Prerequisites for combinatorial biosynthesis: evolution of hybrid NRPS/PKS gene clusters.
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Binding Sites↗
Biomedical subjects
T Oh
Publications and source records attributed to T Oh.
Postoperative pain management after intravenous remifentanil.
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Analgesia↗
Dehydroepiandrosterone sulfotransferase gene induction by bile acid activated farnesoid X receptor.
Dehydroepiandrosterone sulfotransferase (STD) is a hydroxysteroid sulfo-conjugating enzyme with preferential substrate specificity for C-19 androgenic steroids and C-24 bile acids. STD is primarily expressed in the liver, intestine and adrenal cortex. Earlier studies have shown that androgens inhibit the rat Std promoter function through a negative androgen response region located between -235 and -310 base pair positions (Song, C. S., Jung, M. H., Kim, S. C., Hassan, T., Roy, A. K., and Chatterjee, B. (1998) J. Biol. Chem. 273, 21856-21866). Here we report that the primary bile acid chenodeoxycholic acid (CDCA) also acts as an important regulator of the Std gene promoter. CDCA is a potent inducer of the Std gene, and its inducing effect is mediated through the bile acid-activated farnesoid X receptor (FXR), a recently characterized member of the nuclear receptor superfamily. The ligand-activated FXR acts as a heterodimer with the 9-cis-retinoic acid receptor (RXR) and regulates the Std gene by binding to an upstream region at base pair positions -169 to -193. This specific binding region was initially identified by bile acid responsiveness of the progressively deleted forms of the Std promoter in transfected HepG2 hepatoma and enterocyte-like Caco-2 cells. Subsequently, the precise RXR/FXR binding position was established by protein-DNA interaction using in vitro footprinting and electrophoretic mobility shift analyses. Unlike all other previously characterized FXR target genes, which contain an inverted repeat (IR) of the consensus hexanucleotide half-site (A/G)G(G/T)TCA with a single nucleotide spacer (IR-1), the bile acid response element of the Std promoter does not contain any spacer between the two hexanucleotide repeats (IR-0). A promoter-reporter construct carrying three tandem copies of the IR-0 containing -169/-193 element, linked to a minimal thymidine kinase promoter, can be stimulated more than 70-fold in transfected Caco-2 cells upon CDCA treatment. Autoregulation of the STD gene by its bile acid substrate may provide an important contributing role in the enterohepatic bile acid metabolism and cholesterol homeostasis.
Bile Acids and Salts↗
The effect of the Lockit epidural catheter clamp on epidural migration: a controlled trial.
We investigated the efficacy of using the Lockit epidural catheter clamp in preventing epidural catheter migration in patients receiving postoperative analgesia via an epidural infusion after major surgery. Patients were randomly allocated to receive either a standard epidural dressing, a coiled catheter with transparent adhesive dressing and tape (n = 54), or the Lockit epidural catheter clamp (n = 48). There was no movement from the insertion position in 88% of the Lockit group compared with only 28% in the standard group (p < 0.001). Outward migration of > 2 cm occurred in 26% of the standard group compared with just 6% of the Lockit group (p < 0.01). Inward migration of > 1 cm occurred in 17% of the standard group but in none of the Lockit group (p < 0.01). Catheter migration was assessed as being the cause of analgesic failure in 15% of the standard group but in only 4% of the Lockit group (p < 0.05). We conclude that the Lockit epidural catheter clamp significantly reduces catheter migration in a safe and non-invasive fashion.
Analgesia, Epidural↗
The toxR gene of Vibrio (Listonella) anguillarum controls expression of the major outer membrane proteins but not virulence in a natural host model.
To examine the hypothesis that the ancestral role of the toxR gene in the family Vibrionaceae is control of the expression of outer membrane protein (OMP)-encoding genes for adaptation to environmental change, we investigated the role of the toxR gene in Vibrio anguillarum, an important fish pathogen. The toxR gene of V. angullarum (Va-toxR) was cloned from strain PT-87050 isolated from diseased ayu (Plecoglossus altivelis), and the sequence was analyzed. The toxR sequence was 63 to 51% identical to those reported for other species of the family Vibrionaceae. Distribution of the Va-toxR gene sequence in V. anguillarum strains of various serotypes was confirmed by using DNA probe and PCR methods. An isogenic toxR mutant of V. anguillarum PT-24, isolated from diseased ayu, was constructed by using an allelic exchange method. The wild-type strain and the toxR mutant did not differ in the ability to produce a protease(s) and a hemolysin(s) or in pathogenicity for ayu when examined by the intramuscular injection and immersion methods. A 35-kDa major OMP was not produced by the toxR mutant. However, a 46-kDa OMP was hardly detected in the wild-type strain but was produced as the major OMP by the toxR mutant. For the toxR mutant, the MICs of two beta-lactam antibiotics were higher and the minimum bactericidal concentration of sodium dodecyl sulfate was lower than for the wild-type strain. Analysis of the N-terminal amino acid sequences of the 35- and 46-kDa OMPs indicated that these proteins are the porin-like OMPs and are related to the toxR-regulated major OMPs of the family Vibrionaceae. The results indicate that the toxR gene is not involved in virulence expression in V. anguillarum PT-24 and that toxR regulation of major OMPs is universal in the family Vibrionaceae. These results support the hypothesis that the ancestral role of the toxR gene is regulation of OMP gene expression and that only in some Vibrio species has ToxR been appropriated for the regulation of a virulence gene(s).
