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Biomedical subjects

Y Kawamura

Publications and source records attributed to Y Kawamura.

At least 109 records · Page 6Linked to original sources

Reactivation of insertionally inactivated Shiga toxin 2 genes of Escherichia coli O157:H7 caused by nonreplicative transposition of the insertion sequence.

IS1203v is an insertion sequence which has been found in inactivated Shiga toxin 2 genes of Escherichia coli O157:H7. We analyzed the transpositional mechanism of IS1203v in order to investigate whether the Shiga toxin 2 genes inactivated by IS1203v could revert to the wild type. When the transposase activity of IS1203v was enhanced by artificial frameshifting, IS1203v was obviously excised from the Shiga toxin 2 gene in a circular form. The IS1203v circle consisted of the entire IS1203v, but an extra 3-bp sequence (ATC) intervened between the 5' and 3' ends of IS1203v. The extra 3-bp sequence was identical to a direct repeat which was probably generated upon insertion. Moreover, we detected the Shiga toxin 2 gene with a precise excision of IS1203v. In the wild-type situation, the transposition products of IS1203v could be observed by PCR amplification. These results show that IS1203v can transpose in a nonreplicative manner and that the Shiga toxin gene inactivated by this insertion sequence can revert to the wild type.

Bacterial Toxins↗

Swallow syncope associated with complete atrioventricular block and vasovagal syncope.

A 21 year old man presented with multiple, recurrent episodes of complete atrioventricular (AV) block associated with swallowing. Electrophysiological study revealed an AV block with swallowing of carbonated beverages and balloon inflation in the lower oesophagus. Evaluation did not demonstrate any underlying oesophageal or cardiac disease, and the AV block was not induced after intravenous atropine administration. The AV block was probably caused by a hypersensitive vagotonic reflex triggered by mechanical receptors in the lower oesophagus, resulting in suppression of the AV node. Head up tilt test revealed an increase in the high frequency spectrum of heart rate variability before the onset of the syncope. These findings suggest that the amplitude of the continual fluctuations in response to a variety of stimuli and derangement from both intrinsic and extrinsic environments was greater in this patient than in normal subjects. Swallow syncope is an unusual but treatable disorder. These reflexes that become exaggerated to the point of causing illness are poorly understood.

Adult↗

Vascular permeability: quantitative measurement with double-echo dynamic MR imaging--theory and clinical application.

Double-echo dynamic magnetic resonance (MR) imaging was used to evaluate both vascularity and permeability of tissues simultaneously. Vascularity was evaluated on the basis of the T2*-shortening effect due to the intravascular fraction of the contrast agent and permeability on the basis of the T1-shortening effect due to the extravascular fraction. Meningioma was characterized on the basis of higher vascularity and neurinoma on the basis of higher permeability. The proposed method enables better tissue characterization.

Adult↗

Collaborative work to evaluate toxicity on male reproductive organs by repeated dose studies in rats 19). Effects of two-week repeated dosing of enoxacin on the male reproductive organs.

The toxicity of Enoxacin (ENX), a fluoroquinolone antibacterial agent, on the testis and epididymis was studied in rats. ENX was administered to 5 male rats orally once daily for 2 weeks at the dose level of 3000 mg/kg/day. ENX-treated rats showed a marked decrease in body weight, and two of them died on Day 10. At the end of the dosing period, absolute weights of the epididymis were decreased; in contrast, relative weights of testis were increased in the ENX-treated group. On histopathological examination, testis of ENX-treated rats exhibited the following regressive changes: degeneration of spermatids and spermatocytes, retention of Step 19 spermatids, chromatin margination in nuclei of spermatids, multinucleated giant cell formation, and/or vacuolar degeneration of Sertoli cells. Additionally, desquamated cell debris was observed in the epididymis. Degenerative spermatids and spermatocytes were strongly positive by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). From these results, it is concluded that a 2-week treatment is sufficient to detect toxic effects of ENX on reproductive organs in male rats, and that testicular toxicity induced by ENX is associated with germ cell apoptosis.

Administration, Oral↗

Luminescence of Novel Neodymium Sulfonylaminate Complexes in Organic Media.

Luminescence of Nd(III) in an organic solvent having C-H bonds was achieved for the first time by complexing Nd(III) with long-chain perfluoroalkylated ligands such as bis(perfluorooctylsulfonyl)aminate (pos). The complex [Nd(pos)(3)] gave an emission quantum yield of 3.0+/-0.5 % in undeuterated acetone. The bulky pos ligands suppress the excitation of C-H vibrations, energy migration at diffusional collision, and the coordination of acetone molecules (see picture) to the Nd(III) center, which otherwise quench the luminescence.

