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Biomedical subjects

M O'Donnell

Publications and source records attributed to M O'Donnell.

At least 73 records · Page 4Linked to original sources
PubMed2000

Dynamics of beta and proliferating cell nuclear antigen sliding clamps in traversing DNA secondary structure.

Chromosomal replicases of cellular organisms utilize a ring shaped protein that encircles DNA as a mobile tether for high processivity in DNA synthesis. These "sliding clamps" have sufficiently large linear diameters to encircle duplex DNA and are perhaps even large enough to slide over certain DNA secondary structural elements. This report examines the Escherichia coli beta and human proliferating cell nuclear antigen clamps for their ability to slide over various DNA secondary structures. The results show that these clamps are capable of traversing a 13-nucleotide ssDNA loop, a 4-base pair stem-loop, a 4-nucleotide 5' tail, and a 15-mer bubble within the duplex. However, upon increasing the size of these structures (20-nucleotide loop, 12-base pair stem-loop, 28-nucleotide 5' tail, and 20-nucleotide bubble) the sliding motion of the beta and proliferating cell nuclear antigen over these elements is halted. Studies of the E. coli replicase, DNA polymerase III holoenzyme, in chain elongation with the beta clamp demonstrate that upon encounter with an oligonucleotide annealed in its path, it traverses the duplex and resumes synthesis on the 3' terminus of the oligonucleotide. This sliding and resumption of synthesis occurs even when the oligonucleotide contains a secondary structure element, provided the beta clamp can traverse the structure. However, upon encounter with a downstream oligonucleotide containing a large internal secondary structure, the holoenzyme clears the obstacle by strand displacing the oligonucleotide from the template. Implications of these protein dynamics to DNA transactions are discussed.

Base Pairing↗
PubMed2000

Sliding clamps: a (tail)ored fit.

New structural information on the architecture of a DNA replisome provides insights into a number of DNA metabolic processes and their modulation by circular 'sliding damps', which form rings around DNA that play an Important role in processive processes such as replication.

Animals↗
PubMed2000

The effect of maternal exposure to flaxseed on spermatogenesis in F(1) generation rats.

Pregnant Sprague-Dawley rats were exposed to a flaxseed (20 or 40%), flaxmeal (13 or 26%) or standard NIH AIN-93 (0% flaxseed control) diet throughout gestation and until their offspring were weaned. After weaning, F(1) generation males were placed in the same diet treatment groups as their mothers for 70 days. Statistically significant differences were not observed between either low-dose or high-dose flaxseed and flaxmeal-treated animals and the 0% flaxseed control animals for testis weights, homogenization resistant spermatid counts, daily sperm production rates, epididymal weights, seminal vesicle weights, seminiferous tubule fluid testosterone concentrations and the percentage of sperm abnormalities. The following statistically significant differences were observed when treated groups and the 0% flaxseed control groups were compared: (1) increases in serum LH in the 20% and 40% flaxseed treatment groups and in serum LH and testosterone in the 26% flaxmeal treatment group; (2) increases in the cauda epididymal weight from the 20% and 40% flaxseed groups; (3) increases in cauda epididymal sperm numbers/g epididymis from the 20% and 40% flaxseed and the 13% and 26% flaxmeal treatment groups; (4) a decrease in prostatic weight from the 20% flaxseed and 13% and 26% flaxmeal treatment groups. Prostate weight in the 40% flaxseed treatment group was lower but not statistically significantly different than the 0% flaxseed control group. Histological effects on spermatogenesis were not observed in either the control group, flaxmeal or the flaxseed treated groups.

Analysis of Variance↗
PubMed2000

Mechanisms of DNA replication.

Advances in recent years have led to exciting new ideas about the initiation, regulation and coordination of DNA replication. Structural studies have yielded fascinating glimpses of replisome action. In addition, the involvement of replication proteins in other cellular processes has blurred the lines between replication, repair and recombination.

DNA Replication↗
PubMed2000

A tale of toroids in DNA metabolism.

A strikingly large number of the proteins involved in DNA metabolism adopt a toroidal -- or ring-shaped -- quaternary structure, even though they have completely unrelated functions. Given that these proteins all use DNA as a substrate, their convergence to one shape is probably not a coincidence. Ring-forming proteins may have been selected during evolution for advantages conferred by the toroidal shape on their interactions with DNA.

Animals↗
PubMed2000

Static splinting of extensor tendon repairs.

In a prospective review we assessed the results of extensor tendon injuries managed postoperatively with a static splint. Thirty-three patients with 44 injured digits were assessed 4 months after primary tendon repair, using the Strickland-Glogovac criteria. Patients were managed in a static splint, the duration of which was guided by the zone of injury. Four months after repair, excellent or good results were obtained in 95%. Overall it was found that static splinting was an effective and safe method of management after extensor tendon repair.

Adolescent↗
PubMed2000

Squamous cell carcinoma of skin with a rhabdoid phenotype: a case report.

