Effect of pretreatment with coenzyme Q10 and monoclonal antibodies to adhesion molecules on ischemic small intestine preservation.
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Biomedical subjects
Publications and source records attributed to T Maeda.
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Rupture of the aortic valve after a blunt chest trauma is rare. A 63-year-old man was injured in an automobile accident and was hospitalized elsewhere due to acute respiratory failure 4 days after the accident. On admission, he was intubated immediately. The patient did not respond to available medical treatments and was therefore transferred to our hospital. His condition was diagnosed as severe left heart failure, and an emergency operation was performed. The right coronary and non-coronary cusps were perforated and the commissure between the right coronary and non-coronary cusps was torn. The aortic valve was excised and a SJM 25 AEC prosthetic valve was inserted. The patient recovered well postoperatively.
We studied the serial change of 123I-BMIPP SPECT in the process of hypertrophic cardiomyopathy (HCM). In 16 patients with HCM, the imaging data were acquired 15 minutes after injection of 111 MBq 123I-BMIPP. The SPECT image was divided into 17 segments and the severity of the defect in each segment was scored visually using defect score, a 4-point grading system (score 0 = normal, score 1 = mildly decreased uptake, score 2 = moderately decreased uptake, score 3 = severely decreased uptake or defect). Each patient underwent this examination twice with interval of 25 +/- 14 months. In the first examination, we observed reduced uptake in 14 cases (88%), and often found it in septal portion of anterior or posterior wall. In the second examination, compared with the first study, total defect score (TDS: the summation of defect score of 17 segments) had increased in 7 cases (44%) of 16 cases, decreased in only 1 case (6%), and unchanged in 8 cases (50%). The regional defect score often increased in septal portion of anterior and posterior walls, and the high score area widespread toward the septum. There was no difference between TDS unchanged group and TDS worsened group in the point of background (age, family history of HCM), the interval of the two studies, and the findings of echocardiography (Dd, Ds, %F). In TDS worsened group (n = 7), clinically only 1 case became progressed stage. In others there were no progression in each clinical, echocardiographic, or 201Tl scintigraphic findings. This study revealed that myocardial fatty acid metabolic disorder often progresses during short interval in patients with HCM. In conclusion, the serial examination of 123I-BMIPP SPECT may contribute to an estimation of the stage or the prognosis in HCM.
Postnatal development of Ruffini endings was ultrastructurally investigated in the upper incisors of the rat from 1 day to 60 days after birth by means of protein gene product 9.5 (PGP 9.5) immunocytochemistry. The immunostaining with PGP 9.5 antibody clearly demonstrated chronological alterations of the distribution and ultrastructure of the Ruffini endings during postnatal development. At 1 day after birth, the PGP 9.5-positive nerve terminals contained a few mitochondria and vesicles immunonegative for PGP 9.5. Dendritic terminals appeared at 4 days after birth, with a small number of expanded or bulbous portions. These expanded portions possessed morphological features similar to those of the growth cone: several mitochondria and various kinds of vesicles. Typical Ruffini endings with dendritic ramification and expanded portions appeared 7-11 days after birth. At this stage, parts of the axon terminals extended through the slits of Schwann cell covering and formed finger-like projections called axonal spines. These Ruffini endings increased dramatically in number after 24-26 days and were identical in density and morphology to those seen in adult rats. After the commencement of the occlusion between the incisors, the number of large mitochondria increased, in contrast to the decrease of the vesicles in the axon terminals. Moreover, the axonal spines increased both in number and in length. Thus, the periodontal nerve endings showed stage-specific morphological features intimately related in timing to tooth eruption and occlusion. Functional stimuli possibly contribute to the final differentiation and maturation of the periodontal Ruffini endings.
