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1.
Anal Bioanal Chem ; 405(21): 6891-5, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23881364

ABSTRACT

We report a simple, cost-effective, and label-free detection method, consisting of a platelet-derived growth factor (PDGF) binding aptamer and hydrophobic Ru(II) complex as a sensor system for PDGF. The binding of PDGF with the aptamer results in the weakening of the aptamer-Ru(II) complex, monitored by luminescence signal. A substantial enhancement in the luminescence intensity of Ru(II) complex is observed in the presence of aptamer due to the hydrophobic interaction. Upon addition of PDGF, the luminescence intensity is decreased, due to the stronger interaction between the aptamer and PDGF resulting in the displacement of Ru(II) complex to the aqueous solution. Our assay can detect a target specifically in a complex medium such as the mixture of proteins, at a concentration of 0.8 pM.


Subject(s)
Aptamers, Nucleotide/chemistry , Luminescent Measurements/methods , Molecular Probe Techniques , Platelet-Derived Growth Factor/analysis , Protein Interaction Mapping/methods , Ruthenium/analysis , Ruthenium/chemistry , Molecular Probes/analysis , Molecular Probes/chemistry , Platelet-Derived Growth Factor/chemistry , Staining and Labeling
2.
Scott Med J ; 57(4): 247, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22917586

ABSTRACT

A 58-year-old white woman presented with sudden onset of diplopia, headache and vomiting with a history of tiredness and lethargy over the past four to six months. She had smooth, pale, hairless skin and on examination she was found to have left-sided third and sixth nerve palsy. Laboratory tests confirmed pan-anterior hypopituitarism. Computerized tomography scan of head and magnetic resonance imaging appearances were consistent with those of a sphenoid sinus mucocoele. Following adequate replacement with hydrocortisone and thyroxine she underwent sphenoid mucocoele drainage and endoscopic left sphenoethmoidectomy. Her symptoms were relieved over the next few days and she had a near-total recovery of ophthalmoplegia over the following three months. Pituitary function tests showed partial resolution of hypopituitarism with recovery of hypothalamic-pituitary-adrenal axis and hydrocortisone therapy was withdrawn, but she continued to require thyroxine.


Subject(s)
Hypopituitarism/etiology , Mucocele/complications , Diplopia/etiology , Female , Headache/etiology , Humans , Hypopituitarism/drug therapy , Middle Aged , Mucocele/diagnosis , Mucocele/surgery , Ophthalmoplegia/etiology , Sphenoid Sinus , Vomiting/etiology
3.
Nucleic Acids Res ; 33(Database issue): D344-7, 2005 Jan 01.
Article in English | MEDLINE | ID: mdl-15608213

ABSTRACT

The Diatom EST database provides integrated access to expressed sequence tag (EST) data from two eukaryotic microalgae of the class Bacillariophyceae, Phaeodactylum tricornutum and Thalassiosira pseudonana. The database currently contains sequences of close to 30,000 ESTs organized into PtDB, the P.tricornutum EST database, and TpDB, the T.pseudonana EST database. The EST sequences were clustered and assembled into a non-redundant set for each organism, and these non-redundant sequences were then subjected to automated annotation using similarity searches against protein and domain databases. EST sequences, clusters of contiguous sequences, their annotation and analysis with reference to the publicly available databases, and a codon usage table derived from a subset of sequences from PtDB and TpDB can all be accessed in the Diatom EST Database. The underlying RDBMS enables queries over the raw and annotated EST data and retrieval of information through a user-friendly web interface, with options to perform keyword and BLAST searches. The EST data can also be retrieved based on Pfam domains, Cluster of Orthologous Groups (COG) and Gene Ontologies (GO) assigned to them by similarity searches. The Database is available at http://avesthagen.sznbowler.com.