A 67 year old man presented with a polypoid lesion on the temple that had all the light microscopic, immunohistochemical, and ultrastructural features of a rhabdoid tumour. There was an area of intraepidermal carcinoma and invasive squamous carcinoma at the base of the polyp. The tumour progressed aggressively and the patient died five months after primary excision. Cutaneous tumours with a rhabdoid morphology have been described previously and tend to have a very poor prognosis. No previously published report describes a clear squamous histogenesis.

Aged↗
PubMed2000

Pathophysiological role of T lymphocytes in renal ischemia-reperfusion injury in mice.

Mononuclear cell infiltrates are found in human renal ischemia-reperfusion injury (IRI), and peritubular T lymphocytes have been identified in experimental IRI. However, the role of T cells in the pathogenesis of renal IRI is unknown. We hypothesized that T cells are one of the important mediators of renal IRI. To test this hypothesis, we used an established mouse model of renal IRI, and evaluated mice with genetically engineered deficiency of both CD4+ and CD8+ T cells. At 48 h postischemia, CD4/CD8-knockout (KO) mice had marked improvement in renal function compared with control C57BL/6 mice (serum creatinine: 0.7 +/- 0.4 vs. 2.5 +/- 0.3 mg/dl, respectively; P < 0.05). Neutrophil infiltration into postischemic kidney was reduced in CD4/CD8 KO mice, compared with control mice, at both 24 h [polymorphonuclear neutrophils (PMNs)/10 high power fields: 714 +/- 354 vs. 3,514 +/- 660, respectively; P < 0.05] and 48 h (88 +/- 32 vs. 1,979 +/- 209, respectively; P < 0.05). Tubular necrosis score in CD4/CD8 KO mice, compared with control mice, was significantly less at 48 h (0.4 +/- 0.1 vs. 2.4 +/- 0.2, respectively; P < 0.05). Because adhesion between T cells and renal tubular epithelial cells (RTECs) may underlie the pathophysiological role of T cells in renal IRI, we also measured T cell adhesion to primary murine RTECs in vitro. Exposure of RTECs to 2 h of hypoxia followed by 1 h of reoxygenation increased T cell adhesion more than twofold. Phorbol ester treatment, which activates integrins, increased T cell adhesion threefold. These data suggest that T lymphocytes can mediate experimental renal IRI. Moreover, adhesion of infiltrating T cells to renal tubular cells may provide a potential mechanism underlying postischemic tubular dysfunction.

Acute Kidney Injury↗
PubMed2000

Reconstructive ultrasound elasticity imaging for renal transplant diagnosis: kidney ex vivo results.

It may be possible to diagnose and monitor scarring, inflammation and edema in transplant kidney using reconstructive ultrasound elasticity imaging. Kidney elasticity is expected to change dramatically with scar, and to a lesser degree, with acute inflammation and edema. The hypothesis that changes in kidney elasticity can be imaged using a clinical ultrasound scanner was experimentally tested with an ex vivo canine kidney model, and results on a single pair of kidneys are reported in this paper. A cross-linking agent affected kidney elasticity both globally and locally. Elasticity changes were monitored with accurate estimates of internal displacement and strain followed by Young's modulus reconstruction. The results of this study strongly suggest that ultrasound elasticity imaging can detect elasticity changes in complex structures such as the kidney. Moreover, it has the potential to become an important clinical tool for renal transplant diagnosis.

Animals↗
PubMed2000

A dynamic focusing technique for delta-sigma-based beamformers.

Beamformation using oversampling delta-sigma (deltasigma) modulators has been proposed for diagnostic ultrasound. Such a beamformer can reduce the size, complexity and cost of an imaging system while providing adequate signal-to-quantization noise ratio (SQNR). High quality images can also be generated if dynamic receive delays are applied correctly. Several dynamic focusing techniques were previously proposed. Generally, an additional bit or extra compensation circuit is required to preserve the power and frequency distribution of the signal. Without preserving the power and frequency distribution, the image background noise is increased. In this paper, an alternative technique is presented. The new technique exploits the symmetry of focusing delays relative to the center of a transducer array. By properly synchronizing the delays and selecting the inserted values, no noise is added to the beam sum signal and the image background level is not increased. Using real ultrasound data, it is shown that the proposed technique provides the same imaging performance as the previous approaches with reduced system complexity by using only a single bit to encode the output of the deltasigma modulator.

Electronics, Medical↗
PubMed2000

Correlation-based correction of sound velocity inhomogeneities using delta-sigma modulators.

Delta-sigma (deltasigma) modulator based beamformers have been proposed for high-quality ultrasonic imaging. Due to the high sampling rate and single bit data width, the cost and complexity of the receive beamformer can be significantly reduced. It was shown that with proper dynamic focusing controls, equivalent image quality could be achieved with an adequate signal-to-quantization noise ratio (SQNR). In this paper, deltasigma modulator based beamformers are used for correlation-based phase aberrations correction of sound velocity inhomogeneities in the body. It is shown that the correction can be efficiently implemented at a performance level similar to that of a conventional radio frequency (rf) beamformer. In addition, more than 6 dB contrast improvement is demonstrated. The different dynamic focusing techniques are also investigated in the context of phase aberration correction. It is shown that the single bit dynamic focusing approach does not affect the overall performance of phase aberration correction.

