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Biomedical subjects

C Yu

Publications and source records attributed to C Yu.

At least 325 records · Page 18Linked to original sources

Dosimetric evaluation of the conformation of the multileaf collimator to irregularly shaped fields.

PURPOSE: The goal of this study was to evaluate the dosimetric characteristics of geometric MLC prescription strategies and compare them to those of conventional shielding block. METHODS AND MATERIALS: Circular fields, square fields, and 12 irregular fields for patients with cancer of the head and neck, lung, and pelvis were included in this study. All fields were shaped using the MLC and conventional blocks. A geometric criterion was defined as the amount of area discrepancy between the MLC and the prescription outline. The "least area discrepancy" (LAD) of the MLC conformation was searched by selecting the collimator angle, meanwhile keeping a preselected position along the width of the leaf into the prescribed field. Five LAD conventions were studied. These included the LAD-0, LAD-1/3, LAD-1/2, and LAD-2/3 that inserted the leaves at the 0, 1/3, 1/2, and 2/3 of the leaf end into the prescription field, respectively. In addition, the LAD optimization was applied to the transecting (TRN) approach for leaf conformation that prescribed an equal area of overblocking and underblocking under each leaf. Film dosimetry was performed in a 20 cm polystyrene phantom at 10 cm depth 100 cm from source to axis distance (SAD) for both 6 and 18 MV photons with each of the above MLC conformations and conventional blocks. The field penumbra width, defined as the mean of the separation between the 20% and 80% isodose lines along the normal of the prescription field edge, was calculated using both the MLC and conventional block film dosimetry and compared. In a similar way, the d20 is defined as the mean separation between the 20% isodose line and the prescription field edge, and the d80 is defined as the mean separation between the 80% isodose line and the prescription field edge. RESULTS: The field penumbra width for all MLC conventions was approximately 2 mm larger than that of the conventional block. However, there was a larger variation of the separation distribution in the penumbra region of the irregular fields for the MLC, which had a standard deviation of 1 mm (a factor of 5 larger than the conventional block). The dosimetry for the circular fields showed that the LAD-TRN, LAD-1/2, and LAD-1/3 approximated the conventional blocking well in terms of d20 and d80; however, no single convention produced the best conformation for both measures. The dosimetric result of the patient treatment fields was similar for all sites. The LAD-1/3, LAD-1/2, and LAD-TRN strategies conformed to within 1 to 1.5 mm of the d80 of the conventional block for both 6 MV and 18 MV, respectively. The LAD-1/2 and LAD-TRN conformations were virtually identical, although it is proven analytically that the LAD-1/2 convention has the least overall area discrepancy of all conventions. CONCLUSIONS: The five MLC conformation conventions resulted in similar dosimetric penumbrae for all field shapes studied. The LAD-1/3, LAD-TRN, and LAD-1/2 produced the more favorable approximation to conventional block. The field penumbra width, although useful for evaluating irregular field shapes, could not describe the large local variations in the penumbra along the field edge for the MLC. These local variations could be of clinical concern when they appear near vital organs. However, the variation in a local region can potentially be reduced by minimizing the jaggedness of the leaf steps in that local region. The dosimetric results were useful as guidelines for the clinicians in the evaluation and adjustment of MLC leaf positions.

Head and Neck Neoplasms↗

Dual-beam imaging for online verification of radiotherapy field placement.

