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Biomedical subjects

G Porter

Publications and source records attributed to G Porter.

At least 19 recordsLinked to original sources
PubMed1999

Topical aminoglycosides in the management of active mucosal chronic suppurative otitis media.

Debate has currently re-emerged following a renewed warning issued from the Committee on the Safety of Medicines (CSM) regarding the relative risk of ototoxicity from the use of aminoglycoside-containing drops in patients with tympanic membrane perforations. We present the findings of a survey of ENT consultants, questioning their views and current practice, and we add to the debate by means of a review and discussion of the literature.

Administration, Topical↗
PubMed1998

Modulation of quantum yield of primary radical pair formation in photosystem II by site-directed mutagenesis affecting radical cations and anions.

Pigment-protein interactions play a significant role in determining the properties of photosynthetic complexes. Site-directed mutants of Synechocystis PCC 6803 have been prepared which modify the redox potential of the primary radical pair anion and cation. In one set of mutants, the environment of P680, the primary electron donor of Photosystem II, has been modified by altering the residue at D1-His198. It has been proposed that this residue is an axial ligand to the magnesium cation. In the other set, the D1-Gln130 residue, which is thought to interact with the C9-keto group of the pheophytin electron acceptor, has been changed. The effect of these mutations is to alter the free energy of the primary radical pair state, which causes a change in the equilibrium between excited singlet states and radical pair states. We show that the free energy of the primary radical pair can be increased or decreased by modifications at either the D1-His198 or the D1-Gln130 sites. This is demonstrated by using three independent measures of quantum yield and equilibrium constant, which exhibit a quantitative correlation. These data also indicate the presence of a fast nonradiative decay pathway that competes with primary charge separation. These results emphasize the sensitivity of the primary processes of PS II to small changes in the free energy of the primary radical pair.

Alanine↗
PubMed1998

Is local anaesthesia actually beneficial in flexible fibreoptic nasendoscopy?

Although the application of a topical local anaesthetic before fibreoptic nasendoscopy is routine practice in many otolaryngological outpatients, the actual benefit to the patient of this procedure remains in doubt. Eighty-two patients were recruited in this double-blind randomized control trial which compared the patients' experiences of fibreoptic nasendoscopy with nasal preparations of Xylocaine (lidocaine), normal saline, and no spray to the nose and throat. A visual analogue scoring system was used to determine scores for the overall unpleasantness of procedure, receiving a spray, and taste of the spray, and pain. This study has shown significantly worse overall experience (P = 0.001) and pain (P = 0.048) scores for Xylocaine spray versus no spray. It is concluded that the routine use of topical local anaesthetics within the nose before routine fibreoptic nasendoscopy is not only of no value, but actually makes the experience worse for the patient.

Adult↗
PubMed1998

DNA sequence sampling of the Streptococcus pneumoniae genome to identify novel targets for antibiotic development.

We initiated a survey of the Streptococcus pneumoniae genome by DNA sequence sampling. More than 9,500 random DNA sequences of approximately 500 bases average length were determined. Partial sequences sufficient to identify approximately 95% of the aminoacyl tRNA synthetase genes and ribosomal protein (rps) genes were found by comparing the database of partial sequences to known sequences from other organisms. Many genes involved in DNA replication, repair, and mutagenesis are present in S. pneumoniae. Genes for the major subunits of RNA polymerase are also present, as are genes for two alternative sigma factors, rpoD and rpoN. Many genes necessary for amino acid or cofactor biosynthesis and aerobic energy metabolism in other bacteria appear to be absent from the S. pneumoniae genome. A number of genes involved in cell wall biosynthesis and septation were identified, including six homologs to different penicillin binding proteins. Interestingly, four genes involved in the addition of D-alanine to lipoteicoic acid in other gram positive bacteria were found, even though the lipoteicoic acid in S. pneumoniae has not been shown to contain D-alanine. The S. pneumoniae genome contains a number of chaperonin genes similar to those found in other bacteria, but apparently does not contain genes involved in the type III secretion commonly observed in gram negative pathogens. The G+C content of S. pneumoniae genomic DNA is approximately 43 mole percent and the size of the genome is approximately 2.0 Mb as determined by pulsed-field gel electrophoresis. Many of the genes identified by sequence sampling have been physically mapped to the 19 different SmaI fragments derived from the S. pneumoniae genome. The database of random genome sequence tags (GSTs) provides the starting material for determining the complete genome sequence, gene disruption analysis, and comparative genomics to identify novel targets for antibiotic development.

