Biomedical subjects

C G Murphy

Publications and source records attributed to C G Murphy.

At least 19 recordsLinked to original sources

PubMed2000

Mating preference functions of individual female barking treefrogs, Hyla gratiosa, for two properties of male advertisement calls.

A mating preference function describes the relationship between variation in a trait in potential mates and the strength of the preference for that trait. Few studies have measured mating preference functions either at a population level or for individuals. We used two-choice playback experiments to determine the mating preference functions of individual female barking treefrogs for two call characteristics: call-repetition rate and fundamental frequency. We tested each female four times with each pair of stimuli and with three to six pairs of stimuli. Individual females exhibited directional preferences for higher call rates and stabilizing preferences for intermediate fundamental frequency. These individual preference functions were similar to population-level preferences documented in previous studies. Within a stimulus type (call rate or fundamental frequency), nearly all females exhibited the same general preference function. However, females varied in the minimum difference between stimuli necessary to elicit a unanimous choice for the higher call rate, and they differed in both the intermediate fundamental frequency they preferred most and the minimum difference in fundamental frequency that elicited a unanimous choice for one of the two alternatives. The variation we observed among females was not repeatable; repeatabilities were in general low and statistically nonsignificant. The observed variation in the preferences of females, even if unrepeatable, should weaken selection on male traits relative to selection in the absence of such variation.

Animals↗

PubMed2000

Vaccine protection against simian immunodeficiency virus by recombinant strains of herpes simplex virus.

An effective vaccine for AIDS may require development of novel vectors capable of eliciting long-lasting immune responses. Here we report the development and use of replication-competent and replication-defective strains of recombinant herpes simplex virus (HSV) that express envelope and Nef antigens of simian immunodeficiency virus (SIV). The HSV recombinants induced antienvelope antibody responses that persisted at relatively stable levels for months after the last administration. Two of seven rhesus monkeys vaccinated with recombinant HSV were solidly protected, and another showed a sustained reduction in viral load following rectal challenge with pathogenic SIVmac239 at 22 weeks following the last vaccine administration. HSV vectors thus show great promise for being able to elicit persistent immune responses and to provide durable protection against AIDS.

Animals↗

PubMed1999

Glucocorticoids regulate transendothelial fluid flow resistance and formation of intercellular junctions.

The regulation of transendothelial fluid flow by glucocorticoids was studied in vitro with use of human endothelial cells cultured from Schlemm's canal (SCE) and the trabecular meshwork (TM) in conjunction with computer-linked flowmeters. After 2-7 wk of 500 nM dexamethasone (Dex) treatment, the following physiological, morphometric, and biochemical alterations were observed: a 3- to 5-fold increase in fluid flow resistance, a 2-fold increase in the representation of tight junctions, a 10- to 30-fold reduction in the mean area occupied by interendothelial "gaps" or preferential flow channels, and a 3- to 5-fold increase in the expression of the junction-associated protein ZO-1. The more resistive SCE cells expressed two isoforms of ZO-1; TM cells expressed only one. To investigate the role of ZO-1 in the aforementioned Dex effects, its expression was inhibited using antisense phosphorothioate oligonucleotides, and the response was compared with that observed with the use of sense and nonsense phosphorothioate oligonucleotides. Inhibition of ZO-1 expression abolished the Dex-induced increase in resistance and the accompanying alterations in cell junctions and gaps. These results support the hypothesis that intercellular junctions are necessary for the development and maintenance of transendothelial flow resistance in cultured SCE and TM cells and are likely involved in the mechanism of increased resistance associated with glucocorticoid exposure.

Body Fluids↗

PubMed1998

Effect of beta-adrenergic agonists on paracellular width and fluid flow across outflow pathway cells.