PURPOSE: Due to the poor quality of megavoltage (MV) radiographs, detection and assessment of discrepancies in radiation field placement are difficult. Furthermore, the high imaging dose required to produce the megavoltage radiograph prohibits frequent image acquisition, particularly for those fields that require the use of an "open-field" exposure. For these small, or conformal, radiation fields, an alternate method of verifying field placement is required if the out-of-field dose is to be minimized. An open-field image acquired with a kilovoltage (kV) source would (a) deliver a very low patient dose, (b) increase the visibility of bony landmarks, and (c) simplify intercomparison of portal and prescription images. This article describes the development of a dual-beam imaging system that produces diagnostic quality "double-exposure" portal images for verifying radiation field placement. METHODS AND MATERIALS: The dual-beam system consists of a kV x-ray tube mounted on the gantry of a medical linear accelerator. The kV beam shares the same isocenter (+/- 1 mm) as the treatment beam but is at 45 degrees to the central axis. Both the kilovoltage and megavoltage images are collected with a fluoroscopic imaging system that uses a low-noise CCD camera to accumulate the light emitted from a phosphor screen. Two 45 degrees mirrors are used to remove the CCD camera from the x-ray beam. The light integration on the CCD array is controlled by a mechanical shutter, allowing easy synchronization with the radiation exposures. The camera is shielded by a lead housing to reduce the number of x-rays reaching the CCD array. A conventional thickness phosphor screen is used for both the kV and MV exposures. In the dual-beam imaging procedure, an open-field kV radiograph is acquired with the patient in treatment position. Immediately following, a MV image is acquired with the beam-defining blocks in position. Summation of the two images produces an online double-exposure image. The anatomical information in either the kV or MV image can be emphasized by weighting the images appropriately. This system was used to acquire MV and kV images of both a contrast-detail phantom and a Rando head phantom. Dual-beam images were also acquired for a pituitary treatment, demonstrating the feasibility and usefulness of the dual-beam technique. RESULTS: Analysis of the contrast-detail images produced with the MV and kV beams shows the expected advantage of using the kV x-ray beam. Images of a Rando head phantom confirm these results. A clinical demonstration of the dual-beam system for verifying the delivery of a pituitary field is shown. The quality of the dual-beam image is similar to the prescription (simulation) image, contains a larger anatomical region, and delivers a lower integral dose to the patient. In addition, the kV beam also enhances the visibility of small markers implanted in the prostate. CONCLUSIONS: A dual-beam imaging system has been developed for the radiographic verification of small, conformal fields. This development demonstrates the advantages and feasibility of using a kV x-ray beam in combination with the treatment beam to improve the accuracy of detecting patient setup errors.

Humans↗

Genetic strategy for analyzing specificity of dimer formation: Escherichia coli cyclic AMP receptor protein mutant altered in its dimerization specificity.

Many transcriptional regulators function in homo- or heterodimeric combinations. The same protein can carry out distinct regulatory functions depending on the partner with which it associates. Here, we describe a mutant of the Escherichia coli cAMP receptor protein (CRP) that has an altered dimerization specificity; that is, mutant/mutant homodimers form preferentially over wild-type/mutant heterodimers. CRP dimerization involves the formation of a parallel coiled-coil structure, and our CRP mutant bears an amino acid substitution affecting the first "d" position residue within the alpha-helix that mediates CRP dimerization. The genetic strategy we used to isolate this CRP altered dimerization specificity (ADS) mutant is generalizable and could be utilized to isolate ADS mutants of other dimeric transcriptional regulators.

Amino Acid Sequence↗

DLA-identical bone marrow grafts after low-dose total body irradiation: effects of high-dose corticosteroids and cyclosporine on engraftment.

Previous studies found that marrow allografts from DLA-identical littermates resulted in survival of 60% of recipient dogs after an otherwise lethal dose of 450 cGy of total body irradiation (TBI), either because of successful allografts or autologous recovery after rejection of the allografts. Forty percent of dogs died with marrow aplasia after allograft rejection. The current study asked whether allogeneic engraftment could be enhanced and survival improved by treating allograft recipients with high doses of corticosteroids or with cyclosporine (CSP), administered either before or after transplantation. Five dogs in group 1 received corticosteroids beginning on day -5 and ending on day 32 after transplant. The starting dose was 12.5 mg of prednisone per kilogram orally twice daily. All five dogs rejected their allografts; three died early with marrow aplasia and two showed endogenous marrow recovery. Nine dogs received CSP from day -6 to day -1 before transplantation at a dose of 20 mg/kg/d intravenously administered in divided doses. All nine dogs rejected the marrow allograft; six died with marrow aplasia and three survived with endogenous marrow recovery. Seven dogs received CSP after transplantation at a dose of 30 mg/kg/d orally from day -1 to day 35. All seven had sustained allografts (two mixed chimeras and five complete donor-type chimeras) and became healthy long-term survivors without graft-versus-host disease. These results extend previous observations and confirm that grafts of marrow from DLA-identical littermates improved survival of dogs exposed to low but otherwise lethal doses of TBI. Additional therapy with high-dose corticosteroids administered peritransplantation and posttransplantation or CSP administered before transplantation neither enhanced the rate of allogeneic engraftment nor improved survival; however, CSP administered after transplantation resulted in successful allografts and event-free survival in all cases.

Adrenal Cortex Hormones↗

Cysteines 638 and 665 in the hormone binding domain of human glucocorticoid receptor define the specificity to glucocorticoids.