Amino Acyl-tRNA Synthetases↗
PubMed1997

Highly mismatched molecules resembling recombination intermediates efficiently transform mismatch repair proficient Escherichia coli.

The ability of related DNAs to undergo recombination decreases with increased sequence divergence. Mismatch repair has been proposed to be a key factor in preventing homeologous recombination; however, the contribution of mismatch repair is not universal. Although mismatch repair has been proposed to act by preventing strand exchange and/or inactivating multiply mismatched heteroduplexes, there has been no systematic study to determine at what step(s) in recombination mismatch repair acts in vivo. Since heteroduplex is a commonly proposed intermediate in many models of recombination, we have investigated the consequences of mismatch repair on plasmids that are multiply mismatched in heteroduplex structures that are similar to those that might arise during recombination. Plasmids containing multiply mismatched regions were transformed into wild-type and Mut+ Escherichia coli mutants. There was only a 30-40% reduction in transformation of Mut+ as compared to mutS and mutL strains for DNAs containing an 18% mismatched heteroduplex. The products obtained from mutS hosts differed from those obtained from Mut+ hosts in that there were many more colonies containing mixtures of two plasmids, due to survival of both strands of the heteroduplex. There were nearly 10 times more recombinants obtained from the mutS as compared to the wild-type host. Based on these results and those from other studies with E. coli and yeast, we propose that the prevention of recombination between highly diverged DNAs may be at a step earlier than heteroduplex formation.

DNA Repair↗
PubMed1996

Comparison of primary charge separation in the photosystem II reaction center complex isolated from wild-type and D1-130 mutants of the cyanobacterium Synechocystis PCC 6803.

We compare primary charge separation in a photosystem II reaction center preparation isolated from a wild-type (WT) control strain of the cyanobacterium Synechocystis sp. PCC 6803 and from two site-directed mutants of Synechocystis in which residue 130 of the D1 polypeptide has been changed from a glutamine to either a glutamate (mutant D1-Gln130Glu), as in higher plant sequences, or a leucine residue (mutant D1-Gln130Leu). The D1-130 residue is thought to be close to the pheophytin electron acceptor. We show that, when P680 is photoselectively excited, the primary radical pair state P680+Ph- is formed with a time constant of 20-30 ps in the WT and both mutants; this time constant is very similar to that observed in Pisum sativum (a higher plant). We also show that a change in the residue at position D1-130 causes a shift in the peak of the pheophytin Qx-band. Nanosecond and picosecond transient absorption measurements indicate that the quantum yield of radical pair formation (phi RP), associated with the 20-30-ps component, is affected by the identify of the D1-130 residue. We find that, for the isolated photosystem II reaction center particle, phi RP higher plant > phi RP D1-Gln130Glu mutant > phi RP WT > phi RP D1-Gln130Leu mutant. Furthermore, the spectroscopic and quantum yield differences we observe between the WT Synechocystis and higher plant photosystem II, seem to be reversed by mutating the D1-130 ligand so that it is the same as in higher plants. This result is consistent with the previously observed natural regulation of quantum yield in Synechococcus PS II by particular changes in the D1 polypeptide amino acid sequence (Clark, A.K., Hurry, V. M., Gustafsson, P. and Oquist, G. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 11985-11989).

Cyanobacteria↗
PubMed1996

Organizational impact of workaholism: suggestions for researching the negative outcomes of excessive work.