PURPOSE: To determine whether the adrenergic agonists epinephrine and isoproterenol regulate fluid flow across endothelial cells cultured from the human aqueous outflow pathway and to evaluate associated cellular mechanisms. METHODS: Confluent monolayers of human trabecular meshwork (TM) or Schlemm's canal endothelial (SCE) cells were grown on porous filter supports. The monolayers were perfused with media while fluid flow, expressed as hydraulic conductivity (HC = microl/min/mm Hg/cm2), was continuously measured in preparations treated with isoproterenol, epinephrine, or control medium. Morphometric ultrastructural methods were used to measure the area occupied by the intercellular space and by each cell. RESULTS: SCE cells and TM cells exposed to isoproterenol or epinephrine responded with an increase in transendothelial fluid flow. Dose-response curves for both adrenergic agonists showed that HC increased linearly as a function of the log of the isoproterenol and epinephrine concentration. At 10(-4) M isoproterenol, the HC increased threefold, and threshold conditions were reached at 10(-9) M. The increase in HC was apparent after isoproterenol had been applied for 1 hour, reached a peak in 3 to 4 hours, and declined gradually to return to baseline conditions in 10 to 12 hours. Morphometric analyses showed that the area occupied by the intercellular space increased fourfold when isoproterenol was used at 10(-4) M, whereas the cell area decreased as a function of the concentration of adrenergic agonist. Epinephrine's effects on HC and cell morphology were blocked by pretreatment with equimolar concentrations of the nonselective beta-blocker, timolol. CONCLUSIONS: Epinephrine and isoproterenol increase flow through the paracellular pathway of SCE and TM cells through a beta-receptor mediated response that widens the intercellular space and reduces cell area. These findings support the hypothesis that epinephrine decreases the intraocular pressure in glaucoma therapy by promoting fluid flow across the SCE and TM cells lining tissues of the major aqueous outflow pathway.

1-Methyl-3-isobutylxanthine↗

PubMed1995

Combined laparoscopic and endoscopic approach to resection of gastric leiomyoma.

Gastric leiomyoma is an uncommonly found entity. Most leiomyomas are asymptomatic and are found at autopsy or on abdominal exploration for other reasons. Indications for resection include large size and symptoms of compression, obstruction, or hemorrhage. The traditional surgical approach consists of laparotomy and resection. This article describes a technique for minimally invasive resection using a combined laparoscopic and endoscopic approach.

Aged↗

PubMed1995

Requirements for strand- and site-specific cleavage within the oriT region of Tn4399, a mobilizing transposon from Bacteroides fragilis.

Replicons that contain Tn4399, a conjugal mobilizing transposon isolated from Bacteroides fragilis, can be mobilized in the presence of broad-host-range IncP plasmids RP4 and R751 in Escherichia coli to B. fragilis or E. coli recipients (C. G. Murphy and M. H. Malamy, J. Bacteriol. 175:5814-5823, 1993). To identify the initial DNA processing events involved in Tn4399-mediated mobilization in E. coli, plasmid DNA from pCGM328 (a pUC7 vector that contains the mobilization region of Tn4399) was isolated from donor cells following the release of plasmid DNA from the relaxation complex. Site- and strand-specific cleavage within the oriT region of Tn4399 was detected by denaturing gel electrophoresis and Southern hybridization analysis of this DNA in the presence or absence of IncP plasmids. Mutations in either mocA or mocB, two genes which are encoded by Tn4399 and are required for mobilization, significantly decrease the amount of specifically nicked DNA detected. These results suggest roles for the MocA and MocB gene products in specific processing of Tn4399-containing plasmid DNA prior to mobilization. By isolation of the nicked strand and primer extension of this template, we mapped the precise 5' end of the single-stranded cleavage reaction. The nucleotide position of nicTn4399 is adjacent to two sets of inverted repeats, a genetic arrangement similar to those of previously characterized oriT regions. Two site-directed mutations which remove nicTn4399 (oriT delta 1 and oriT delta 2) cannot be mobilized to recipients when they are present in trans along with functional MocA and MocB proteins and an IncP mobilizing plasmid; they are cis-dominant loss-of-function mutations.

Bacterial Proteins↗

PubMed1994

Detection of herpes simplex viral DNA in the iridocorneal endothelial syndrome.

OBJECTIVE: To test the hypothesis that the iridocorneal endothelial (ICE) syndrome has a viral origin by comparing the incidence of viral DNA in corneal specimens from patients with the ICE syndrome and from controls. DESIGN: Thirty-one corneas obtained from 25 patients with the ICE syndrome and six with chronic herpetic keratitis (n = 31) were compared with 30 control specimens obtained from 15 healthy donors and from 15 patients with other, nonviral chronic corneal diseases. METHODS: Primer pairs and polymerase chain reaction methods were used to identify and amplify either a segment of the DNA polymerase gene in the case of the herpes simplex and zoster viruses or a region of the nuclear antigen gene for the Epstein-Barr virus. The oligonucleotide amplified by polymerase chain reaction was fully characterized with the use of restriction enzyme, hybridization, and sequence analyses to determine that it contained the expected base pair sequence. RESULTS: Sixteen of 25 ICE syndrome specimens and four of six herpetic keratitis specimens were positive for herpes simplex virus (HSV) DNA. All nine ICE syndrome specimens tested were negative for the presence of DNA from the herpes zoster or the Epstein-Barr viruses. Controls were uniformly negative for HSV DNA whether they were obtained from ostensibly normal corneas (n = 15) or from corneas with intestinal keratitis, aphakic bullous keratopathy, or keratoconus (n = 15). Tissue samples cut from positive ICE syndrome specimens yielded negative results when retested after the endothelial layer was removed. These findings indicate that localization of HSV DNA is within the endothelium, the tissue primarily involved in the pathogenesis of the ICE syndrome. CONCLUSIONS: Polymerase chain reaction evidence shows that HSV DNA is present in a substantial percentage of ICE syndrome corneal specimens and that HSV-DNA is absent in normal corneas and in corneas from patients with three other chronic corneal diseases. These results provide direct evidence to support our hypothesis that the ICE syndrome has a viral origin. We discussed clinical implications, including possible therapeutic interventions.

