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Biomedical subjects

H Ueyama

Publications and source records attributed to H Ueyama.

At least 37 records · Page 2Linked to original sources

An increased number of CD40-high monocytes in patients with Crohn's disease.

OBJECTIVE: CD40-CD40 ligand (CD40L) interaction is essential for the T-lymphocyte-dependent immune response. This interaction may be operational in the pathogenesis of inflammatory bowel diseases (IBD). The present study examined the expression of CD40 in peripheral blood mononuclear cells (PBMNCs) and tissue specimens, and CD40-stimulated interleukin (IL)-12 release from PBMNCs in IBD. METHODS: The expression of CD40 in PBMNCs and tissue inflammatory cells was examined by flowcytometry and immunohistochemistry, respectively. IL-12 release was measured in cultured media of PBMNCs by an enzyme-linked immunosorbent assay. RESULTS: Most peripheral blood B-lymphocytes expressed CD40 in all subjects. However, in ulcerative colitis (UC) patients, a significantly increased mean fluorescence intensity (MFI) of CD40 on B-lymphocytes was detected, compared with control subjects and patients with Crohn's disease (CD). In contrast, both the percentage positivity and MFI of CD40 on monocytes of active CD subjects were significantly increased, compared with the other groups. In active CD patients, a high level of IL-12 release from PBMNCs was observed by CD40 stimulation, compared with those of the other groups. When primed with IFN-gamma, PBMNCs from inactive CD patients released a significantly high level of IL-12, probably via stimulation by the CD40 monoclonal antibody. In the affected mucosa of CD, numerous CD40-positive cells were demonstrated, and they were also CD68-positive, suggesting these double CD40/ CD68-positive cells are tissue macrophages. CONCLUSIONS: These results suggest that the examination of CD40 expression in PBMNCs might enable the differentiation of CD from UC. CD40-high monocytes in CD patients may play a role in the pathogenesis of CD.

Adolescent↗

[Anesthetic management of heart transplantation].

We anesthetized a 47-yr-old man with end-stage hypertrophic cardiomyopathy for heart transplantation. This is the first case of heart transplantation from a patient with brain death, since the organ transplantation law had become valid in Japan. Anesthesia was induced and maintained with fentanyl and diazepam. Aseptic technique was used in inserting and securing all catheters. The patient was assisted by left ventricular assist system, and hemodynamic suppression at anesthetic induction was trivial. Since complete AV block was present at the termination of cardiopulmonary bypass (CPB), VVI pacing and infusion of isoproterenol were started. In addition, nitroglycerin was given for pulmonary vasodilation. The cardiovascular support used for weaning from CPB included dobutamine, isoprote-renol, dopamine and milrinone. Following weaning from CPB sinus rhythm appeared spontaneously and function of the transplanted heart was satisfactory. When the patient was transported to ICU reduction in doses of catecholamines was possible, and dopamine and milrinone were infused. The patient was extubated 10 hours after admission to ICU.

Anesthesia, Inhalation↗

[A case of acute quadriplegic myopathy].

A 22-year-old man developed unconsciousness, severe quadriplegia and muscle atrophy, and had markedly elevated serum creatine kinase levels after using the high-dose steroid and nondepolarizing neuromuscular blocking agents during the course of sepsis and DIC. On neurological examination, he was lethargic. The patient had generalized muscle weakness and wasting, and diminished deep tendon reflexes. He weakly responsed to painful stimuli on the legs. The motor nerve conduction study demonstrated decreased CMAP (compound muscle action potential) amplitudes. Motor and sensory nerve conduction velocities and their distal latencies were normal. Muscle biopsy revealed marked muscle fiber atrophy predominantly in type 2 fibers and numerous basophilic and a few necrotic fibers. Some atrophic fibers had decreased to absent myosin adenosine triphosphatase activity in their center. Accordingly, he was diagnosed as having acute quadriplegic myopathy (AQM), which has been reported mainly in Western countries. The mechanism of muscle fiber degradation in this myopathy is still unknown. On immunohistochemical analysis to our patient, enzyme activities of various proteases such as calpain, cathepsin B, and proteasomes were increased in the sarcoplasm, especially in the atrophic fibers. We suggest that lysosomal cathepsin, nonlysosomal calpain, and ATP-ubiquitin-proteasome proteolytic pathways participate in muscle fiber degradation in AQM.

