Biomedical subjects

J A Morgan

Publications and source records attributed to J A Morgan.

At least 37 records · Page 2Linked to original sources

PubMed2000

Determination of metabolic rate-limitations by precursor feeding in Catharanthus roseus hairy root cultures.

Precursors from the terpenoid and tryptophan branches were fed to Catharanthus roseus to determine which of the two branches limits metabolic flux to indole alkaloids. The feeding of tryptophan at 17 days of the culture cycle produced auxin-like effects. Addition of low levels of auxin or tryptophan resulted in significant increases in flux to the indole alkaloids. Conversely, feeding higher levels of auxin or tryptophan resulted in increased branching and thickening of the hairy root cultures. A dramatic reduction in flux to the alkaloids was also observed. However, feeding tryptamine or terpenoid precursors had no effect. Therefore, neither pathway tested revealed to be rate-limiting during the late growth phase. Feeding of either geraniol, 10-hydroxygeraniol, or loganin at 21 days each resulted in significant increases in the accumulation of tabersonine. The addition of tryptophan or tryptamine had no effect during the stationary phase of the growth cycle. Thus, during the early stationary phase of growth the terpenoid pathway appears to be rate-limiting. Combined elicitation with jasmonic acid and feeding either loganin or tryptamine did not further enhance the accumulation of indole alkaloids.

Culture Media↗

PubMed2000

Effects of buffered media upon growth and alkaloid production of Catharanthus roseus hairy roots.

The influence of buffered media upon the growth and alkaloid productivity of Catharanthus roseus hairy root culture was examined. As expected, the buffers minimized shifts in the pH of the media and had slightly negative effects upon growth. The growth of the hairy roots remained optimal in unbuffered media. The specific yield of lochnericine was significantly lower in response to the addition of buffers, while tabersonine was significantly higher. In contrast, the specific yields of ajmalicine, serpentine, and horhammericine remained unchanged.

Alkaloids↗

PubMed2000

Isolation and characterization of 23 carbofuran-degrading bacteria from soils from distant geographical areas.

The aim of this work was to isolate, identify and type carbofuran-degrading bacteria from two geographically distant soils. Restriction Fragment Length Polymorphism (RFLP) patterns of the 16S rRNA gene and partial 16S rRNA sequence analysis were used to classify the 23 isolates obtained. Nine of them showed high similarity to Pseudomonas strains, seven showed similarity to the Flexibacter/Cytophaga/Bacteroides group and the remainder showed similarity to other bacterial genera. Isolates within the same group were sub-typed by comparing partial 16S rRNA sequences and SDS-PAGE analysis of their total protein profiles. Many of the UK isolates showed similarity to the Pseudomonas genera, while most of the Greek isolates showed similarity to the Flexibacter/Cytophaga/Bacteroides group. Only two Chrysobacterium strains isolated from both the UK and Greek soils were identical.

Bacteria↗

PubMed2000

Plasmid transfer between the Bacillus thuringiensis subspecies kurstaki and tenebrionis in laboratory culture and soil and in lepidopteran and coleopteran larvae.

Plasmid transfer between Bacillus thuringiensis subsp. kurstaki HD1 and B. thuringiensis subsp. tenebrionis donor strains and a streptomycin-resistant B. thuringiensis subsp. kurstaki recipient was studied under environmentally relevant laboratory conditions in vitro, in soil, and in insects. Plasmid transfer was detected in vitro at temperatures of 5 to 37 degrees C, at pH 5.9 to 9.0, and at water activities of 0.965 to 0.995, and the highest transfer ratios (up to 10(-1) transconjugant/donor) were detected within 4 h. In contrast, no plasmid transfer was detected in nonsterile soil, and rapid formation of spores by the introduced strains probably contributed most to the lack of plasmid transfer observed. When a B. thuringiensis subsp. kurstaki strain was used as the donor strain, plasmid transfer was detected in killed susceptible lepidopteran insect (Lacanobia oleracea) larvae but not in the nonsusceptible coleopteran insect Phaedon chocleriae. When a B. thuringiensis subsp. tenerbrionis strain was used as the donor strain, no plasmid transfer was detected in either of these insects even when they were killed. These results show that in larger susceptible lepidopteran insects there is a greater opportunity for growth of B. thuringiensis strains, and this finding, combined with decreased competition due to a low initial background bacterial population, can provide suitable conditions for efficient plasmid transfer in the environment.