Base Sequence↗

PubMed1993

Comparison of efficacy of longest, average, and shortest axial length measurements with a solid-tip ultrasound probe in predicting intraocular lens power.

The accuracy of intraocular lens power predictions was calculated for 93 eyes using the shortest, average, and longest of multiple solid-tip probe ultrasound axial length readings for each eye. Averaging the readings reduced the effect of corneal indentation. The single longest and average readings gave comparable results.

Anthropometry↗

PubMed1993

Characterization of a "mobilization cassette" in transposon Tn4399 from Bacteroides fragilis.

Derivatives of nonconjugal plasmids that carry Tn4399, a transposon isolated from Bacteroides fragilis, can be mobilized for transfer by the broad-host-range IncP plasmids pRK231 or R751 in Escherichia coli. To characterize regions of Tn4399 involved in mobilization, we have isolated and analyzed subcloned fragments of Tn4399 in E. coli, as well as mutations within the element. We have identified a "mobilization cassette" within a 2.8-kb region of Tn4399 which, when cloned into mobilization-deficient plasmids, allows these plasmids to be mobilized in trans by the IncP plasmids pRK231 and R751. The 2.8-kb region has been sequenced, and several open reading frames have been identified. Mutants defective in two genes, designated mocA and mocB, coding for deduced products of 36.4 and 16.4 kDa, respectively, cannot be mobilized by either IncP plasmid; these mutants can be complemented in the presence of the respective wild-type genes in trans. This suggests that the putative MocA and MocB proteins have a role in the mobilization process. The 36.4-kDa MocA protein contains a 14-amino-acid sequence which is closely related to a highly conserved motif within DNA relaxases encoded by a wide variety of conjugal or mobilizable plasmids. Subcloning experiments also lead to the localization of an oriT region within a 199-bp fragment, internal to the mobilization cassette.

Amino Acid Sequence↗

PubMed1992

Outflow obstruction in pigmentary and primary open angle glaucoma.

To localize the site of outflow obstruction in glaucoma, we evaluated the trabecular meshwork tissues by morphometric methods. Thirty-three specimens from 27 patients with primary open angle glaucoma (n = 13), pigmentary glaucoma (n = 4), and pigment dispersion syndrome (n = 2), and from nonglaucomatous normal subjects (n = 8) were compared. In these specimens, the extent of aqueous channels and the area occupied by these channels where they terminate in cul-de-sacs were measured. In 32 nonglaucomatous normal specimens (six of the eight mentioned plus an additional 26), we discovered that 94% of the surface area of the cul-de-sacs is lined by trabecular cells. These measurements were used to calculate the resistance to aqueous outflow offered by cul-de-sacs. Three new concepts are advanced in this report: (1) the cul-de-sacs provide a major portion of the normal outflow resistance, (2) the cul-de-sac area is markedly reduced in pigmentary glaucoma and primary open angle glaucoma, accounting for a major portion of the increase in resistance in these conditions, and (3) macrophages are the major cell type responsible for trabecular meshwork clearance of pigment and debris. A common pathophysiologic sequence of events is proposed for the development of glaucoma in pigmentary glaucoma and primary open angle glaucoma.

Adult↗

PubMed1992

Juxtacanalicular tissue in pigmentary and primary open angle glaucoma. The hydrodynamic role of pigment and other constituents.

