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Biomedical subjects

A Ito

Publications and source records attributed to A Ito.

At least 415 records · Page 23Linked to original sources

Cyclooxygenase inhibitors augment the production of pro-matrix metalloproteinase 9 (progelatinase B) in rabbit articular chondrocytes.

Matrix metalloproteinase 9 (MMP-9/gelatinase B) has recently been proposed to participate in the destruction of articular cartilage. Here, we report that interleukin 1 (IL-1) enhances the production of the precursor of MMP-9 in rabbit articular chondrocytes in primary culture, and this IL-1-mediated production of proMMP-9 is greatly augmented by cyclooxygenase inhibitors such as diclofenac and indomethacin, whereas the constitutive production of proMMP-2 (progelatinase A) is not modulated by IL-1 and/or cyclooxygenase inhibitors. Exogenous prostaglandin (PG) E1 and PGE2 suppress the proMMP-9 production in a dose-dependent manner. Similar results are also obtained with cultured rabbit synoviocytes. These results provide the first evidence that PGE down-regulates the production of proMMP-9 in chondrocytes and synoviocytes. Thus, cyclooxygenase inhibitors probably exert undesirable catabolic actions on the maintenance of articular cartilage under inflammatory conditions.

Alprostadil↗

Plasma (1-->3)-beta-D-glucan measurement in diagnosis of invasive deep mycosis and fungal febrile episodes.

(1-->3)-beta-D-glucan is a characteristic fungal cell-wall constituent. To assess the clinical usefulness of this glucan in screening for invasive fungal infection or fungal febrile episodes, we measured the plasma concentration at the time of routine blood culture in 202 febrile episodes by means of factor G, a horseshoe-crab coagulation enzyme that is extremely sensitive to this polysaccharide. With a plasma cut-off value of 20 pg/mL, 37 of 41 episodes of definite fungal infections (confirmed at necropsy or by microbiology) had positive results (sensitivity 90%). All of 59 episodes of non-fungal infections, tumour fever, or collagen diseases had concentrations below the cut-off value (specificity 100%). Of 102 episodes of fever of unknown origin, 26 had plasma glucan concentrations of more than 20 pg/mL. With those 102 cases taken as non-fungal infections, the positive predictive value of the test was estimated as 59% (37/63), the negative predictive value as 97% (135/139), and the efficiency as 85% (172/202). The positive predictive value should improve if there were a sensitive gold standard that could discriminate fungal from non-fungal infections. Causative fungi included candida, aspergillus, cryptococcus, and trichosporon. Determination of plasma (1-->3)-beta-D-glucan with factor G is a highly sensitive and specific test for invasive deep mycosis and fungal febrile episodes, and will substantially benefit immunocompromised patients.

Adolescent↗

In vitro biocompatibility, mechanical properties, and corrosion resistance of Ti-Zr-Nb-Ta-Pd and Ti-Sn-Nb-Ta-Pd alloys.

There is much discussion about the toxic effect of vanadium and aluminum contained in Ti-6Al-4V alloy for prosthetic implants. The goal of the present investigation was to develop new titanium alloys with sufficient mechanical properties using more biocompatible alloying elements: zirconium, tin, niobium, tantalum, and palladium. The relative growth rates of L929 and MC3T3-E1 cells were significantly higher when cultured with the extraction of Ti-10Zr-8Nb-2Ta-0.2Pd or Ti-15Zr-4Nb-2Ta-0.2Pd alloys than when cultured with the extraction of Ti-6Al-4AV ELI alloy. The tensile strength, elongation, and reduction of area for Ti-15Sn-4Nb-2Ta-0.2Pd alloy were 989 MPa, 14.4%, and 49.3%, respectively, surpassing Ti-6Al-4V ELI alloy. (ASTM F138-84); those for Ti-15Zr-4Nb-2Ta-0.2Pd alloy were 725 MPa, 23.6% and 54.9%, respectively. More than 15% addition of tin as well as zirconium deteriorated the tensile properties. Titanium release into a 5% hydrochloric acid solution from the new titanium alloys was 20-50 micrograms/cm2 per day, though that from Ti-6Al-4V ELI alloy was 1300 micrograms/cm2 per day. The optimum alloy compositions are Ti-15Zr-4Nb-2Ta-0.2Pd and Ti-15Sn-4Nb-2Ta-0.2Pd, judging from cytocompatibility, corrosion resistance, and mechanical properties. The former is characterized by its higher level cytocompatibility and corrosion resistance, while the latter is characterized by mechanical properties.

