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Biomedical subjects

K Yamaguchi

Publications and source records attributed to K Yamaguchi.

At least 685 records · Page 38Linked to original sources

Penicillin-binding protein sensitive to cephalexin in sporulation of Bacillus cereus.

Cephalexin, cefaclor, cefadroxil, and cefotaxime strongly inhibited sporulation of Bacillus cereus ts-4 at 1 microgram/ml. Cephalexin was most inhibitory on the sporulation of B. cereus when the antibiotic was added at 3 h after induction of sporulation by nutrient downshift technique. Examination of 4',6-diamidino-2-phenylindole-stained cells by fluorescence-phase contrast microscopy showed that cephalexin inhibited the formation of asymmetric septum. By using [3H]penicillin, eight penicillin-binding proteins (PBPs) were detected from the cells of B. cereus ts-4. Among them, four PBPs were also detected in sporulating cells. Affinity of cephalexin to PBPs were measured indirectly by competition for subsequent binding of radioactive penicillin G. Cephalexin strongly bound to PBP 4 with molecular weight of 72,000 in sporulating cells.

Bacillus cereus↗

Differentiation of cultured neuroblastoma induced by staurosporine and cyclic AMP: methods for assessing a neuronal phenotype.

In cultured neuronal cells, neurite outgrowth provides a prominent morphological marker for assessing neurotrophic activity derived from different sources. Permanent cell lines are often used for such studies; however, the process of differentiation should be evaluated biochemically or electrophysiologically to determine whether the affected cells acquire a neuronal phenotype. We have reported that low-dose staurosporine acts synergistically with cyclic AMP to induce neuronal differentiation in a human neuroblastoma cell line. This protocol describes a method for examining phenotypic changes in cultured cells.

Cell Differentiation↗

Kawaguchipeptin B, an antibacterial cyclic undecapeptide from the cyanobacterium Microcystis aeruginosa.

Kawaguchipeptin B, an antibacterial cyclic undecapeptide, was isolated from the cultured cyanobacterium Microcystis aeruginosa (NIES-88). Its structure was elucidated as 1 on the basis of 2D NMR data and chemical degradation. Kawaguchipeptin B (1) inhibited the growth of the Gram-positive bacterium Staphylococcus aureus at a concentration of 1 microgram/mL (MIC).

Anti-Bacterial Agents↗

Magnetic resonance imaging in acute optic neuropathy by sphenoidal mucocele.

A 45-year-old man suffered sudden visual loss with Marcus-Gunn pupil and a central scotoma in the right eye as evaluated by visual field examination. The patient's right visual acuity was 0.02 (0.08 x -4.5 D). T1-weighted magnetic resonance imaging (MRI) demonstrated a cystic lesion of high intensity in the right sphenoid sinus. Short T1 inversion recovery (STIR) images presented a cystic lesion of high intensity in the right sphenoid sinus. In addition, a ring-shaped high intensity signal was seen surrounding the right optic nerve. Post-contrast fat suppression T1-weighted images did not demonstrate any enhancement within the right optic nerve. After intranasal sphenoidectomy, good recovery of the right visual acuity was noted. On subsequent MRI, the cystic lesion in the sphenoid sinus disappeared. Also, the ring of high intensity surrounding the right optic nerve became unclear. It was thought that the ring of hyperintensity seen on the STIR images demonstrated cerebrospinal fluid in the dilated perioptic subarachnoid space. Therefore, these MRI findings suggested that the acute visual loss resulted from a disturbed blood supply to the optic nerve due to compression by the sphenoidal mucocele, and not due to an inflammatory process.

Humans↗

The atypical antipsychotic profile of NRA0045, a novel dopamine D4 and 5-hydroxytryptamine2A receptor antagonist, in rats.