Yeast cells can respond and adapt to osmotic stress. In our attempt to clarify the molecular mechanisms of cellular responses to osmotic stress, we cloned seven cDNAs for hyperosmolarity-responsive (HOR) genes from Saccharomyces cerevisiae by a differential screening method. Structural analysis of the clones revealed that those designated HOR1, HOR3, HOR4, HOR5 and HOR6 encoded glycerol-3-phosphate dehydrogenase (Gpd1p), glucokinase (Glk1p), hexose transporter (Hxt1p), heat-shock protein 12 (Hsp12p) and Na+, K+, Li(+)-ATPase (Ena1p), respectively. HOR2 and HOR7 corresponded to novel genes. Gpd1p is a key enzyme in the synthesis of glycerol, which is a major osmoprotectant in S. cerevisiae. Cloning of HOR1/GPD1 as a HOR gene indicates that the accumulation of glycerol in yeast cells under hyperosmotic stress is, at least in part, caused by an increase in the level of GPDH protein. We performed a series of Northern blot analyses using HOR cDNAs as probes and RNAs prepared from cells grown under various conditions and from various mutant cells. The results suggested that all the HOR genes are regulated by common signal transduction pathways. However, the fact that they exhibited certain distinct responses indicated that they might also be regulated by specific pathways in addition to the common pathways. Ca2+ seemed to be involved in the signaling systems. In addition, Hog1p, one of the MAP kinases in yeast, appeared to be involved in the regulation of expression of HOR genes, although its function seemed to be insufficient for the overall regulation of expression of these genes.
STUDY DESIGN: The present study investigated the distribution of sensory and autonomic nerves to the posterior longitudinal ligament of the rat lumbar vertebra. OBJECTIVE: The results were analyzed to provide a new insight into the pathogenesis of low back pain. SUMMARY OF BACKGROUND DATA: Previous studies that have descriptively reported on posterior longitudinal ligament innervation have attributed important roles to the sinuvertebral nerve as an output of nociceptive sensation. Meanwhile, development of immunohistochemistry has made it possible to identify numerous neurotransmitters, and thus various suggested pathophysiologic roles of the nerve fibers in the posterior longitudinal ligament. However, such microscopic identification of the nerve fibers has not been able to reveal comprehensive distributions of the various fibers, which are presumed to be derived from the sinuvertebral nerve. METHODS: Rat posterior longitudinal ligaments of whole thickness was immunohistochemically stained for calcitonin gene-related peptide and tyrosine hydroxylase, which have been considered markers of nociceptive and noradrenergic fibers respectively. Light-microscopic and electron-microscopic observation was reinforced with another experimental study in which resection of the dorsal root ganglia induced characteristic denervation patterns to the rat posterior longitudinal ligament. RESULTS: The present study revealed that the lumbar. PLL was dually innervated by two distinctive systems of nociceptive fibers. One of the systems was polysegmentally innervated and closely associated with autonomic innervation, whereas the other was unisegmentally innervated and not associated with autonomic fibers. CONCLUSION: Pathogenesis of low back pain can be better understood if the presence of the two distinctive nerve systems innervating the posterior longitudinal ligament is taken into account.
The oviduct fluid mainly derived from the oviduct epithelium is reported to provide the environment necessary for embryonic development. To elucidate the origin of glucose in the oviduct fluid, we examined the expression and localization of glucose transporter 1 (GLUT1) in the rat oviduct by Northern blot analysis, immunoblot analysis and immunohistochemistry using both light and electron microscopy. Northern blot and immunoblot analyses both showed the presence of the GLUT1 mRNA and protein. Specific staining for GLUT1 was observed in the ampulla and the isthmus, but only slightly in the fimbria and the utero-tubal junction. Staining was confined to the luminal surface of the epithelial mucosa. Immunoelectron microscopic analysis revealed that GLUT1 was observed only on the surface of the microvilli in non ciliated secretory cells, but not in ciliated cells. These findings suggest that GLUT1 plays an important role in the glucose transfer from the oviduct epithelium into the lumen and in maintaining the adequate glucose concentration of the oviduct fluid for embryonic development in rat oviduct.