Amino Acid Sequence↗
[A case of mediastinal esophago-bronchogenic cyst associated with high serum level of CA 19-9].
We report a case of mediastinal esophago-bronchogenic cyst in a patient with a high serum level of CA 19-9. A 49-year-old man presented with a complaint of dysphagia. Chest X-ray, CT, and MRI showed posterior mediastinal cystic shadow which was diagnosed preoperatively. The serum level of CA 19-9 was 158 U/ml. We underwent extirpation of the cyst through video-assisted thoracoscopic surgery (VATS) and confirmed to be a bronchogenic cyst histologically. The serum level of CA 19-9 returned to normal after the operation.
Bronchogenic Cyst↗
In vitro activity of DU-6681a, an active form of the new oral carbapenem compound DZ-2640, in comparison with that of R-95867, faropenem and oral cephalosporins.
We compared the in vitro antibacterial activity of DU-6681a against Gram-positive and Gram-negative bacteria with that of R-95867, faropenem and oral cephalosporins such as cefcapene, cefotiam and cefpodoxime. DU-6681a is an active form of the new oral carbapenem compound DZ-2640, which is an ester-type prodrug, and R-95867 is an active form of the oral carbapenem CS-834. Against most Gram-positive bacteria, DU-6681a was as active as or two- to 16-fold more potent than R-95867 and faropenem in terms of MIC(90), and comparable to or two- to 64-fold more effective than the cephalosporins. Against most Gram-negative bacteria, the activity of DU-6681a was the same as or two- to 16-fold more potent than that of R-95867, and comparable to or two- to 2048-fold higher than that of faropenem and the cephalosporins.
Administration, Oral↗
Analysis of gyrB and toxR gene sequences of Vibrio hollisae and development of gyrB- and toxR-targeted PCR methods for isolation of V. hollisae from the environment and its identification.
Isolation of Vibrio hollisae strains, particularly from the environment, is rare. This may be due, in part, to the difficulty encountered when using conventional biochemical tests to identify the microorganism. In this study, we evaluated whether two particular genes may be useful for the identification of V. hollisae. The two genes are presumed to be conserved among the bacterial species (gyrB) or among the species of the genus Vibrio (toxR). A portion of the gyrB sequence of V. hollisae was cloned by PCR using a set of degenerate primers. The sequence showed 80% identity with the corresponding Vibrio parahaemolyticus gyrB sequence. The toxR gene of V. hollisae was cloned utilizing a htpG gene probe derived from the V. parahaemolyticus htpG gene, which is known to be linked to the toxR gene in V. hollisae. The coding sequence of the cloned V. hollisae toxR gene had 59% identity with the V. parahaemolyticus toxR coding sequence. The results of DNA colony hybridization tests using the DNA probes derived from the two genes of V. hollisae indicated that these gene sequences could be utilized for differentiation of V. hollisae from other Vibrio species and from microorganisms found in marine fish. PCR methods targeting the two gene sequences were established. Both PCR methods were shown to specifically detect the respective target sequences of V. hollisae but not other organisms. A strain of V. hollisae added at a concentration of 1 to 10(2) CFU/ml to alkaline peptone water containing a seafood sample could be detected by a 4-h enrichment incubation in alkaline peptone water at 37 degrees C followed by quick DNA extraction with an extraction kit and 35-cycle PCR specific for the V. hollisae toxR gene. We conclude that screening of seafood samples by this 35-cycle, V. hollisae toxR-specific PCR, followed by isolation on a differential medium and identification by the above htpG- and toxR-targeted PCR methods, can be useful for isolation from the environment and identification of V. hollisae.
Animals↗
Rapid mini-scale plasmid isolation for DNA sequencing and restriction mapping.
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Bacteria↗
Staphylococcus haemolyticus lipase: biochemical properties, substrate specificity and gene cloning.
Lipase of Staphylococcus haemolyticus L62 was purified from culture supernatant and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for the hydrolysis of olive oil was 28 degrees C and pH 8.5, respectively. The enzyme was stable up to 50 degrees C in the presence of Ca(2+)and over the pH range 5-11. It had high hydrolytic activity against tributyrin, tripropionin, and trimyristin among various triglycerides. The gene encoding the lipase was cloned in Escherichia coli. Sequence analysis showed an open reading frame of 2136 bp, which encodes a preproenzyme of 711 amino acids. The preproenzyme is composed of a signal peptide (60 aa), a pro-peptide (259 aa), and a mature enzyme (392 aa). The mature enzyme has 49-67% amino acid sequence homology with other staphylococcal lipases.