1. The atypical antipsychotic profile of (R)-(+)-2-amino-4-(4-fluorophenyl)-5-[1-[4-(4-fluorophenyl)-4-oxobutyl] pyrrolidin-3-yl] thiazole (NRA0045), a potent dopamine D4 and 5-hydroxytryptamine (5-HT)2A receptor antagonist, was examined in rats. 2. Spontaneous locomotor activity was decreased dose-dependently with i.p. administration of clozapine (ED50 3.7 mg kg-1), haloperidol (ED50 0.1 mg kg-1) and chlorpromazine (ED50 0.9 mg kg-1), whereas inhibition of this type of behaviour induced by i.p. administration of NRA0045, at doses up to 10 mg kg-1, did not exceed 50%. 3. Locomotor hyperactivity induced by methamphetamine (MAP, 2 mg kg-1, i.p.) in rats (a model of antipsychotic activity) was dose-dependently antagonized by NRA0045 (ED50 0.4 mg kg-1, i.p., and 0.3 mg kg-1, p.o., respectively), clozapine (ED50 0.3 mg kg-1, i.p. and 0.8 mg kg-1, p.o., respectively), haloperidol (ED50 0.02 mg kg-1, i.p. and 0.1 mg kg-1, p.o., respectively), chlorpromazine (ED50 0.3 mg kg-1, i.p. and 3.3 mg kg-1, p.o., respectively). In contrast, the MAP (3 mg kg-1, i.v.)-induced stereotyped behaviour in rats (a model of extrapyramidal symptoms) was not affected by NRA0045 or clozapine, at the highest dose given (30 mg kg-1, i.p.). Haloperidol (ED50 0.3 mg kg-1, i.p.) and chlorpromazine (ED50 4.8 mg kg-1, i.p.) strongly blocked the MAP-induced stereotyped behaviour. NRA0045 and clozapine selectively blocked behaviour associated with activation of the mesolimbic/mesocortical dopamine neurones rather than nigrostriatal dopamine neurones. 4. Extracellular single-unit recording studies demonstrated that MAP (1 mg kg-1, i.v.) decreased the firing rate in the substantia nigra (A9) and ventral tegmental area (A10) dopamine neurones in anaesthetized rats. NRA0045 completely reversed the inhibitory effects of MAP on A10 dopamine neurones (ED50 0.1 mg kg-1, i.v.), whereas the inhibitory effects of MAP on A9 dopamine neurones were not affected by NRA0045, in doses up to 1 mg kg-1 (i.v.). Clozapine completely reversed the inhibitory effects of MAP on A10 dopamine neurones (ED50 1.9 mg kg-1, i.v.) and on A9 dopamine neurones (ED50 2.5 mg kg-1, i.v.). Haloperidol completely reversed the inhibitory effects of MAP on A10 (ED50 0.03 mg kg-1, i.v.) and on A9 dopamine neurones (0.02 mg kg-1, i.v.). NRA0045, like clozapine, was more potent in reversing the effects of MAP on A10 than A9 dopamine neurones. 5. Prepulse inhibition (PPI) is impaired markedly in humans with schizophrenia. The disruption of PPI in rats by apomorphine (0.5 mg kg-1, s.c.) was reversed significantly by NRA0045 (3 mg kg-1, i.p.), clozapine (3 mg kg-1, i.p.) and haloperidol (0.3 mg kg-1, i.p.). 6. Phencyclidine (PCP) elicits predominantly psychotic symptoms in normal humans and in schizophrenics. NRA0045 (0.03-0.3 mg kg-1, i.p.) and clozapine (0.1-1 mg kg-1, i.p.) significantly and dose-dependently shortened the PCP(1.25 mg kg-1, i.p.)-induced prolonged swimming latency in rats in a water maze task, whereas haloperidol (0.01-0.1 mg kg-1, i.p.) did not significantly alter swimming latency. 7. These findings suggest that NRA0045 may have unique antipsychotic activities without the liability of motor side effects typical of classical antipsychotics.

Animals↗

Mechanical agitation induces gene expression of NOR-1 and its closely related orphan nuclear receptors in leukemic cell lines.

NOR-1, NGFI-B and Nurr1 are closely related transcription factors that constitute a distinct subfamily within the nuclear receptor superfamily. Genes for these proteins are immediate-early genes, and are inducible in diverse cell types by various stimuli. In the present study, we investigated the effect of mechanical agitation on the gene expression of these transcription factors in cultured suspension cells by the quantitative reverse transcription-polymerase chain reaction. We found that mechanical agitation transiently induced NOR-1, NGFI-B and Nurr1 mRNAs in several leukemic cell lines in a dose-dependent manner. This induction was most pronounced in the HL-60 promyelocytic leukemia cell line, but also occurred to a lesser extent in other cell lines including KG-1, THP-1 and U937 cells. The induction was attenuated by serum or albumin, which are shear stress protectants for suspension culture cells. These reagents did not suppress forskolin-induced NOR-1 gene expression. These findings suggest the involvement of fluid shear stress in agitation-induced immediate-early gene expression. Since even moderate agitation could cause the induction, investigators should be cautious when evaluating the expression of immediate-early genes in some leukemic cell lines.