Subject(s)
Databases, Nucleic Acid , Diatoms/genetics , Expressed Sequence Tags/chemistry , DNA, Algal/chemistry , Sequence Analysis, DNA , Systems Integration
4.
Cornea ; 20(6): 597-602, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11473159

ABSTRACT

PURPOSE: To report the acute effects of laser in situ keratomileusis (LASIK) on the corneal endothelium. METHODS: Twenty eyes of 10 consecutive patients (mean age, 38.1 +/- 10.84 years) underwent bilateral simultaneous LASIK for myopic astigmatism (spherical equivalent ranging from -1.75 to -7.13 diopters) without any complications. Each eye was evaluated by slit-lamp biomicroscopy and noncontact specular microscopy preoperatively, within 15 minutes after LASIK and 1 day after surgery. Specular microscopy images were then analyzed to calculate endothelial cell density (ECD), coefficient of variation (CV) of cell size, and percentage of hexagonal cells. RESULTS: All corneas demonstrated marked alterations in endothelial cell morphology by slit-lamp biomicroscopy within 15 minutes after surgery that resolved by the first postoperative day. Central corneal endothelial analysis by noncontact specular microscopy confirmed pleomorphism with definite loss of hexagonality. Mean ECD was calculated to be 2,816.3 +/- 286.02 cells/mm(2) preoperatively, 2,750.85 +/- 327.95 cells/mm(2) on day 0 (p = 0.395), and 2,810.55 +/- 218.48 cells/mm(2) on day 1 (p = 0.461). Mean CV was 32.65 +/- 7.29 preoperatively, 34.4 +/- 6.19 on day 0 (p = 0.412), and 30.9 +/- 5.54 on day 1 (p = 0.067). Mean percentage of hexagonal cells was 63.35 +/- 10.76 preoperatively, 47.55 +/- 9.69 on day 0 (p = 0.000009), and 60 +/- 9.3 on day 1 (p = 0.00003). CONCLUSION: Qualitative and quantitative changes in endothelial cell morphology (i.e., decreased endothelial cell hexagonality) demonstrate that LASIK does induce an acute effect on the corneal endothelium that may represent transient endothelial cell edema.


Subject(s)
Astigmatism/surgery , Endothelium, Corneal/pathology , Keratomileusis, Laser In Situ/adverse effects , Myopia/surgery , Postoperative Complications/pathology , Acute Disease , Adult , Astigmatism/pathology , Cell Count , Corneal Edema/etiology , Corneal Edema/pathology , Female , Humans , Male , Middle Aged , Myopia/pathology , Prospective Studies
5.
Ophthalmology ; 108(7): 1203-8, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11425676

ABSTRACT

OBJECTIVE: To document the corneal astigmatism that occurs with macular translocation after scleral infolding surgery. DESIGN: Retrospective case series of a nonrandomized clinical trial. PARTICIPANTS: Eight consecutive age-related macular degeneration patients (eight eyes) with choroidal neovascularization who underwent macular translocation with scleral infolding at the Duke University Eye Center from December 1998 through October 1999. METHODS: We retrospectively reviewed the charts of eight consecutive patients who underwent macular translocation surgery involving scleral infolding in the superotemporal quadrant. Two patients subsequently underwent release of scleral infolding. MAIN OUTCOME MEASURES: After surgery, these eyes were evaluated for corneal astigmatism with manifest refraction, keratometry, and computerized corneal topography. RESULTS: All eight eyes of eight patients revealed marked degrees of corneal astigmatism. Measurement of astigmatism via manifest refraction, keratometry, and corneal topography confirmed postoperative astigmatism corresponding to the axis of the scleral infolding. The amount of corneal astigmatism ranged from 1.75 to 7.37 diopters (D; mean, 4.60 D), with steepening along the axis of scleral infolding in the superotemporal quadrant of each eye (mean, 42.50 degrees from vertical; range, 24 degrees -66 degrees from vertical). Release of scleral infolding in two patients resulted in significant reduction of corneal astigmatism. CONCLUSIONS: Scleral shortening procedures used in macular translocation surgery may induce large amounts of corneal astigmatism. These patients should be assessed with keratometry and corneal topography to determine the accurate amount and axis. Thereafter, contact lens fitting or scleral infolding release may be considered as therapeutic options for large amounts of astigmatism persisting after surgery.