Acute Disease↗

[A case of chronic hypertrophic cranial pachymeningitis causing cranial polyneuropathy and unilateral central retinal vein occlusion].

We presented a 70-year-old woman who developed unilateral visual loss due to central retinal vein occlusion caused by chronic hypertrophic cranial pachymeningitis. She had developed right blindness with optic atrophy due to chronic intracranial pachymeningitis one year before admission. In June 1999, she noticed visual loss of the left eye. On admission, neurological examination revealed left visual loss and sensory impairment in the first branch area of bilateral trigeminal nerves. Ophthalmological examination revealed central retinal vein occlusion of the left eye. Brain MRI showed dural thickening with gadolinium enhancement of the cavernous sinus near the left optic nerve and dilatation of the left supraorbital vein. There was no compression of the left optic nerve. We suggest that central retinal vein occlusion may be caused by compression of the supraorbital vein by dural thickening. This is the first case report of central retinal vein occlusion associated with chronic hypertrophic cranial pachymeningitis.

Aged↗

[A case of welder presenting with parkinsonism after chronic manganese exposure].

A 56-year-old welder working for 30 years developed postural instability and writing clumsiness since October, 1998. Neurologic findings revealed dystonia of the bilateral shoulders and distal four limbs as well as parkinsonism such as masked face, bradykinesia, rigidity, and retropulsion. Brain MRI showed hyperintensity lesions on T1-weighted images in the bilateral globus pallidus, midbrain, pontine tegmentum, dentate nucleus and cerebral white matter, which reduced in size and density after ten months. The diagnosis of manganese poisoning was made by the high manganese levels of both serum and urine, and by the marked elevated urinary manganese level after administration of the cheleting agent. We pointed out the diagnostic significance of brain MRI in patients with chronic manganese exposure.

Brain↗

Molecular cloning and expression of a mouse thiamin pyrophosphokinase cDNA.

Thiamin pyrophosphokinase (EC 2.7.6.2) catalyzes the pyrophosphorylation of thiamin with adenosine 5'-triphosphate to form thiamin pyrophosphate. A mouse thiamin pyrophosphokinase cDNA clone (mTPK1) was isolated using a combination of mouse expressed sequence tag database analysis, a two-step polymerase chain reaction procedure, and functional complementation screening with a Saccharomyces cerevisiae thiamin pyrophosphokinase-deficient mutant (thi80). The predicted protein contained 243 amino acid residues with a calculated molecular weight of 27,068. When the intact mTPK1 open reading frame was expressed as a glutathione S-transferase fusion protein in Escherichia coli lacking thiamin pyrophosphokinase, marked enzyme activity was detected in the bacterial cells. The corresponding 2.5-kilobase pair mRNA was expressed in a tissue-dependent manner and was found at relatively high levels in the kidney and liver, indicating that the mode of expression of mTPK1 genes differs with cell type. The expression of mTPK1 genes in cultured mouse neuroblastoma and normal liver cells was unaffected by the thiamin concentration in the medium (10 microM versus 3.0 nM). This is the first report on identification of the primary sequence for mammalian thiamin pyrophosphokinase.

Amino Acid Sequence↗

Tau expression in denervated rat muscles.

We studied whether denervation affects the expression of tau, in particular phosphorylated tau, and how it is degraded in rat soleus muscles. Immunoblot analysis showed a high molecular weight, approximately 110 kDa (big tau), in normal muscle. Tau levels increased significantly in denervated muscles treated with chloroquine (a lysosomotrophic agent) and in untreated ones, as compared to levels of similarly treated contralateral, innervated muscles. Most of the tau in the innervated and denervated muscles was phosphorylated. Immunohistochemically, tau and beta-tubulin colocated in the sarcoplasm of innervated, saline-treated (intact) muscle, but the staining intensities were very weak. Both proteins, however, were expressed extensively in these areas in the denervated muscles from saline-treated rats. In the denervated muscle of chloroquine-treated rats there were numerous autophagic vacuoles in the sarcoplasm, and phosphorylated-tau accumulation was marked within these vacuoles, indicative that tau first was taken into autophagic, vacuoles by nonselective autophagy then degraded via the lysosomal as well as the nonlysosomal calpain system. Our findings suggest that phosphorylated big tau accumulates with beta-tubulin in denervated muscular atrophy, possibly in order to maintain or preserve the integrity of the muscle fiber during progressive atrophy or regeneration.