DNA-Binding Proteins↗

Combined effects of prolonged prostaglandin E1-induced hypotension and haemodilution on human hepatic function.

Combined effects of prolonged prostaglandin E1 (PGE1)-induced hypotension and haemodilution on hepatic function were studied in 30 patients undergoing hip surgery. The patients were randomly allocated to one of three groups; those in group A (n = 10) were subjected to controlled hypotension alone, those in group B (n = 10) to haemodilution alone and those in group C (n = 10) to both controlled hypotension and haemodilution. Haemodilution in groups B and C was produced by withdrawing approximately 1000 mL of blood and replacing it with the same amount of dextran solution, and final haematocrit values were 21 or 22%. Controlled hypotension in groups A and C was induced with PGE1 to maintain mean arterial blood pressure at 55 mmHg for 180 min. Measurements included arterial ketone body ratio (AKBR, aceto-acetate/3-hydroxybutyrate) and clinical hepatic function parameters. AKBR and biological hepatic function tests showed no change throughout the time course in groups A and B. In group C, AKBR showed a significant decrease at 120 min (-40%) and at 180 min (-49%) after the start of hypotension and at 60 min (-32%) after recovery of normotension, and SGOT, SGPT, LDH and total bilirubin showed significant increases after operation. The results suggest that a prolonged combination of more than 120 min of PGE1-induced hypotension and moderate haemodilution would cause impairment of hepatic function.

Adult↗

Differential expression of NGFI-B and RNR-1 genes in various tissues and developing brain of the rat: comparative study by quantitative reverse transcription-polymerase chain reaction.

NGFI-B and RNR-1 are closely related transcription factors that constitute a distinct subclass within the steroid/thyroid hormone receptor superfamily. They have been implicated in neuronal differentiation, neuroendocrine regulation of adrenocortical function and T-cell apoptosis. In this study, we measured and compared NGFI-B and RNR-1 mRNA levels in various adult rat tissues and in the developing rat brain by means of the quantitative reverse transcription-polymerase chain reaction. The use of RNA standards synthesized in vitro allowed direct comparison of the amount of the transcripts of these two genes. We demonstrated that the transcripts of both genes were present in all tissues examined although the expression levels widely varied. We found the highest constitutive expression of both genes in the pituitary. High levels of NGFI-B were also expressed in the cerebral cortex, muscle, ventral prostate, thymus and adrenal glands, whereas high levels of RNR-1 expression were restricted to the pituitary and cerebral cortex. These findings were consistent with the notion that NGFI-B and RNR-1 are involved in various signal transduction systems in diverse cell types. The amount of NGFI-B mRNA was greater than that of RNR-1 mRNA in all adult rat tissues, with the highest ratio of NGFI-B relative to RNR-1 expression in the muscle and leukocytes. In contrast, fetal rat brain showed relatively high RNR-1 gene expression. These findings suggested that the NGFI-B and RNR-1 genes are differentially expressed in a tissue-specific and developmentally regulated manner.

Adrenal Glands↗

Human T-cell lymphotropic virus type 1 can infect primary rat retinal glial cells and induce gene expression of inflammatory cytokines.