Alloys↗

Intraspecific variation of isoenzymes in Taenia taeniaeformis.

The technique of isoenzyme electrophoresis was applied to Japanese wild populations of Taenia taeniaeformis (isolated from Norway rats) and three laboratory reared isolates (KRN isolated from a Malaysian Norway rat, BMM from a Belgian house mouse and ACR from a Japanese gray red-backed vole). The average heterozygosities of Japanese wild populations were fairly small and total genetic variability was 0.0499. The genetic make-up of T. taeniaeformis in Norway rats was rather uniform in the whole of Japan. In KRN isolate, each of all 10 loci examined possessed the allele which was predominant in Japanese wild populations. Similarly, each of 9 loci in BMM isolate possessed the same alleles, but one of 2 alleles at HK locus was different from that in the others. T. taeniaeformis parasitizing house mice and rats were considered to be genetically closely related to each other. In ACR isolate, 7 out of 10 loci possessed different alleles from those in the other populations. It was considered that ACR isolate was genetically distant and its phylogenetic origin in Japan should be different from worms parasitizing Norway rats.

Animals↗

A simple method for collecting eggs of taeniid cestodes from fresh, frozen or ethanol-fixed segments.

A simple method was devised for collecting eggs of Taenia taeniaeformis and T. saginata. All gravid segments, either fresh or frozen or 70% ethanol-fixed, were gently scraped using a pestle on a 150 mesh stainless steel sieve. Eggs and tissue debris were washed out all together with mouse tonicity phosphate buffered saline (MTPBS) through the 150 mesh sieve into a glass beaker. Egg suspension with a huge amount of tissue debris in MTPBS was centrifuged 5 min at 3000 r.p.m. (x 1600 g) and the pellet of eggs and tissue debris was resuspended with 1 vol. of MTPBS and 2 vol. of Percoll (Pharmacia) and centrifuged 60 min at 3000 r.p.m. More than 90% of eggs sedimented in the pellet. The supernatant covered with tissue debris was decanted, and the egg pellet was resuspended and centrifuged several times with MTPBS to remove Percoll. It is suggested that this simple method may prove useful for preparation of eggs of biohazardous taeniid cestodes, such as Taenia solium and Echinococcus spp.

Animals↗

A case of allergic aspergillus sinusitis with acute onset.

Reports of allergic aspergillus sinusitis have increased in recent years and diagnostic criteria have also come to be established, yet this disease remains a rare disorder. The disorder is generally reported to follow a chronic course. Here we report a case of allergic aspergillus sinusitis with a short morbid term of 3 weeks.

Adult↗

Effect of hemorrhagic hypotension on blood flow to the basal and upper turns of the cochlea.

The effect of hemorrhagic hypotension on blood flow in the cochlea was studied in guinea pigs. Using two laser Doppler probes, simultaneous measurements were made from the basal and upper turns of the cochlea. Hypotension induced by removal of blood resulted in reduction of cochlear blood flow. Cochlear blood flow, however, did not reduce the same extent as the extent of hypotension and the reduction of blood flow was observed to be more remarkable in the upper turns than in the basal turn.

Animals↗

The minimum effective dose of praziquantel in treatment of Hymenolepis diminuta in rats.

To determine the minimum effective dose of praziquantel against Hymenolepis diminuta in rats, 5.0 mg/kg, 2.5 mg/kg, 1.0 mg/kg, 0.5 mg/kg, 0.1 mg/kg, or 0.05 mg/kg praziquantel were given to each of five experimentally infected rats in six groups. Faecal samples from each rat were examined on ten consecutive days following treatment. The rats were killed and examined for worms on day 10. Based on the results of faecal examination and autopsy, the minimum effective dose of praziquantel against Hymenolepis diminuta in rats was determined to be 0.5 mg/kg.

