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Biomedical subjects

J W Barton

Publications and source records attributed to J W Barton.

At least 19 recordsLinked to original sources

Reductive transformation of methyl parathion by the cyanobacterium Anabaena sp. strain PCC7120.

Organophosphorus compounds are toxic chemicals that are applied worldwide as household pesticides and for crop protection, and they are stockpiled for chemical warfare. As a result, they are routinely detected in air and water. Methods and routes of biodegradation of these compounds are being sought. We report that under aerobic, photosynthetic conditions, the cyanobacterium Anabaena sp. transformed methyl parathion first to o,o-dimethyl o-p-nitrosophenyl thiophosphate and then to o,o-dimethyl o-p-aminophenyl thiophosphate by reducing the nitro group. The process of methyl parathion transformation occurred in the light, but not in the dark. Methyl parathion was toxic to cyanobacteria in the dark but did not affect their viability in the light. Methyl parathion transformation was not affected by mutations in the genes involved in nitrate reduction in cyanobacteria.

Aerobiosis↗

Influence of high biomass concentrations on alkane solubilities.

Alkane solubilities were measured experimentally for high-density biomass. The resulting Henry's law constants for propane were found to decrease significantly for both dense yeast suspensions and an actual propane-degrading biofilm consortium. At the biomass densities of a typical biofilm, propane solubility was about an order of magnitude greater than that in pure water. For example, a dense biofilm had a propane Henry's law constant of 0.09+/-0.04 atm m(3) mol(-1) compared to 0.6+/-0.1 atm m(3) mol(-1) measured in pure water. The results were modeled with mixing rules and compared with octanol-water mixtures. Hydrogels (agar) and salts decreased the alkane solubility. By considering a theoretical solubility of propane in dry biomass, estimates were made of intrinsic Henry's law constants for propane in pure yeast and biomass, which were 13+/-2 and 5+/-2 atm kg biomass mol(-1) for yeast and biofilm consortium, respectively.

Agar↗

Determination of effective transport coefficients for bacterial migration in sand columns.

A well-characterized experimental system was designed to evaluate the effect of porous media on macroscopic transport coefficients which are used to characterize the migration of bacterial populations. Bacterial density profiles of Pseudomonas putida PRS2000 were determined in the presence and absence of a chemical attractant (3-chlorobenzoate) gradient within sand columns having a narrow distribution of particle diameters. These experimental profiles were compared with theoretical predictions to evaluate the macroscopic transport coefficients. The effective random motility coefficient, used to quantify migration due to a random process in a porous medium, decreased nearly 20-fold as grain size in the columns decreased from 800 to 80 (mu)m. The effective random motility coefficient (mu)(infeff) was related to the random motility coefficient (mu), measured in a bulk aqueous system, according to (mu)(infeff) = ((epsilon)/(tau))(mu) with porosity (epsilon) and tortuosity (tau). Over the times and distances examined in these experiments, bacterial density profiles were unaffected by the presence of an attractant gradient. Theoretical profiles with the aqueous phase value of the chemotactic sensitivity coefficient (used to quantify migration due to a directed process) were consistent with this result and suggested that any chemotactic effect on bacterial migration was below the detection limits of our assay.

Journal Article↗

Quantitative Doppler assessment of acute scrotal inflammation.

PURPOSE: To establish quantitative Doppler criteria for acute unilateral epididymitis and/or orchitis. MATERIALS AND METHODS: Triplex ultrasonography (US) was used to prospectively evaluate 31 patients with acute hemiscrotal pain and 15 asymptomatic control subjects. Morphology and perfusion were assessed, and peak systolic velocities (PSVs) were recorded. Ratios of PSV were calculated by using values from the right and left sides. RESULTS: PSVs in control subjects differed significantly from those in patients (P < .0001). A PSV > or = 15 cm/sec produced diagnostic accuracy of 90% for orchitis and 93% for epididymitis: Five of six false-negative findings were in patients younger than age 15 years. Epididymal PSV ratios > or = 1.7 or testicular ratios > or = 1.9 were diagnostic of acute inflammation (P < .0001). In three cases, morphologic and color Doppler findings were normal and only the PSV ratios were diagnostic. CONCLUSION: PSV and PSV ratios provide diagnostic criteria for acute unilateral epididymitis and/or orchitis that are more accurate than morphologic evaluation and color flow imaging either alone or in combination.

Acute Disease↗

Purification of, generation of monoclonal antibodies to, and mapping of phosphoribosyl N-formylglycinamide amidotransferase.