PURPOSE: To examine whether or not retinal glial cells can be infected by human T-cell lymphotropic virus type 1 (HTLV-1) and test the possibility that HTLV-1-infected retinal glial cells are involved in the pathogenesis of HTLV-1 uveitis (HU). METHODS: We tested infection of HTLV-1 by a standard coculturing method using WKAH rat retinal glial cells and irradiated MT-2, a human T cell line that produces HTLV-1. Infection was confirmed by detecting the integrated HTLV-1 provirus, using polymerase chain reaction (PCR), viral gene expression, using reverse transcriptase-PCR (RT-PCR) and HTLV-1 p19 ELISA, and by identifying the HTLV-1-infected glial cells by immunofluorescence cytochemistry and in situ hybridization. Changes in cytokine gene expression were studied by RT-PCR. RESULTS: Using a semiquantitative PCR of HTLV-1 provirus sequence, we found that 2.6% of the retinal glial cells were infected at 3 days after infection, followed by a gradual decrease in the percentage with an extended period of culture up to 4 weeks. This time course of infection was also verified by RT-PCR and ELISA studies that detect viral mRNA expression and protein production, respectively. Expression of HTLV-1 gag protein and tax mRNA was detected in a part of glial cells by indirect immunofluorescence cytochemistry and in situ hybridization, respectively. RT-PCR analysis of cytokine gene expression revealed that gene expression of IL-6, CINC-1 (Gro, KC), and TNF-alpha were induced in these cells, with a peak at 3 weeks after infection. CONCLUSION: These results provided supportive evidence for the theory that the infection of retinal glial cells by HTLV-1 and subsequent production of inflammatory cytokines could be one contributing factor for the development of the unique clinical features of HU. A better understanding of the specific roles of the inflammatory cytokines in the pathogenesis of HU would be beneficial in the treatment and control of this disease.

Animals↗

Different characteristics of gustatory responses between the greater superficial petrosal and chorda tympani nerves in the rat.

The integrated responses to gustatory stimuli applied to the soft palate were recorded from the greater superficial petrosal nerve (GSP) and were compared with those from the chorda tympani nerve (CT) innervating the anterior part of the tongue in the rat. Stimuli included various concentrations of NaCl, sucrose, HCl and quinine hydrochloride, and 0.5 M of six sugars. The inhibitory effects of amiloride on the responses to sodium salts, including various concentration of NaCl, 0.1 M sodium acetate and 0.01 M sodium saccharin, were also tested. Both the phasic and tonic responses to sugars in the GSP were significantly larger than those in the CT, whereas both responses to NaCl in the GSP were significantly smaller than those in the CT. Although amiloride at 50 microM significantly depressed the phasic and tonic responses to NaCl with a wide range of concentration in the CT, little inhibitory effect was observed in the GSP. The tonic response to sodium acetate, when dissolved in amiloride solution, was depressed to 15% of the control in the CT, and slightly but significantly depressed to 70% in the GSP. These response characteristics of the GSP may play important roles in the processing of gustatory information.

Amiloride↗

New murine model of bronchopneumonia due to cell-bound Haemophilus influenzae.

This murine model of nontypeable (unencapsulated) Haemophilus influenzae (NTHI) bronchopneumonia used organisms bound to mouse fetal lung (MFL) cells as an inoculum. Pretreatment of the mice with 40 microL of 1% formalin 3 days before intranasal instillation of the bacteria was necessary to allow infection. The number of NTHI recovered from the lungs plus trachea on day 7 after instillation was >100 times the number originally inoculated. Later, however, the number of recovered bacteria diminished gradually, and by day 14 it was almost identical to the original inoculum size. Serum IgM also peaked on day 7 after infection, after which IgG increased while IgM decreased. Histologically, bronchoalveolar infiltration of neutrophils was observed on day 3 after inoculation and continued at least for the following 4 days. The present experiment demonstrates that MFL cells can protect bacteria that have invaded the cells from the opsonizing and killing activities of host humoral defense mechanisms.

Animals↗

Antibiotic MICs and short time-killing against Helicobacter pylori: therapeutic potential of kanamycin.

We compared the susceptibility of Helicobacter pylori to several antibiotics, expressed as MICs and as bactericidal effectiveness in short (3 h) time-killing studies. Of the antimicrobial agent tests, clarithromycin and amoxycillin had the lowest MIC50, 0.063 and 0.125 mg/L respectively, for 24 strains of H. pylori. Minocycline, levofloxacin and lansoprazole followed, with MIC50s of 0.5, 1, and 2 mg/L, respectively. Three-hour time-killing studies using a standard strain demonstrated a different pattern. At 4 x MIC, kanamycin, metronidazole and clarithromycin produced 4.4, 2.6 and 2.1 log decreases in viability, whereas the remaining seven antibiotics (including amoxycillin) were less bactericidal. Amoxycillin's lack of bactericidal activity during brief incubations was confirmed by examining several different clinically isolated H. pylori strains. Clarithromycin's effect, on the other hand, was strain- and concentration-dependent. Kanamycin was the most potent antibiotic in short time-killing studies, with concentrations of 1 x MIC and 4 x MIC producing a reduction of more than 2 and 4 log respectively in all ten strains. Our data suggest that the MIC of antimicrobial agents against H. pylori does not necessarily predict their activity in short time-killing studies. Furthermore, our short time-kill data suggest kanamycin as a potential therapeutic choice for H. pylori infection, even though this agent's MIC would suggest limited activity.