Animals↗

Expression of genes for estrogen receptors alpha and beta in human articular chondrocytes.

OBJECTIVE: To investigate the gene expression of estrogen receptor (ER) alpha and ERbeta in human articular chondrocytes. METHODS: 16 articular cartilage specimens were obtained from 15 patients during surgery. Three of the specimens were from men and 13 from women; three from hip joints and 13 from knee joints; four were normal and 12 showed osteoarthritic cartilage. Total RNA was extracted from the articular chondrocytes and the expression of both ERalpha and ERbeta genes was investigated by the reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Gene expressions of ERalpha were detected in all specimens and those of ERbeta were found in 15 specimens by the RT-PCR method. There was a significant correlation between the amounts of ERalpha and ERbeta. Expression levels of both genes were significantly higher in men than in women. There were no significant differences in the expression levels of both ER genes between the hip and knee joint sites, nor between normal and osteoarthritic tissues. CONCLUSION: This study is to our knowledge the first to demonstrate the gene expression of both ERalpha and ERbeta in human articular chondrocytes. Since there are some functional differences between the two receptors, the effects of estrogen on cartilage metabolism should be elucidated by two different receptor mechanisms.

Adult↗

Effects of crystalloid and colloid preload on blood volume in the parturient undergoing spinal anesthesia for elective Cesarean section.

BACKGROUND: The role of crystalloid preloading to prevent hypotension associated with spinal anesthesia in parturients during cesarean section has been challenged. Direct measurement of blood volume should provide insight regarding the volume-expanding effects. The aim of the current study was to clarify the effects of volume preload with either crystalloid or colloid solution on the changes in blood volume of parturients undergoing spinal anesthesia for cesarean section. METHODS: Thirty-six healthy parturients scheduled for elective cesarean section during spinal anesthesia were allocated randomly to one of three groups receiving 1.5 l lactated Ringer's solution (LR; n = 12), 0.5 l hydroxyethylstarch solution, 6% (0.5 l HES; n = 12), and 1.0 l hydroxyethylstarch solution, 6% (1.0 l HES; n = 12), respectively. Blood volume and cardiac output were measured before and after volume preloading with indocyanine green (ICG), and the indocyanine green blood concentrations were monitored by noninvasive pulse spectrophotometry. RESULTS: After volume preload, the blood volume significantly increased in all three groups (P < 0.01). The volume of infused solution remaining in the vascular space in the LR, 0.5-l HES, and 1.0-l HES groups were 0.43+/-0.20 l, 0.54+/-0.14 l, and 1.03+/-0.21 l, respectively, corresponding to 28% of lactated Ringer's solution and 100% of hydroxyethylstarch solution infused. Significant increases in cardiac output were observed in the 0.5-l and 1.0-l HES groups (P < 0.01). A significant correlation between the percentage increase in blood volume and that of cardiac output was observed by volume preloading (r2 = 0.838; P < 0.001). The incidence of hypotension was 75% for the LR group, 58% for the 0.5-l HES group, and 17% for the 1.0-l HES group, respectively. CONCLUSIONS: The incidence of hypotension developed in the 1.0-l HES group was significantly lower than that in the LR and 0.5-l HES groups, showing that greater volume expansion results in less hypotension. This result indicates that the augmentation of blood volume with preloading, regardless of the fluid used, must be large enough to result in a significant increase in cardiac output for effective prevention of hypotension.

Adult↗

Polymorphic CAG repeats of the androgen receptor gene and rheumatoid arthritis.

