Identification of Escherichia coli ubiB, a gene required for the first monooxygenase step in ubiquinone biosynthesis.

It was recently discovered that the aarF gene in Providencia stuartii is required for coenzyme Q (CoQ) biosynthesis. Here we report that yigR, the Escherichia coli homologue of aarF, is ubiB, a gene required for the first monooxygenase step in CoQ biosynthesis. Both the P. stuartii aarF and E. coli ubiB (yigR) disruption mutant strains lack CoQ and accumulate octaprenylphenol. Octaprenylphenol is the CoQ biosynthetic intermediate found to accumulate in the E. coli strain AN59, which contains the ubiB409 mutant allele. Analysis of the mutation in the E. coli strain AN59 reveals no mutations within the ubiB gene, but instead shows the presence of an IS1 element at position +516 of the ubiE gene. The ubiE gene encodes a C-methyltransferase required for the synthesis of both CoQ and menaquinone, and it is the 5' gene in an operon containing ubiE, yigP, and ubiB. The data indicate that octaprenylphenol accumulates in AN59 as a result of a polar effect of the ubiE::IS1 mutation on the downstream ubiB gene. AN59 is complemented by a DNA segment containing the contiguous ubiE, yigP, and ubiB genes. Although transformation of AN59 with a DNA segment containing the ubiB coding region fails to restore CoQ biosynthesis, transformation with the ubiE coding region results in a low-frequency but significant rescue attributed to homologous recombination. In addition, the fre gene, previously considered to correspond to ubiB, was found not to be involved in CoQ biosynthesis. The ubiB gene is a member of a predicted protein kinase family of which the Saccharomyces cerevisiae ABC1 gene is the prototypic member. The possible protein kinase function of UbiB and Abc1 and the role these polypeptides may play in CoQ biosynthesis are discussed.

Topical pharyngeal anesthesia does not improve upper gastrointestinal endoscopy in conscious sedated patients.

OBJECTIVE: We undertook this study to determine whether topical pharyngeal anesthesia with conscious sedation is superior to conscious sedation alone, with respect to procedure performance or tolerance in patients undergoing diagnostic upper gastrointestinal endoscopy. METHODS: Ninety-five patients undergoing diagnostic upper endoscopy with conscious sedation were randomized to receive either topical pharyngeal anesthesia with 2% tetracaine/14% benzocaine spray or no pharyngeal anesthesia. Conscious sedation was achieved in all patients using intravenous midazolam and meperidine. Patients were asked to rate their pretest anxiety, comfort during endoscopy, recollection of the procedure, and willingness to undergo subsequent examinations using a 100-mm visual analog scale. Additionally, they were asked to estimate procedure duration and rate their tolerance for topical pharyngeal anesthesia. All examinations were performed by two endoscopists who were blinded to whether or not patients had received pharyngeal anesthesia. Endoscopists were asked to determine whether they believed that patients had received topical pharyngeal anesthesia and to estimate ease of esophageal intubation and procedure performance using a 100-mm visual analog scale. Procedure duration and doses of midazolam and meperidine were measured. RESULTS: The two groups did not differ with respect to age, gender, and previous endoscopic history. There were no significant differences between the two groups with respect to pretest anxiety, procedural comfort, and willingness to undergo subsequent examinations. Patients receiving topical pharyngeal anesthesia rated it as moderately unpleasant. Endoscopists were able to discriminate patients who received pharyngeal anesthesia from those who did not with a sensitivity of 0.73 and a specificity of 0.59. There were no significant differences between the two groups with respect to ease of intubation, procedure performance, procedure duration, and dosing of midazolam or meperidine. CONCLUSIONS: In patients undergoing diagnostic upper endoscopy using intravenous midazolam and meperidine, the use of topical pharyngeal anesthesia does not improve patient tolerance or procedure performance. Elimination of this agent in the performance of diagnostic upper endoscopy will save time and money without adversely affecting patient care or outcomes.

Biological activity of 26-succinylbryostatin 1.

Bryostatin 1, a macrocyclic lactone, has undergone phase I trials as an anticancer agent. Because of the lipid solubility of this compound it must be delivered either in ethanol or in a PET formulation. During the trial, these vehicles caused a large number of treatment-related side effects. We have synthesized the triethanolamine salt of 26-succinylbryostatin 1 and find that this compound is approx. 100-fold more water soluble than bryostatin 1. Because of the potential for clinical use, we have evaluated the biologic activity of this compound. We find that in a concentration-dependent manner 26-succinylbryostatin 1 is capable of activating protein kinase C (PKC) in vitro and displacing [3H]PDBu from PKC. However, at all concentrations tested the activity was less than the parent compound bryostatin 1. Addition of bryostatin 1 but not 26-succinylbryostatin 1 to U937 leukemic cells in culture stimulated a drop in cytosolic PKC, secondary to translocation of PKC to the membrane. Although 26-succinylbryostatin 1 did not stimulate a drop in the cytosolic levels of PKC, addition to U937 cells activated transcription from an AP-1 enhancer construct and c-Jun protein phosphorylation in a similar fashion to bryostatin 1 and differentiation of U937 cells. Unlike bryostatin 1, 26-succinylbryostatin 1 was unable to cause aggregation of human platelets. Although injection of bryostatin-1 into mice carrying B16 melanoma inhibits tumor growth, there was no significant inhibition of melanoma growth when identical doses of 26-succinylbryostatin 1 were injected. Therefore, 26-succinylbryostatin 1 shares some but not all of the pharmacologic properities of bryostatin 1. This compound can activate protein phosphorylation without lowering cytosolic levels of PKC.

