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Biomedical subjects

H Imanaka

Publications and source records attributed to H Imanaka.

At least 73 records · Page 4Linked to original sources

Construction of a 7-aminocephalosporanic acid (7ACA) biosynthetic operon and direct production of 7ACA in Acremonium chrysogenum.

We have used cDNA encoding D-amino acid oxidase, and genomic DNA encoding cephalosporin acylase from Fusarium solani and Pseudomonas diminuta, respectively, to construct a novel hybrid 7-aminocephalosporanic acid (7ACA) biosynthetic operon under the control of regulatory elements from the alkaline protease gene of Acremonium chrysogenum. Transformants of A. chrysogenum BC2116, a high cephalosporin-producing strain, containing this operon, synthesized and secreted low levels of 7ACA. Although the amounts are not yet commercially significant, this represents the first microbial production of 7ACA and demonstrates the feasibility of introducing new biosynthetic capabilities into industrial microorganisms by combining fungal and bacterial genes.

Acremonium↗

Cloning and nucleotide sequencing of a novel 7 beta-(4-carboxybutanamido)cephalosporanic acid acylase gene of Bacillus laterosporus and its expression in Escherichia coli and Bacillus subtilis.

A strain of Bacillus species which produced an enzyme named glutaryl 7-ACA acylase which converts 7 beta-(4-carboxybutanamido)cephalosporanic acid (glutaryl 7-ACA) to 7-amino cephalosporanic acid (7-ACA) was isolated from soil. The gene for the glutaryl 7-ACA acylase was cloned with pHSG298 in Escherichia coli JM109, and the nucleotide sequence was determined by the M13 dideoxy chain termination method. The DNA sequence revealed only one large open reading frame composed of 1,902 bp corresponding to 634 amino acid residues. The deduced amino acid sequence contained a potential signal sequence in its amino-terminal region. Expression of the gene for glutaryl 7-ACA acylase was performed in both E. coli and Bacillus subtilis. The enzyme preparations purified from either recombinant strain of E. coli or B. subtilis were shown to be identical with each other as regards the profile of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were composed of a single peptide with the molecular size of 70 kDa. Determination of the amino-terminal sequence of the two enzyme preparations revealed that both amino-terminal sequences (the first nine amino acids) were identical and completely coincided with residues 28 to 36 of the open reading frame. Extracellular excretion of the enzyme was observed in a recombinant strain of B. subtilis.

Amidohydrolases↗

Cloning and nucleotide sequences of the complementary and genomic DNAs for the alkaline protease from Acremonium chrysogenum.

Complementary DNA encoding Ac. chrysogenum alkaline protease (Alp) was isolated from the Ac. chrysogenum ATCC11550 cDNA library by express-blot assay. The genomic DNAs encoding Ac. chrysogenum Alp were isolated from the Ac. chrysogenum genomic DNA library using the cloned cDNA as a probe. The 3150 nucleotides of the gene were sequenced. The prepro-Alp consists of 402 amino acids and two intervening sequences are found within the coding region. The amino acid sequence of Ac. chrysogenum Alp has 57% homology to that of Aspergillus oryzae Alp. The entire cDNA, encoding Ac. chrysogenum Alp, when introducing into the yeast Saccharomyces cerevisiae, directed the secretion of enzymatically active Alp into the culture medium.

Acremonium↗

Measurement of functional residual capacity during high-frequency oscillatory ventilation (HFOV) by argon washout method without interruption of HFOV.

A modified indicator gas washout method was developed to measure functional residual capacity (FRC) during high-frequency oscillatory ventilation (HFOV) without interruption of HFOV. A hot-wire flowmeter and medical gas analyzer measured the flow rate and argon concentration, respectively, at the expiratory end of the respiratory circuit. Upstream of the hot-wire flowmeter, two heat-and-moisture exchangers for resistance and a rubber balloon for capacitance were placed to convert the oscillating expiratory flow to an almost continuous flow. This made it possible to measure FRC during HFOV without interrupting HFOV. To measure the volume of the entire respiratory circuit, a 10 percent argon in 90 percent oxygen gas mixture was initially used as a bias flow, and after equilibration, the test gas was switched to 100 percent oxygen. By electrical integration of the product of the expiratory flow rate and argon concentration, the total amount of argon equilibrated in the entire respiratory circuit was calculated. The volume of the circuit was calculated by dividing the total amount of argon by the initial argon concentration. Functional residual capacity plus the volume of the respiratory circuit was similarly calculated and the difference was estimated as FRC. The accuracy and reproducibility of our method were evaluated by using a one-compartment lung model. There was a high correlation between the volume setting of the model lung and the estimated FRC. This method can be used to estimate FRC in a one-compartment lung model during HFOV, and it is potentially useful in clinical situations.

Argon↗

Studies on new dehydropeptidase inhibitors. III. Biological properties of WS1358A1.

