PubMed HealthSearch

Biomedical subjects

Y Sakakibara

Publications and source records attributed to Y Sakakibara.

At least 19 recordsLinked to original sources

Novel Escherichia coli mutant, dnaR, thermosensitive in initiation of chromosome replication.

A newly isolated Escherichia coli mutant thermosensitive in DNA synthesis had an allele named dnaR130, which was located at 26.3 minutes on the genetic map. The mutant was defective in initiation of chromosome replication but not in propagation at a high temperature. This mutant was capable of growing in the absence of the rnh function at the high temperature by means of a dnaA-independent replication mechanism. In the mutant exposed to the high temperature, an oriC plasmid was able to replicate, although at a lower rate than at the low temperature. The plasmid replication at the high temperature depended on the dnaA function essential for the initiation of replication from oriC. The mutant lacking the rnh function persistently maintained the oriC plasmid at the high temperature in a dnaA-dependent manner. Thus, the dnaR function was required for initiation of replication of the bacterial chromosome from oriC but not the oriC plasmid. This result reveals that a dnaR-dependent initiation mechanism that is dispensable for oriC plasmid replication operates in the bacterial chromosome replication.

Bacterial Proteins

dnaR function of the prs gene of Escherichia coli in initiation of chromosome replication.

A new Escherichia coli mutant named dnaR, which was temperature sensitive in initiation of DNA replication, has been characterized through identification of the mutant gene. A 1.65 x 10(3) base-pair chromosomal DNA fragment isolated from wild-type cells, but not the corresponding fragment from the dnaR mutant, exhibited an activity that reversed temperature-sensitive growth of the mutant. The DNA fragment was found to include the entire prs-coding sequence and specify a 34,000 M(r) protein with phosphoribosylpyrophosphate synthetase activity. The dnaR mutation resided within the prs-coding segment and made the synthetase thermolabile. The coding segment for the dnaR product was determined, by introduction of various mutations into the cloned fragment, to be the same as that for the synthetase. The dnaR function of the prs gene product in DNA replication is discussed on the basis of an observation that thermal treatment of the dnaR mutant caused a delay in initiation of chromosome replication after the downshift, despite the presence of the synthetase activity at the preheat level.

Base Sequence

Laser therapy for early upper gastrointestinal carcinoma.

An invasive procedure generally is the first choice of therapy for the treatment of gastric cancer. The development of improved microimaging and innovative engineering, however, has established a new field of endoscopic laser therapy. This article discusses the use of high-power and low-power lasers in the treatment of early gastric cancer.

Carcinoma

Characterization of the sodium current in single human atrial myocytes.

Patch-clamp recording techniques have permitted measurement of the fast Na+ current (INa) in isolated cardiac cells from a number of species in recent years. However, there is still only very little information concerning human cardiac INa. The purpose of this study was to describe the kinetics of INa in normal-appearing, Ca(2+)-tolerant, enzymatically isolated human atrial myocytes using whole-cell voltage-clamp techniques. Atrial specimens were obtained from 46 patients undergoing open heart surgery. Cs+ was substituted for K+ in both pipette and external solutions and F- was added to the former. The reversal potential of the rapid inward current varied approximately 57 mV at 17 +/- 1 degrees C with a 10-fold change in [Na+]o, and the current was completely blocked by 100 microM tetrodotoxin, findings typical of the fast cardiac Na+ current. The tetrodotoxin dose-response curve was best fitted by an equation describing binding to high- and low-affinity sites. INa was activated at a voltage threshold of -70 to -60 mV, and peak inward current was obtained at approximately -30 mV (holding potential, -140 mV). The inactivation time course was voltage dependent and was fitted best by the sum of two exponentials. The relation between voltage and steady-state availability (h infinity) was sigmoidal with the half-inactivation at -95.8 +/- 0.9 mV and a slope factor of 5.3 +/- 0.1 mV (n = 46), and we did not observe a significant difference with disease and age. The overlap of the h infinity and activation curves suggested the presence of a Na+ "window" current. Recovery from inactivation also was voltage dependent and best fitted by a model describing the sum of two exponentials. Recovery occurred after an initial delay at potentials positive to -140 mV, suggesting that inactivation of human atrial INa is a multistate process. We conclude that INa of normal-appearing, Ca(2+)-tolerant human atrial myocytes is similar to that of other mammalian cardiac cells with the possible exception of having two tetrodotoxin binding sites.

Adolescent

Effect of estradiol and ethynylestradiol on microtubule distribution in Chinese hamster V79 cells.