Excessive work can be viewed as an addictive behavior, and, as such, it will have a negative impact on the setting in which it occurs, as well as on the individual. This change in perspective is needed to address dysfunctional behavior patterns that interfere with organizational operations. Similarities with other addictions include identity issues, rigid thinking, withdrawal, progressive involvement, and denial. These factors influence decision making and goals of the workaholic. They also interfere with effectiveness by distorting interpersonal relations. Suggestions for further research build on established organizational topics that coincide with characteristics of an addictive pattern.

Behavior, Addictive↗
PubMed1996

Homologous and homeologous intermolecular gene conversion are not differentially affected by mutations in the DNA damage or the mismatch repair genes RAD1, RAD50, RAD51, RAD52, RAD54, PMS1 and MSH2.

Mismatch repair (MMR) genes or genes involved in both DNA damage repair and homologous recombination might affect homeologous vs. homologous recombination differentially. Spontaneous mitotic gene conversion between a chromosome and a homologous or homeologous donor sequence (14% diverged) on a single copy plasmid was examined in wild-type Saccharomyces cerevisiae strains and in MMR or DNA damage repair mutants. Homologous recombination in rad51, rad52 and rad54 mutants was considerably reduced, while there was little effect of rad1, rad50, pms1 and msh2 null mutations. DNA divergence resulted in no differential effect on recombination rates in the wild type or the mutants; there was only a five to 10-fold reduction in homeologous relative to homologous recombination regardless of background. Since DNA divergence is known to affect recombination in some systems, we propose that differences in the role of MMR depends on the mode of recombination and/or the level of divergence. Based on analysis of the recombination breakpoints, there is a minimum of three homologous bases required at a recombination junction. A comparison of Rad+ vs. rad52 strains revealed that while all conversion tracts are continuous, elimination of RAD52 leads to the appearance of a novel class of very short conversion tracts.

Amino Acid Sequence↗
PubMed1995

Mediation by HLA-DM of dissociation of peptides from HLA-DR.

Human leukocyte antigen (HLA)-DM is an unconventional major histocompatibility complex (MHC) class II heterodimer that is important for B-cell-mediated antigen processing and presentation to MHC class II-restricted T cells. HLA-DM is encoded by two genes, DMA and DMB, which map to the MHC class II region, and shares some homology with MHC class I and class II proteins. Here we define the biochemical role of HLA-DM. Recombinant soluble HLA-DM heterodimers have been purified from culture supernatants of insect cell transformants. At pH 5.0, they induce the dissociation of a subset of peptides bound to HLA-DR, including a nested set of class-II-associated invariant chain peptides (CLIP). This process liberates HLA-DR and leads to the enhanced binding of exogenous peptides.

Amino Acid Sequence↗
PubMed1995

Thrombolysis in acute myocardial infarction: reducing in hospital treatment delay.

AIM: Thrombolytic treatment when given early in acute myocardial infarction is beneficial. This study was initiated to firstly, document the existing time delays in the administration of thrombolysis at Auckland Hospital, and secondly, prospectively assess the effect of a staff education programme to reduce in-hospital delay. The goal was a "door to needle time" of less than 30 minutes. METHODS: The time delays in the administration of thrombolysis to patients with acute myocardial infarction admitted to the coronary care unit at Auckland Hospital were established. This was done with a retrospective chart review over a six month period January to June 1993. This was followed by a staff education programme to fast track the management of patients eligible for thrombolysis. A prospective assessment was performed from February to May 1994 to audit the effectiveness of the programme. RESULTS: Most of the delay in the administration of thrombolysis occurred in the community prior to arrival at hospital (median delay 2.5 hours). However there was still a significant delay in hospital with a median door to needle time of 59 minutes in 1993. Following the education programme in 1994 the median door to needle time was reduced by 32% to 40 minutes (p = 0.03). The proportion of patients with a door to needle time of less than 30 minutes doubled from 13% in 1993 to 27% in 1994 (p = 0.18). CONCLUSION: Staff training and thrombolysis guidelines are effective in reducing in-hospital treatment delay but additional strategies may be warranted. Thrombolysis should be administered in the coronary care unit or emergency department to avoid delay. Ongoing assessment of standards will be required in the general hospital setting as a quality indicator.