Amino Acid Sequence↗
Total Synthesis of (+/-)-Gelsemine.
A complex molecular reorganization (1-->2), a sequential anionic aza-Cope rearrangement and Mannich cyclization, and an unprecedented intramolecular Heck reaction of the tetrasubstituted double bond of a vinylogous carbamate are key steps in a new total synthesis of (+/-)-gelsemine (3). MOM=methoxymethyl, DBU=1,8-diazabicyclo[5.4.0]undec-7-ene.
Journal Article↗
Thermodynamics of the reconstitution of tuna cytochrome c from two peptide fragments.
Two peptide fragments from tuna cytochrome c (cyt c), N-fragment (residues 1-44 containing the heme) and C-fragment (residues 45-103), combine to form a 1:1 fragment complex. This was clearly proved by ion-spray mass spectrometry. It was found from CD and NMR spectra that the structure of the fragment complex formed is similar to that of an intact cyt c, although each isolated fragment itself is unstructured. Binding constants and enthalpies upon the complex formation were directly observed by isothermal titration calorimetry. Thermodynamic parameters (deltaG(o)b, deltaHb, deltaS(o)b, and deltaC(b)p)) associated with the complex formation were determined at various pHs and temperatures. DeltaHb was found to be almost independent of pH values. The change in heat capacity accompanying the complex formation (deltaC(b)p) was directly determined from the temperature dependence of deltaHb. In addition, the change in heat capacity and enthalpy upon tuna cyt c unfolding were determined by differential scanning calorimetry. Thermodynamic parameters for the unfolding/dissociation process of the fragment complex were compared with those for cyt c unfolding at pH 3.9 and 303 K. In a comparison of two unfolding processes, the heat capacity change of each was very close to the other, while both the unfolding enthalpy and entropy of the fragment complex were larger than those of tuna cyt c. These thermodynamic data suggest that the internal interactions between polar groups (hydrogen bonding) and nonpolar groups (van der Waals interactions) are preserved in the fragment complex as well as in the native state of cyt c.
Animals↗
In-vitro and in-vivo antibacterial activities of CS-834, a new oral carbapenem.
We evaluated the in-vitro and in-vivo antibacterial activities of R-95867 and CS-834, a new oral carbapenem which is an ester-type prodrug of R-95867. Against Gram-positive bacteria, R-95867 was as active or two to 256 times more active than cefpodoxime, cefdinir, cefditoren and ofloxacin, while its activity was similar to or two to eight times lower than that of imipenem. Against most Gram-negative bacteria it was as active or two to 1024 times more active than the other compounds tested. Against Helicobacter pylori it was two to 64 times more active than orally active anti-H. pylori agents, i.e. amoxycillin, clarithromycin and lansoprazole. It also showed potent bactericidal activity against Staphylococcus aureus and Escherichia coli. R-95867 induced a spherical form in E. coli and showed high affinity for PBP 2 in E. coli. Against systemic infections in mice caused by various bacteria, CS-834 showed an excellent protective effect and its in-vivo efficacy correlated well with the in-vitro activity of R-95867. These results suggest that CS-834 may be effective in the therapy of various bacterial infections.
Administration, Oral↗
Gastric adenocarcinoma in patients 40 years of age or younger.
BACKGROUND: Gastric adenocarcinoma is considered a disease of the middle aged and elderly and has been infrequently reported in patients under 40 years of age. The purpose of this study was to determine the proportion of young patients diagnosed with gastric adenocarcinoma and to compare the demographic, clinical and pathologic features of younger and older patients with gastric adenocarcinoma. METHODS: A retrospective cohort study using tumor registry records of all patients with gastric adenocarcinoma diagnosed from 1982 through 1996 at a public teaching hospital. Demographic, clinical, and pathologic comparisons were made between patients younger than 41 years of age and race- and sex-matched older patients with gastric adenocarcinoma. RESULTS: Thirty of 203 (15%) cases of gastric adenocarcinoma were diagnosed in patients less than 41 years (range 23 to 40). Male to female ratio was 1:1. Young patients were more likely to be black (33% versus 17%, P = 0.04) Both younger and older patients presented with advanced disease, with nearly half of each group having metastases. Twelve of 29 (41%) younger patients were operated on without a histologic diagnosis of gastric adenocarcinoma in contrast to only 1 older patient (P < 0.001). One of 30 (3%) young patients is alive 39 months following gastrectomy. Twenty patients died and the remaining 9 were lost to follow-up, all with known residual or recurrent disease. Six-month survival of young patients (23%) was less than older patients (42%) (P = 0.14). Young patients were more likely to have diffuse histology (80% versus 55%, P = 0.12). Overt infection with Helicobacter pylori was uncommon in both groups. CONCLUSIONS: Young patients accounted for an unusually high proportion of patients with gastric adenocarcinoma diagnosed at our public teaching hospital. Young patients were significantly more likely to be black and less likely to have an accurate preoperative histologic diagnosis. Both young and older patients presented with advanced disease and had poor survival. Young patients were more likely to have diffuse histology and had poorer 6-month survival, suggesting a more aggressive variety of the disease in this group.