Amoxicillin↗

Treatment of minute medullary thyroid carcinoma in multiple endocrine neoplasia 2A families first diagnosed by DNA analysis of RET proto-oncogene mutations: a case report.

Multiple endocrine neoplasia (MEN) 2A is an inherited disease characterized by the development of medullary thyroid carcinoma, pheochromocytoma and hyperparathyroidism. It has recently been shown to be associated with germ-line mutations in the RET proto-oncogene. We describe a 21-year-old man from a MEN 2A family who was found by DNA analysis to be a gene carrier of MEN 2A and then was diagnosed, using a stimulated calcitonin test, as having presymptomatic medullary thyroid carcinoma. His morbidity seems to have been cured by total thyroidectomy as postoperative calcitonin levels after stimulation are normal. It is concluded that direct genetic analysis for mutations in the RET proto-oncogene should be the diagnostic test of choice for identifying family members at risk for MEN 2A.

Adrenal Gland Neoplasms↗

A nationwide clinical survey of patients with multiple endocrine neoplasia type 2 and familial medullary thyroid carcinoma in Japan.

MEN (multiple endocrine neoplasia) type 2 syndrome is an inherited disease characterized by medullary thyroid carcinoma, pheochromocytoma, hyperparathyroidism and/or developmental anomalies. Germ-line mutations of the RET proto-oncogene have recently been identified as the underlying cause of the syndrome. Accordingly, several investigators have advocated prophylactic total thyroidectomy for medullary thyroid carcinoma at an early age in MEN 2 gene carriers identified by DNA analysis. Before applying this strategy in Japan, the biological behavior of each category of tumor in MEN 2 syndrome, and medullary thyroid carcinoma in particular, should be well understood. We conducted a nationwide questionnaire survey to clarify the clinicopathological features of MEN 2 in Japan, obtaining data for 230 patients diagnosed as having MEN 2. They included 84 males and 146 females, with a median age of 37.5 years (range 5-83). Patients were categorized as 179 with MEN 2A, 17 with MEN 2B, 12 with familial medullary thyroid carcinoma and 22 'other'. Medullary thyroid carcinoma, pheochromocytoma and parathyroid lesions occurred in 224 (97%), 132 (57%) and 25 (11%) patients respectively. Twelve patients (5.2%) died of medullary thyroid carcinoma and 11 patients died of other or unknown causes. Of 163 patients for whom follow-up data were obtained, 82 (50%) experienced recurrences of medullary thyroid carcinoma, including symptomatic recurrent tumors in 24 patients and elevated calcitonin levels alone in 54. In the era of RET mutational analysis for screening relatives of patients with MEN 2, these data provide useful information about surgical management for patients with MEN 2 in Japan.

Adolescent↗

Multiple endocrine neoplasia type 1 presented with manic-depressive disorder: a case report with an identified MEN1 gene mutation.

We report a case of multiple endocrine neoplasia type 1 who had repeated hypoglycemic episodes and had previously been diagnosed with bipolar manic-depressive disorder. The patient had a positive family history of multiple endocrine neoplasia type 1 and had multiple pancreatic endocrine tumors, hyperparathyroidism and possibly a pituitary tumor. The pancreatic tumors were resected by subtotal pancreatectomy and examined by histochemical staining and gene analysis. The tumor cells were positive for immunoreactive insulin and glucagon. A microsatellite polymorphism analysis revealed loss of heterozygosity on 11q13 in the tumors. By polymerase chain reaction-based nucleotide sequencing, we identified a germline mutation 483del2 of the MEN1 gene in the normal pancreatic tissue of the patient. This mutation causes a shift of the reading frame of menin mRNA at codon 125. It seems that the wild type allele of the MEN1 gene had been lost in the tumor cells whereas the mutant allele remained intact. This is the first identified MEN1 gene mutation in Japanese families and is different from all MEN1 gene mutations reported previously.