Journal Article↗

[Coagulation/fibrinolysis disorders in the elderly].

To establish reference intervals for the interpretation of hemostatic tests in the elderly, 11 hemostatic tests were performed on 120 elderly individuals(26 males and 94 females) aged 78.6 years in average. The subjects lived in hostel for the elderly, walked without assistance, and were independent in activities in daily living. In 25.8% to 93.3% of the subjects, the results of the tests, except for prothrombin time and thrombin-antithrombin III complex, deviated from reference intervals obtained from young healthy subjects. This suggests the necessity of setting up reference intervals for the elderly. Fibrinolysis was more activated than coagulation, although coagulation had been considered to be more activated because thrombosis is common in the elderly. Wilcoxon test on the test values of the early elderly(65 to 74 years of age) and late elderly(75 to 92 years of age) as well as the correlation coefficient for age and test values revealed that separate reference intervals for antithrombin III, protein C, D-dimer, and thrombomodulin should be established for the early elderly(65 to 74 years) and late elderly(75 years or over). Our proposed reference intervals for the elderly are appropriate for the diagnosis of disseminated intravascular coagulation.

Aged↗

Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside.

BACKGROUND: : Many different methods have been used to assay amylase activity, using nitrophenylated oligosaccharides as substrate; however, the hydrolysis steps in these methods are complex. METHODS: : We developed a new continuously monitoring assay for amylase activity in biological fluids, using 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside (GalG2CNP) as the substrate; this assay was used with anti-human salivary amylase monoclonal antibodies for specific determination of the pancreatic isoenzyme. Amylase converted GalG2CNP into beta-D-galactopyranosylmaltose and 2-chloro-4-nitrophenol, which was measured at 405 nm. RESULTS: : GalG2CNP was cleaved between 2-chloro-4-nitrophenol and beta-D-galactopyranosylmaltose and did not undergo transfer reactions. The within-assay CVs (n = 20) for total amylase (T-AMY) and pancreatic amylase (P-AMY) were 0.6-1.6% and 0.5-2.5%, respectively; and day-to-day CVs (n = 10) for T-AMY and P-AMY were 0.8-3.7% and 0.6-4.1%, respectively. T-AMY and P-AMY activities in serum or urine obtained by the proposed method correlated well with those determined by the 2-chloro-4-nitrophenyl 4-O-beta-D-galactopyranosyl-beta-maltotetraoside method or the modified IFCC method. CONCLUSIONS: : This novel assay for T-AMY and P-AMY measures both activities stoichiometrically, directly, and easily, and may be suitable for routine procedures.

Amylases↗

[Effect of radiofrequency catheter ablation on autonomic tone in patients with common atrial flutter: difference depending on the site of ablation].

OBJECTIVES: Radiofrequency catheter ablation (RFCA) of the inferior vena cava-tricuspid annulus isthmus has become an effective procedure for radical treatment of common atrial flutter. This site is close to the right atrial posteroseptal region, so vagal denervation after slow pathway ablation in patients with atrioventricular nodal reentrant tachycardia can also occur after the procedure. To confirm this hypothesis, we tested whether autonomic disturbance can occur after RFCA in 16 patients with common atrial flutter. METHODS: Parameters of the autonomic function were measured using several components of the time domain analyses and power spectra of heart rate variability calculated from Holter recordings and the baroreflex sensitivity. Patients were divided into 2 groups based on the location of the ablated area: the isthmus group (9 patients) with the ablated area of the exact isthmus, and the posteroseptal group (7 patients) with the ablated area slightly shifted toward the right atrial posteroseptal position. All values after RFCA were compared with those before RFCA, and the ratio (after/before) was compared between the groups. The correlation between the ratio and cumulative delivered energy was also studied for each parameter. RESULTS: After RFCA, the heart rate increased, and the SD, rMSSD and pNN50 decreased significantly in the posteroseptal group, whereas no significant changes occurred in the isthmus group. The high frequency of the power spectra decreased after RFCA only in the posteroseptal group, and the ratio was significantly smaller in the posteroseptal group compared to the isthmus group. A significant reverse correlation between the ratio of the high frequency and cumulative energy was observed only in the posteroseptal group. The baroreflex sensitivity tended to be diminished after RFCA only in the posteroseptal group. CONCLUSIONS: Vagal denervation can occur after RFCA of common atrial flutter when the ablation site is shifted toward the right atrial posteroseptal region.