The effects of estradiol (E2) and ethynylestradiol (EE2) on the chromosome number and cellular microtubule architecture of Chinese hamster V79 cells were studied using fluorescent anti-tubulin antibody. Treatment with 20 microM E2 for 48 h induced only a small amount of tetraploid cells, but the normal microtubule network was disrupted completely by only 3 h of treatment. This data reveals that E2 has higher microtubule-disruptive activity than diethylstilbestrol in V79 cells.

Animals

Effects of 3,3'-dihydroxy-alpha,beta-diethylstilbene and 3,3',4,5'-tetrahydroxystilbene on microtubule assembly in vitro, aneuploidy induction, and cellular microtubule and actin networks.

We examined the inhibitory activities of 3,3'-dihydroxy-alpha,beta-diethylstilbene (DDS) and 3,3',4,5'-tetrahydroxystilbene (THS) on microtubule assembly in vitro and their effects on chromosome number and cellular microtubule networks in Chinese hamster V79 cells. DDS showed half the inhibitory activity of diethylstilbestrol (DES) on microtubule assembly in vitro, while THS had none of the inhibitory activity. DDS induced tetraploid at 30 microM, whereas THS was found to be inactive. Furthermore, DDS disturbed cellular microtubule networks at 100 microM. We also examined the effects of DES, DDS and THS on cellular actin networks in mouse BALB 3T3 cells. DES induced a change of actin stress fiber distribution and THS had similar activity, while DDS showed no activity.

Actins

Effects of 32-oxygenated lanosterol derivatives on 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and cholesterol biosynthesis from 24,25-dihydrolanosterol.

The effects of 32-oxygenated lanosterol derivatives on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and cholesterol biosynthesis from [24,25-3H]24,25-dihydrolanosterol were studied. Among the derivatives, 3 beta-hydroxylanost-7-en-32-oic acid was the most active in depressing HMG-CoA reductase activity (IC50: 0.7 microM) and cholesterol biosynthesis (IC50: 0.4 microM) from 24,25-dihydrolanosterol.

Animals

Effects of (+)-, (-)- and (+/-)-indenestrols A and B on microtubule polymerization.

Indenestrol A (IA) is a metabolite of diethylstilbestrol (DES), and indenestrol B (IB) is an analog of IA. IA was simply obtained from E,E-dienestrol in the presence of dilute sulfuric acid, and a mixture of IA and IB was formed by thermal cyclization of E,E-dienestrol. In order to elucidate the effects of optically active IA and IB on microtubule assembly, the IA and IB enantiomers were separated to greater than 99% purity by high-pressure liquid chromatography using a chiral column. The di(4-bromobenzoate) of (-)-IB was analyzed by X-ray crystallography and its absolute structure was determined as C(3)-S. The (+)-, (-)-, and (+/-)-indenestrols A and B were shown to be inhibitors of microtubule assembly in vitro using microtubule proteins from porcine brain. (+/-)-IB is more active than (+/-)-IA, and the order of inhibitory activity of the enantiomers on microtubule assembly was (+)-IB greater than (+)-IA greater than (-)-IA greater than (-)-IB.

Animals

Effect of verapamil on ventilatory and circulatory responses to hypoxia and hypercapnia in normal subjects.

Recent investigations have shown that the calcium channel blocker verapamil attenuated the hypoxic ventilatory chemosensitivity of carotid body in animals. To determine whether this is also the case in humans, transient physiological chemodenervation by O2 breaths (withdrawal test) during sustained hypoxia (N = 7), and ventilatory and circulatory responses to progressive hypoxia and hypercapnia (N = 8) were examined after oral administration of verapamil. During sustained hypoxia after verpamil, there was a significant reduction of withdrawal response from 5th to 25th min value (p < 0.01), but not after placebo. On the other hand, no significant difference in ventilatory responses to progressive hypoxia and hypercapnia was observed after verapamil. Verapamil run reveals similar features with placebo run in circulatory parameters except blood pressure response, which tended to be suppressed by verapamil. We conclude that verapamil attenuates peripheral chemoreceptor activity with time during sustained mild hypoxia in normal adult humans and this may be explained by delayed depletion in intracellular Ca2+ for chemotransduction of the peripheral chemoreceptors.

Administration, Oral

Post-translational modification of protein by tyrosine sulfation: active sulfate PAPS is the essential substrate for this modification.