The three-dimensional solution structure of omega-conotoxin MVIIA, a 25-mer peptide antagonist of N-type calcium channels, was determined by two-dimensional 1H NMR spectroscopy with simulated annealing calculations. A total of 13 converged structures of omega-conotoxin MVIIA were obtained on the basis of 273 experimental constraints, including 232 distance constraints obtained from nuclear Overhauser effect (NOE) connectivities, 22 torsion angle (phi, chi 1) constraints, and 19 constraints associated with hydrogen bonds and disulfide bonds. The atomic root mean square difference about the averaged coordinate positions is 0.47 +/- 0.08 A for the backbone atoms (N, C alpha, C) and 1.27 +/- 0.14 A for all heavy atoms of the entire peptide. The molecular structure of omega-conotoxin MVIIA is composed of a short triple-stranded antiparallel beta-sheet. The overall beta-sheet topology is +2x, -1, which is the same as that reported for omega-conotoxin GVIA, another N-type calcium channel blocker. The orientation of beta-stranded structure is similar to each other, suggesting that the conserved disulfide bond combination is essential for the molecular folding. We have recently determined by using alanine substitution analyses that Tyr 13 is essential for the activity of both toxins. On the basis of functional and structural analysis, it is shown that both omega-conotoxin MVIIA and GVIA retain a similar conformation to locate Tyr 13 in the appropriate position to allow binding to N-type calcium channels. These results provide a molecular basis for understanding the mechanism of calcium channel modulation through the toxin-channel interaction and insight into the discrimination of different subtypes of calcium channels.
The role of mitogen-activated protein (MAP) kinase cascades in integrating distinct upstream signals was studied in yeast. Mutants that were not able to activate PBS2 MAP kinase kinase (MAPKK; Pbs2p) at high osmolarity were characterized. Pbs2p was activated by two independent signals that emanated from distinct cell-surface osmosensors. Pbs2p was activated by MAP kinase kinase kinases (MAPKKKs) Ssk2p and Ssk22p that are under the control of the SLN1-SSK1 two-component osmosensor. Alternatively, Pbs2p was activated by a mechanism that involves the binding of its amino terminal proline-rich motif to the Src homology 3 (SH3) domain of a putative transmembrane osmosensor Sho1p.
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Whether or not IL-8 attracts T lymphocytes and activates neutrophils in vivo remains unclear. Most studies on function of IL-8 in vivo have been done on human IL-8 in heterologous animals. To elucidate the role of IL-8 in vivo, we injected homologous IL-8 into rabbit knee joints and investigated the inflammatory response. Injection of 10 micrograms of rabbit IL-8 induced a massive accumulation of neutrophils. IL-8 attracts T lymphocytes in vitro; however, rabbit IL-8 induced no appreciable lymphocyte accumulation in rabbits. Although human IL-8 was reported not to induce cartilage destruction when injected into heterologous animals, we observed that rabbit IL-8 did provoke a release of neutrophil elastase, leading to cartilage destruction, when injected into rabbits. An inhibitor against neutrophil elastase (ONO-5046) prevented destruction of the cartilage. Injection of rabbit IL-8 induced bioactive and immunoreactive IL-1 beta and IL-1 receptor antagonist (IL-1Ra) in the joint cavity. Immunohistochemistry showed that IL-1 beta and IL-1Ra positive cells were infiltrating leukocytes. In neutrophil-depleted rabbits, rabbit IL-8 induced far lesser concentrations of IL-1 beta and IL-1Ra and no cartilage destruction compared with findings in normal rabbits. Thus, the infiltrating neutrophils are the main producers of these cytokines and are responsible for the cartilage destruction. In addition to neutrophil chemotactic activity, IL-8 proved to have a neutrophil-activating capability in vivo, with respect to release of neutrophil elastase and induction of IL-1 beta and IL-1Ra.