Animals↗

Em18 and Em16, new serologic marker epitopes for alveolar echinococcosis in western blot analysis, are the only two epitopes recognized by commercially available weak positive (cut off) sera for Em2plus-ELISA.

The assay system for antibody responses against Em2, the most specific antigen for serodiagnosis of alveolar echinococcosis (AE), has been established by enzyme-linked immunosorbent assay (ELISA) but not by Western blot assay, since Em2 antigen is not protein but carbohydrate in nature. Recently we reported that previously undescribed protein epitopes, designated Em18 and Em16 due to their molecular weights, were good serologic markers for AE by Western blot analysis. It has been shown that Em18 and Em16 are the only two epitopes recognized by commercially available weak positive (cut off) sera for the Em2plus-ELISA.

Antibodies, Helminth↗

Co-culture of human breast adenocarcinoma MCF-7 cells and human dermal fibroblasts enhances the production of matrix metalloproteinases 1, 2 and 3 in fibroblasts.

No measurable amounts of matrix metalloproteinases (MMPs) were produced by human breast adenocarcinoma cell lines MCF-7 and BT-20 in culture. When MCF-7 cells were co-cultured with human dermal fibroblasts enhanced production of precursors of MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), MMP-3 (stromelysin 1) and tissue inhibitor of metalloproteinase type 1 (TIMP-1) was observed. Immunohistochemical studies indicated that these pro-MMPs originated primarily from the fibroblasts, suggesting that MCF-7 cells have a stimulatory effect on stromal cells to produce at least three pro-MMPs and TIMP-1. BT-20 cells also enhanced the production of pro-MMP-2 and TIMP-1 in the dermal fibroblasts, but not of pro-MMP-1 and pro-MMP-3. Normal mammary epithelial cells promoted only TIMP-1 production. To investigate further the stimulatory factors from MCF-7 cells, the conditioned medium and the cell membrane were prepared and examined. The cell membrane fraction enhanced the production of pro-MMP-1 and -3 and TIMP-1, but not of pro-MMP-2. The conditioned medium, on the other hand, augmented the production of all four proteins in the fibroblasts. These observations suggest that breast adenocarcinoma MCF-7 cells in culture produce both soluble and membrane-bound factor(s) which stimulate the production of pro-MMPs and TIMP-1 in neighbouring stromal cells, but the factor(s) released into the medium and that associated with cell membranes are probably different. Such communication between the normal and malignant cell types may, in part, assist the cancer cells to invade and metastasise.

Adenocarcinoma↗

Differential effects of hypoxia on ligand binding properties of nicotinic and muscarinic acetylcholine receptors on cultured bovine adrenal chromaffin cells.

Hypoxia is known to disturb neuronal signal transmission at the synapse. Presynaptically, hypoxia is reported to suppress the release of neurotransmitters, but its postsynaptic effects, especially on the function of neurotransmitter receptors, have not yet been elucidated. To clarify the postsynaptic effects, we used cultured bovine adrenal chromaffin cells as a model of postsynaptic neurons and examined specific binding of L-[3H]nicotine (an agonist for nicotinic acetylcholine receptors: nAChRs) and 22Na+ flux under control and hypoxic conditions. Experiments were performed in media preequilibrated with a gas mixture of either 21% O2/79% N2 (control) or 100% N2 (hypoxia). Scatchard analysis of the specific binding to the cells revealed that the KD under hypoxic conditions was twice as large as that under control conditions, whereas the Bmax was unchanged. When the specific [3H]nicotine binding was kinetically analyzed, the association constant (k1) but not the dissociation constant (k-1) was decreased to 40% of the control value by hypoxia. When the binding assay was performed using the membrane fraction, these changes were not observed. Nicotine-evoked 22Na+ flux into the cells was suppressed by hypoxia. In contrast, specific [3H]quinuclidinyl benzilate binding to the intact cells was unaffected by hypoxia. These results demonstrate that hypoxia specifically suppresses the function of nAChRs (and hence, neuronal signal transmission through nAChRs), primarily by acting intracellularly.