Animals↗

PubMed1999

Breakpoints of the t(11;18)(q21;q21) in mucosa-associated lymphoid tissue (MALT) lymphoma lie within or near the previously undescribed gene MALT1 in chromosome 18.

Lymphomas arising in mucosa-associated lymphoid tissue (MALT) are indolent B-cell tumors that have a predilection for epithelial sites and often develop in a setting of chronic inflammation or autoimmunity. As many as 50% of low-grade MALT lymphomas contain an (11;18)(q21; q21) chromosomal translocation. Using fluorescence in situ hybridization, we have analyzed the position of recombination within chromosome 18 DNA in three examples of MALT lymphoma bearing this translocation. In all three cases, the breakpoint maps to DNA in BAC b357H2, covering about 150 kb of sequence. A previously undescribed, ubiquitously expressed gene, which we refer to as MALT1, was identified within this sequence and was found to be broken in one case for which we have definitively located the position of recombination between chromosomes 18 and 11. The sequence of this gene indicates the presence of two immunoglobulin-like C2 domains and a region of partial homology to caspases, suggesting a possible role for MALT1 in the regulation of apoptosis.

Amino Acid Sequence↗

PubMed1999

Inhibitor studies of tabersonine metabolism in C. roseus hairy roots.

The conversion of tabersonine to lochnericine and hörhammericine was investigated in C. roseus hairy root cultures. The accumulation of lochnericine and hörhammericine, like tabersonine, was associated with growth. Through the use of oxygenase inhibitors, 1-aminobenzotriazole (ABT), clotrimazole (CLOT), and 2.5-pyridinedicarboxylic acid (PCA), details of the metabolic pathway around tabersonine in hairy roots of C. roseus were elucidated. ABT specifically inhibited the formation of hörhammericine, while CLOT inhibited the accumulation of lochnericine. Using jasmonic acid in combination with the inhibitors suggests an inducible P-450 enzyme responsible for the formation of hörhammericine. The inhibitor study also revealed that both lochnericine and hörhammericine are 'turned over' in hairy root cultures.

Alkaloids↗

PubMed1999

Endovascular graft repair of ruptured aortoiliac aneurysms.

BACKGROUND: The feasibility of endovascular graft (EVG) repair of ruptured aortoiliac aneurysms (AIAs) has yet to be demonstrated. There are inherent limitations in EVG repair, including the need for preoperative measurements of the aneurysmal and adjacent arterial anatomy to determine the appropriate size and type of graft and the inherent delay to obtain proximal occlusion. We developed an EVG system with broad versatility that largely eliminates these problems. STUDY DESIGN: Between 1993 and 1998, within an experience of 134 endovascular AIA repairs, 12 ruptured AIAs were treated using EVGs that facilitated intraoperative customization and eliminated the need for preoperative measurements. The EVGs consisted of either a Palmaz stent and a PTFE graft deployed by a compliant balloon (n = 9) or a self-expanding covered stent graft (n = 3). Both grafts were cut to the appropriate length intraoperatively. The mean age of the patients was 72 years (range 40 to 86 years). The mean size of the aneurysms was 7.6 cm (range 3 to 16 cm). Preoperative symptoms were present in all patients and included abdominal or back pain (n = 9), syncope (n = 4), and external bleeding (n = 2). All patients were high surgical risks because of comorbid disease (n = 10) or previous abdominal operations (n = 6), and nine experienced hypotension. RESULTS: All EVGs were inserted successfully and excluded the aneurysms from the circulation. The mean operating time was 263 minutes, the mean blood loss was 715 mL, and the mean length of hospital stay was 6.5 days. There were two deaths (16%), one from the preexisting acute myocardial infarction and one from multiple organ failure. There were three minor complications (25%). Two patients required evacuation of an intraabdominal hematoma from the initial rupture. All but one of the grafts was functioning at a mean followup of 18 months. CONCLUSIONS: This study demonstrates the feasibility of EVG repair for ruptured AIAs using a graft that can be customized intraoperatively for each patient. Such repairs currently are valuable in patients with ruptured AIAs and serious comorbidities and may be applicable in other circumstances as well.

Adult↗

PubMed1999

Comparison of flagellin genes from clinical and environmental Pseudomonas aeruginosa isolates.