5'-Phosphoribosyl N-formylglycinamide (FGAR) amidotransferase (EC 6.3.5.3) catalyzes the fourth reaction in the de novo synthesis of purines, that is, the conversion of FGAR to 5'-phosphoribosyl N-formylglycinamidine (FGAM). This is the only step of the pathway for which a vertebrate gene has not been cloned. FGAR amidotransferase has been highly purified from Chinese hamster ovary (CHO) cells, and this preparation has been used to generate monoclonal antibodies in mice. Two of these antibodies, designated BD4 and DD2, have been shown to recognize a 150-kDa protein in CHO-K1 cells that is of very low abundance in Ade-B cells, a CHO line in which FGAR amidotransferase activity is undetectable. Furthermore, the protein recognized by these antibodies is 5-10-fold more abundant in Azr cells. The CHO Azr cell line was made resistant to azaserine, a potent inhibitor of FGAR amidotransferase, and displays a 5-10-fold increase in FGAR amidotransferase activity over the parental K1 line. FGAR amidotransferase activity and the 150-kDa protein recognized by both monoclonal antibodies were found to immunoprecipitate concomitantly using antibody BD4. Monoclonal antibody DD2 cross-reacted with a human protein of identical molecular mass. A number of Ade-B/human hybrid cells were generated by somatic cell fusion and subsequent 5-bromo-2-deoxyuridine segregation. Analysis of these lines, together with two independently generated human/mouse hybrid cell lines, by both cytogenetics and immunoblotting with antibody DD2 revealed that the human FGAR amidotransferase gene is located on chromosome 17p.

Animals↗

MR diagnosis of tears of anterior cruciate ligament of the knee: importance of ancillary findings.

OBJECTIVE: Detection of tears of the anterior cruciate ligament with MR is usually based on the appearance of the ligament. However, other MR findings may be useful to establish the diagnosis. We assessed the utility of these ancillary MR findings for detecting tears and for differentiating partial from complete tears of the anterior cruciate ligament. MATERIALS AND METHODS: MR images of 68 patients who had arthroscopic correlation were retrospectively reviewed by two radiologists who did not know the arthroscopic findings. The reviewers noted the presence and location of bone bruises (nonlinear low signal in the bone marrow on T1-weighted images), assessed posterior displacement of the posterior horn of the lateral meniscus, and measured the posterior cruciate ligament angle. At arthroscopy, 39 patients had tears of the anterior cruciate ligament (24 complete, 15 partial) and 29 had intact anterior cruciate ligaments. RESULTS: The presence of bone bruising in the posterolateral tibial plateau had sensitivities of 50% and 46%, with corresponding specificities of 97% and 97% for the two reviewers. Posterior displacement of the posterior horn of the lateral meniscus had a sensitivity of 56% for both reviewers, with 100% and 97% specificities. An angle of the posterior cruciate ligament less than 105 degrees had sensitivities of 72% and 74%, with corresponding specificities of 79% and 86%. The prevalence of these findings was similar in patients with partial tears and those with complete tears. CONCLUSION: Ancillary MR imaging findings may be helpful for diagnosing tears of the anterior cruciate ligament when diagnosis based on the MR appearance of the ligament is equivocal. Ancillary findings cannot be used to differentiate patients with partial tears from those with complete tears. The presence of bone bruising in the posterior aspect of the tibial plateau and posterior displacement of the posterior horn of the lateral meniscus are highly specific for a torn anterior cruciate ligament; therefore, the presence of either of these findings should be considered strong evidence of a tear of the anterior cruciate ligament.

Adult↗

Pelvic MR imaging findings in gestational trophoblastic disease, incomplete abortion, and ectopic pregnancy: are they specific?

Thirty-nine patients with abnormally elevated levels of serum beta subunit of human chorionic gonadotropin (beta-hCG) were studied to determine whether findings at magnetic resonance (MR) imaging are specific for primary molar disease, persistent gestational trophoblastic disease (GTD), incomplete abortion, and ectopic pregnancy. Among the latter three groups, the only significant differences were a higher prevalence of endometrial distention in the group with incomplete abortion (P < .0035) and the absence of junctional zone disruption in the group with ectopic pregnancy (P < .05). In the group with primary molar disease, total intrauterine volume was significantly increased (P < .001), and endometrial distention and presence of an endometrial mass had a significantly higher prevalence than that in the persistent GTD groups with (P < .04) or without (P < .001) metastases. Myometrial or extrauterine disease was identified in 65% of the patients with persistent disease and a beta-hCG level greater than 500 mIU/mL (500 IU/L). Thus, although MR imaging findings in persistent GTD, incomplete abortion, and ectopic pregnancy are relatively nonspecific, MR imaging can depict invasive disease that may alter therapeutic management in patients with documented GTD.

