S-transferase) fusion proteins of the 60-kDa intermediate and the 20- and 45-kDa fragments were constructed. Neither the GST-20-kDa fusion protein (GST-20) nor the GST-45-kDa fusion protein (GST-45) was actively toxic against mosquito larvae of Culex pipiens, whereas the GST-60-kDa intermediate fusion protein (GST-60) exhibited significant toxicity. However, when the two fusion proteins GST-20 and GST-45 coexisted, significant toxicity was observed. The coprecipitation experiment demonstrated that the two fragments associated with each other. Therefore, it is strongly suggested that the two fragments formed an active complex of apparently 60 kDa. A mutant of the 60-kDa protein which was apparently resistant to the intramolecular cleavage with the midgut extract of C. pipiens larvae had toxicity slightly lower than that of GST-60.
Amino Acid Sequence↗
SP-A enhances viral clearance and inhibits inflammation after pulmonary adenoviral infection.
Surfactant protein A (SP-A) is a member of the collectin family of host defense molecules expressed primarily in the epithelial cells of the lung. To determine the role of SP-A in pulmonary adenoviral infection, SP-A-deficient (SP-A -/-) mice were intratracheally infected with a replication-deficient recombinant adenovirus, Av1Luc1. Lung inflammation was markedly increased in SP-A -/- compared with SP-A +/+ mice and was associated with increased hemorrhage and epithelial cell injury. Polymorphonuclear cells in bronchoalveolar lavage fluid (BALF) were increased in SP-A -/- mice after administration of adenovirus. Coadministration of adenovirus and purified human SP-A ameliorated adenoviral-induced lung inflammation in SP-A -/- mice. Concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, and IL-1beta were increased in BALF of SP-A -/- mice. Likewise, TNF-alpha, IL-6, macrophage inflammatory protein (MIP)-1alpha, monocyte chemotactic protein-1, and MIP-2 mRNAs were increased in lung homogenates from SP-A -/- mice 6 and 24 h after viral administration. Clearance of adenoviral DNA from the lung and uptake of fluorescent-labeled adenovirus by alveolar macrophages were decreased in SP-A -/- mice. SP-A enhances viral clearance and inhibits lung inflammation during pulmonary adenoviral infection, providing support for the importance of SP-A in antiviral host defense.
Adenoviridae↗
Nonspecific inflammation inhibits adenovirus-mediated pulmonary gene transfer and expression independent of specific acquired immune responses.
Replication-deficient adenovirus vectors (Avs) have shown high-efficiency gene transfer in a variety of animal models, but demonstrated lower than expected efficiency in the intensely inflammatory milieu of the respiratory tract of individuals with cystic fibrosis (CF). Specific acquired immune responses directed at adenovirus capsid proteins are known to limit the duration of transgene expression and the effectiveness of vector readministration. In these models, however, nonspecific inflammation is also frequently noted to accompany specific immune responses. Because inflammation can occur early after Av administration, we hypothesized that inflammation may block Av-mediated gene transfer in the lung independent of specific immune responses. To evaluate this hypothesis, we measured pulmonary gene transfer and expression in the absence or presence of the potent antiinflammatory agent dexamethasone. To address and eliminate concerns over the potentially confounding effects of systemic, vector-specific acquired immune responses, evaluations were confined to a 3-day period following Av administration and were carried out, in parallel, in normal and immunodeficient (athymic) mice. Dexamethasone significantly reduced Av-associated inflammation in all animals as measured by a significant reduction of blinded, quantitative lung histopathology scores and by reduced proinflammatory cytokine release. Concomitant with reduced inflammation, gene transfer efficiency was significantly increased in both normal and immunodeficient animals as measured by transgene product activity (beta-galactosidase) in total lung homogenates 3 days after vector administration. This finding could not be explained by a direct effect of dexamethasone on transgene specific activity. To begin to understand the molecular mechanisms of Av-induced inflammatory responses, lung levels of the chemoattractive chemokines MIP-2, MIP-1alpha, and MCP-1 were quantified. All were elevated significantly in Av-exposed animals. Dexamethasone reduced levels of MCP-1 and MIP-1alpha, but not MIP-2, consistent with the observed pattern of inflammatory cell changes. Expression of several proinflammatory cytokines including TNF-alpha, IL-6, IL-1beta, and IFN-gamma were also elevated in Av-exposed animals and modulated by dexamethasone. These observations demonstrate that nonspecific inflammation is an important determinant of the efficiency of in vivo pulmonary gene transfer and expression independent of specific immune responses and may have important implications for human gene therapy for diseases of the lung.
Adenoviridae↗
Regional localization of agmatine in the rat brain: an immunocytochemical study.
The distribution of agmatine (decarboxylated arginine) was mapped in the central nervous system (CNS) in the rat. Agmatine-like immunoreactivity was identified by light microscopy, exclusively in the cytoplasm of neuronal perikarya. Immunoreactive neurons were present in the cerebral cortex, predominantly within laminae VI and V and, to a lesser extent, III and mainly in retrosplenial, cingulate, primary somatosensory and auditory cortices, and the subiculum. In the lower brainstem, immunoreactivity was selectively localized to visceral relay nuclei: the nucleus tractus solitarii and pontine parabrachial complex, and periventricular areas including the laterodorsal nucleus, locus coeruleus and dorsal raphe. In the midbrain, immunolabeled cells were concentrated in the ventral tegmental area and periaqueductal gray. In the forebrain, subcortical neurons were labeled predominantly in the preoptic area, amygdala, septum, bed nucleus of the stria terminalis, midline thalamus, and the hypothalamus. Ultrastructural analysis of layer V of the somatosensory cortex demonstrated agmatine-immunoreactivity in neurons, primarily in large dense-core vesicles located in the cytoplasm. Agmatine immunoreactivity was also affiliated with endoplasmic reticulum and the plasmalemma. Cortical neurons and the subiculum were labeled in animals not administered the axonal transport inhibitor, colchicine; thus, may normally contain higher concentrations of the amine than other brain regions. The central distribution of agmatine is consistent with the hypothesis that the amine may be a novel neurotransmitter of neurons involved in behavioral and visceral control.