WS1358A1, a novel inhibitor of renal dehydropeptidase (DHP), augmented the urinary recovery of a carbapenem antibiotic imipenem and improved its protective effect in experimental infections when simultaneously administered to mice with the antibiotic. WS1358A1 was a competitive DHP inhibitor with a Ki value of 1.6 x 10(-7) M.

Animals↗

FR112123, a new oligopeptide antibiotic from Streptomyces viridochromogenes. Taxonomy, fermentation, isolation, physico-chemical properties, structure and biological activity.

FR112123 is a new oligopeptide antibiotic produced by Streptomyces viridochromogenes No. 7587. The structure of FR112123 is elucidated as N-(N6-(N2-glycyl-L-glutaminyl)-D-lysyl)-D-alanine (1) by spectroscopic and chemical evidence. It resembles a partial structure of peptidoglycan in bacteria. The compound has a superior activity against an Escherichia coli mutant sensitive to inhibitors of cell wall synthesis, although it has a weak activity against the parent strain. These suggest that FR112123 might act on the biosynthesis of bacterial cell wall.

Animals↗

[Measurement of flow-resistive work of heat and moisture exchangers during simulated active breathing].

The performance of seven commercially available heat and moisture exchangers (HME) was evaluated in the dynamic flow state. A piston pump was used as a simulator with operative conditions with stroke volume of 500 ml and with respiratory rates of 20 and 40 cycles.min-1. A pressure-volume loop was produced for each HME and the flow-resistive work calculated. Each HME was studied under both dry and wet conditions. It was found that the flow-resistive work of the Humid-Vent 2 was the smallest and that of the Breathaid the largest under both conditions. The results suggest that application of HME causes a significant increase in resistance and demands a greater breathing effort from patients. Its use should thus be avoided in marginal patients.

Anesthesiology↗

High-frequency oscillation for persistent fetal circulation after repair of congenital diaphragmatic hernia.

A female neonate who had been diagnosed as having congenital diaphragmatic hernia by ultrasonography was delivered by cesarean section. After the hernia was repaired, she developed hypoxemia and hypercapnia, probably due to persistent fetal circulation (PFC). Neither conventional mechanical ventilation (CMV) nor manual ventilation improved the respiratory status. High-frequency oscillation (HFO) successfully improved pulmonary gas exchange, but we failed to wean the patient from HFO by using intermittent HFO. The patient was again placed on CMV for weaning and was extubated on the 12th ICU day. We conclude that HFO can be an alternative respiratory modality in patients with respiratory failure due to PFC after the repair of congenital diaphragmatic hernia.

Female↗

Effects of norepinephrine on renal function in septic patients with normal and elevated serum lactate levels.

Effects of iv norepinephrine (NE) on renal function were investigated retrospectively in 15 patients with hyperdynamic septic shock. All patients had either a low systolic BP less than 80 mm Hg, and/or oliguria less than 0.5 ml/kg-h. We examined their serum creatinine level (SCr), daily urine flow (UF), 24-h creatinine clearance (Ccr), and hemodynamic indices before and during NE infusion. Before NE administration, the patients were divided into those with with a serum lactate level (Lac) less than 20 mg/dl (group A, n = 9) and greater than 20 mg/dl (group B, n = 6). NE was infused continuously at rates between 0.05 and 0.24 microgram/kg.min which increased systolic BP by greater than or equal to 20 mm Hg. Cardiac index was not significantly changed in either group. In group A, NE increased both UF (p less than .05), and systemic vascular resistance index (SVRI) (p less than .01), but did not affect Ccr. In group B, NE did not increase UF nor SVRI, and decreased Ccr significantly (p less than .05). It is concluded that NE increased UF and SVRI only when Lac was in normal range; otherwise, NE reduced renal function. Thus, when administering NE to increase UF, both Lac and renal function should be monitored carefully.

Adult↗

Jet flow-regulated expiratory resistance to maintain constant CPAP during the entire respiratory phase.

We have developed a new continuous positive airway pressure (CPAP) device that consists of a microcomputer, a pressure transducer, and a pair of electronic interface valves. One of these valves creates the inspiratory demand flow, and the other creates the opposing jet flow by acting as an expiratory valve to maintain a constant CPAP. By controlling the two electronic interface valves, the airway pressure can be kept constant during the entire respiratory cycle. We compared our device with CPAP systems supplied with commercially available ventilators: the Puritan-Bennett 7200a, the Bear 5, the Servo 900C, and the CV 2000. A two-chambered spring loaded model lung was used to simulate inspiration and a piston pump model lung to simulate active exhalation. We compared both the inspiratory triggering work (WWIt) and expiratory flow-resistive work (WE) of each ventilator while in CPAP mode by calculating the corresponding areas of the pressure-volume loops using electrical integration. The WWIt of our apparatus and demand-flow ventilators was much smaller than that of the CV 2000. In our device, WE was also much smaller than those of the others. These results indicate that our device can be used for CPAP without causing airway pressure fluctuation, and therefore, without imposing an extra workload on the patient.

