Role of leuX in Escherichia coli colonization of the streptomycin-treated mouse large intestine

Escherichia coli F-18, a normal human fecal isolate, is an excellent colonizer of the streptomycin-treated mouse large intestine. E. coli F-18 Col-, a derivative of E. coli F-18 that no longer makes the E. coli F-18 colicin, colonizes the mouse large intestine as well as E. coli F-18 when fed alone, but is eliminated when fed together with E. coli F-18. Recently, a random bank of E. coli F-18 DNA was transformed into E. coli F-18 Col-, the resultant population was fed to streptomycin-treated mice, and the intestine was used to select the best colonizer. In this fashion, a 6.5 kb E. coli F-18 DNA fragment was isolated. This fragment was shown to enhance E. coli F-18 Col- mouse large intestinal colonizing ability and survival during stationary phase in intestinal mucus in vitro, as well as stimulate the synthesis of type-1 fimbriae. Here, we present evidence that the gene responsible for the enhanced E. coli F-18 Col- colonizing ability and survival during stationary phase in vitro is leuX. This gene encodes a rare leucine tRNA specific for the UUG codon. In addition, we show that the presence of a functional leuX gene is necessary for E. coli K-12 intestinal colonization and for survival in stationary phase.     

Longitudinal trends in total serum cholesterol levels in a Japanese cohort, 1958-1986

This study is a preliminary examination of the reliability of adolescent self-reported pretreatment alcohol and other drug (AOD) use frequency. Assessments of self-reported pretreatment AOD use were conducted at admission and discharge (approximately a 1-month time period) at an adolescent drug misuser treatment program. The sample consisted of 197 male and female adolescents. There were statistically significant increases between admission and discharge assessments of pretreatment AOD use frequency. The greatest discrepancy was found for alcohol use, in which three-fourths (76%) of the sample reported a higher level of pretreatment alcohol use frequency at discharge assessment as compared to their admission assessment. Over one-third (35%) of the sample was found to have a significantly higher level of pretreatment alcohol use frequency at discharge assessment. The cause of this response discrepancy is unknown, but if it represents underreporting at admission, it may cause diagnostic and referral errors, as well as attenuate effect sizes in treatment outcome studies.     

Protamine sulfate enhances lipid-mediated gene transfer

OBJECTIVE: To analyze myoelectric activity of the cecum and proximal loop of the ascending colon (PLAC) in cows after spontaneous cecal dilatation/dislocation (CDD) and compare it with that in healthy cows after surgical evacuation of the cecum. ANIMALS: 12 cows with spontaneous CDD and 6 healthy cows (group C). Cows with spontaneous CDD were retrospectively assigned to 2 groups: delayed recovery from surgery or recurrence (group A; n = 3), and normal recovery (group B; n = 9). PROCEDURE: After surgical evacuation of the cecum, 8 pairs of bipolar, retrievable electrodes were implanted in the ileum, cecum, and PLAC. Cows were evaluated daily from postoperative day 1 to 7, using routine clinical methods and computer-based analysis of myoelectric activity of the cecum, and PLAC. Parameters of myoelectric activity included rate of spike bursts, duration of individual spike bursts, duration of overall spike burst activity per electrode, rate of propagated spike burst sequences, and ratio of orally propagated spike burst sequences. RESULTS: Rate of spike bursts, duration of cecocolic spike burst activity, and ratio of orally to aborally propagated spike burst sequences did not vary among groups during the 7-day recording period. However, cows with delayed recovery had a typical, uniform pattern of myoelectric activity of the cecum and PLAC at days 1 and 2 after surgery that consisted of repeated, propagated spike burst sequences, made up of spike bursts of significantly (P < 0.05) increased duration at postsurgical day 1 and substantially prolonged duration at postsurgical day 2, interrupted by periods of little or no activity. CONCLUSION: Delayed recovery and recurrence of CDD in cows after spontaneous CDD is not caused by hypomotility of the cecum and PLAC. CLINICAL RELEVANCE: Postoperative treatment, intended to reduce recurrence of CDD or delayed recovery after surgical evacuation of the cecum, should address propagation of digesta in the spiral colon.     

Cardioprotection by orotic acid: metabolism and mechanism of action

The pyrimidine base, orotic acid (OA), improves the function of recently infarcted hearts subjected to global ischemia but its mechanism of action is unclear. Our aims were to examine (i) in normal rats, the effect of OA on pyrimidine levels in plasma, liver and heart; (ii) in rats with normal or infarcted hearts, the effect of OA on adenine nucleotide metabolism and mechanical function, before and after global ischemia. To investigate the metabolism of OA, normal rats received 100 mg/kg OA, and changes in plasma and tissue concentrations of pyrimidines were examined. The effects of OA were also studied in rats receiving OA for 2 days after coronary ligation or sham operations, and plasma and tissue pyrimidine concentrations examined. Their hearts were isolated and perfused, then subjected to 30 min of global ischemia. Mechanical function and adenine nucleotide content were assessed pre- and post-ischemia. In normal, unoperated rats, administration of 100 mg/kg OA significantly increased hepatic uracil and cytosine nucleotide concentrations, then increased plasma uridine (+124%) and cytidine (+55%), and transiently increased myocardial uracil nucleotides (+21%). Infarction significantly reduced recovery of cardiac work after global ischemia (sham=62%; infarct=26%; P<0.05), and OA treatment in infarcted hearts increased post-ischemic work by 192% (P<0.05), but not in shams. Pre-ischemic ATP was reduced in the surviving myocardium of infarcted hearts from 21.7+/-0.8 to 14.7+/-0. 7 micromol/g dry weight (P<0.001) and total adenine nucleotides (TAN) from 30.3+/-0.8 to 22.4+/-1.1 micromol/g dry weight (P<0.001). OA treatment prevented these reductions in infarcted hearts (ATP 20. 7+/-0.5; TAN, 29.1+/-0.6 micromol/g dry weight). We conclude that OA protects the infarcted heart against global ischemia by enhancing hepatic release of pyrimidine nucleosides into the plasma, which then prevent depletion of adenine nucleotides in the surviving myocardium.     