We tested the hypothesis that obstruction of the juxtacanalicular tissues, by melanin granules in pigmentary glaucoma and by other impermeable material in primary open angle glaucoma, leads to the development of a chronic glaucomatous condition. The distribution and concentration of melanin and other impermeable materials in the juxtacanalicular tissues and elsewhere in the trabecular meshwork was determined in 13 specimens. Six specimens were from patients with pigmentary glaucoma, two from patients with pigment dispersion syndrome, and three from patients with primary open angle glaucoma, as well as two from normal subjects. The effect of these materials on flow resistance was estimated using two hydrodynamic models. In model A, the electron-lucent spaces of the juxtacanalicular tissue were assumed to be open spaces, while in model B, these spaces and spaces filled with ground substance were assumed to be gel filled. In pigmentary glaucoma, 3.5% of the pigment was found in the juxtacanalicular tissue, while 96.5% was found in the corneoscleral and uveoscleral tissues. Permeabilities calculated according to model A were much higher than those expected from estimates of outflow facility in all groups, in agreement with the previous report of Ethier et al. The gel-filled spaces available for fluid flow, as determined by model B, showed no statistically demonstrable differences (pigmentary glaucoma, 32.9%; primary open angle glaucoma, 36.6%; pigment dispersion syndrome, 43.4%; normal, 44.1%). Furthermore, the amount of pigment present in the juxtacanalicular tissue was determined to have a negligible influence on permeability. Thus, the development of the chronic glaucomatous condition cannot be directly attributed to pigment accumulation in the juxtacanalicular tissue in pigmentary glaucoma.

Adult↗

PubMed1992

Minimizing anisometropia in bilateral pseudophakia.

One hundred twenty patients who had bilateral posterior chamber intraocular lens implantation were analyzed for postoperative anisometropia. All pairs of lenses had the same A constant and similar designs. Several parameters were analyzed to identify patients at higher risk for clinically significant anisometropia upon implantation of the second eye and determine whether the results of the first eye could be used to modify the implant power selected for the second eye to reduce the risk of anisometropia. In most cases, simply using the value of the linear regression prediction for emmetropia in the second eye without modifications minimized anisometropia.

Anisometropia↗

PubMed1992

Infection after injury: association with blood transfusion.

This study was undertaken to evaluate the association between red blood cell transfusions and infections in an easily stratified, homogenous group of injured adults. All received their initial transfusions upon arrival to the emergency department. Over 5 years, 390 uncross-matched trauma patients received type "O" red blood cells (RBCs) during initial resuscitation. One hundred fifty-four (39%) died within 7 days because of injuries sustained: 236 (61%) survived at least 7 days. Of these 236, clear differences could be seen between those receiving 6 or fewer or 7 or more units of RBCs. When adjusted for age, sex, and severity of injury (Champion Trauma Score, Injury Severity Score, TRISS), the risk of infection was higher in those receiving 7 or more units of RBCs. Similarly, risk of infection was related to units of RBCs transfused in a dose-related fashion. Blood transfusions should be avoided, if possible. Arbitrary "trigger points" for transfusions should be abandoned.

ABO Blood-Group System↗

PubMed1992

Postoperative ascitic leaks: the ongoing challenge.

BACKGROUND: The leak of ascitic fluid from surgical incisions is thought to be associated with a very high mortality rate. There have been few reports, however, focusing on the clinical characteristics, management, or mortality rates of this condition. METHODS: During a 10-year period, 18 patients with postoperative ascitic fluid leaks were treated. All patients had ascites before surgery and all had liver disease; in 13 of the 18 patients alcoholic liver disease was the cause of ascites. RESULTS: Ten of the 18 patients died (56%). Midline incisions were more often associated with recalcitrant leaks and fatal complications than were transverse incisions. CONCLUSIONS: Early consideration of fascial dehiscence and prompt repair is emphasized. The most effective predictor of survival was cessation of the leak.

Ascites↗

PubMed1991

A Bacillus subtilis dipeptide transport system expressed early during sporulation.

Two previously identified Bacillus subtilis DNA segments, dciA and dciB, whose transcripts accumulate very rapidly after induction of sporulation, were found in the same 6.2 kb transcription unit, now known as the dciA operon. Analysis of the sequence of the dciA operon showed that its putative products are homologous to bacterial peptide transport systems. The product of the fifth gene, DciAE, is similar to peptide-binding proteins from Escherichia coli and Salmonella typhimurium (DppA and OppA) and B. subtilis (OppA). A null mutation in dciAE abolished the ability of a proline auxotroph to grow in a medium containing the dipeptide Pro-Gly as sole proline source, suggesting that the dciA operon encodes a dipeptide transport system.

Amino Acid Sequence↗

PubMed1990

Treatment of candidosis in severely injured adults with short-course, low-dose amphotericin B.