OBJECTIVE: In view of the possible role of androgens in the pathogenesis of rheumatoid arthritis (RA), this study investigated the association between repeat lengths of CAG microsatellites of the androgen receptor (AR) gene and RA. METHODS: The number of CAG repeats in exon 1 of the AR gene was determined in 90 men and 276 women with RA, as well as in 305 male and 332 female controls. RESULTS: The male RA patients tended to have shorter repeats than the male controls (22.5 versus 23.1, p=0.07), whereas the female RA patients had similar repeats to the female controls (22.7 versus 22.9, p=0.17). Patients of both sexes were divided into younger and older age at onset groups, and compared with younger and older controls. Younger onset male RA patients had significantly shorter CAG repeat lengths than the younger male controls (21.8 versus 23.2, p=0.007) or the older onset male RA patients (21.8 versus 23.2, p=0.04). Older onset male RA and both younger and older onset female RA patients had similar CAG repeat lengths when compared with their controls. Neither seropositivity nor rheumatoid nodule positivity had a significant relation with CAG repeat lengths. CONCLUSION: Shorter CAG repeats of the AR gene, presenting high levels of transactivation activity, are related to younger age onset male RA, suggesting the possible role of androgens as a modulating factor.

Adult↗

[Two adult siblings with Chédiak-Higashi syndrome presenting as hyperpigmentation of the skin and the iris].

We report two adult siblings with Chédiak-Higashi syndrome presenting as hyperpigmentation of the skin and the iris. Patient 1 was a 30-year-old man who had generalized hyperpigmentation from one month of age, developed mental deterioration at age 9 years, and gait difficulty at age 20 years. On admission, he showed hyperpigmentation of the skin and the iris without partial albinism. Skin pigmentation was predominated at the face and the extremities. Neurologic examinations revealed mental dysfunction with IQ of 60 by WAIS, cerebellar ataxia, pyramidal signs, extrapyramidal signs, and polyneuropathy. Hematologic examination revealed peroxidase-positive giant granules in leukocytes and decreased activity of natural killer cells, leading to the diagnosis of Chédiak-Higashi syndrome. Patient 2, a younger brother of patient 1, was a 28 year-old man who also had hyperpigmentation of the skin and the iris with similar neurologic findings with patient 1. Both had no episodes of systemic infections. In Japanese cases of Chédiak-Higashi syndrome, more than 50% of them showed hyperpigmentation of the skin from the early stage of the disease. We pointed out that hyperpigmentation of the skin may be a good diagnostic help of Chédiak-Higashi syndrome in Japanese cases.

Adult↗

Modulation of apoptosis by endogenous Bcl-xL expression in MKN-45 human gastric cancer cells.

This study was designed to clarify the role of endogenous Bcl-xL expression in modulating apoptosis of malignant cells. Administration of bcl-x-antisense oligonucleotides decreased Bcl-xL protein levels in the MKN-45 human gastric cancer cell line. The decrease in Bcl-xL protein content resulted in increased cell death induced by serum deprivation or Fas-antibody administration. Flow cytometric analysis revealed that the increased apoptotic cell death was more prominent in bcl-x-antisense-treated cells as compared to control cells, bcl-x-sense-treated cells, or bcl-x-nonsense-treated cells. To inhibit the effect of intrinsic Bcl-xL protein, we overexpressed Bak, which binds Bcl-xL and inhibits the anti-apoptotic effect of Bcl-xL, by transfection into MKN-45 cells. Bak-overexpressing cells showed increased apoptotic cell death induced by Fas-antibody when compared to parent cells and MKN-neo-transfected cells. Bak-overexpressing cells also showed greater sensitization to 5-fluorouracil and cisplatin than parent cells and MKN-neo-transfected cells. In conclusion, we demonstrated that administration of bcl-x-antisense oligonucleotides or overexpression of Bak protein induces sensitization to apoptosis in MKN-45 gastric cancer cells, suggesting that endogenous Bcl-xL expression in cancer cells is an important modulator of apoptosis.

Antibodies, Monoclonal↗

The expression of beta-microseminoprotein gene is regulated by cAMP.