To understand the function of cysteines, we have substituted cysteines 638, 643, and 665 by serine in the hormone-binding domain (HBD) of the human glucocorticoid receptor (hGR). In hormone-binding assays using [3H]dexamethasone, hGR C643S and hGR C665S exhibited wild type receptor Kd of 2.5 nM and hGR C665SM666L displayed a Kd of 3.7 nM, while hGR C638S exhibited a Kd of 162 pM, a 15-fold higher affinity. The affinity of hGR C638S for RU486 was 10-fold higher, and the mutants C643S and C665S bound RU486 with a 10-fold lower affinity when compared to wild type GR. While C665S bound aldosterone with very high relative affinity, the double mutant C665SM666L failed to bind aldosterone. The expression of wild type, mutant, and truncated hGRs in vitro showed an identical level of expression of the cloned receptors. Similar levels of expression of the receptors were observed in transfected cells, both by immunoprecipitation and by Western blotting. Transcription activation of the chimeric reporter gene mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) with hGR C638S was 4-fold higher than the level observed with wild type hGR in the presence of dexamethasone. In the presence of RU486, hGR C638S induced MMTV-CAT 25-fold compared to the highest levels observed with wild type hGR and RU486. Even though the hGR C665S stimulated transcription with aldosterone, hGR C665SM666L did not. DNA-receptor interaction analyses by gel mobility shift assay demonstrated that the increased transactivation potential of hGR C638S was due to its intense interaction with DNA. These findings suggest that C638 and C665 are involved in maintaining specificity to glucocorticoids.

Animals↗

Activation of cyclin E-dependent kinase by DNA-damage signals during apoptosis.

Preexposure of HL-60 cells to a DNA-damaging agent, cytosine arabinoside (Ara-C), dramatically induced the levels of H1 kinase activities associated with cyclin E (CycE-H1K) but not cyclin A. This induction was cell cycle-independent and accompanied by loss of cell viability, a late event in apoptosis. When an Ara-C-resistant variant of HL-60 cells were treated with Ara-C at a low concentration, neither CycE-H1K nor apoptosis were observed. Both events were induced in the resistant cells but only after treatment with Ara-C at a much higher concentration for a longer period. The DNA-damage-induced CycE-H1K is proposed to be involved in a late apoptosis checkpoint.

Antimetabolites, Antineoplastic↗

Synthesis and evaluation of terbenzimidazoles as topoisomerase I inhibitors.

The synthesis and pharmacological activity of a series of terbenzimidazoles are described. The ability of these derivatives to induce DNA cleavage in the presence of topoisomerase I was evaluated in vitro. These analogs were also assayed for their cytotoxicity in RPMI 8402 cells and the camptothecin-resistant CPT-K5 cells. In addition the potential for these compounds to serve as substrates for MDR1 was also determined. Several terbenzimidazoles exhibited similar cytotoxicity against variants of human tumor cells that either overexpress MDR1 or are camptothecin-resistant.

Animals↗

Site-directed mutation in conserved anionic regions of guinea pig liver transglutaminase.

Transglutaminases (EC 2.3.2.13) catalyze the formation of epsilon-(gamma-glutamyl) lysine cross-links and the substitution of primary amines for the gamma-carboxamide groups of protein-bound glutamine residues. There are conserved anionic regions in transglutaminases, some of which are thought to be possible calcium-binding sites. By site-directed mutagenesis, three mutant forms of recombinant guinea-pig liver transglutaminase, in which some acidic amino acid residues in two conserved regions became nonionic, were expressed in Escherichia coli: TGM1, with Asp-231 and -232 changed to Asn; TGM2, with Glu-445, -448, -449, -450, and -452 changed to Gln; and TGM3, with the mutations of both TGM1 and TGM2. The size and level of synthesis of the mutant proteins were unchanged when monitored by immunoblotting. All mutants retained enzyme activity, and their apparent Km values for substrates during histamine incorporation into acetyl alpha s1-casein were similar to those of the wild-type enzyme, but their Vmax values were smaller. The deamidation rate of glutamine residues in the acetyl alpha s1-casein was unaffected, but the rate of protein cross-linking catalyzed by these mutants was very low. All mutations caused with the enzyme a decrease in the sensitivity to activation by calcium and an increase in the sensitivity to inhibition by GTP. These results indicated that the negative charges of some acidic amino acid residues in the two conserved anionic regions of transglutaminase are not essential for its activity but the loss of their negative charges affects some catalytic properties.

Amino Acid Sequence↗

Nerve growth factor immunoreactivity and sympathetic sprouting in the rat hippocampal formation.