Evaluation of a patient teaching skills course disseminated through staff developers.

Effective Patient Teaching (EPT), a course designed to improve health professionals' and health professions students' teaching skills, reliably produces gains in participants' skills when presented by its developers. The objective of this dissemination research study was to investigate whether, using a 'training of trainers' approach, seven nurses with staff development responsibilities in five different sites could teach EPT with similar effectiveness. The evaluation included pre- and post-course analysis of audiotaped patient education sessions conducted by 48 health professional participants who took EPT from one of the trainers in their home institutions. Post-course participant satisfaction surveys were also administered. EPT resulted in teaching skill improvements in four of five sites, and overall teaching skills scores improved significantly (P < 0.01). EPT can improve participants' teaching skills when taught by health professional trainers with staff development responsibilities who have recently received EPT training.

Peptides from conserved regions of paramyxovirus fusion (F) proteins are potent inhibitors of viral fusion.

The synthetic peptides DP-107 and DP-178 (T-20), derived from separate domains within the human immunodeficiency virus type 1 (HIV-1) transmembrane (TM) protein, gp4l, are stable and potent inhibitors of HIV-1 infection and fusion. Using a computer searching strategy (computerized antiviral searching technology, C.A.S.T.) based on the predicted secondary structure of DP-107 and DP-178 (T-20), we have identified conserved heptad repeat domains analogous to the DP-107 and DP-178 regions of HIV-1 gp41 within the glycoproteins of other fusogenic viruses. Here we report on antiviral peptides derived from three representative paramyxoviruses, respiratory syncytial virus (RSV), human parainfluenza virus type 3 (HPIV-3), and measles virus (MV). We screened crude preparations of synthetic 35-residue peptides, scanning the DP-178-like domains, in antiviral assays. Peptide preparations demonstrating antiviral activity were purified and tested for their ability to block syncytium formation. Representative DP-178-like peptides from each paramyxovirus blocked homologous virus-mediated syncytium formation and exhibited EC50 values in the range 0.015-0.250 microM. Moreover, these peptides were highly selective for the virus of origin. Identification of biologically active peptides derived from domains within paramyxovirus F1 proteins analogous to the DP-178 domain of HIV-1 gp4l is compelling evidence for equivalent structural and functional features between retroviral and paramyxoviral fusion proteins. These antiviral peptides provide a novel approach to the development of targeted therapies for paramyxovirus infections.

Measuring outcomes in psychiatry: an inpatient model.

BACKGROUND: This article describes a system for measuring outcomes recently implemented in the department of psychiatry of Baptist Memorial Hospital, a 78-bed inpatient and day treatment unit that represents one service line of a large, urban teaching hospital in Memphis. In June 1993 Baptist Hospital began a 15-month pilot test of PsychSentinel, a measurement tool developed by researchers in the Department of Community Medicine at the University of Connecticut. The hospital identified the following four primary goals for this pilot project: provide data for internal hospital program evaluation, provide data for external marketing in a managed care environment, satisfy requirements of the Joint Commission on Accreditation of Health Care Organizations, and generate studies that add to the literature in psychiatry and psychology. DESCRIPTION OF MEASURE: PsychSentinel is based on the standardized diagnostic criteria in the Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV). The outcome measure assesses the change in the number of symptoms of psychopathology that occurs between admission and discharge from the hospital. Included in the nonproprietary system are risk adjustment factors, as well as access to a national reference database for comparative analysis purposes. Data collection can be done by trained ancillary staff members, with as much or as little direct physician involvement as desired. The system has proven to be both time effective and cost effective, and it provides important outcome information both at the program level and at the clinician level. RESULTS: After the pilot test, the staff at Baptist Memorial Hospital determined that the system met all initial objectives identified and recently adopted the system as an ongoing measure of quality patient care in the department of psychiatry.

A reliable high-performance liquid chromatography assay for high-throughput routine cyclosporin A monitoring in whole blood.

We report here a reliable high-performance liquid chromatography-ultraviolet assay for routine assay of cyclosporin A (CsA) in whole blood using solid-phase extraction. This assay is linear, between 20 and 2,000 micrograms/L, with correlation coefficients > 0.998 for five consecutive standard curves. All coefficients of variation (CV) were < 8% at CsA concentrations of 45, 480, and 1,800 micrograms/L, with the exception of the between-day CV at 45 micrograms/L, which was < 15%. The relative accuracy of the method is > 94% at 45, 480, and 1,800 micrograms/L. The mean recoveries for CsA and cyclosporin D (internal standard) were 38.2 +/- 4.8% (n = 45) and 40.1 +/- 6.7% (n = 45), respectively. This method has proven to be reliable and robust in a high-throughput therapeutic drug monitoring laboratory.