Thirty-three (0.7%) of 4,818 trauma patients admitted between January 1, 1987, and July 1, 1989, developed invasive candidosis requiring IV antifungal therapy. All patients were seriously traumatized. Before developing candidosis, all patients had documented bacterial infections. These infections were generally polymicrobial and were treated with multiple broad-spectrum antibiotics (an average of 5.4 antibiotics for 17.2 days). Twenty-eight (85%) of 33 patients received enteral feedings for an average of 11 days +/- 1.5 (SEM) before developing candidosis and 24 (73%) received NG/oral nystatin for an average of 7.6 days +/- 0.9 before developing candidosis. All patients with candidosis were treated with intravenous amphotericin B: cumulative dose of 157.3 mg +/- 31.3 mg given over 10 days +/- 1.1. One patient developed recurrent candidosis despite NG/oral prophylaxis and enteral feedings. Six patients (18%) died due to sepsis and multiple organ failure. The patients who died did not objectively differ from the survivors. Candidosis is an infrequent infection in severely injured patients. Candidosis was invariably preceded by treatment with multiple broad-spectrum antibiotics for a variety of polymicrobial bacterial infections. NG/oral nystatin and enteral feedings did not prevent candidosis, in contrast to widely accepted beliefs. Amphotericin B therapy was safe. Recurrent candidosis was unusual. Candida infections had a high mortality rate associated with multiple blood transfusions and prolonged hospitalization. Candidosis represents a sign of severe injury and illness but can be amenable to prompt, aggressive treatment.

Adult↗

PubMed1989

Kinetics of phagocytosis in trabecular meshwork cells. Flow cytometry and morphometry.

Confluent human trabecular meshwork (HTM) cells from three different donors and at various stages of serial passage were fed fluorescein-labeled polystyrene beads. Phagocytosis was monitored for up to 6 days using flow cytometry, fluorescence microscopy, and morphometric calculations from comprehensive electron microscopic observations at key time points. During the first 4 hr after initiation of phagocytosis, the confluent endothelial monolayer lost its cohesiveness and became segregated into separate cells. During the first 3 days the cells underwent marked and progressive changes in shape and size. After 4 days, some cells detached from the dish, as necrotic debris and degenerative changes appeared. The kinetics of phagocytosis in this stable, confluent monolayer showed that recruitment (the percentage of cells which had ingested at least one bead) proceeded semilogarithmically, with 50% of the cells recruited by 8 hr and 97% by 96 hr. The time course of phagocytosis (ie, the average number of beads phagocytosed per cell) is described by a sigmoidlike curve, reaching half-maximum at 40 hr and maximum (about 500 beads per cell) at 96 hr. The rate of uptake (ie, the first derivative of the average number of beads per cell) reached a peak (nine beads per cell per hr) at 24 hr and then decelerated slowly over the next 5 days. Cytochalasin B treatment, as a control, reduced phagocytosis by approximately 70%. Flow cytometry, when combined with electron microscopy, should provide a useful tool to examine phagocytosis in HTM cells exposed to steroids and other hormones and drugs.

Adult↗

PubMed1989

Proteins secreted by human trabecular cells. Glucocorticoid and other effects.

The capacity of cultured human trabecular meshwork (HTM) cells to secrete an extracellular matrix was studied by indirect immunofluorescence. Synthesis of nine extracellular matrix (ECM) proteins known to be present in the normal trabecular meshwork was assessed in three HTM cell lines. Fourteen primary antibodies were used and cultures were labeled two and four weeks after confluence. The HTM cell lines showed consistent labelling patterns for the normal extracellular connective tissue constituents including collagens (types I, III, IV, V and VI), glycoproteins (laminin and fibronectin) and a basement membrane-associated proteoglycan. These antigens were localized to the basal cell surface in an extracellular reticular pattern corresponding to cell margins. Dextran addition at confluence helped to intensify the staining of these components, but ascorbate had no apparent effect. Interestingly, elastin, another normal component of the trabecular meshwork, was not identified under standard conditions, or after addition of ascorbate or dextran. However, elastin could be detected intracellularly following dexamethasone treatment for three days, and extracellularly in punctate deposits when this treatment was used for 1 or 2 weeks. Our findings indicate that HTM cells may be responsible for the secretion and maintenance of all the major ECM constituents of the trabecular meshwork. The elastin results suggest a possible mechanism contributing to obstruction of outflow in steroid glaucoma if increased amounts of elastin are also produced in vivo. This approach can also serve as a useful baseline for comparison with HTM cell lines treated with glaucoma medications or obtained from patients with glaucoma.

Antibodies↗