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Biomedical subjects

N Hamada

Publications and source records attributed to N Hamada.

At least 19 recordsLinked to original sources

Effects of an oil spill and discharge of domestic sewage on the insect fauna of Cururu stream, Manaus, AM, Brazil.

Assemblages of aquatic and edaphic insects in three streams in Central Amazonia were used to evaluate the impact on one of the streams (Cururu) caused by an oil spill that occurred in August 1999. The Cururu stream had already been impacted by domestic sewage. These three streams were evaluated during four different periods of the regional hydrological cycle (falling-, low-, rising- and high-water), from September 2000 to May 2001. An Eckman dredge, an aquatic entomological net and a manual auger were used to collect insects, which were used to estimate the taxonomic richness (at the family level) and insect density in each stream. Anoxic and eutrophic conditions were determined in each stream based on the mean concentration of dissolved oxygen, total nitrogen and phosphorus. Richness and abundance of insect fauna were higher in the Cristalino stream (not impacted) than in the Bom Jardim stream (impacted by domestic sewage) and in the Cururu stream, with the exception of samples collected along the stream banks (littoral fauna). The mean dissolved oxygen was lower in the Cururu stream than in the other two, while the mean concentrations of total nitrogen and phosphorus were higher. These elements showed intermediate values in the Bom Jardim stream, while the opposite pattern was observed in the Cristalino stream, with higher concentrations of dissolved oxygen and lower concentrations of total nitrogen and phosphorus. The concentrations of these variables reflect the modifications resulting from anthropogenic eutrophication caused by the discharge of domestic sewage into Bom Jardim and Cururu and by the oil spill in Cururu, which negatively affected the richness and abundance of insects in these streams.

Animals↗

Restraint stress enhances alveolar bone loss in an experimental rat model.

BACKGROUND AND OBJECTIVE: The purpose of the present study was to examine the effects of restraint stress on periodontal breakdown resulting from Porphyromonas gingivalis-challenged periodontitis in rats. MATERIAL AND METHODS: To examine the influence of restraint stress on periodontal breakdown, rats were orally challenged with the periodontal pathogen P. gingivalis. Twenty male, specific pathogen-free (SPF) 3-wk-old, Sprague-Dawley rats were divided into four groups: group A (controls), group B (exposed to restraint stress for 12 h/d for 22 d), group C (orally challenged with P. gingivalis), and group D (exposed to restraint stress for 12 h/d for 22 d and orally challenged with P. gingivalis). After 22 d, all animals were killed. The distance from the alveolar bone crest to the cemento-enamel junction was determined, concentrations of adrenocorticotropic hormone were measured as stress markers, and atrophy of the thymus and spleen were assessed. In addition, the furcation area of the maxillary molars was examined histologically, while gingival cytokine gene expression was assessed by mRNA using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In the restrained group, all stress markers were elevated, and the thymus and spleen were atrophied. Combined restraint stress and oral challenge with P. gingivalis resulted in significantly higher bone loss, and osteoclasts were observed. RT-PCR analysis revealed low cytokine gene expression in the restrained groups. CONCLUSION: These results suggest that the presence of restraint stress significantly enhances the progression of P. gingivalis-challenged periodontitis in rats.

Alveolar Bone Loss↗

The p53-Mdm2 association in epithelial cells in idiopathic pulmonary fibrosis and non-specific interstitial pneumonia.

BACKGROUND: Wild-type p53 is increased during cellular responses to various stresses. Mdm2, which is induced by p53, regulates p53 protein concentrations through the ubiquitin-proteasome pathway. AIM: To investigate whether the Mdm2 mediated ubiquitination of p53 is associated with epithelial cell apoptosis in idiopathic pulmonary fibrosis (IPF). METHODS: Immunohistochemistry and western blot analysis were carried out on lung samples obtained by lung biopsy from patients with IPF and non-specific interstitial pneumonia (NSIP). RESULTS: The expression of p53, phosphorylated p53, Mdm2, p21, and Bax was upregulated in epithelial cells from patients with IPF and NSIP compared with normal lung parenchyma. Except for p21, there was a significant increase in the expression of these factors in IPF compared with NSIP. In addition, the number of apoptotic cells and the number of p53 and Bax positive cells was increased compared with controls. p53 conjugated with Mdm2 was decreased in IPF compared with NSIP and controls. Ubiquitinated p53 was increased in both IPF and NSIP compared with controls. CONCLUSIONS: Signalling molecules associated with p53 mediated apoptosis may participate in epithelial cell apoptosis, and the attenuation of p53-Mdm2 conjugation and of p53 degradation may be involved in the epithelial cell apoptosis seen in IPF. Augmented epithelial apoptosis in IPF may lead to the poor prognosis compared with NSIP.