Abortion, Incomplete↗

Peripheral transcatheter embolization with platinum microcoils.

Embolotherapy with platinum microcoils delivered through the Tracker-18 microcatheter (Target Therapeutics, San Jose, Calif) was performed in 16 patients when peripheral superselective catheterization with standard angiographic catheters was not possible. The Tracker-18 catheter could be directed distally into small peripheral vessels for delivery of the microcoils. These microcoils, with attached fiber strands, resulted in vascular occlusion within a few minutes in all cases. Embolotherapy was technically successful in all patients. The procedures were clinically successful in 15 of 16 patients (94%), and no complications were encountered. Embolization with platinum microcoils through the Tracker-18 catheter is useful when standard methods of embolization are not possible.

Adolescent↗

Long-term outcome of pediatric sarcoidosis with emphasis on pulmonary status.

Sixty-one pediatric and adolescent patients (age < or = 16 years) with sarcoidosis proved by biopsy specimen were identified during the period 1957 to 1976; 19 patients with elapsed time from diagnosis of 8 to 35 years (mean, 21 years) were reexamined in 1985. Age at onset of disease ranged from 4 to 16 years (mean, 12.5 years). Sex distribution was equal; 68 percent of individuals were black. At follow-up, clinical evaluation, chest roentgenograms, pulmonary function tests, electrocardiograms (ECGs), echocardiograms (ECHO), and angiotensin-converting enzyme (ACE) activity were performed on each subject. In addition, complete blood cell counts, erythrocyte sedimentation rates, serum calcium, immunoglobulin levels, B- and T-cell enumerations, and intradermal skin tests for delayed hypersensitivity were also obtained. Although all initial pediatric chest roentgenograms were abnormal, at follow-up only 37 percent were abnormal. Pulmonary function test results were available for ten children, and 90 percent were decreased. In 1985, 68 percent of the adults had abnormal lung function; furthermore, eight patients had reduced diffusing capacities, one had hypoxemia, and two had elevated ACE activity. Specific abnormalities were noted on two ECGs and 12 ECHOs. One individual had an elevated sedimentation rate, while another had an increased serum calcium level. Six patients had elevated IgA values, two had elevated IgM values, and two had depressed IgM values; IgG values were normal in all subjects. B- and T-cell percentages were unremarkable in all patients tested. Four individuals were anergic to skin test antigens. Long-term pulmonary morbidity was observed in four patients; in addition, one of these and four others suffered nonpulmonary sequelae. These results are in agreement with those of other investigators, but insufficient data still exist on the long-term effects of sarcoidosis on the pediatric host.

Adolescent↗

Isolation of a human cDNA encoding amidophosphoribosyltransferase and functional complementation of a CHO Ade-A mutant deficient in this activity.

We report here the isolation of a human cDNA encoding the first step in de novo purine biosynthesis, amidophosphoribosyltransferase (PRAT). The human PRAT cDNA was isolated by complementation of a Saccharomyces cerevisiae ade4 mutant deficient in PRAT enzymatic activity. The identity of the isolated cDNA, designated pAdeA-3, was confirmed by several independent methods. Genomic DNA sequences homologous to pAdeA-3 show coordinate segregation with the hypoxanthine nutritional requirement in Chinese hamster ovary (CHO) cell Ade-A-human hybrids, segregants of these hybrids, and irradiation reduction hybrids. The PRAT cDNA after insertion into a mammalian expression vector was capable of correcting the PRAT cDNA after insertion into a mammalian expression vector was capable of correcting the PRAT enzyme deficiency in CHO Ade-A mutants. This correction was monitored by both cell-free PRAT assays and in vivo phosphoribosylformylglycinamide (FGAR) accumulation studies. FGAR accumulation is a classic method for assessment of the early steps of purine nucleotide biosynthesis. Two of the isolated transformants, designated PRAT-1 and PRAT-2, exhibited 22% and 53%, respectively, of wild-type CHO K1 PRAT enzymatic activity using a cell-free enzyme assay. These same two transformants plus an additional transformant, designated PRAT-13, showed FGAR accumulations of 150%, 260%, and 140%, respectively, compared to the levels of accumulation seen in CHO K1. Transformants PRAT-1 and PRAT-2 both contained a mRNA species recognized by the PRAT cDNA of identical size to a mRNA species in human fibroblasts homologous to the PRAT cDNA. This observation, along with the functionality of the cDNA in both yeast and CHO cells deficient in PRAT activity, suggests the isolated cDNA is full length.