Adult↗

Postural and gait disturbance correlated with decreased frontal cerebral blood flow in Alzheimer disease.

In a group of 45 patients with Alzheimer disease (AD) and 15 control subjects, we studied the relationship between parameters showing postural and gait disturbance and regional cerebral blood flow (rCBF) measured with N-isopropyl-p-[123I]iodoam-phetamine single photon emission computed tomography at different clinical stages. Patients with AD with mild dementia exhibited only increased postural sway associated with a reduced mean value of rCBF in the cortex. At a moderate stage, the reduced mean values of rCBF in the cortex and in the frontal lobe were associated with increased postural sway and stride length variability and with decreased stride length. At a severe stage, reduced rCBF in the basal ganglia and in the frontal lobe additionally were associated with increased postural sway, double support time, and stride length variability, and with decreased walking speed and stride length. In multiple regression analysis, there was a high contribution by rCBF in the frontal lobe to account for postural and gait disturbance in AD. We propose that impaired frontal lobe circulation is an important factor causing postural and gait disturbance as AD progresses.

Aged↗

Biceps activity during shoulder motion: an electromyographic analysis.

Electromyographic responses in 44 shoulders from 30 subjects were examined. Fourteen shoulders from 13 patients had documented rotator cuff tears. The remaining volunteers had normal cuff integrity by history and examination. Electromyographic responses were recorded from the long head of the biceps, brachioradialis (elbow control), and from the supraspinatus (shoulder control). Elbow related biceps activity was minimized by using a brace locked in neutral forearm rotation and 100 degrees flexion. Analysis of normal and rotator cuff deficient data was performed in a masked fashion and electromyographic activity normalized as a percent of maximal muscle contraction during 10 shoulder motions based on the scapular plane. Normal shoulders in all ranges of active motion exhibited significant supraspinatus activity (20%-50% maximum muscle contraction). The response followed patterns expected for a shoulder stabilizer. In contrast, with every normal shoulder, biceps and brachioradialis activity remained insignificant (1.7%-3.6% maximum muscle contraction) and did not follow a patterned response. In patients with rotator cuff tears, biceps activity remained low (1.6%-4.4% maximum muscle contraction). As opposed to previous studies using electromyography about the shoulder, this trial examined shoulder specific biceps activity by relaxing the elbow. No significant biceps activity was observed in any shoulder, including patients with rotator cuff tears. Given these findings, any function of the long head of the biceps in shoulder motion does not involve active contractions.

Adult↗

Cloning and characterization of pejerrey mitochondrial DNA and its application for RFLP analysis

Probes were cloned, characterized, and developed for all regions of the mitochondrial DNA (mtDNA) of pejerrey Odontesthes bonariensis to provide the basis for the study of genetic diversity of South American atherinopsinii and to enable species identification from small amounts of tissue. The mtDNA was extracted from liver and cleaved with Eco RI, producing four fragments (7·4, 3·4, 3·1 and 2·9 kb) which were cloned using pUC118 plasmid vectors. Sequence analysis from both ends of the fragments showed that they encode tRNA (Asp, Phe, and Ser-TGA), 12 S rRNA, cytochrome oxidase (CO) II, NADH 4, 5, and 6, and the d-loop, and that the relative positions of these genes are identical to those in the mtDNA of other teleosts. A comparison of homology with carp mtDNA nucleotide sequences revealed that tRNA (Phe and Ser-TGA) and CO II were relatively conserved, whereas the d-loop region was highly divergent. The cloned mtDNA probes detected mtDNA fragments from about 800 ng of total DNA extracted from liver, muscle, and single embryos of O. bonariensis, and were effective for restriction length fragment polymorphism (RFLP) analysis of Patagonina hatcheri, the most distant atherinopsine relative of pejerrey. The cloned mtDNA probes may be useful for the analysis of genetic diversity and non-destructive species identification, including the examin-ation of eggs, larvae and juveniles. The mtDNA sequences reported here provide the basis for the design of primers for PCR-based RFLP analysis. 1997 The Fisheries Society of the British Isles

Journal Article↗