Electronics, Medical↗
PubMed2000

Detection of cytomegalovirus (CMV) antigens in kidney biopsies and transplant nephrectomies as a marker for renal graft dysfunction.

Chronic rejection accounts for the greatest loss of renal allografts. HLA mismatching has been minimised by organ allocation and new immunosuppressive drugs have been employed, but the average cadaveric graft survival still does not exceed 12 years. Though the aetiology is multifactorial, one contributory factor for this condition is cytomegalovirus (CMV). Detection of CMV in kidney biopsies and sera can diagnose and monitor this inflammatory event and define its role in chronic nephropathy. Twenty five biopsies taken at the time of transplantation, 10 biopsies for graft dysfunction and tissue blocks from 20 explanted kidney grafts were collected and investigated for CMV antigens by immunohistochemistry. Tissue samples were snap frozen and cryostat sections were incubated with monoclonal antibodies for CMV antigens followed by immunoperoxidase staining. In 12 out of 20 transplant nephrectomies CMV antigens were found. Only two of these patients had clinical CMV disease. Time 0 biopsies from CMV seronegative donors (n = 11) and CMV seropositive donors (n = 14) were negative for CMV antigens. The prevalence of CMV antigens in grafts lost due to chronic rejection was 60%. These antigens were not found within the time 0 biopsies, but were detected in 30% of biopsies taken at the time of clinical graft dysfunction. CMV appears to contribute to chronic rejection even without clinical disease.

Antigens, Viral↗
PubMed2000

Effect of a boil water notice on behaviour in the management of a water contamination incident.

In late 1998, the water supply of 878 households was affected by possible sewage contamination. A notice was issued to advise residents of the need to boil their water. This provided an opportunity to assess to what extent the boil water notice led people to avoid activities that might put them at risk of waterborne infection. A postal questionnaire sent to 350 randomly selected households in the affected area asked about timing and mode of receipt of the notice, risk behaviour (boiling water, brushing teeth, washing dishes, drinks for pets, preparation of food), and subsequent changes in drinking water consumption habits. Eighty-one per cent of the households surveyed engaged in behaviour likely to increase the risk of waterborne infection. Comments were collected from consumers on how to improve the management of future water contamination incidents.

Health Behavior↗
PubMed2000

Acceptability of health status questionnaire screening and vaccination among Kosovar refugees.

During 1999, following the war in Kosovo, thousands of Kosovar refugees were received by different countries. The public health reception protocols for these refugees varied widely between and within reception countries. We surveyed a group of Kosovar refugees received in Bolton to assess the acceptability of vaccination and health status questionnaire screening. We also examined the refugees' opinions about the confidentiality of their medical history and their willingness to take part in research. Fifteen out of 29 people responded. Most respondents were happy with the interventions.

Adult↗
PubMed1999

Multiple competition reactions for RPA order the assembly of the DNA polymerase delta holoenzyme.

Processive extension of DNA in eukaryotes requires three factors to coordinate their actions. First, DNA polymerase alpha-primase synthesizes the primed site. Then replication factor C loads a proliferating cell nuclear antigen (PCNA) clamp onto the primer. Following this, DNA polymerase delta assembles with PCNA for processive extension. This report shows that these proteins each bind the primed site tightly and trade places in a highly coordinated fashion such that the primer terminus is never left free of protein. Replication protein A (RPA), the single-stranded DNA-binding protein, forms a common touchpoint for each of these proteins and they compete with one another for it. Thus these protein exchanges are driven by competition-based protein switches in which two proteins vie for contact with RPA.

Binding, Competitive↗
PubMed1999

Division of labor--sequential ATP hydrolysis drives assembly of a DNA polymerase sliding clamp around DNA.

The beta sliding clamp encircles DNA and enables processive replication of the Escherichia coli genome by DNA polymerase III holoenzyme. The clamp loader, gamma complex, assembles beta around DNA in an ATP-fueled reaction. Previous studies have shown that gamma complex opens the beta ring and also interacts with DNA on binding ATP. Here, a rapid kinetic analysis demonstrates that gamma complex hydrolyzes two ATP molecules sequentially when placing beta around DNA. The first ATP is hydrolyzed fast, at 25-30 s(-1), while the second ATP hydrolysis is limited to the steady-state rate of 2 s(-1). This step-wise reaction depends on both primed DNA and beta. DNA alone promotes rapid hydrolysis of two ATP molecules, while beta alone permits hydrolysis of only one ATP. These results suggest that beta inserts a slow step between the two ATP hydrolysis events in clamp assembly, during which the clamp loader may perform work on the clamp. Moreover, one ATP hydrolysis is sufficient for release of beta from the gamma complex. This implies that DNA-dependent hydrolysis of the other ATP is coupled to a separate function, perhaps involving work on DNA. A model is presented in which sequential ATP hydrolysis drives distinct events in the clamp-assembly pathway. We also discuss underlying principles of this step-wise mechanism that may apply to the workings of other ATP-fueled biological machines.

Adenosine Triphosphate↗