Pseudomonas aeruginosa, an important opportunistic pathogen, was isolated from environmental samples and compared to clinically derived strains. While P. aeruginosa was isolated readily from an experimental mushroom-growing unit, it was found only rarely in other environmental samples. A flagellin gene PCR-restriction fragment length polymorphism analysis of the isolates revealed that environmental and clinical P. aeruginosa strains are not readily distinguishable. The variation in the central regions of the flagellin genes of seven of the isolates was investigated further. The strains used included two strains with type a genes (998 bp), four strains with type b genes (1,258 bp), and one strain, K979, with a novel flagellin gene (2,199 bp). The route by which flagellin gene variation has occurred in P. aeruginosa is discussed.

Agaricales↗

PubMed1999

Vocal cord dysfunction in patients with exertional dyspnea.

STUDY OBJECTIVES: To evaluate patients for vocal cord dysfunction (VCD) in a military population presenting with symptoms of exertional dyspnea. DESIGN: Cross-sectional, controlled study. SETTING: Pulmonary disease clinic at an army tertiary care center. PATIENTS: Forty military patients with complaints of exertional dyspnea and 12 military asymptomatic control subjects. INTERVENTION: Patients underwent direct visualization of vocal cords with flexible laryngoscopy before and after exercise to evaluate for presence of inspiratory vocal cord adduction. MEASUREMENTS AND RESULTS: Complete evaluation for all patients consisted of spirometry with flow-volume loops, lung volumes, diffusing capacity, and maximum voluntary ventilation at rest; chest radiograph; methacholine bronchoprovocation testing; and a maximal cardiopulmonary exercise test with expiratory gas analysis. Fifteen percent of patients studied prospectively were found to have VCD, whereas all control subjects were negative for VCD. There was minimal difference in pulmonary function testing between VCD-positive and VCD-negative patients, whereas control subjects had higher spirometric values. Twenty percent of VCD-positive patients had abnormal flow-volume loops compared with 14% of patients without VCD, but after methacholine, 60% of VCD-positive patients developed abnormal flow-volume loops. In the VCD-positive group, 60% had a positive methacholine response, but there was less decrease in FEV(1)/FVC ratio compared with either VCD-negative patients or control subjects. CONCLUSIONS: Paradoxical inspiratory vocal cord closure is a frequent occurrence in patients with symptoms of exertional dyspnea and should be strongly considered in their evaluation.

Adolescent↗

PubMed1998

Transient studies of light-adapted cultures of hairy roots of Catharanthus roseus: growth and indole alkaloid accumulation.

Cultures of C. roseus transgenic ("hairy") root clones LBE-6-1 and LBE-4-2 were adapted with periodic daily illumination to investigate the effect of light on growth and nutrient utilization, and the accumulation of the indole alkaloids. Light-adapted roots appeared green and had radially thickened morphology compared with dark-grown controls. Their growth rates were higher than dark-grown controls, with 45% lower doubling times: LBE-6-1, 3.6 days; LBE-4-2, 2.8 days. Relative to dark-grown controls, light-adapted growth increased the biomass (DW) of LBE-6-1 by 25%, but had no effect on the DW of LBE-4-2. The macronutrients NH4+, NO3-, Pi, and sugars, were depleted completely by light-adapted root cultures in that order. The specific and total levels of the indole alkaloid serpentine was enhanced and of tabersonine was lowered in both root clones, while the overall trends of growth and non-growth association of tabersonine and serpentine, respectively, remained unaltered by light adaptation. Ajmalicine accumulation was enhanced in LBE-6-1, but lowered in LBE-4-2; its accumulation was growth-associated in dark-grown LBE-6-1, but appeared non-growth associated in light-adapted cultures. The accumulation of tabersonine-related compounds, lochnericine, and hörhammericine exhibited growth-associated trends, and were either negatively affected or unaffected by light adaptation of LBE-6-1. Neither vindoline nor its precursor, deacetylvindoline, was detected.

Adaptation, Physiological↗

PubMed1998

Trematode and monogenean rRNA ITS2 secondary structures support a four-domain model.