Subject(s)
Astigmatism/etiology , Choroidal Neovascularization/surgery , Corneal Diseases/etiology , Macula Lutea/transplantation , Macular Degeneration/surgery , Postoperative Complications , Sclera/surgery , Astigmatism/physiopathology , Corneal Diseases/physiopathology , Corneal Topography , Humans , Refraction, Ocular , Retrospective Studies , Transplantation, Autologous/adverse effects , Visual Acuity
6.
Am J Respir Crit Care Med ; 162(2 Pt 1): 617-22, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10934096

ABSTRACT

Alveolar surfactant is known to exist in several morphologic forms or subtypes which have been separated from bronchoalveolar lavage fluid (BAL) by two types of methods-differential centrifugation (DC) and equilibrium buoyant density gradient centrifugation (EBDC). DC separates BAL into large aggregates (LA) and small aggregates (SA); EBDC separates BAL into three peaks called ultraheavy (UH), heavy (H), and light (L). We compared these two separation methods by subjecting replicates of the same pools of BALF from groups of mice to DC and EBDC in parallel assays. We found that each method was highly internally consistent, but that the amount of phospholipid in the LA fraction of DC was consistently and substantially less (by 33 to 43%) than that found in the UH + H fractions of EBDC. This appeared to be due to failure of DC to sediment all of the phospholipid that banded as UH or H in EBDC despite adjustments in the time and g-force of DC. In experiments where differentially labeled purified H and L subtypes were subjected to DC over a wide range of g-force and time conditions, cross-contamination of the DC pellet and supernatant with heterologous subtypes was always present (4 to 33% cross-contamination). Addition of extraneous serum proteins to the BAL, as a model of lung damage, resulted in further inconsistencies in DC but not EBDC. Investigators may wish to bear these considerations in mind when planning or interpreting the results of experiments bearing on surfactant subtype analysis.


Subject(s)
Pulmonary Surfactants/chemistry , Animals , Blood Proteins/analysis , Bronchoalveolar Lavage Fluid/chemistry , Centrifugation/methods , Female , Mice , Phospholipids/analysis
7.
Am J Physiol Lung Cell Mol Physiol ; 278(1): L19-24, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10645886

ABSTRACT

Convertase has homology with carboxylesterases, but its substrate(s) is not known. Accordingly, we determined whether dipalmitoylphosphatidylcholine (DPPC), the major phospholipid in surfactant, was a substrate for convertase. We measured [(3)H]choline release during cycling of the heavy subtype containing [(3)H]choline-labeled DPPC with convertase, phospholipases A(2), B, C, and D, liver esterase, and elastase. Cycling with liver esterase or peanut or cabbage phospholipase D produced the characteristic profile of heavy and light peaks observed on cycling with convertase. In contrast, phospholipases A(2), B, and C and yeast phospholipase D produced a broad band of radioactivity across the gradient without distinct peaks. [(3)H]choline was released when natural surfactant containing [(3)H]choline-labeled DPPC was cycled with yeast phospholipase D but not with convertase or peanut and cabbage phospholipases D. Similarly, yeast phospholipase D hydrolyzed [(3)H]choline from [(3)H]choline-labeled DPPC after incubation in vitro, whereas convertase, liver esterase, or peanut and cabbage phospholipases D did not. Thus convertase, liver esterase, and plant phospholipases D did not hydrolyze choline from DPPC either on cycling or during incubation with enzyme in vitro. In conclusion, conversion of heavy to light subtype of surfactant by convertase may require a phospholipase D type hydrolysis of phospholipids, but the substrate in this reaction is not DPPC.