The promoter of the human gene for beta-microseminoprotein (MSP) was analyzed by transient transfection assay using LNCaP cells. The region between -2738 and -207 was not essential for the basal promoter activity, but regions from -206, -108, and -83 were necessary. Forskolin induced promoter activity to 3- to 14-fold when constructions containing the possible cAMP response element (CRE, -60 to -53) were used, but did not do so when it was deleted from them. In a gel-mobility shift assay using a probe corresponding to -68 to -48, the shifted band was competed with a consensus CRE, and supershifted when the anti-CREB-1 antibody was included. Forskolin added to LNCaP cells increased MSP mRNA to 6-fold within 12 h. These results suggest that the expression of MSP gene is regulated by cAMP via the CRE present in the promoter. Hormones such as (-)-epinephrine or vasoactive intestinal polypeptide also increased MSP mRNA to 2- to 3-fold.

Colforsin↗

Purification and characterization of a tumor lipid-mobilizing factor.

Cancer patients with weight loss showed urinary excretion of a lipid-mobilizing factor (LMF), determined by the ability to stimulate lipolysis in isolated murine epididymal adipocytes. Such bioactivity was not detectable in the urine of cancer patients without weight loss or in normal subjects. The LMF was purified using a combination of ion exchange, exclusion, and hydrophobic interaction chromatographies to give a single component of apparent Mr 43,000, which showed homology in amino acid sequence with human plasma Zn-alpha2-glycoprotein. Both substances showed the same mobility on denaturing and nondenaturing gels and the same chymotrypsin digestion pattern, both stained heavily for carbohydrate, and they showed similar immunoreactivity. Polyclonal antisera to human plasma Zn-alpha2-glycoprotein was also capable of neutralization of the bioactivity of human LMF in vitro. Using competitive PCR to quantify expression of Zn-alpha2-glycoprotein, we found that only those tumors that were capable of producing a decrease in carcass lipid expressed mRNA for Zn-alpha2-glycoprotein. These results provide strong evidence to suggest that tumor production of Zn-alpha2-glycoprotein is responsible for the lipid catabolism seen in cancer patients.

Adipocytes↗

Expression of three calpain isoform genes in human skeletal muscles.

Calpain is thought to be involved in muscular degradation in progressive muscular dystrophy (PMD), especially Duchenne and Becker muscular dystrophies. To assess the expression of calpain genes in skeletal muscles of patients with myopathies, we examined mRNA levels of three calpain isoforms by the quantitative reverse transcriptase-polymerase chain reaction method in biopsied muscles from control, PMD and amyotrophic lateral sclerosis (ALS) patients. There was a statistically significant increase in calpain 1 and calpain 2 mRNA levels in PMD and ALS patients as compared to controls. In contrast, there was a decrease in expression of calpain 3 mRNA in PMD, but it was not statistically significant. Expression of calpain 1 and calpain 2 positively correlated with each other, but not with calpain 3. These results indicate that expression of calpain 1 and calpain 2, but not calpain 3, are upregulated in diseased human muscles, likely playing a regulatory role in the process of myofibrillar degradation at the transcriptional as well as posttranslational level.

Adolescent↗

Localized muscle wasting as an initial symptom of skeletal muscle lymphoma.

A 25-year-old man who developed painless muscle wasting and weakness of only the right thigh, was eventually shown to have systemic lymphoma with muscle infiltration. The patient was initially misdiagnosed as focal inflammatory myopathy, and he had a partial response to steroid therapy. Immunohistochemical analysis of the biopsied muscle and the subcutaneous tumor revealed the infiltrating cells with a positive macrophage-associated marker (CD68) and negative T- or B-cell-associated antigens, leading to the final diagnosis of true histiocytic lymphoma. Although skeletal muscle lymphoma is extremely rare, it should be considered in the differential diagnosis of localized muscle wasting.

Adult↗

Effect of chloroquine-induced myopathy on rat soleus muscle sarcoplasm and expression of clathrin.

Clathrin-coated vesicles are involved in receptor-mediated intracellular transport pathways related to lysosomal proteolysis. Clathrin levels were significantly elevated in denervated soleus muscles from chloroquine- and saline-treated rats as compared with their contralateral, innervated muscles. No difference was found in the clathrin levels of the denervated muscles in both groups. The accumulation of autophagic vacuoles was marked only in chloroquine-treated muscles after denervation. These findings suggest that chloroquine does not inhibit intracellular trafficking of clathrin-coated vesicles during the overdevelopment of autophagic vacuoles.

Animals↗