Atrial Flutter↗

Choroid plexus changes after temporal lobectomy.

BACKGROUND AND PURPOSE: Postoperative contrast-enhanced MR imaging of the brain is routinely used when evaluating for residual or recurrent brain tumor. It is imperative to be aware of morphologic changes and imaging features that typically occur in response to surgical manipulation at the postoperative site to avoid misinterpretation of imaging findings. Our purpose was to determine normal postoperative changes and alterations in the choroid plexus among patients who had undergone temporal lobectomy in order to distinguish this appearance from pathologic changes that may be seen in the presence of infection or recurrent tumors. METHODS: We reviewed 159 MR scans from 95 patients with hippocampal sclerosis or gliosis who underwent temporal lobectomy for treatment of intractable epilepsy. Choroid plexus location and size were assessed on contrast-enhanced T1-weighted images. RESULTS: After temporal lobectomy, the choroid plexus enlarged and sagged into the resection site. Increase in the size of the choroid plexus occurred in 58% of cases overall. The degree of enhancement also increased after surgery, sometimes resulting in a nodular pattern of enhancement. The changes were most marked during the 1st week after temporal lobectomy, and showed an enlarged, markedly enhancing choroid plexus on 86% of the scans. CONCLUSION: Postoperative changes of the choroid plexus after temporal lobectomy include sagging into the resection site, an increased size, and an increased degree of enhancement. Normal postoperative morphologic characteristics may mimic neoplastic enhancement pattern. Familiarity with this appearance is important to avoid a pitfall in diagnosis of recurrent postoperative temporal lobe neoplasms.

Adolescent↗

[A case of intravascular malignant lymphomatosis presenting as cerebral infarction].

A case of intravascular malignant lymphomatosis (IML) presenting as progressive cerebral infarction is reported. A 62-year-old previously healthy male developed progressive dementia. MRI of the brain at the nearest hospital revealed multiple infarcts with unknown etiology. His level of consciousness deteriorated rapidly, and then he was transferred to our hospital for further evaluation. High grade fever, raised serum C reactive protein (CRP), and raised lymphoma markers (serum LDH and soluble IL-2 receptor (sIL-2R)) were observed. Repeated brain MRI disclosed progression of multifocal cerebral infarctions. We considered IML most likely, and we performed muscle biopsy. However muscle biopsy didn't demonstrate any proliferation of neoplastic cells of lymphoid origin within small vessels. Thereafter IML was diagnosed by brain biopsy. The patient underwent chemotherapy, but died of pneumonia due to severe myelosuppression. IML is a rare disease but most commonly shows neurological symptomatology as its clinical manifestation. Dementia is the most common neurological symptom, and progressive multiple infarction is the most common of the MRI findings. Rapidly progressive dementia associated with multiple infarction, when elevated CRP, LDH and sIL-2R are observed in the laboratory data, is suggestive of IML.

Biomarkers, Tumor↗

[Ambulatory blood pressure monitoring in hypertensive CAPD patients].

The periodic structure of 24-hour blood pressure variation(circadian rhythm of blood pressure by ambulatory blood pressure monitoring(ABPM) in hypertensive CAPD patients was investigated by a new method of analysis based upon the maximum entropy method(MEM). In addition, this method allows the adequacy of antihypertensive therapies to be evaluated in such patients. The results were as follows; 1) The frequency of non-dipper type hypertension was 88%(36/41 cases), and the remaining 12% (5/41) were dipper type hypertension patients. The rise in morning blood pressure(morning surge: MS) was noted in 64% of the former. 2) Night time systolic blood pressure(182 +/- 22 mmHg, n = 36) was higher in patients with non-dipper type hypertension than in those with the dipper type(151 +/- 17 mmHg, n = 5, p < 0.01). 3) The standardized level of systolic blood pressure(SLSBP) calculated by MEM analysis in patients with non-dipper type hypertension(177 +/- 7 mmHg) was comparable with that in those with dipper type hypertension(168 +/- 13 mmHg, ns). 4) Treatment with long-acting Ca antagonist alone significantly reduced both SLSBP and the area over the SLSBP from 188 +/- 18 mmHg to 160 +/- 7 mmHg(p < 0.01, n = 8), and area over the SLSBP from 2,735 +/- 340 mmHg.hr to 1,945 +/- 298 mmHg.hr(p < 0.01, n = 8). 5) In addition to long-acting Ca antagonist, administration of alpha 1-blocker given at bed time was significantly efficacious in reducing the rise in morning blood pressure, MS. The present study using MEM analysis of ABPM suggests that the blood pressure profile of hypertensive CAPD patients is characterized by a non-dipper type dominance and a frequent morning surge. Furthermore, the combined therapy with long-acting Ca antagonist and alpha 1-blocker was substantially effective both in reducing the overall blood pressure level, and in inhibiting the MS. This combined antihypertensive therapy may be potentially useful to prevent CAPD patients from the future development of cardiovascular complications.