Subject(s)
1,2-Dipalmitoylphosphatidylcholine/metabolism , Carboxylic Ester Hydrolases/biosynthesis , Animals , Choline/metabolism , Esterases/metabolism , Liver/enzymology , Mice , Phospholipase D/metabolism , Pulmonary Surfactants/metabolism , Rats , Substrate Specificity
8.
Lung ; 177(3): 127-38, 1999.
Article in English | MEDLINE | ID: mdl-10192760

ABSTRACT

The influence of phospholipids on the ultrastructure and metabolism of reconstituted surfactants has not been well defined. The aim of this study was to determine if changes in the phospholipid composition of reconstituted surfactants altered their biophysical properties, ultrastructure, and conversion to light subtype by cycling. We prepared various surfactants containing radiolabeled dipalmitoylphosphatidylcholine ([14C]DPPC). The addition of phosphatidylglycerol (PG) or dipalmitoylphosphatidic acid (PA) to DPPC increased conversion to light subtype. In contrast, the addition of dipalmitoylphosphatidylglycerol (DPPG) to DPPC markedly reduced conversion to light subtype on cycling. DPPC and DPPC+PG produced large liposomes ( approximately 1,000 nm), whereas DPPC+PA or DPPC+DPPG formed multilamellar membranes. Mixtures of DPPC and PA were highly surface active in vitro, whereas the surface activity of DPPC+DPPG was similar to that of DPPC. In conclusion, the ultrastructure, metabolism, and surface active properties of DPPC+PG mixtures were influenced markedly by alterations in the fatty acid composition or polar head group of PG.


Subject(s)
1,2-Dipalmitoylphosphatidylcholine/analysis , Phospholipids/analysis , Pulmonary Surfactants/pharmacology , 1,2-Dipalmitoylphosphatidylcholine/pharmacology , Carbon Radioisotopes , Humans , Microscopy, Electron , Phosphatidylglycerols , Pulmonary Surfactants/chemistry
9.
Am J Physiol ; 275(5): L969-75, 1998 11.
Article in English | MEDLINE | ID: mdl-9815115

ABSTRACT

We recently reported the purification and partial amino acid sequence of "surfactant convertase," a 72-kDa glycoprotein involved in the extracellular metabolism of lung surfactant (S. Krishnasamy, N. J. Gross, A. L. Teng, R. M. Schultz, and R. Dhand. Biochem. Biophys. Res. Commun. 235: 180-184, 1997). We report here the isolation of a cDNA clone encoding putative convertase from a mouse lung cDNA library. The cDNA spans a 1,836-bp sequence, with an open reading frame encoding 536 amino acid residues in the mature protein and an 18-amino acid signal peptide at the NH2 terminus. The deduced amino acid sequence matches the four partial amino acid sequences (68 residues) that were previously obtained from the purified protein. The deduced amino acid sequence contains an 18-amino acid residue signal peptide, a serine active site consensus sequence, a histidine consensus sequence, five potential N-linked glycosylation sites, and a COOH-terminal secretory-type sequence His-Thr-Glu-His-Lys. Primer-extension analysis revealed that transcription starts 29 nucleotides upstream from the start codon. Northern blot analysis of RNA isolated from various mouse organs showed that convertase is expressed in lung, kidney, and liver as a 1,800-nucleotide-long transcript. The nucleotide and amino acid sequences of putative convertase are 98% homologous with mouse liver carboxylesterase. It thus may be the first member of the carboxylesterase family (EC 3.1.1.1) to be expressed in lung parenchyma and the first with a known physiological function.


Subject(s)
Gene Expression Regulation, Enzymologic , Lung/enzymology , Serine Endopeptidases/genetics , Amino Acid Sequence , Animals , Base Sequence , Carboxylic Ester Hydrolases/chemistry , Cloning, Molecular , Female , Gene Library , Humans , Male , Mice , Molecular Sequence Data , Organ Specificity , Polymerase Chain Reaction , Rats , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Sequence Alignment , Sequence Homology, Amino Acid , Serine Endopeptidases/biosynthesis , Serine Endopeptidases/chemistry
10.
Biochem Biophys Res Commun ; 244(3): 712-9, 1998 Mar 27.
Article in English | MEDLINE | ID: mdl-9535730