Seasonal change in irradiance: a zeitgeber for circannual rhythms in ground squirrels.

1. Zeitgebers for circannual rhythms have been elusive. Demonstration that an external factor is a zeitgeber requires proof of a phase-shift that endures for several years. 2. The California ground squirrel (Spermophilus beecheyi) is an ideal subject. Many features of behavior have circannual rhythms of which change in mass is the easiest to measure. The squirrels thrive in captivity for up to 10 years. The squirrels were kept in individual cages in an air conditioned room, fed lab chow ad lib, and weighed twice a month. They were exposed to a 6-month phase shift of (a) length of day (b) seasonal change in temperature, (c) both, (d) seasonal cycle of irradiance. 3. The squirrels maintained circannual rhythms for up to 9 years. Entrainment was evident only by squirrels exposed to seasonal change in irradiance.

Septic arthritis due to gram negative bacilli: older patients with good outcome.

Previous studies of gram negative bacillary septic arthritis conclude it is an uncommon disease of older patients with poor outcome, or of younger patients with good outcome. We reviewed 22 cases of gram negative bacillary septic arthritis in a rural population. The mean age was 61 years, symptoms were of short duration (6.5 days mean), and a chronic illness was frequently present (77%). The knee was involved in 73% and E. coli was the usual causative organism (32%). Antibiotics plus aspiration was a successful initial therapy. Overall survival was 95% and a good outcome was achieved in 68%. Gram negative bacillary septic arthritis affects older patients, and the outcome appears good.

Cricoarytenoid arthritis: a cause of laryngeal obstruction.

Cricoarytenoid arthritis, principally as a result of long-standing rheumatoid arthritis, is a disorder that may present to the emergency physician with a number of symptoms and signs referable to the larynx. The presentation of this disorder as acute laryngeal obstruction and collapse is uncommon. It is important to recognize early cricoarytenoid joint involvement and not mistakenly diagnose mild cases as asthma or psychoneurosis. The use of steroids in mild cases has been beneficial at times, but progressive airway obstruction and fatalities have occurred during their administration.

Regulation of human population in northern France and adjacent lands in the Middle Ages.

"In northern France and Europe the population trebled after 1000 and then ceased to increase about 1300, before the Black Death caused an abrupt decline. The birthrate, as indicated by number of surviving children, increased and then declined. Death rates increased after 1250 and, of course, during the Black Death. Migration...was high after 1100 but ceased by about 1250. The evidence about resources, disease, and internecine strife is compatible with this scenario which can be predicted based on other animal populations."

Tuberculosis of peripheral joints: an often missed diagnosis.

Eleven elderly patients with tuberculous arthritis of 12 peripheral joints were studied retrospectively. The disease usually presented as a monoarticular arthritis often in the absence of other signs of tuberculosis. A delay in diagnosis was a common feature and synovial biopsies provided the highest diagnostic yield.

Aortic arch arteritis in the elderly. An important manifestation of giant cell arteritis.

Five elderly patients with aortic arch syndrome secondary to large-vessel vasculitis all presented with upper-extremity claudication and absence of blood pressure and pulses in the affected extremity. Diagnosis was suspected by a markedly elevated erythrocyte sedimentation rate associated with constitutional symptoms and angiograms showing changes consistent with arteritis of the aortic arch vessels. Treatment with high-dose corticosteroids resulted in rapid resolution of constitutional symptoms and improved functional use of the upper extremity. Recognition of this steroid-responsive disease is essential to avoid unnecessary reconstructive surgery and to prevent catastrophic events such as aortic dissection or visual loss, which may result from the underlying disease process.

Septic arthritis. Clinical approach to the 'hot joint'.

Joint space infections continue to be among the most common forms of arthritis. Diagnosis can be difficult, particularly in the presence of underlying joint disease. Septic arthritis should be suspected in all patients who present with an acutely swollen joint. Immediate joint aspiration with Gram's stain and culture of the synovial fluid is essential. Intensive, long-term antibiotic therapy is necessary to prevent a high rate of morbidity and, possibly, death.

Sarcoid arthritis in a North American Caucasian population.

Thirty-two of 150 Caucasian patients (21%) with sarcoidosis studied had articular symptoms. Twenty-one had acute sarcoid arthritis with bilateral ankle arthritis/periarthritis. Joint symptoms resolved within 4 months and none developed chronic arthropathy or progressive systemic sarcoidosis. Eleven patients developed articular involvement during the course of chronic pulmonary sarcoidosis manifested by bilateral hilar adenopathy in 8 and parenchymal disease in 9. Joint symptoms were generally transient, however, persistent synovitis in excess of one year was seen in 2 patients. Acute sarcoid arthritis confers a favorable prognosis in North American Caucasians. Differences from previous American series likely reflects the racial compositions of the population studied. Sarcoid arthritis less often evolves during the course of chronic systemic sarcoidosis and even then rarely progresses to a chronic arthropathy.