Adult↗

Apoptosis signaling pathways in lung diseases.

Evidence that apoptosis plays an important role in the pathophysiology of lung diseases has been accumulated. Apoptosis signaling is classically composed of two principle pathways. One is a direct pathway from death receptor ligation to caspase cascade activation and cell death. Death receptor ligation triggers recruitment of the precursor form of caspase-8 to a death-inducing complex, through the adaptor protein FADD, which leads to caspase-8 activation. The other pathway triggered by stimuli such as drugs, radiation, infectious agents and reactive oxygen species is initiated in mitochondria. After cytochrome c is released into the cytosol from the mitochondria, it binds to Apaf1 and ATP, which then activate caspase-9. Recently, endoplasmic reticulum has also been shown to be the organelle to execute apoptosis. Further understanding of molecular mechanisms of apoptosis and its regulation by novel drugs may lead to the development of effective strategies against lung diseases. We overview the signaling pathways of apoptosis and discuss the involvement of apoptosis in the pathophysiology of various lung diseases.

Animals↗

Retinoic acid fails to reverse emphysema in adult mouse models.

BACKGROUND: Previous work has shown that all-trans-retinoic acid reverses elastase induced emphysema in rats. Since there is currently no effective treatment for pulmonary emphysema, the effect of retinoic acid should be further investigated in other adult species. A study was undertaken using two murine models of emphysema to evaluate the effect of retinoic acid. METHODS: The models used were an elastase induced emphysema model for acute alveolar destruction and a tumour necrosis factor (TNF)-alpha transgenic mouse which exhibits chronic air space enlargement, loss of elastic recoil, increased lung volume, and pulmonary hypertension comparable to human pulmonary emphysema. All-trans-retinoic acid (2 mg/kg) was injected for 12 successive days after the establishment of emphysema. The effects of treatment were evaluated using physiological and morphometric analyses. RESULTS: In contrast to the rat, administration of all-trans-retinoic acid in these murine models did not improve the emphysema. Moreover, worsening of emphysema was observed in TNF-alpha transgenic mice treated with all-trans-retinoic acid. The level of keratinocyte chemoattractant (KC), a CXC chemokine, in bronchoalveolar lavage fluid was increased in TNF-alpha transgenic mice following retinoic acid treatment. These data raise the possibility that retinoic acid causes deterioration of emphysema by promoting inflammation in this model. CONCLUSIONS: In these models, retinoic acid did not show positive effects on emphysema. The effect of retinoic acid in the treatment of pulmonary emphysema remains controversial, and further studies are required to determine its physiological effects under a variety of experimental conditions.

Animals↗

Direct observation of charge transfer in double-perovskite-like RbMn[Fe(CN)6].

The charge density distribution has been determined for a transition metal cyanide, RbMn[Fe(CN)(6)], by means of the maximum entropy-Rietveld method combined with the highly angularly resolved synchrotron radiation x-ray powder diffraction at SPring-8 BL02B2. We directly observed a charge transfer from the Mn site to the Fe site in the low-temperature phase. On the basis of a local density approximation calculation, we discuss the origin for the anisotropic bonding electron distribution around the Mn3+ ion in the low-temperature phase.

Journal Article↗

Charge-ordered state in single-crystalline CaFeO3 thin film studied by x-ray anomalous diffraction.

X-ray anomalous diffraction, together with a band-structure calculation, was employed to obtain a quantitative understanding of the charge-ordering state in a single-crystalline CaFeO3 thin film. The experimental result shows a characteristic energy dispersion of the nearly inhibited reflection at 150 K, implying Fe atoms split into two distinct states. The energy dispersion is in good agreement with the calculated spectrum based on the LDA+U scheme. The calculation also reveals an electronic structure of the system where holes in the oxygen orbital surround one of the distinct Fe atoms, in spite of the total electron number in both Fe atoms remaining unchanged.

Journal Article↗

In vitro expansion of human basophils by interleukin-3 from granulocyte colony-stimulating factor-mobilized peripheral blood stem cells.