In vitro tyrosine sulfation of recombinant proteins would be a valuable tool in converting those proteins expressed in prokaryotic vectors to their natural form. For this purpose tyrosylprotein sulfotransferase (TPST), the enzyme responsible for tyrosine sulfation of proteins, was characterized from a bovine liver Golgi preparation. TPST was active in a acidic environment with a pH optimum of 6.25, and displayed a stimulation by the Mn2+, with the optimum activity in the presence of 5mM MnCl2. TPST was able to sulfate recombinant hirudin variant 1 (rHV-1) expressed in Escherichia coli and the C-terminal hirudin fragment 54-65 but not the N-terminal hirudin fragment 1-15 by using 3'-phosphoadenosine 5'-phosphosulfate (PAPS), indicating its specificity for the naturally sulfated tyrosine 63. Comparison of the reaction kinetics on synthetic peptides showed that the bovine liver TPST has a higher affinity and reaction rates for those peptides with a aspartyl residue on the N-terminal side of the tyrosine when compared with a glutamyl residue.

Amino Acid Sequence

[A successful surgical treatment of chronic traumatic mid-descending thoracic aortic aneurysm].

A 48-year-old woman with chronic mid-descending thoracic aortic aneurysm was successfully repaired. She received a blunt chest trauma due to automobile accident at July in 1988. Left upper lobectomy was performed for her lung contusion. March 1990, she admitted our hospital with abnormal shadow revealed by a chest roentgenogram. A computed tomogram of the chest and an aortogram revealed two false aneurysms. One of them was located at mid-descending thoracic aorta and another was aortic isthmus. Under a partial femoral veno-arterial bypass, a Dacron graft replacement of mid-descending thoracic aorta was performed and aneurysm of isthmus was wrapped by Teflon mesh after the left pneumonectomy. 2 months after the operation, bronchopulmonary fistula occurred at the left bronchial stump. The fistula was successfully covered with major omentum. The mid-descending thoracic aortic aneurysm due to blunt chest trauma is rare. The traumatic aortic aneurysms commonly occur aortic isthmus or ascending aorta. Initial diagnosis of traumatic mid-descending thoracic aortic aneurysm is often missed or delayed. Careful follow up is need and when an abnormality is revealed by chest roentgenogram, computed tomogram and aortogram should be obtained to make diagnosis of chronic traumatic aneurysm.

Aorta, Thoracic

[A case report of poststernotomy mediastinitis treated with the pectoralis major muscle flap after protracted wound irrigation].

A 8-year-old boy developed mediastinitis after direct closure of atrial septal defect. Methicillin-resistant Staphylococcus aureus (MRSA) was detected from the wound. Intermittent wound irrigation with povidone-iodine solution and vancomycin solution was protracted for two months. After the remission of MRSA infection, implantation of pectoralis major muscle flap was performed. Without recurrence of infection the wound was healed completely. Although pectoralis major muscle flap is an effective method of choice for mediastinitis, an appropriate timing of enforcing this method should be investigated hereafter.

Child

[Biventricular heart assist using centrifugal pump and roller pump in profound ventricular failure after open heart surgery: a case report].

A forty-seven-year-old female with profound biventricular heart failure probably caused by coronary artery spasm after open mitral commissurotomy was successfully survived by mechanical circulatory support. Centrifugal and roller pumps were applied to left and right side circulatory assist respectively. Recovery of cardiac function was evaluated by transesophageal echocardiography and hemodynamic parameters during intentional low flow assist. Finally, she was weaned from assist after 136 hours. No systemic thromboembolism was recognized after pump removal. Combination of centrifugal and roller pumps could be applicable to biventricular heart assist without crucial complication.

Cardiac Output, Low

[Surgical managements of the compression of trachea and/or bronchus associated with congenital cardiovascular anomalies].

From 1987 through 1990, 17 cases with tracheal and/or bronchial compression due to congenital cardiovascular malformations were experienced at our institutes. Respirator was required preoperatively in 10 patients (59%) due to severe respiratory symptoms by airway compression and narrowing. In all cases, bronchoscopy was carried out pre- or intra-operatively for the precise diagnosis and for the decision of operative procedures. Postoperatively 12 cases were successfully extubated and remained free of respiratory distress. We investigated the relationships among pre- and post-operative pressure ratio, preoperative flow ratio, requirement of preoperative respirator and indication for plication and suspension of pulmonary artery in the group of the patients, who had airway compression by the dilated pulmonary artery due to large left to right shunt. As a result of this investigation, these factors have no significant correlation. The severity of associated tracheomalacia might be a most susceptible cause, which required the preoperative managements with respirator and the surgical interventions to the dilated pulmonary artery. Infant with patent ductus arteriosus (PDA) could be surgically treated by the division of PDA through left lateral thoracotomy. If the left main bronchus shows complete obstruction after division of PDA, the additional surgical intervention like suspension of aortic arch is needed. Two cases were lost with reoperation and these suggested not only the priority of simultaneous repair of trachea and cardiovascular anomaly, but the necessity of minor intervention against adhesion (insertion of Gore-Tex sheet between suspended pulmonary artery and sternum) in the first-step palliative operation.(ABSTRACT TRUNCATED AT 250 WORDS)