Adrenergic alpha-Antagonists↗

Expression of the mRNA for two isoforms of neural plakophilin-related arm-repeat protein/delta-catenin in rodent neurons and glial cells.

Neural plakophilin-related arm-repeat protein (NPRAP) is a mammalian brain protein of the armadillo family. Here, in an attempt to elucidate its function, we determined the mouse brain regions and cell types expressing the mRNAs for two NPRAP isoforms (the longer and the shorter isoforms), and examined the regulation of expression of the NPRAP mRNAs during the differentiation of P19 cells into neurons. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that both variants were expressed in various mouse brain regions, but the mRNA for the short isoform was predominant in most regions. Primary cultures of both neurons and glial cells exhibited high expression levels of both the mRNAs, indicating that NPRAP mRNA expression is not neuron-specific. The expression of the two NPRAP mRNA variants was dramatically induced even prior to the terminal neuronal and glial differentiation of P19 cells after retinoic acid treatment. These data suggest that the two NPRAP isoforms function not only in neurons and glial cells in the brain, but also play a role in the differentiation of precursor cells into neurons and glial cells.

Animals↗

Traumatic injury in vitro induces IEG mRNAs in cultured glial cells, suppressed by co-culture with neurons.

Glial changes following traumatic injury to glial monolayers as well as to neuronal-glial co-culture systems in vitro were examined with a focus on the expression of mRNAs coding for the immediate early genes (IEG) c-fos, c-jun and zif/268, demonstrated using in situ hybridization. Glial cells along scratch wound lines extended cytoplasmic processes as early as 10 min post-injury and the whole wound was covered with gliosis by 24 h. For complete restoration in the case of glial cells co-cultured with neurons, this required 48 h. Induction of the three IEG mRNAs was eminent along the edges of scratch wound, peaking at 30-60 min post-injury and subsiding by 3 h. The peak expression of IEG mRNAs was delayed to 1-3 h post-injury and became undetectable at 6 h in neuronal-glial co-cultures. The data suggest that mechanical injury to glial cells causes gliosis and the expression of IEG mRNAs, which are suppressed by co-culture with neurons, indicating some influence of neuronal-glial interactions.

Animals↗

Purification and characterization of brevinase, a heterogeneous two-chain fibrinolytic enzyme from the venom of Korean snake, Agkistrodon blomhoffii brevicaudus.

A fibrinolytic enzyme, designated as brevinase, was purified from the venom of Korean snake, Agkistrodon blomhoffii brevicaudus. Brevinase cleaved both the Aalpha- and Bbeta-chains of fibrinogen but did not affect the gamma-chain. It showed beta-fibrinogenase activity devoid of fibrinogen clotting and caseinolytic activity. The fibrinolytic activity was completely inhibited by PMSF, DFP, Pefabloc, and DTT, indicating brevinase is a serine protease requiring disulfide bridge(s) for its activity. It kept 80% of the initial activity after heating at 100 degrees C for 3 min, showed an equal maximum activity in the pH range from 5.5 to 8.5, and was inactivated by Zn(2+). Brevinase consists of two polypeptide chains of 16.5 and 17 kDa linked by disulfide bridge(s). The N-terminal amino acid sequences of 16.5 and 17 kDa chains showed homology to the N-terminal and the internal (central region) amino acid sequences of single-chain fibrinolytic enzymes in snake venom, respectively.

Amino Acid Sequence↗

Implications of presenilin 1 mutations in Alzheimer's disease.