Adrenal Glands↗

Ceramide prevents neuronal programmed cell death induced by nerve growth factor deprivation.

We examined the ability of ceramide and sphingomyelinase (SMase) to prevent neuronal programmed cell death (PCD). We found that a cell-permeable ceramide analogue prevented neuronal PCD when applied to established sympathetic neuron primary cultures at the time of nerve growth factor (NGF) deprivation. Other amphiphilic lipids such as oleic acid failed to prevent cell death. Exogenous SMase also showed the same effect, probably by raising the intracellular ceramide level by sphingomyelin (SM) breakdown. Phosphocholine, another hydrolytic product of SM by SMase, did not prevent cell death. Other phospholipases, such as phospholipase C and phospholipase A2, could not prevent cell death. Given the recent findings that the SM cycle is activated to increase the intracellular ceramide level on NGF binding to the low-affinity NGF receptor (LNGFR) and that NGF binding to LNGFR suppresses apoptosis in neuronal cell lines, our results suggest the possibility of the SM cycle as a signaling mechanism transducing the PCD-preventing activity of NGF.

Animals↗

Immunoblot evaluation of IgG and IgG-subclass antibody responses for immunodiagnosis of human alveolar echinococcosis.

Antigen binding of total-IgG and IgG-subclass antibodies from patients with alveolar or cystic echinococcosis (AE and CE) was assessed by immunoblotting. Antigen extracts were prepared from Echinococcus multilocularis protoscoleces (EmP) or from homogenized E. multilocularis metacestode tissue (EmCH). Antigens of approximately 44, 35, 21, 17.5 and 16.5 were recognized by total-IgG and IgG1- and IgG4-subclass antibodies in some of 50 human AE sera from China, Japan or France. The 44- and 35-kDa polypeptides, present in both EmP and EmCH extracts, were recognized by total-IgG antibodies in sera from 82% and 66% of the AE patients, respectively. However, over 30% cross-reactivity occurred between these two antigens and sera from CE and Taenia solium cysticercosis patients. The immunoblot specificities of the 27-, 21- and 17.5-kDa antigens in EmP for E. multilocularis infection were 73%, 88% and 93%, respectively. Recognition of the 17.5-kDa antigen in the EmP immunoblot was much higher for the Japanese AE cases (11/13; 85%) than for the French (9/19; 47%) or Chinese (9/18; 50%) AE cases. None of the CE cases from Uruguay or Libya, where human AE has not been reported, was seropositive for the 17.5-kDa antigen. Antibodies from three (7.3%) of the 41 Chinese CE cases recognized the 17.5-kDa antigen. Within the 13 Japanese AE sera, the combined detection by IgG1, IgG4 and total-IgG antibodies of the 27-, 21- and 17.5-kDa antigens in either EmP or EmCH immunoblots was greater than that by each class/subclass alone, increasing the overall sensitivity for AE patients. A combined ELISA/immunoblot approach, including IgG-subclass detection using E. multilocularis protocolex or cyst extracts, could be useful for the differential diagnosis of human alveolar echinococcosis. An algorithm for such an approach is given.

Algorithms↗

Type I and type II interferons upregulate functional type I interleukin-1 receptor in a human fibroblast cell line TIG-1.

The regulation of type I interleukin-1 receptor (IL-1R) expression by type I, interferon (IFN)-alpha A/D, and type II IFN, IFN-gamma, in a human fibroblast cell line TIG-1 was investigated. After 2 h stimulation with human IFN-alpha A/D or IFN-gamma, the levels of type I IL-1R mRNA increased. We previously reported that IL-1 upregulates transcription and cell surface molecules of type I IL-1R in TIG-1 cells through induction of prostaglandin (PG) E2 and cAMP accumulation. However, indomethacin was unable to inhibit the effect of IFNs, indicating that IFNs augment IL-1R expression through a pathway distinct from that of IL-1. The augmentation was also observed in other fibroblast cell lines. Nuclear run-on assays and studies of the stability of mRNA suggested that the increase in IL-1R mRNA was a result of the enhanced transcription of IL-1R gene. Binding studies using 125I-IL-1 alpha revealed that the number of cell surface IL-1R increased with no change in binding affinity by treatment with these IFNs. Pretreatment of the cells with IFNs enhanced IL-1-induced IL-6 production, indicating that IFNs upregulate functional IL-1R. IL-1 and IFNs are produced by the same cell types, as well as by the adjacent different cell types, and are concomitantly present in lesions of immune and inflammatory reactions. These results therefore suggest that IFNs exhibit synergistic effects with IL-1 through upregulation of IL-1R. Augmented production of IL-6 may also contribute to the reactions.