BACKGROUND: A number of studies support the belief that human basophils play an important role in allergic inflammation. The exact mechanism of basophil activation at the site of allergic inflammation, however, has not been well understood, mainly due to their low number in blood and difficulty in obtaining a sufficient number of highly purified basophils for investigation. OBJECTIVE: The purpose of this study is to expand human basophils in vitro with high yield and purity by culturing peripheral blood stem cells (PBSCs). METHODS: We collected PBSC-rich mononuclear cells containing CD34+ cells (0.15-4.9%) by leukapheresis from patients with malignant lymphoma and lung cancer during haematopoietic recovery after chemotherapy plus granulocyte colony-stimulating factor-induced mobilization. PBSC-rich mononuclear cells were cultured in the presence of IL-3. RESULTS: When PBSC-rich mononuclear cells containing more than 1% of CD34+ cells were cultured, 20.0-83.3% of the cells, mostly with a yield of >10%, were metachromatic cells after 3 weeks of culture. These cells resembled mature peripheral blood basophils morphologically when examined by light and electron microscopy. Flow cytometric analysis showed that they expressed both FcepsilonRI and FcgammaRII. FcepsilonRI cross-linking resulted in intracellular calcium mobilization, histamine release and synthesis of cysteinyl leukotrienes. The intracellular histamine content and the release of these chemical mediators triggered by anti-IgE antibodies were comparable to those of peripheral blood basophils. CONCLUSION: These findings suggest that PBSC-derived basophils expanded in vitro are morphologically and functionally mature and will be a useful tool for the analysis of basophil functions.

Adolescent↗

Gonadotropin-releasing hormone increases melatonin release in the pineal gland of the female rat in vitro.

To evaluate the effect of gonadotropin-releasing hormone (GnRH) on melatonin ( N-acetyl-5-methoxytryptamine) release and its synthesizing enzyme activities in pineal glands, pineals from adult female rats during diestrus were organ-cultured in a medium containing 10 -12, 10 -10, or 10 -8 M GnRH for 6 h. Melatonin release increased significantly in pineals cultured with 10 -10 and 10 -8 M GnRH compared to controls. However, in pineal glands that were organ-cultured in a medium containing 10 -12 to 10 -8 M GnRH, the activity of arylalkylamine N-acetyltransferase, which is the key regulatory enzyme in melatonin biosynthesis, showed no significant difference from controls. Likewise, GnRH at these concentrations had no significant effect on the activity of pineal hydroxyindole- O-methyltransferase, which catalyzes the final step of melatonin biosynthesis. These results show that GnRH stimulates pineal melatonin release, but suggest that GnRH does not affect its melatonin synthesis.

Acetylserotonin O-Methyltransferase↗

Intraductal papillary mucinous tumors of the pancreas. Comparison of helical CT and MR imaging.

PURPOSE: To compare the effectiveness of thin-section helical CT and MR imaging with gadolinium-enhanced dynamic technique and MR cholangiopancreatography (MRCP) in the examination of patients with intraductal papillary mucinous tumors. MATERIAL AND METHODS: Helical CT, dynamic MR imaging, and MRCP of 25 intraductal papillary mucinous tumors were compared with ERCP and surgical findings. RESULTS: The duodenal papilla was identified by helical CT and dynamic MR imaging in 11 (44%) and 20 (80%) of the 25 patients, respectively (p<0.05). The main pancreatic duct was visualized on helical CT, dynamic MR imaging, and MRCP in all patients (100%): 25 (96.2%), 24 (92.3%), and 26 (100%) cystic lesions were depicted, respectively. A communicating duct between the main pancreatic duct and the cystic lesion was visualized on helical CT, dynamic MR imaging, and MRCP in 14 (53.8%), 11 (42.3%), and 15 (55.7%) lesions, respectively. The papillary projections corresponding to 3 mm or larger papillary neoplasms were depicted on helical CT and MR imaging in 7 patients (25%). CONCLUSION: MR imaging was equal or slightly superior to thin-section helical CT in the evaluation of intraductal papillary mucinous tumors.

Adult↗

Oxidative stress in lung epithelial cells from patients with idiopathic interstitial pneumonias.

Lung epithelial cells are a primary target for reactive oxygen species (ROS). ROS can cause oxidative deoxyribonucleic acid modification, such as 8-hydroxy-deoxyguanosine (8-OHdG). A human homologue of the MutT protein (hMTH1) prevents this modification. Mitochondria are the most important cellular source of ROS and may be susceptible to oxidative damage. The purpose of this study is to investigate oxidative stress and mitochondrial damage in lung epithelial cells from idiopathic interstitial pneumonias (IIPs). The authors analysed 8-OHdG, hMTH1, and mitochondrial proteins on lung specimens from 13 patients with IlPs consisted of eight patients with usual interstitial pneumonia and five patients with nonspecific interstitial pneumonia using Western blot analysis and immunohistochemistry. Immunoreactivity for 8-OHdG and hMTH1 was significantly increased in the lung epithelial cells from patients with IIPs compared with controls. The expression of hMTH1 was localised in the nuclear and cytoplasmic, but not the mitochondrial, fraction of lung homogenates. Immunoreactivity for mitochondrial protein and cytochrome c oxidase complex subunit IV was increased in the lung epithelial cells from patients with IIPs compared with controls. The current study concludes that oxidative stress may participate in epithelial cell damage in idiopathic interstitial pneumonia, and that increased mitochondrial mass may associate with increased reactive oxygen species production in idiopathic interstitial pneumonia.