Mutations in presenilin 1 (PS1) and presenilin 2 (PS2) are the most common genetic factors underlying the development of early-onset familial Alzheimer's disease (FAD). To investigate the pathogenic mechanism of PS1 mutations linked to FAD, we established inducible mouse neuroblastoma (Neuro 2a) cell lines expressing the human wild-type (wt) or mutated PS1(M146L or deltaexon 10) under the control of the Lac repressor. Using this inducible PS1 system, the influence of PS1 mutations on the generation of endogenous murine Abeta species was assessed using a highly sensitive immunoblotting technique. The induction of mutated PS1 resulted in an increase in the extra- and intracellular levels of two distinct Abeta species ending at residue 42, Abeta1-42 and its N-terminally truncated variant(s), Abetax-42. In addition, the intracellular generation of these Abeta42 species was completely blocked by brefeldin A. In contrast, it exhibited differential sensitivities to monensin such that there was an increased accumulation of intracellular Abetax-42 but an inhibition of intracellular Abeta1-42 generation. These data strongly suggest that Abetax-42 is generated in a proximal Golgi compartment, whereas Abeta1-42 is generated in a distal Golgi and/or a post-Golgi compartment. Thus, it appears that PS1 mutations enhance the degree of 42-specific gamma-secretase cleavage which occurs (i) in the ER or the early Golgi apparatus prior to gamma-secretase cleavage, or (ii) in the distinct sites where Abetax-42 and Abeta1-42 are generated. To date, the site of Abeta42 generation has not been firmly established. Our data provide new information regarding the site of Abeta42 generation mediated by the FAD-linked mutant PS1.

Alzheimer Disease↗

Identification of a human cDNA encoding a novel protein structurally related to the yeast membrane-associated metalloprotease, Ste24p.

Recently, a novel membrane-associated metalloprotease, designated Ste24p, has been identified in Saccharomyces cerevisiae [K. Fujimura-Kamada, F.J. Nouvet, S. Michaelis, J. Cell Biol. 27 (1997) 271-285]. We cloned a human brain cDNA encoding a protein homologous to Ste24p (designated Hs Ste24p). The predicted 475-amino acid product of its open reading frame exhibited 62% similarity to Ste24p, and contained a zinc metalloprotease motif (HEXXH) and multiple predicted membrane spans. Northern blot analysis showed that this gene was expressed in most tissues. Immunofluorescence analysis of epitope-tagged Hs Ste24p constructs suggested that it is localized in the ER and possibly also in the Golgi compartment. A search of the expression sequence tag database identified a fragment of DNA encoding a segment homologous to the segment of Hs Ste24p containing the HEXXH motif in insects and nematodes. Thus, Hs Ste24p could be a member of a new family of Ste24p-like membrane-associated metalloproteases which are widely conserved in eukaryotes.

Amino Acid Sequence↗

Quantitative analysis of cerebral microvascular hemodynamics with T2-weighted dynamic MR imaging.

The purpose of this study was to quantify cerebral microvascular hemodynamics with T2-weighted dynamic susceptibility contrast-enhanced magnetic resonance imaging (DSC-MRI) using a half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequence. We performed T2-weighted DSC-MRI with HASTE sequence in 19 normal subjects. After bolus injection of gadopentetate dimeglumine, HASTE images of two sections were acquired for the simultaneous creation of concentration-time curves in the internal carotid artery and in brain tissue. Absolute regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), and mean transit time (MTT) values of brain tissue were calculated on a base of the indicator dilution theory, and all values were corrected on the assumption that rCBF of white matter is constant in 22 mL/100 g tissue/min without age-dependent alteration. A decrease in rCBV and rCBF of gray matter was age dependent, while rCBV of white matter did not show significant change with aging. The mean rCBF value in gray matter was 37.3 +/- 8.4 mL/100 g tissue/min. The mean rCBV value was 4.1 +/- 0.8 mL/100 g tissue in gray matter and 2.9 + 0.4 mL/100 g tissue in white matter. The rCBV and rCBF values of gray and white matter obtained from T2-weighted DSC-MRI with HASTE sequence were slightly lower than the published data calculated by gradient-echo sequence. We were able to perform absolute quantifications of the capillary blood volume and flow, using a HASTE sequence, which would not have been possible with a gradient-echo sequence. This technique provides a new method for estimating cerebral microvascular hemodynamics.

Adult↗

Identification of an insertion sequence, IS1203 variant, in a Shiga toxin 2 gene of Escherichia coli O157:H7.

An insertion sequence composed of 1310 bp was found in Shiga toxin 2 genes of some isolates of Escherichia coli O157:H7. This insertion sequence showed extremely high homology with IS1203 of E. coli O111:H(-). This IS1203 variant was inserted in the region encoding the amino-terminus of the B subunit with a duplication of 3 bp at the target site, resulting in inactivation of the Shiga toxin 2 gene.

Journal Article↗