ABSTRACT

Surfactant convertase is required for conversion of heavy density (H) natural surfactant to light density (L) subtype during cycling in vitro, a technique that reproduces surfactant metabolism. To study mechanisms of H to L conversion, we prepared liposomes of dipalmitoylphosphatidylcholine (DPPC) and phosphatidylglycerol (PG), or the phospholipids (PL) in combination with either surfactant protein A (SP-A), surfactant protein B (SP-B), or both SP-A and SP-B. Phospholipids alone showed time-dependent conversion from heavy to light subtype on cycling in the absence of convertase, which was decreased by adding SP-B, but not SP-A, to phospholipids (p < 0.01 for PL+SP-B, or PL+SP-A+SP-B vs. PL, or PL+SP-A). The ultrastructure, surface activity, buoyant density, and L subtype generation on cycling PL+SP-A+SP-B with partially purified convertase or with phospholipase D were similar to those of natural TM. In conclusion, a reconstituted surfactant mimics the behavior of natural surfactant on cycling, and reveals that interaction of SP-B with phospholipids decreases L subtype generation. In addition, esterase/ phospholipase D activity is required for conversion of heavy to light subtype on cycling.


Subject(s)
Biological Products , Proteins , Pulmonary Surfactants/metabolism , Animals , Female , Mice , Myelin Sheath , Phospholipase D/metabolism , Phospholipids/metabolism , Proteolipids/metabolism , Proteolipids/ultrastructure , Pulmonary Surfactant-Associated Protein A , Pulmonary Surfactant-Associated Proteins , Pulmonary Surfactants/classification , Pulmonary Surfactants/ultrastructure , Serine Endopeptidases/metabolism , Specific Gravity
11.
Neurosurgery ; 41(5): 1028-36; discussion 1036-8, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9361056

ABSTRACT

OBJECTIVE: This study used quantitative radiological imaging to determine the effect of surgical resection on postoperative survival of patients with malignant astrocytomas. Previous studies relied on the surgeons' impressions of the amount of tumor removed, which is a less reliable measure of the extent of resection. METHODS: Information concerning possible prognostic factors was collected for 75 patients undergoing magnetic resonance imaging or computed tomography preoperatively and within 10 days postoperatively. Image analysis of the neuroradiological studies was conducted to quantify pre- and postoperative total tumor volumes and enhancing volumes. Univariate and multivariate proportional hazards models were used to analyze the regression of survival regarding 22 covariates that might affect survival. The covariates that were entered included age, gender, tumor grade, cumulative radiation dose, chemotherapy, seizures as a first symptom, Karnofsky performance status at presentation, pre- and postoperative total and enhancing tumor volumes, ratio of pre- to postoperative total and enhancing tumor volumes, tumor location, surgeon's impression of the degree of resection, and subsequent surgery. RESULTS: There were 23 patients with anaplastic astrocytomas and 52 with glioblastomas multiforme. The estimated mean survival time was 27 months for patients undergoing gross total resection, 33 months for subtotal resection, and 13 months for open or stereotactic biopsy. Five factors that were significant predictors of survival in multivariate analysis were tumor grade, age, Karnofsky performance status, radiation dose, and postoperative complications (P < 0.05). In univariate analysis, tumor grade, radiation dose, age, Karnofsky status, complications, presence of enhancing tumor in postoperative imaging, and postoperative volume of enhancing tumor were significantly associated with survival (P < 0.05). CONCLUSION: We conclude that the most important prognostic factors affecting survival of patients with anaplastic astrocytomas and glioblastomas multiforme are tumor grade, age, preoperative performance status, and radiation therapy. Postoperative complications adversely affect survival. Aggressive surgical resection did not impart a significant increase in survival time. Surgical resection may improve survival, but its importance is less than that of other factors and may be demonstrable only by larger studies.