The secondary structure of rRNA internal transcribed spacer 2 is important in the process of ribosomal biogenesis. Trematode ITS sequences are poorly conserved and difficult to align for phylogenetic comparisons above a family level. If a conserved secondary structure can be identified, it can be used to guide primary sequence alignments. ITS2 sequences from 39 species were compared. These species span four orders of trematodes (Echinostomiformes, Plagiorchiformes, Strigeiformes, and Paramphistomiformes) and one monogenean (Gyrodactyliformes). The sequences vary in length from 251 to 431 bases, with an average GC content of 48%. The monogenean sequence could not be aligned with confidence to the trematodes. Above the family level trematode sequences were alignable from the 5' end for 139 bases. Secondary structure foldings predicted a four-domain model. Three folding patterns were required for the apex of domain B. The folding pattern of domains C and D varies for each family. The structures display a high GC content within stems. Bases A and U are favored in unpaired regions and variable sites cluster. This produces a mosaic of conserved and variable regions with a structural conformation resistant to change. Two conserved strings were identified, one in domain B and the other in domain C. The first site can be aligned to a processing site identified in yeast and rat. The second site has been found in plants, and structural location appears to be important. A phylogenetic tree of the trematode sequences, aligned with the aid of secondary structures, distinguishes the four recognized orders.

Animals↗

PubMed1998

Mitochondrial ND1 gene sequences used to identify echinostome isolates from Australia and New Zealand.

Echinostomes were collected in Australia and New Zealand as cercariae, metacercariae or adults. Using DNA sequences from the mitochondrial ND1 gene Echinostoma revolutum and Echinostoma paraensei were discovered in Australia. The presence of a further five, as yet unidentified, echinostome species was inferred in Northern Australia and a further isolate, closely allied to E. revolutum, occurs in New Zealand. ND1 sequences of species within the genus diverge from each other by 9.6-30.8%. Sequence divergence levels among strains within a single species are 0-3.6%. The phylogenetic tree produced from the Australasian isolates, in addition to species described previously, identifies the 37-collar-spine species as a well supported monophyletic group. The five unidentified Australian species cluster away from the 37-collar-spine group. These unidentified species appear to divide further into > 37-collar-spine and < 37-collar-spine clusters. Three strains of E. revolutum, collected as metacercariae from snails, were identified from two ponds located 6 km apart. Two of these strains may be cycling through a planorbid snail, Glyptophysa sp., as first intermediate host; however, this hypothesis could not be confirmed as specimens could not be obtained to match sequences between larvae and adults.

Animals↗

PubMed1998

Relative merits of nuclear ribosomal internal transcribed spacers and mitochondrial CO1 and ND1 genes for distinguishing among Echinostoma species (Trematoda).

Cryptic species, belonging to the 37 collar-spine Echinostoma group, were distinguished using nuclear rDNA ITS (884 bases) and mtDNA CO1 (257 bases) and ND1 (530 bases) sequences. Sequences were obtained from five 37 collar-spine species, Echinostoma trivolvis, E. paraensei, E. caproni, E. revolutum and E. sp.I, a parthenogenetic isolate from Africa. Three geographic isolates of E. caproni were compared. Average sequence divergence among the 37 collar-spine species range from 2.2% in the rDNA ITS through 8% for the CO1 and 14% for the ND1. In addition, genes were sequenced from 2 non 37 collar-spine species, E. hortense and an undescribed Australian species, E. sp. (Aus). For each gene, distances of terminals from a predicted ancestral sequence were calculated. These indicated that ND1 is diverging significantly faster than the other 2 regions. In the CO1 gene most substitutions are synonymous and saturation has been reached for the majority of pairwise comparisons. The ND1 gene exhibits greater pairwise divergence but less evidence of saturation due to weaker conservation of first and second codon positions. The ITS has no amino acid coding constraints and displays no evidence of saturation. Although all 3 regions successfully distinguished the nominal species, ND1 appears to be the most informative region for investigating relationships within the 37 collar-spine group.

Animals↗

PubMed1998

Variation in flagellin genes and proteins of Burkholderia cepacia.

The majority of isolates of Burkholderia cepacia, an important opportunistic pathogen associated with cystic fibrosis, can be classified into two types on the basis of flagellin protein size. Electron microscopic analysis indicates that the flagella of strains with the larger flagellin type (type I) are wider in diameter. Flagellin genes representative of both types were cloned and sequenced to design oligonucleotide primers for PCR amplification of the central variable domain of B. cepacia flagellin genes. PCR-restriction fragment length polymorphism analysis of amplified B. cepacia flagellin gene products from 16 strains enabled flagellin type classification on the basis of product size and revealed considerable differences in sequence, indicating that the flagellin gene is a useful biomarker for epidemiological and phylogenetic studies of this organism.