The Himar1 mariner transposase cloned in a recombinant adenovirus vector is functional in mammalian cells

Mariner transposons belong to the mariner /Tc1 superfamily of class II, DNA-mediated elements. One of these transposons, Himar1 , isolated from the horn fly, is independent of host-specific factors that would limit transfer between different species, making it an ideal candidate for gene transfer technology development. To determine the activity of Himar1 transposase in mammalian cells, we introduced the Himar1 transposase gene into an adenovirus (Ad) vector under control of the phage T7 RNA polymerase promoter. Mammalian cells infected with the Ad vector carrying the Himar1 gene efficiently expressed the Himar1 transposase in the presence of T7 polymerase. In in vitro inter-plasmid transposition reactions, Himar1 transposase expressed by the Ad vector mediated precise cut-and-paste transposition and resulted in a characteristic duplication of TA at the integration site of the target plasmid. Further studies showed that this transposase was capable of catalyzing transposition between twoplasmids co-transfected into 293T7pol cells, which express T7 RNA polymerase. Combining the integration capability of mariner transposons with the transduction efficiency of Ad vectors is expected to provide a powerful tool for introducing transgenes into the host chromosome.     

Acute and chronic effects of bilateral lung transplantation without cardiopulmonary bypass on the first transplanted lung

BACKGROUND: Bilateral lung transplantation (BLT) without cardiopulmonary bypass (CPB) may exacerbate reperfusion injury to the initially engrafted lung because of increases in pulmonary flow during implantation of the second graft. METHODS: In a retrospective review of 23 BLT patients, we hypothesized that BLT without CPB injures the first transplanted lung measured by acute and late graft dysfunction compared to the second transplanted lung. Of the 23 BLT, 19 underwent transplantation without CPB while 4 patients were placed on CPB secondary to hemodynamic instability. RESULTS: Acute graft function was assessed by radiographic scoring of lung quadrants (blinded radiologist; 0 = no infiltrate; 1 = infiltrate; maximum = 2 per lung) and by arterial/alveolar oxygen tension ratios (PaO2/ FiO2) ratios. Late graft function was evaluated by quantitative perfusion scan. Lung perfusion was graded as abnormal if less than 50% on the right or less than 45% on the left (Fisher's exact). Radiographic scores were not different between first and second implanted lungs at 1 and 24 hours, PaO2/FiO2 ratios at 1 and 24 hours were 273+/-26 and 312+/-23, respectively, and perfusion scans at 3 and 12 months revealed normal differential blood flow. CONCLUSIONS: These findings suggest no acute or chronic differences occur between the first or second transplanted lung completed without CPB.     

The effects of state intergovernmental transfers on local public mental health services

The provision of public mental health services is shifting increasingly from states to local areas. Yet state governments continue to bear financial responsibility for the majority of these services. One implication of this trend is that the success of state policies become dependent on a state's ability to influence the behavior of local areas. This paper discusses the different options states have in designing intergovernmental grant contracts with local areas, and describes likely impacts of the different strategies. These propositions are then tested using data from the Ohio state mental health system from 1989-1993. This study finds that the design of grants affects public expenditures, local revenue generation, and the mix of services provided at the local level.     

Heart transplantation in children and young adults: early and intermediate-term results

The purpose of this article is to report our short- and intermediate-term follow-up of cardiac transplantation for congenital heart disease and cardiomyopathy in children (age greater than 6 months), adolescents, and young adults. Thirty patients (ages 8 months to 24 years) with end-stage heart failure have undergone cardiac transplantation in our program: 12 (40%) for postoperative end-stage heart failure, 9 (30%) as primary treatment for congenital heart disease, 5 (17%) for dilated cardiomyopathy, and 4 (13%) for restrictive/hypertrophic cardiomyopathy. Nineteen patients (63%) had undergone prior operations; 4 patients received transplants for failed Fontan procedures. Induction therapy with antithymocyte therapy was used routinely, and long-term immunosuppression was by cyclosporine and azathioprine alone. Rejection surveillance/diagnosis was based on echocardiographic criteria. Posttransplantation follow-up ranges from 3 to 78 months. Operative mortality was 3.3% (1/30). No patients have been diagnosed with either accelerated allograft atherosclerosis or posttransplantation lymphoproliferative disease. We conclude that cardiac transplantation may be performed with excellent early and intermediate-term results.