Humans↗

FK409, a novel vasodilator isolated from the acid-treated fermentation broth of Streptomyces griseosporeus. I. Taxonomy, fermentation, isolation, and physico-chemical and biological characteristics.

FK409, a novel vasodilator with anti-platelet aggregation activity, has been isolated from the acid-treated fermentation broth of Streptomyces griseosporeus No. 16917, which was cultured on a medium containing NaNO3 for 4 days. FK409 was purified from the culture-filtrate by extraction with ethyl acetate after adjusting the pH to 3.0 with HC1, followed by silica gel chromatography. The molecular formula of this compound was determined to be C8H13N3O4. In vitro, FK409 showed a potent relaxation activity on noradrenaline induced contraction of rat aorta. In addition to the vasodilating activity, this compound also showed potent anti-aggregation activities towards rabbit platelets. In vivo, intravenously administered FK409 resulted in marked blood pressure lowering in rats.

Animals↗

FK409, a novel vasodilator isolated from the acid-treated fermentation broth of Streptomyces griseosporeus. II. Structure of FK409 and its precursor FR-900411.

In a purification study of novel vasodilator FK409 from the fermentation broth of Streptomyces griseosporeus, we found the existence of FR-900411, a precursor of FK409 in the broth, suggesting that FK409 was derived from FR-900411 via a chemical reaction. We therefore isolated FR-900411 from the broth. The structure of FK409 and FR-900411 were determined to be 1 and 2 respectively, on the basis of spectroscopic and chemical evidence.

Alkadienes↗

FK409, a novel vasodilator isolated from the acid-treated fermentation broth of Streptomyces griseosporeus. III. Reaction mechanism and synthesis.

FK409 (1), a novel vasodilator, is a semi-artificial fermentation product of Streptomyces griseosporeus No. 16917, which was cultured in a medium containing nitrate. And the acid-treatment of the broth is essential for the generation of FK409. FK409 was considered to be formed via a novel synchronous nitrosation-nitration reaction of FR-900411 (2) which was produced by the strain as a precursor of FK409. The conversion of FK-900411 to FK409 proceeded under acidic conditions with nitrite formed by microbial reduction of nitrate. Total synthesis of FK409 was achieved starting from (E)-2-ethyl-2-butenal (3) via a nitrosation reaction of FR-900411 (2) as a key step.

Alkadienes↗

Gravity-dependent atelectasis. Radiologic, physiologic and pathologic correlation in rabbits on high-frequency oscillation ventilation.

The authors identify the radiologic features of progressive atelectasis induced under conditions of reduced lung volume. Control (n = 5) and experimental (n = 7) animals were placed on high-frequency oscillation (HFO) ventilation (mean airway pressure: 3 cm H2O) for 6 hours. In the experimental animals, lung volume was artificially reduced by pneumoperitoneum during HFO ventilation. Computed tomography scans and chest radiographs were obtained every hour, and arterial blood gases analyzed. No changes were detected in the control animals. In the experimental animals, in which hypoxemia developed, homogeneous opacity in the dependent lung was found on CT images, and chest radiographs showed a diffuse homogenous shadow with loss of lung volume. Study of pathologic sections from the lung showed that the roentgenographic findings represented atelectasis. The lung was divided into three zones, from dependent to nondependent regions: severe atelectasis, mild atelectasis, and normal lung. Hyperinflations eliminated atelectasis seen on the CT images and alleviated hypoxemia; however an undesirable effect that causes barotrauma also was observed.

Animals↗

[Quantitative comparison of ventilator-induced work during simulated CPAP in eight demand-flow valve ventilators].

The ventilator-induced work during continuous positive airway pressure (CPAP) mode in demand-valve ventilators was evaluated by using a piston pump as a simulator for active breathing. A piston pump delivered and withdrew a stroke volume of 500 ml at rates of 10, 20 and 40 cycle.min-1 with a sinusoidal waveform. A hot-wire flowmeter and a differential pressure transducer were interposed between the pump and ventilators and their signals were fed to a microcomputer to display a pressure-volume loop. The area of the loop was divided into the four parts. Inspiratory work associated with the opening of the demand valve was represented by the area of baseline airway pressure (BPa) during inspiration. The remaining area during inspiration reflected the work done by the ventilators. Expiratory work for overcoming the flow resistance of the expiratory apparatus was represented by the area above BPa during exhalation. The fourth was the area below the baseline during exhalation. The Puritan-Bennett 7200a, the Bear 5, the Siemens Servo 900C, the Hamilton Veolar, the Bird 6400ST, the Engström Erica, the Dräger EV-A, and the CPU-1 were examined at varying CPAP and pressure support levels. Because of demand valve oscillation throughout inspiration, the inspiratory workload of the Bear 5, the Siemens Servo 900C, the Hamilton Veolar, the Bird 6400ST, and the Dräger EV-A could not be calculated. Expiratory flow-resistive work was higher in the Siemens Servo 900C and the Bird 6400ST than the others. The present system can assess the entire performance of ventilators, and may serve to compare ventilators' performance.

Evaluation Studies as Topic↗