Adult↗
PubMed1994

Does slow energy transfer limit the observed time constant for radical pair formation in photosystem II reaction centers?

We have used spectrally photoselective femtosecond transient absorption spectroscopy on photosystem II reaction centers to show that there are at least two pools of chlorin molecules/states which can transfer excitation energy to P680, the primary electron donor in photosystem II. It has previously been shown that one chlorin pool equilibrates with P680 in 100 fs [Durrant et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11632-11636], and we report here the observation of energy transfer from a second more weakly coupled chlorin pool. The effect of the weakly coupled pool is to increase the apparent time constant for radical pair formation from 21 ps when P680 is selectively excited to 27 ps when the accessory chlorins are excited. We conclude that it is possible to observe both radical pair formation somewhat slowed by an energy transfer step and radical pair formation not limited by this slow energy transfer, depending upon which chromophores are initially excited. These observations provide evidence that when using photoselective excitation of P680, the observed 21 ps time constant for radical pair formation is not limited by a slow energy transfer step.

Energy Transfer↗
PubMed1994

Transformation-associated recombination between diverged and homologous DNA repeats is induced by strand breaks.

Rearrangements within plasmid DNA are commonly observed during transformation of eukaryotic cells. One possible cause of rearrangements may be recombination between repeated sequences induced by some lesions in the plasmid. We have examined the mechanisms of transformation-associated recombination in the yeast Saccharomyces cerevisiae using a plasmid system which allowed the effects of physical state and/or extent of homology on recombination to be studied. The plasmids contain homologous or diverged (19%) repeats of the URA3 genes (from S. cerevisiae or S. carlsbergensis) separated by the genetically detectable ADE2 colour marker. Recombination during transformation for covalently closed circular plasmids was over 100-fold more frequent than during mitotic growth. The frequency of recombination is partly dependent on the method of transformation in that procedures involving lithium acetate or spheroplasting yield higher frequencies than electroporation. When present in the repeats, unique single-strand breaks that are ligatable, as well as double-strand breaks, lead to high levels of recombination between diverged and identical repeats. The transformation-associated recombination between repeat DNAs is under the influence of the RAD52 and RAD1 genes.

DNA Damage↗
PubMed1994

Urinary enzymes as biomarkers of renal injury in experimental nephrotoxicity of immunosuppressive drugs.

Urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) and of alanine-aminopeptidase (AAP) was studied after administration of cyclosporine A (CSA A), FK 506, or the corresponding vehicles to salt-depleted rats. On days 7, 14, and 28 after treatment for CSA and day 14 after treatment for FK 506, measurements of the urinary enzymes, serum creatinine (SCr), creatinine clearance (ClCr), and blinded renal histology were done. After 1 week on CSA there was a dramatic increase of 489% in the urinary excretion of AAP (162.6 IU/g Cr, CSA vs. 27.6 IU/g Cr control, p < .03), a significant decrease of 32% in ClCr, a significant increase of 41% in SCr, and mild proximal tubular atrophy and vacuolization. After 2 or 4 weeks of CSA treatment there were no more differences in the urinary AAP between CSA and control rats, but the urinary excretion of NAG was increased: 29.6 IU/g Cr, CSA vs. 20.9 IU/g Cr, control, p < .03 on day 14 and 26.9 IU/g Cr, CSA vs. 21.5 IU/g Cr, control, p < .008 on day 28. At the same time there was a progressive decline of the ClCr, a progressive increase in the SCr, and an increase in the severity of the histological lesion. After 14 days of treatment with FK 506 we observed a striking elevation in urinary AAP (62.6 IU/g Cr, FK 506 vs. 36.0 IU/g Cr, control, p < .01) consistent with a significant decrease in ClCr, a significant increase in SCr, and a moderate proximal tubular vacuolization and atrophy.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylglucosaminidase↗
PubMed1993

Rate of oxidation of P680 in isolated photosystem 2 reaction centers monitored by loss of chlorophyll stimulated emission.