The integrin alpha v beta 3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. We have explored molecular recognition by alpha v beta 3 through selection of ligands from large random libraries of peptides displayed on phage. Ligands bound by alpha beta 3 consisted primarily of RGD peptides; however, these peptides showed considerable heterogeneity with respect to the identities of amino acids flanking RGD. The tolerance of alpha v beta 3 for RGD peptides of diverse composition is consistent with its role in vivo as a versatile receptor for RGD-containing extracellular matrix proteins. Peptide ligands for alpha v beta 3 also included a novel binding sequence, identical to a tetrapeptide found in vitronectin, which is a candidate for a synergistic site in this adhesive protein that may act in concert with RGD to promote molecular recognition.
The localization of low-affinity nerve growth factor receptor (LNGFR) in the human urinary bladder was examined immunohistochemically using the mouse monoclonal antibody (ME20-4) against human LNGFR. LNGFR immunoreactivity was present in the human urinary bladder. The distribution of LNGFR-positive fibers was more abundant in the mucosa than in the muscle layer. Results also showed that some LNGFR-positive fiber bundles contained tyrosine hydroxylase immunoreactivity. Electron microscopic examination revealed that LNGFR immunoreactivity was located on the surface of Schwann cells, and frequently on the interface of axons and Schwann cells.
We present two cases of patients experiencing needle track seeding after undergoing percutaneous ethanol injection therapy (PEIT) for small hepatocellular carcinoma, who were treated by surgical resection of the metastases. One patient demonstrated metastatic tumors due to needle track seeding 6 months after the beginning of PEIT (a total of 7 injection sessions for 2 tumors measuring < 20 mm in diameters), whereas the other developed a metastatic tumor 20 months after beginning PEIT (a total of 30 injection sessions for 3 tumors measuring < 20 mm in diameter). In the two cases, both the primary and metastatic tumors histologically revealed moderately differentiated hepatocellular carcinoma. Moreover, the histological findings of the metastatic foci did not always appear to be more aggressive than those of the primary tumors. Therefore, in order to detect such metastasis as early as possible, more careful attention should be paid to the appearance of needle track seeding after performing PEIT for hepatocellular carcinoma, even if the target of such PEIT is small HCC.
A total of 128 surgically resected small hepatocellular carcinomas, measuring less than or equal to 3 cm in diameter, were studied by both macroscopic and histologic examinations. In 95 single nodular-type tumors of the 128 lesions, eight tumors were associated with the cancerous areas of well differentiated hepatocellular carcinoma around the nodule. These surrounding cancerous areas went undetected by both the preoperative radiological examinations and the gross findings of resected specimens. Based on the immunohistochemical findings, the labeling index, both of the proliferating cell nuclear antigen (PCNA) and of the Ki-67 in the surrounding cancerous areas, were lower than that of the main nodules but higher than in the nontumorous liver parenchyma in seven of eight cases. These results suggest that the main nodule was generated from the surrounding cancerous area, supporting the hypothesis of a stepwise progression of HCC. Even if the tumor seems to be a small and single nodular type, it is recommended that its surrounding areas should be closely examined and the surgical cutting margin should be made more than 1.0 cm away from the main nodule at hepatic resection.