Amino Acid Sequence↗

PubMed1998

Recovery of cellular functions following oxidant injury.

This study investigated the recovery of renal proximal tubule cellular (RPTC) functions following oxidant-induced sublethal injury. tert-Butylhydroperoxide (TBHP) treatment resulted in 24% cell death and loss 4 h following the exposure. The remaining sublethally injured RPTC proliferated, and monolayer DNA content returned to control values on day 4 following TBHP exposure. Basal oxygen consumption (Qo2) and ATP content in sublethally injured RPTC were decreased 64 and 63%, respectively, at 4 h and returned to control values on day 6. Net lactate consumption decreased 71% at 4 h and returned to control values on day 4. In contrast, net glutamine consumption increased 2.7-fold at 4 h and returned to control values on day 6. Ouabain-sensitive Qo2, Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) activity, and Na(+)-coupled glucose transport were inhibited 77, 88, and 83%, respectively, at 4 h and recovered to control values on day 6. These data show that 1) mitochondrial function, Na(+)-K(+)-ATPase activity, active Na+ transport, and Na(+)-coupled glucose transport are decreased in sublethally injured RPTC following oxidant exposure and are repaired over time; 2) monolayer regeneration precedes the recovery of mitochondrial and transport functions, and 3) sublethal injury and subsequent regeneration are associated with alterations in metabolic substrate utilization. These results suggest that oxidant-induced sublethal injury to RPTC may contribute to renal dysfunction and that RPTC can repair and regain cellular functions following oxidant injury.

Animals↗

PubMed1997

The flagellin gene as a stable marker for detection of Pseudomonas fluorescens SBW25.

Flagellin gene central regions from 111 isolates of Pseudomonas fluorescens SBW25 obtained from soil during a field release experiment were analysed using a combined PCR/RFLP technique to look for variation. In addition, a 858 bp flagellin gene sequence from the original strain and the last isolate obtained from the release site were compared. There was no variation in flagellin gene sequences indicating that the gene was stable over the period of the release, and that the flagellin gene is a suitable marker for use in the detection of bacteria in release experiments. A comparison of Ps. fluorescens SBW25 flagellin with other sequenced flagellins revealed closest homology to the flagellin of Ps. putida PRS2000.

Base Sequence↗

PubMed1996

Flagellin gene and protein variation amongst clinical isolates of Pseudomonas aeruginosa.

Flagellin gene sequences from 64 clinical isolates of the opportunistic pathogen Pseudomonas aeruginosa were amplified by PCR and subjected to RFLP analysis by using seven restriction enzymes to digest the amplified products. Using this approach the isolates were assigned to one of 13 groups. The method was rapid, reproducible and applicable to all isolates. In contrast, serotyping failed to satisfactorily resolve 49% of the strains tested. The vast majority of clinical isolates generated amplified products of 1.02 kb (type a) or 1.25 kb (type b). Electron microscopical analysis revealed evidence fax some. flagellar structural variation between P. aeruginosa strains. This study provides further evidence that the flagellin gene is a widely applicable and useful genetic marker for studying genetic variation within populations of closely related bacteria.

Base Sequence↗

PubMed1996

Association of p300 and CBP with simian virus 40 large T antigen.

p300 and the CREB-binding protein CBP are two large nuclear phosphoproteins that are structurally highly related. Both function, in part, as transcriptional adapters and are targeted by the adenovirus E1A oncoprotein. We show here that p300 and CBP interact with another transforming protein, the simian virus 40 large T antigen (T). This interaction depends on the integrity of a region of T which is critical for its transforming and mitogenic properties and includes its LXCXE Rb-binding motif. T interferes with normal p300 and CBP function on at least two different levels. The presence of T alters the phosphorylation states of both proteins and inhibits their transcriptional activities on certain promoters. Although E1A and T show little sequence similarity, they interact with the same domain of p300 and CBP, suggesting that this region exhibits considerable flexibility in accommodating diverse protein ligands.

Amino Acid Sequence↗