Subject(s)
Astrocytoma/surgery , Brain Neoplasms/surgery , Glioblastoma/surgery , Adult , Aged , Aged, 80 and over , Analysis of Variance , Astrocytoma/diagnostic imaging , Astrocytoma/mortality , Astrocytoma/pathology , Brain Neoplasms/diagnostic imaging , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Female , Glioblastoma/diagnostic imaging , Glioblastoma/mortality , Glioblastoma/pathology , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Multivariate Analysis , Postoperative Complications/epidemiology , Prognosis , Proportional Hazards Models , Radiography , Recurrence , Retrospective Studies , Seizures , Survival Rate , Time Factors
12.
Biochem Biophys Res Commun ; 235(1): 180-4, 1997 Jun 09.
Article in English | MEDLINE | ID: mdl-9196059

ABSTRACT

The extracellular conversion of lung surfactant from tubular myelin to the small vesicular form has previously been shown to require a serine-active enzyme called "surfactant convertase." In the present study, a 72kD serine-active enzyme previously identified in mouse lung alveolar lavage and having convertase activity was partially sequenced. Sixty-eight residues obtained from amino acid sequencing of this protein show that it is a new member of the mouse carboxylesterase family (EC 3.1.1.1). The 72kD lung protein also has esterase activity. A commercial esterase of the same family was able to reproduce surfactant convertase bioactivity in vitro, unlike several serine proteinases previously tested. We conclude that surfactant convertase is a carboxylesterase which mediates a biochemical step in the extracellular metabolism of surfactant.


Subject(s)
Carboxylic Ester Hydrolases/chemistry , Lung/enzymology , Pulmonary Surfactants/metabolism , Serine Endopeptidases/chemistry , Amino Acid Sequence , Animals , Carboxylic Ester Hydrolases/isolation & purification , Carboxylic Ester Hydrolases/metabolism , Centrifugation, Density Gradient , Electrophoresis, Polyacrylamide Gel , Enzyme Inhibitors/pharmacology , Female , Isoflurophate/metabolism , Isoflurophate/pharmacology , Liver/enzymology , Lung/metabolism , Mice , Mice, Inbred Strains , Molecular Sequence Data , Sequence Analysis , Serine Endopeptidases/isolation & purification , Serine Endopeptidases/metabolism , Swine
13.
Plant Mol Biol ; 35(4): 471-81, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9349270

ABSTRACT

Glyoxalase II is part of the glutathione-dependent glyoxalase detoxification system. In addition to its role in the detoxification of cytotoxic 2-oxo-aldehydes, specifically methylglyoxal, it has been suggested that the glyoxalase system may also play a role in controlling cell differentiation and proliferation. During the analysis of a T-DNA-tagged mutant of Arabidopsis we identified the gene for a glyoxalase II isozyme (GLY1) that appears to be mitochondrially localized. The cDNA encoding a glyoxalase II cytoplasmic isozyme (GLY2) was also isolated and characterized. Southern blot and sequence analyses indicate that glyoxalase II proteins are encoded by at least two multigene families in Arabidopsis. Escherichia coli cells expressing either GLY1 or GLY2 exhibit increased glyoxalase II activity, confirming that they do, in fact, encode glyoxalase II proteins. Northern analysis shows that the two genes are differentially expressed. Transcripts for the mitochondrial isozyme are most abundant in roots, while those for the cytoplasmic isozyme are highest in flower buds. The identification of glyoxalase II isozymes that are differentially expressed suggests that they may play different roles in the cell.


Subject(s)
Arabidopsis/genetics , Genes, Plant/genetics , Thiolester Hydrolases/genetics , Amino Acid Sequence , Arabidopsis/enzymology , Cloning, Molecular , DNA, Complementary/genetics , DNA, Plant/genetics , Escherichia coli/genetics , Gene Expression Regulation, Plant/physiology , Isoenzymes/genetics , Isoenzymes/metabolism , Mitochondria/enzymology , Molecular Sequence Data , Multigene Family/genetics , RNA, Messenger/analysis , RNA, Plant/analysis , Recombinant Fusion Proteins , Sequence Analysis, DNA , Thiolester Hydrolases/metabolism
14.
Cancer Invest ; 13(5): 453-9, 1995.
Article in English | MEDLINE | ID: mdl-7552809