We investigated the role of natural killer (NK) cells in the development of autoantibody production in which (C57BL/6 x DBA/2) F1 (BDF1) hybrid mice were injected intravenously with spleen cells (SC) from parental DBA/2 mice (treated BDF1 mice). Treated BDF1 mice began to show an increase in serum anti-dsDNA antibody 2 weeks after injection, while the NK activity of their SC transiently increased initially in the first 1 to 2 weeks after injection, but subsequently decreased dramatically. Flow cytometric analysis suggested that this sequential change in NK activity correlated with the absolute number of host-derived NK1.1+ cells in SC from treated BDF1 mice. We demonstrated that the level of anti-dsDNA in serum is directly influenced by the level of NK activity in treated BDF1 mice. Depletion of NK cells by administration of anti-NK1.1 mAb accelerated the development of autoantibody production, whereas augmentation of NK activity by administration of poly-(I:C) inhibited the development of autoantibody production. This inhibitory effect of poly(I:C) was abolished by prior depletion of NK cells. Interestingly, suppression of autoantibody production was seen only when poly(I:C) was administrated within 1 week after injection of parental SC. Last, we demonstrate that adoptive transfer of interleukin-2 (IL-2)-activated NK cells had a protective effect against the development of autoantibody production. These findings imply that NK cells may have a protective role in lupus-like disease especially in its early stage. In addition, it opens up the possibility that adoptive immunotherapy with IL-2-activated NK cells can delay or even prevent the development of autoimmune disease.
Lipopolysaccharide (LPS) triggers a wide range of cellular responses in mammalian cells. Several proteins, including CD14, have been reported to possess LPS binding capacity. However, the signal transduction molecule(s) and pathway(s) through which LPS induces cellular responses have not been identified. A rabbit antiserum (5299), which had been made against the N-terminal 20-amino-acid sequence of the cytokine, soluble immune response suppressor, unexpectedly activated the human monocyte cell line THP-1 as assessed by tumor necrosis factor (TNF alpha) production. Normal rabbit serum or IgG caused no TNF production, while antiserum 5299 induced TNF at a 1:10(4) dilution. The activity of antiserum 5299 was not due to endotoxin contamination, since antiserum 5299 contained less endotoxin than required for activation of THP-1 and the TNF-inducing activity disappeared when the antiserum was boiled. Moreover, the activity was recovered in purified immunoglobulin G fraction from the antiserum (5299 IgG). These results suggest that antiserum 5299 contains an antibody which recognizes a signal-transducing molecule. THP-1 desensitized by incubation with LPS for 24 hr could not respond to LPS or antiserum 5299 while responsiveness to phorbol ester remained, suggesting that the signals induced by the antiserum 5299 and LPS are transduced through at least a partially common pathway. Western blotting with antiserum 5299 showed a 69-kDa protein which rapidly appeared in Triton X-100-insoluble fraction after stimulation with LPS or 5299 IgG, suggesting an association of 69-kDa protein with the cytoskeleton. Furthermore, two mutant cell lines were isolated (T-15 and T-25) from gamma-irradiated THP-1 which lack responsiveness to LPS. In these mutant cell lines, the 69-kDa protein was not observed in Triton X-100-insoluble fraction after stimulation with LPS. It is proposed that this protein may play an important role in LPS-stimulated signal transduction.
Interleukin-6 (IL-6) has been shown to promote the attachment of rabbit corneal epithelial cells to fibronectin-coated substratum and ex vivo migration of the cells on the corneal stroma. To examine whether IL-6 promotes cell attachment through up-regulation of expression of integrin alpha 5 beta 1, i.e., the major cell surface fibronectin receptor, we quantified the levels of both alpha 5 and beta 1 subunit transcripts by reverse transcription-polymerase chain reaction in cultured rabbit corneal epithelial cells pretreated with various concentrations of IL-6. The levels of both alpha 5 and beta 1 mRNAs were dose-dependently elevated by IL-6, attaining 1.5- and 1.8-fold increases, respectively, at 10 ng/ml. The stimulatory effect of IL-6 was transient; the levels of both subunit mRNAs reached a maximum 1 h after the addition of IL-6 and returned to the basal levels after 6 h. The IL-6-induced up-regulation of integrin alpha 5 and beta 1 mRNAs was also confirmed by Northern blot analysis. These results indicate that the increased attachment of corneal epithelial cells to fibronectin and enhanced ex vivo migration on corneal stroma by IL-6 is, at least in part, due to the temporal up-regulation of integrin alpha 5 beta 1 expression in corneal epithelial cells.