We have continued our studies of the primary photochemistry of isolated photosystem 2 reaction centers using femtosecond transient absorption spectroscopy. Experiments were performed over a wide range of excitation and probe wavelengths, using several data collection time scales. This has enabled us to resolve five different lifetimes ranging between 100 fs and 200 ps plus a nanosecond component. We demonstrate here and elsewhere [e.g., Durrant, J.R., Hastings, G., Joseph, D. M., Barber, J., Porter, G., & Klug, D. R. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11632-11636] that the kinetic spectra associated with all but two of these lifetimes are clearly distinguishable. We have previously reported that a 21-ps lifetime is associated with pheophytin reduction [Hastings, G., Durrant, J. R., Hong, Q., Barber, J., Porter, G., & Klug, D. R. (1992) Biochemistry 31, 7638-7647]. In this paper, we show that it is possible to spectrally and temporally resolve stimulated emission from PS2 reaction centers with great accuracy and that this stimulated emission is largely unaffected by those kinetic components which are faster than 21 ps. The observation of a distinct stimulated emission band allows us to distinguish charge-separated states from chlorin singlet states. In this way, we are able to show that the proportion of charge-separated states prior to the 21-ps component is between 0% and 25%. We also show that the shape of the spectrum which we obtain for the state P680+Ph- is essentially invariant between 100 ps and 9 ns, and is the same as that previously reported for P680+Ph- by other researchers.(ABSTRACT TRUNCATED AT 250 WORDS)

Chlorophyll↗
PubMed1993

Recognition by HLA-A2-restricted cytotoxic T lymphocytes of endogenously generated and exogenously provided synthetic peptide analogues of the influenza A virus matrix protein.

Experiments were carried out to determine whether complexes between MHC class I molecules and synthetic peptides are representative of those formed under more physiologically relevant conditions, with peptides derived intracellularly from processed antigens. Lysis of cells sensitized with exogenously provided and endogenously generated peptide analogues of the optimal nonameric peptide 58-66 (GILGFVFTL; derived from the influenza virus matrix protein) was compared. Endogenous loading was accomplished by expressing minigene DNA coding for alanine-substituted analogues of peptide 58-66 in HLA-A2-positive cells. Susceptibility to lysis by HLA-A2-restricted, peptide-specific cytotoxic lymphocytes was compared with lysis of cells sensitized with the same synthetic peptides. Although results were quite comparable, differences were observed. The endogenously presented analogues 58-66L60A, G61A, T65A, and L66A were recognized more efficiently than the corresponding exogenously presented analogues. This difference in recognition was most striking for peptide 58-66G61A. These results indicate the need for caution in using synthetic peptides in defining peptide binding motifs. Additional experiments with endogenously expressed analogues of 58-66 with substitutions other than alanine were carried out to define the interaction between this peptide and HLA-A2. Results are compatible with the interpretation that residues 58, 59, and 60 interact with pockets A, B, and D, respectively, in the HLA-A2 binding groove and that these interactions contribute to peptide binding.

Amino Acid Sequence↗
PubMed1993

A clinical and radiological study of back pain in rheumatoid arthritis.

Five hundred and three patients with RA were questioned about the symptom of back pain. Chronic back pain, lasting more than 3 months, occurred in 33 per cent of the group. A group of 100 back pain patients were studied in more detail using a structured questionnaire, clinical examination and radiology. Ninety-four of these patients had low back pain. Particular clinical patterns (such as that of the facet syndrome) were sought but no clear characteristics were found. Fifty-two lumbar spine X-rays were available from the RA population and these were compared to 52 age and sex matched X-rays from outpatients with chronic mechanical low back pain. Significant differences between these groups radiologically were a higher frequency of osteoporosis and a higher frequency of disc narrowing without associated osteophytes in the RA population. This study differs from previous reports which found other characteristic radiological features of RA of the lumbar spine (spondylolisthesis, facet erosions, and vertebral fracture), a discrepancy possibly resulting from the use of a control group having low back pain.

Adult↗