Blotting, Western↗

Human reticulon 1-A and 1-B interact with a medium chain of the AP-2 adaptor complex.

Human reticulon family gene 1 (RTN1) is expressed predominantly in neuroendocrine tissues, and produces three proteins termed RTN1-A, RTN1-B, and RTN1-C. Yeast two-hybrid screening indicated that RTN1-A and RTN1-B interacted with AP50, a component of the AP-2 adaptor complex involved in endocytosis. In contrast, RTN1-C did not interact with AP50. Three RTN1 proteins contain the same C-terminal domain. In addition, RTN1-A and RTN1-B share N-terminal 168 amino acid region, suggesting that the 168 amino acid region might play a role in regulating the endocytic process. Although no apparent morphological change of the endocytic organelles was observed, the association of AP50 with the internalized clathrin-coated vesicles was moderately affected when CV-1 cells were directed to express stably RTN1-A or RTN1-B.

Adaptor Protein Complex 2↗

Self-regeneration of a Pd-perovskite catalyst for automotive emissions control.

Catalytic converters are widely used to reduce the amounts of nitrogen oxides, carbon monoxide and unburned hydrocarbons in automotive emissions. The catalysts are finely divided precious-metal particles dispersed on a solid support. During vehicle use, the converter is exposed to heat, which causes the metal particles to agglomerate and grow, and their overall surface area to decrease. As a result, catalyst activity deteriorates. The problem has been exacerbated in recent years by the trend to install catalytic converters closer to the engine, which ensures immediate activation of the catalyst on engine start-up, but also places demanding requirements on the catalyst's heat resistance. Conventional catalyst systems thus incorporate a sufficient excess of precious metal to guarantee continuous catalytic activity for vehicle use over 50,000 miles (80,000 km). Here we use X-ray diffraction and absorption to show that LaFe(0.57)Co(0.38)Pd(0.05)O(3), one of the perovskite-based catalysts investigated for catalytic converter applications since the early 1970s, retains its high metal dispersion owing to structural responses to the fluctuations in exhaust-gas composition that occur in state-of-the-art petrol engines. We find that as the catalyst is cycled between oxidative and reductive atmospheres typically encountered in exhaust gas, palladium (Pd) reversibly moves into and out of the perovskite lattice. This movement appears to suppress the growth of metallic Pd particles, and hence explains the retention of high catalyst activity during long-term use and ageing.

Journal Article↗

Cytokine production induced by a 67-kDa fimbrial protein from Porphyromonas gingivalis.

Fimbriae have been reported to play an important role in the adherence of Porphyromonas gingivalis to oral surfaces and possibly in triggering host responses. P. gingivalis ATCC 33277 has two distinctly different fimbriae expressed on the cell surface. The 67-kDa fimbriae differ in size and antigenicity from the earlier reported FimA, a major 41-kDa fimbrial component of P. gingivalis. Expression of the 67-kDa fimbriae on the cell surface of a fimA mutant was investigated by electron microscopy. The 67-kDa fimbrial protein was purified from the fimA mutant by sonication, precipitation, and chromatography on a DEAE Sepharose CL-6B column. The N-terminal amino acid sequence of the 67-kDa fimbrillin was distinct from that of the 41-kDa fimbrillin. Moreover, we have found that the 67-kDa fimbrial protein from P. gingivalis ATCC 33277 induced IL-1alpha, IL-beta, IL-6 and TNFalpha cytokine expression in mouse peritoneal macrophages. These results suggest that P. gingivalis 67-kDa fimbriae may play a part in the inflammatory response during the development of periodontal diseases.

Amino Acid Sequence↗

Stimulatory effects of LH on release of melatonin and activities of its synthesizing enzymes NAT and HIOMT in organ-cultured pineal glands of female rats.