Adenocarcinoma↗
Poststreptococcal glomerulonephritis with the nephrotic range of proteinuria.
In order to characterize the clinical features in poststreptococcal glomerulonephritis (PSGN) in relation to the range of proteinuria, 27 patients with PSGN were divided into 3 groups according to the amount of urinary protein excretion; Group I: with proteinuria over 3.5 g/day (n = 8), Group II: with proteinuria between 1.0 and 3.4 g/day (n = 9) and Group III: with proteinuria less than 1.0 g/day (n = 10). Bed-rest was ordered until proteinuria decreased to less than 1.0 g/day. The serum creatinine levels in Group I were significantly higher than in Group III both on admission and at discharge, although the duration of hospitalization was longer in the former than in the latter group. Furthermore, the durations of proteinuria and hypocomplementaemia were longer in the former than in the latter. In addition, the former showed a higher systolic blood pressure and a greater expanded extracellular fluid volume expressed as body weight change during hospitalization. However, none of the patients in this study demonstrated any persistent proteinuria or renal function impairment. In conclusion, bed-rest in the acute phase of PSGN might improve the short-term prognosis of PSGN.
Adult↗
Evaluating the value, accuracy, and operational feasibility of DUE criteria.
The purpose of this investigation was to evaluate the value, accuracy, and operational feasibility of indicators/criteria in a drug use evaluation (DUE) examining IV histamine2-receptor antagonists (H2-RA). Pharmacists in 40 hospitals collected DUE data concurrent with hospital stays on a total of 1,200 patients. After completing the DUE, pharmacist data collectors were asked to evaluate each DUE criterion. Most reported that criteria relating to indications for use, therapeutic drug monitoring, and adverse drug reactions/drug interactions should be included in a DUE, but some criteria were not easy to collect or were inaccurate. The data suggest the need to carefully select DUE criteria that can be used for continuous improvements that meet JCAHO accreditation requirements.
Data Collection↗
A pharmacoeconomic analysis of IV H2-receptor antagonist use in 40 hospitals.
The objectives of this study were to determine (1) the expenditures of hospitals for IV histamine2-receptor antagonists (H2-RA), and (2) the cost savings that might be realized if only a single IV H2-RA was purchased for use. Forty hospitals provided data about purchase prices for each IV H2-RA dosage form purchased (cimetidine, ranitidine, and famotidine), the number of each dosage form used during the 12-month study period, purchase price and extent of usage for supplies, labor costs for preparing and administering IV H2-RAs, and IV H2-RA dosage schedules. The study showed that most hospitals were spending more money for IV H2-RAs than necessary given the pricing structures of the three products available to them at the time of this study. Also, that significant cost savings could be realized if a single H2-RA was used exclusively.
Cimetidine↗
Use of i.v. cimetidine, ranitidine, and famotidine in 40 hospitals.
The drug experiences of 1200 patients receiving i.v. histamine H2-receptor antagonists were studied. Forty hospitals in five southeastern states participated in a drug-use evaluation (DUE). In addition to supplying background information, a pharmacist at each hospital used a standardized form to perform a concurrent review of 30 consecutive adult patients who had been started on i.v. cimetidine, ranitidine, or famotidine. In addition to the patient's age and the prescriber's medical specialty, specific DUE criteria included the reason given in the medical record for use; dosage regimen and adjustments made on the basis of the patient's renal function; other GI drugs taken concurrently; pharmacist intervention; simultaneous use of oral medications; occurrence of adverse events; H2-antagonist use with specified drug products known to affect serum drug concentrations of one or both medications; and use of gastric pH monitoring. Therapy with i.v. H2 antagonists was usually started by internal medicine specialists or surgeons, and most of the evaluated patients received such therapy for prevention of stress-related mucosal damage. According to estimated creatinine clearance, 34% of patients were in need of dosage adjustments, but such adjustments were made in only 49% of these. Forty percent of patients began therapy while taking at least one medication orally, and almost one fourth of patients were receiving an additional drug for the treatment of an acid-peptic or related indication. The occurrence of adverse events was similar for the three H2 antagonists studied. The findings identify several problematic areas in the way H2 antagonists are used in clinical practice.
Adolescent↗