Several lines of evidence support a role for nerve growth factor (NGF) in the sympathetic sprouting response that occurs following septal cholinergic denervation of the rat hippocampal formation. The present study was undertaken to compare the distribution of NGF-like immunoreactivity and the topography of sympathetic sprouting in rats receiving medial septal lesions. Comparisons were made using adjacent sections of the hippocampal formation stained either for NGF-like immunoreactivity or for NGF receptor-immunoreactivity (p75, to visualize sympathetic fibers). p75-immunoreactive sympathetic axons were localized within the same regions exhibiting NGF-like staining, i.e., the hilus of the dentate gyrus and stratum lucidum in the CA3 area. Furthermore, the sympathetic fibers that invaded the hippocampal formation exhibited NGF-like immunostaining. These results provide additional evidence in support of NGF's role in this collateral sprouting response in the mature rat CNS.

Animals↗

Identification of 'molten globule'-like state in all beta-sheet protein.

The cardiotoxin analogue III (CTX III), isolated from the Taiwan Cobra venom (Naja naja atra), is a sixty amino acid, all beta-sheet protein. The 2,2,2-trifluoro ethanol (TFE) induced unfolding of CTX III is studied under acidic conditions (pH 2.5). Using circular dichroism, 1-anilino-8-napthalene sulphonic acid binding and NMR experiments, it is shown that stable, partially structured state(s) ['molten globule'-like state] is formed between 50 and 80% TFE concentrations. The protein was found to exist in an unfolded state in 80% TFE containing 2M urea. The TFE induced unfolding process is shown to be completely reversible. In the 'molten globule' state of CTX III in 80% TFE, though portion(s) of the backbone of the protein assume helical conformation, most of the original beta-sheet secondary structural elements in the protein are intact. In our opinion, this is the first report of the identification of a 'molten globule'-like state in the unfolding pathway of an all beta-sheet monomeric protein.

Anilino Naphthalenesulfonates↗

In situ detection, by spin trapping, of hydroxyl radical markers produced from ionizing radiation in the tumor of a living mouse.

Hydroxyl radicals are thought to be responsible for the toxicity associated with ionizing radiation in tissues. Measurements of hydroxyl radicals generated by ionizing radiation in cellular systems have failed thus far to elucidate higher-level homeostatic responses to this and other reactive oxygen species. Careful assessment of prior indirect hydroxyl radical assays in living tissues indicates that they are prone to a variety of artifacts, making all but the most qualitative relationships difficult to establish. This paper describes the detection of hydroxyl radicals produced during radiation in the leg tumor of a living mouse, where the free radicals evolve; detection uses low-frequency electron paramagnetic resonance in combination with in vivo spin trapping. To our knowledge, this is the first report of such a direct measurement of free radical production in the tissues of a living animals.

Animals↗

Synthesis and application of hepatitis E virus peptides to diagnosis.

Based on computer analysis of hydrophobicity and prediction of secondary structures for the full-length putative proteins encoded by open reading frame-1 (ORF-1), ORF-2 and ORF-3 of hepatitis E virus (HEV), we selected antigenic regions with hydrophilicity, beta-turn, and beta-sheet, and synthesized 7 peptides of possible epitope-containing regions of the polypeptide encoded by all 3 ORFs of HEV genomic RNA by Merrifield's method of solid-phase synthesis. The synthetic peptides were screened and identified by solid-phase enzyme-linked immunosorbent assay (ELISA). Three of the peptides (EH174 from ORF-1, EH286 from ORF-2 and EH362 from ORF-3) showed antigenic activity and possible application for the development of anti-HEV test kits (the peptide-based ELISA). The laboratory experiments and clinical trials showed that the kits, using a set of 3 synthetic HEV peptides as coating antigens, were of high specificity and exhibited good reproducibility. The small-scale seroepidemiological survey indicated high seroprevalence (14.3%) of anti-HEV in Tibetan populations. Additionally, the results also demonstrated good agreement with clinical findings, suggesting that the test kits will be of major use for immunodiagnosis and seroepidemiological surveys of HEV infection.

Amino Acid Sequence↗

Effects of field size on the survival of pig epidermal colony-forming in situ after electron irradiation.

The survival of colony-forming cells in pig epidermis after irradiation was measured using different electron field sizes. The sensitivity of the colony-forming cells was characterized by D(o) = 2.5-3.0 Gy. There was an effect of field size, described approximately by: Dose (to give five colonies/cm2) (Gy) = (31 +/- 2) x Area-(0.048 +/- 0.015). The effect of field size was less than found previously using tolerance to skin reactions (area exponent = 0.16). This work indicates for the first time that the effects of different large field sizes in skin can be detected at the level of colony-forming cell survival.

Animals↗