The pineal hormone melatonin (N-acetyl-5-methoxytryptamine) exerts antigonadotropic effects in some mammalian species. To evaluate the effect of luteinizing hormone (LH) on melatonin release and its synthesizing enzyme activities in pineal glands, pineals of adult female rats undergoing diestrus were organ-cultured in a medium containing 10(-12), 10(-10) or 10(-8) M LH for 6 h. Melatonin release increased significantly in pineals cultured with 10(-12) and 10(-10) M LH, as compared to control values. Similarly, the activity of arylalkylamine N-acetyltransferase (NAT), the key regulatory enzyme in melatonin biosynthesis, was significantly higher in pineals cultured with 10(-12) and 10(-10) M LH for 6 h, while LH at 10(-8) M had no effect. Although LH at 10(-10) M increased pineal hydroxyindole-O-methyltransferase (HIOMT) activity, which catalyzes the final step of melatonin biosynthesis, LH at 10(-12) and 10(-8) M had no effect. These results demonstrate that at relatively low physiological levels, LH stimulates pineal melatonin synthesis and release, mainly by increasing NAT activity.

Acetylserotonin O-Methyltransferase↗

Immunoproteasome assembly and antigen presentation in mice lacking both PA28alpha and PA28beta.

Two members of the proteasome activator, PA28alpha and PA28beta, form a heteropolymer that binds to both ends of the 20S proteasome. Evidence in vitro indicates that this interferon-gamma (IFN-gamma)-inducible heteropolymer is involved in the processing of intracellular antigens, but its functions in vivo remain elusive. To investigate the role of PA28alpha/beta in vivo, we generated mice deficient in both PA28alpha and PA28beta genes. The ATP-dependent proteolytic activities were decreased in PA28alpha(-/-)/beta(-/-) cells, suggesting that 'hybrid proteasomes' are involved in protein degradation. Treatment of PA28alpha(-/-)/beta(-/-) cells with IFN-gamma resulted in sufficient induction of the 'immunoproteasome'. Moreover, splenocytes from PA28alpha(-/-)/beta(-/-) mice displayed no apparent defects in processing of ovalbumin. These results are in marked contrast to the previous finding that immunoproteasome assembly and immune responses were impaired in PA28beta(-/-) mice. PA28alpha(-/-)/beta(-/-) mice also showed apparently normal immune responses against infection with influenza A virus. However, they almost completely lost the ability to process a melanoma antigen TRP2-derived peptide. Hence, PA28alpha/beta is not a prerequisite for antigen presentation in general, but plays an essential role for the processing of certain antigens.

Adenosine Triphosphate↗

A low-pH culture condition enhances the radiosensitizing effect of wortmannin.

PURPOSE: The radiosensitizing effect of wortmannin on human tumor cells in a low-pH microenvironment was compared with that in a neutral-pH environment. METHODS AND MATERIALS: A172 human glioblastoma cells, A549 human lung adenocarcinoma cells, and HMV-1 human melanoma cells were treated with 20 microM wortmannin 2 h before irradiation, and cell survival was examined. A low-pH microenvironment was simulated by exposing cells to low-pH culture medium for 24 h before wortmannin treatment. The effects of wortmannin on the repair of DNA double-strand breaks (dsbs) after 50-Gy irradiation in both low- and neutral-pH conditions were measured by pulsed-field gel electrophoresis. Expression of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) in low-pH conditions was also compared with that in neutral-pH conditions by Western blot analysis. RESULTS: The radiosensitizing effect of wortmannin was greater in low-pH cultures than in neutral-pH cultures for all cell lines. The fast-rejoining component of DNA dsb repair was inhibited more strongly in low-pH than in neutral-pH conditions, although there was little difference in DNA-PKcs expression between groups. CONCLUSIONS: The low-pH culture condition, which was designed to mimic the microenvironment of the central tumor mass in actively proliferating solid tumors, enhanced the radiosensitizing effect of wortmannin by inhibiting the fast-rejoining component of DNA dsb repair and by prolonging the retention of nonrejoined DNA dsbs.

Adenocarcinoma↗

Reduced fimbria-associated activities of Porphyromonas gingivalis induced by recombinant fimbrial expression.

The adhesion properties of the recombinant fimbriae (r-fimbriae) recovered from a YH522 transformant of Porphyromonas gingivalis which harbors a chimeric plasmid, pYHF2, containing the fimA gene of strain 381 were compared with those of the endogenous fimA fimbriae of strain 33277. The adhesion level of the r-fimbriae to Actinomyces viscosus was clearly lower than that of the endogenous fimbriae. In addition, the r-fimbriae were shown to lack some minor components detectable in the endogenous fimbriae. The plasmid pYHF2 prepared from the YH522 transformant was then transformed into six different P. gingivalis strains and the resultant pYHF2-containing strains were examined for their fimbrial expression. In spite of the presence of a considerable diversity in the expression level of the r-fimbriae among these transformants, it was evident that the strains expressing higher levels of the r-fimbriae exhibited a greater decrease in adhesion activity to other bacteria and to oral epithelial cells, as well as in self-aggregation.

Actinomyces viscosus↗