Amidophosphoribosyltransferase↗

Mapping of a locus correcting lack of phosphoribosylaminoimidazole carboxylase activity in Chinese hamster ovary cell Ade-D mutants to human chromosome 4.

The human phosphoribosylaminoimidazole (AIR) carboxylase locus has been until this report one of the genes encoding purine biosynthetic enzymes that had not been assigned to an individual human chromosome. Characterization of Chinese hamster ovary (CHO) cell mutant Ade-D showed that the cell line was unable to produce IMP and accumulated AIR. CHO Ade-D cells were fused with normal human lymphocytes utilizing inactivated Sendai virus and the resulting hybrid cell lines were selected for purine prototrophy. Cytogenetic analysis showed a 100% concordance value for chromosome 4. Two of the isolated subclones contained only the long arm of chromosome 4 translocated onto a CHO chromosome, providing evidence for a regional assignment of the Ade-D gene to the long arm of chromosome 4. Two of the subclones containing chromosome 4 were subjected to the BrdU visible light segregation. All of the isolated purine auxotrophic cell lines showed a loss of the q arm of chromosome 4. The localization of the Ade-D locus to the long arm of chromosome 4 may reveal further clustering of the mammalian purine genes since the Ade-A locus has previously been regionally assigned to 4pter-q21.

Animals↗

Liver transplantation for hepatoblastoma in a child with congenital absence of the portal vein.

Congenital absence of the portal vein with systemic visceral venous return in a 8-year-old girl with oculoauriculovertebral dysplasia (Goldenhar Syndrome) had been previously reported following its discovery during preoperative evaluation of a liver mass which was diagnosed as focal nodular hyperplasia after open biopsy. Subsequently, an enlarging diffuse hepatic neoplasm developed with associated elevated alpha feto-protein levels. Repeat biopsy and imaging showed the tumor to be a hepatoblastoma involving both lobes of the liver. The patient was treated by hepatic resection and orthotopic liver transplantation and is doing well at 18 months follow-up.

Abnormalities, Multiple↗

Generation of deletions in the 3'-flanking sequences of the Escherichia coli crp gene that induce cyclic AMP suppressor functions.

The crp structural gene and its 3'-flanking sequences were subcloned into M13mp8, and in vitro deletions were constructed in both the 5' and 3' ends of the gene by using Bal 31 nuclease. Deletions ranged in size from 24 to 250 base pairs at the 5' end of crp. Sixteen deletions generated at the 3' end of the gene ranged in size from 133 to 675 base pairs. The majority of deletions extended into the crp structural gene. Another class of deletions, i.e., delta crp-4, delta crp-17, and delta crp-2, had endpoints extending in the 3'-flanking sequences external to the crp structural gene. Deletions were subcloned into pBR322 and transformed into the Escherichia coli cya crp deletion strain NCR438. Transformants containing plasmid pBM4 with the delta crp4 mutation, a deletion of 133 base pairs, were cyclic AMP independent. Strain NCR440 harboring this plasmid expressed beta-galactosidase and threonine dehydratase activities and fermented lactose, ribose, arabinose, and xylose in the absence of exogenous cyclic AMP. The delta crp-4 mutation also caused strain NCR440 to be hypersensitive to exogenous cyclic AMP. The cylic AMP receptor protein expressed in maxicells from pBM4 carrying the delta crp-4 mutation comigrated with the wild-type protein on electrophoretic gels. The delta crp-4 mutation demonstrates that sequences distal to the crp structural gene can mediate cyclic AMP suppressor functions.

Bacterial Proteins↗

Reductive microbial dechlorination of indigenous polychlorinated biphenyls in soil using a sediment-free inoculum.

In laboratory experiments, unagitated soil slurry bioreactors inoculated with micro-organisms extracted from polychlorinated biphenyl-contaminated (PCBs) sediments from the Hudson River were used to anaerobically dechlorinate PCBs. The onset of dechlorination activity was accelerated by the addition of certain organic acids (pyruvate and maleate) and single congeners (2,3,6-trichlorobiphenyl). Dechlorination was observed under several working conditions after 19 weeks of incubation with PCB-contaminated soil and nutrient solution. Best results showed a drop in average chlorine content from 4.3 to 3.6 chlorines per biphenyl due to a loss of m-chlorines. Soil used for these experiments was obtained from a PCB-contaminated (weathered Aroclor 1248) site at an electric power substation. Dechlorination was observed with no sediment particles or other matrix being added.

Anaerobiosis↗