Cell Line↗

Regions of the molecule responsible for substrate specificity of monoamine oxidase A and B: a chimeric enzyme analysis.

To examine regions of the monoamine oxidase (MAO, EC 1.4.3.4) molecule responsible for substrate recognition, a series of these enzymes, in which discrete regions in one molecule were substituted by corresponding sequences of the other, were constructed from the cDNAs of rat liver MAO A and MAO B and were expressed in yeast, Saccharomyces cerevisiae. Substrate specificities of the original and chimeric enzymes were examined in terms of the maximum activity (Vmax) and the affinity (Km) for serotonin, beta-phenylethylamine (PEA), and benzylamine. Chimeric enzymes with the amino-terminal portion (about 220 residues) and the amino-terminal and middle portions (about 400 residues) of MAO A and MAO B, respectively, exhibited substantially the same Km values as those of the parent enzymes. Extension of the substitution in the middle portion of a chimeric enzyme to the second half of the amino-terminal portion resulted in conversion of the Km values for serotonin to those of the counterpart. Data on relative Vmax values of the chimeric enzymes for the three substrates revealed that the relative catalytic activities were mainly determined by the presence of the middle portion. We conclude from these observations that the region between about residues 120-220 and about residues 50-400 is responsible for determination of the substrate specificity of MAO A and MAO B, respectively, while the middle portion, of about residues 220-400, may relate to the relative catalytic activity towards substrates.

Base Sequence↗

A putative metal-binding site in the beta subunit of rat mitochondrial processing peptidase is essential for its catalytic activity.

Mitochondrial processing peptidase (MPP) consists of alpha- and beta-subunits (alpha-MPP and beta-MPP). beta-MPP has a putative metal-binding sequence (HXXEH). To determine whether the sequence of beta-MPP is essential for the enzymatic activity, we individually mutated the histidines and glutamic acid to arginines and glutamine, respectively. The wild-type and mutated beta-MPPs were co-expressed with alpha-MPP in Escherichia coli. All three mutants had completely lost the activity, whereas the lost activity was recovered on the addition of wild-type beta-MPP. The activity of the wild-type enzyme was reduced by the mutant beta-MPPs. We conclude from these observations that the HXXEH region is involved in the formation of the active site and that beta-MPP is the catalytic subunit of MPP.

Adrenodoxin↗

Possible role of osteopontin in deposition of calcium phosphate in human pilomatricomas.

Human pilomatricomas are benign epidermal appendage tumors composed of hair matrix-like basaloid cells and keratinized remnant cells referred to as shadow cells. Deposition of calcium phosphate usually occurs in the shadow cell nests. Because osteopontin is believed to be involved in the deposition of calcium phosphate in bones, we asked whether osteopontin messenger RNA also is expressed in pilomatricomas. Using Northern blotting and in situ hybridization we detected osteopontin messenger RNA in pilomatricoma tissues but not in normal skin tissue. By the combination of in situ hybridization and immunohistochemistry, osteopontin messenger RNA-expressing cells were identified as CD68-positive macrophages surrounding the shadow cell nests. Immunohistochemistry of anti-human osteopontin antibodies revealed that the localization of osteopontin protein was consistent with that of calcium phosphate. The present results suggest that osteopontin produced by macrophages may play a significant role in the deposition of calcium phosphate in the shadow cell nests of pilomatricomas.

Amino Acid Sequence↗