Agmatine↗
Single midline thalamic neurons projecting to both the ventral striatum and the prefrontal cortex in the rat.
The midline thalamic nuclei have been known to send projection fibres to the ventral striatum and the autonomic/limbic-associated areas of the prefrontal cortex. In the present study, we sought to determine whether or not single midline thalamic neurons project both to the ventral striatum and to the cerebral cortical areas. Experiments were performed on chloral hydrate-anaesthetized male Sprague Dawley rats; two fluorescent retrograde tracers were centred on the medial or lateral part of the nucleus accumbens--the major part of the ventral striatum--and the medial or lateral prefrontal viscerolimbic cortex. Our retrograde double-labelling study revealed that a subset of midline thalamic neurons send projection fibres to both the nucleus accumbens and the cerebral cortex. Such neurons projecting to both targets were principally identified in the paraventricular thalamic nucleus. The majority of the dually-labelled neurons in the paraventricular thalamic nucleus projected to the lateral part of the nucleus accumbens and the medial wall of the prefrontal cortex. Dually-labelled neurons were additionally found in other midline nuclei, including the paratenial, intermediodorsal, rhomboid, and reuniens nuclei, as well as in the medial part of the parafascicular thalamic nucleus. Dually-projecting neurons identified in the present study may represent a potential link between the limbic striatum and the viscerolimbic-associated cortex, thus suggesting that non-discriminative information relayed to the prefrontal cortex might exert an influence through the same neurons on the nucleus accumbens implicated in affective behaviour.
Animals↗
Artificial surfactant (Surfactant TA) modulates adherence and superoxide production of neutrophils.
Neutrophils cause lung injuries by releasing proteases and active oxygen radicals in patients with acute respiratory distress syndrome (ARDS). Artificial surfactant is used to replace native surfactant whose functions are deteriorated by serum-derived inhibitors in these patients. We investigated potential interactions between exogenous surfactant (Surfactant TA) and neutrophils in in vivo and in vitro experimental models. Neutrophil alveolitis was induced in hamster lungs by the intratracheal administration of bleomycin (5 mg/kg) on Day 0. Some of the animals were followed by replacement with Surfactant TA (5 and 10 mg/100 g body weight) on Day 1. Alveolar cells were harvested by lung lavage on Day 2. The numbers of the neutrophils obtained from the lungs treated with bleomycin and Surfactant TA were unchanged, but the superoxide production from these cells was significantly decreased when compared with control animals (no Surfactant TA). From the in vitro experiments, Surfactant TA was shown to inhibit adherence and superoxide production of human neutrophils. These effects were derived from the heat-resistant components of Surfactant TA and were mimicked by treatment with liposomes of dipalmitoyl phosphatidylcholine. Surfactant-TA-treated neutrophils were demonstrated to have picnotic nuclei and to express Fas antigens, which were characteristic of apoptotic cells. These results suggest that exogenous Surfactant TA may play an important role not only in improving surfactant functions but in preventing neutrophils from further activation, probably through enhancing apoptosis.
1,2-Dipalmitoylphosphatidylcholine↗
Radioimmunoassay for Aquaporin-2.
OBJECTIVE: To develop radioimmunoassay for aquaporin-2 (AQP-2). METHODS: Anti-AQP-2 antiserum has been raised in New Zealand white rabbits immunized with a conjugate of synthetic AQP-2 peptide (257-271) with bovine serum albumin. Radioiodination of synthetic peptide (tyrosine-AQP2 (257-271) was performed by chloramine T method, followed by purification of radioiodinated material on Sephadex G-25 column. RESULTS: The obtained antibody did not crossreact with vasopressin, pituitary hormones, hypothalamic hormones and neuropeptides. The assay was performed with a double antibody system. The values are expressed as an equivalent of synthetic AQP-2 peptide (257-271). The dilution curve of high AQP-2 urine in radioimmunoassay system was parallel to the standard curve. The recovery percentage of AQP-2 added to urine was about 100 % in this assay system. Intra-assay and inter-assay variation was 4.5 % and 7.2 %, respectively. Mean urinary excretion of AQP-2 was 1.16 ng equivalent of AQP-2 (257-271)/mg creatine and was lower in patients with diabetes insipidus. CONCLUSION: These data suggest that his assay system is a suitable to measure AQP-2 in urine.
Journal Article↗
Epitope mapping of murine monoclonal antibodies against human immunodeficiency virus type 1 Nef.
It has shown that the human immunodeficiency virus type 1 (HIV-1) Nef protein has the high antigenicity in HIV-1 seropostive individuals. We newly obtained seven monoclonal antibodies (mAbs). To identify the antigenic determinants of HIV-1 Nef protein against murine, epitope mapping of the mAbs was performed by enzyme-linked immunosorbent assay (ELISA) by using several recombinant truncated Nef fusion proteins, that were expressed in Esherichia coli, and synthetic peptides. The results showed that mAbs A6, A7, F2, F3, F4, F8 and E5 recognized epitopes on Nef protein located at amino acid residues 18-26, 28-45, 115-137, 128-137, 115-126, 128-137, and 170-181, respectively.
Animals↗