Aorta

[Long-term clinical results after aortic valve replacement with mechanical heart valves and mitral valve replacement with porcine valves].

Long-term clinical results of aortic valve replacement (AVR) with mechanical heart valves and mitral valve replacement (MVR) with porcine valves were analysed. Sixty-three patients received isolated AVR and 48 received isolated MVR. Sixty-eight patients with MVR including double or triple valve replacement were also added in order to evaluate the primary tissue failure (PTF). The patients with operative deaths were excluded. Survival rate at 11 years in AVR was 68 +/- 10% and 67 +/- 15% in MVR without statistical difference. At 11 years, 76 +/- 8% of the patients in AVR were free from valve-related complications in contrast with the poor result of 34 +/- 31% in MVR (p less than 0.01). Main cause of this poor result in MVR was PTF as indicated in following event free rates; 83 +/- 9% at 7 years, 61 +/- 25% at 10 years and 49 +/- 31% at 13 years. There was no statistical difference between patients of above 50 years and below 49 years in PTF. Valve-related death event free was 93 +/- 5% in AVR and 86 +/- 11% in MVR at 11 years (not significant), however, there was statistical difference in re-operation event free rate as 94 +/- 5% in AVR and 76 +/- 11% in MVR at 11 years (p less than 0.001). These results suggest that the use of porcine valves in mitral position is confined to the selected patients.

Adult

[Differentiation therapy of acute promyelocytic leukemia: two successful cases of remission induction by all-trans retinoic acid].

We described two pediatric patients with acute promyelocytic leukemia (APL) who were successfully induced into complete remission with all-trans retinoic acid (ATRA, 45 mg/m2 per day) after failing on conventional chemotherapy. Initial response was observed as correction of DIC within a week of treatment. Hematologically, initial increase of maturing leukocytes reached a peak peripheral WBC count on the 16th and 20th day, respectively. However, these seemingly differentiated leukocytes retained Auer body and dysplastic features and there was no concomitant recovery of erythroid and megakaryocytic lineages at this point. A sudden drop of leukocyte counts after this peak made a brief period of leukopenia before the complete remission was finally attained morphologically in 4-5 weeks. Thus, remission of APL by ATRA therapy consisted of a two-phase course. In one patient, we observed an increase of histiocytes phagocytizing leukocytes in the marrow during the recovery from leukopenia. It is, therefore, postulated that the two-phase course of recovery may reflect the differentiation of leukemic cells by ATRA and subsequent clearance of senescent cells by the reticuloendothelial system followed by regeneration and differentiation of residual normal hematopoietic stem cells.

Adolescent

Action site of circulating interleukin-1 on the rabbit brain.

Interleukin-1 (IL-1) is now generally accepted as an endogenous pyrogenic mediator of fever induction. IL-1 induces fever by means of activation of arachidonate metabolism in the brain. However, whether circulating IL-1 enters the brain or not, further, the question of where is the action site of circulating IL-1 on the brain, have not been clearly demonstrated. In the present study, to investigate the site of action of circulating IL-1 on the brain, recombinant rabbit interleukin-1 beta (IL-1 beta) was labeled with colloidal gold, injected into the carotid artery of anesthetized rabbits and traced in the brain tissue by means of electron microscopy. Dose-dependent increase in colonic temperature was induced by intravenous injection of used IL-1 in rabbits, while heated IL-1 beta lost the pyrogenicity. Injection of solution of colloidal gold per se did not affect the colonic temperature of afebrile conscious rabbits. However, the IL-1/gold conjugates induced fever; IL-1 beta retained its pyrogenic potency even after labeling with gold. By electron microscopy, the IL-1/gold conjugates were observed on the surface and in the vesicle of endothelial cells of the capillary in the region of the anteroventral third ventricle. This implies that circulating IL-1 acts, as the initial step to induce fever, on the endothelium in that region.

Animals