ABSTRACT

The survival rate for patients with malignant gliomas is poor. We describe the results of a prospective study using concomitant chemoradiotherapy, neutron boost, and adjuvant chemotherapy for patients with malignant gliomas. Forty-two patients with anaplastic astrocytoma (AA) and glioblastoma multiforme (GBM) were treated with postoperative photon radiation 45 Gy/25 fraction (fxs) with concomitant continuous intravenous infusion of 5-fluorouracil at 300 mg/m2/day x 5 days and hydroxyurea 0.5 g orally every 12 hr for 6 days for 5 consecutive weeks, followed by a neutron boost of 450 N cGy/6 fxs delivered twice weekly. Adjuvant chemotherapy with procarbazine, CCNU, and vincristine (PCV) was given up to 1 year or until tumor progression. Thirty-four patients (81%) had GBM and 8 patients (19%) had AA. Sixteen patients (38%) were ineligible for the neutron boost because of large tumors or poor performance status and instead received a photon boost with concomitant chemotherapy for a total dose of 60-65 Gy to the tumor. The overall median survival is 68 weeks at a median follow-up of 203 weeks (range 166-302 weeks for the 11 patients remaining alive); 7/8 patients with AA are alive, 2 of these with progressive disease. For AA the median survival is not reached at a median follow-up of 203 weeks (range 166-302 weeks for the 7 patients alive with AA). Time to tumor progression for the 1 dead patient with AA was 35 weeks and the other 2 patients failed at 171 weeks and 179 weeks following treatment. The median survival for the 34 patients with GBM was 62 weeks; 4/34 patients with GBM are alive at 285, 238, 216, and 206 weeks. Multivariate survival analysis in the 34 patients with GBM revealed age and Karnofsky performance status as important prognostic factors. Extent of surgery and neutrons did not affect survival. Concomitant chemoradiotherapy was well tolerated by all patients. The only toxicities observed were mucositis < or = grade II in 3 patients (7%) and mild myelosuppression in 1 patient (2.4%). Adjuvant PCV was well tolerated. Continuous concomitant chemoradiotherapy was well tolerated by all patients with acceptable side effects. The survival rate for the patients with GBM suggests no significant impact on the prognosis for these patients. Patients with AA did well; however, the patient numbers are small.


Subject(s)
Astrocytoma/therapy , Glioblastoma/therapy , Neutrons/therapeutic use , Adolescent , Adult , Aged , Astrocytoma/mortality , Chemotherapy, Adjuvant , Combined Modality Therapy , Female , Glioblastoma/mortality , Humans , Male , Middle Aged , Prospective Studies , Survival Rate
15.
Plant Mol Biol ; 26(3): 935-46, 1994 Nov.
Article in English | MEDLINE | ID: mdl-8000006

ABSTRACT

The mitochondrial DNA of plants containing the male sterility-causing Ogura cytoplasm of radish contain a novel gene, orf138, that is transcribed as part of a bicistronic mRNA. Genetic studies have previously linked male sterility with the orf138 locus. To determine if orf138 is expressed at the protein level, and investigate the effect of fertility restoration on ORF138 levels, we have raised antibodies to an ORF138-glutathione S-transferase fusion protein. Anti-ORF138 antibodies detect a 20 kDa protein that is associated with the mitochondrial membrane of sterile Ogura radish plants. Nuclear restoration is accompanied by a dramatic reduction in the amount of this protein in mitochondria of flowers and leaves, but not roots of fertile Ogura radish plants. The presence or absence of fertility restoration genes has no detectable effect on the size, abundance, or RNA editing patterns of orf138 transcripts. These results support genetic studies that have implicated orf138 in Ogura cytoplasmic male sterility and suggest that the restorer genes may be affecting either the translation or stability of ORF138.


Subject(s)
Extrachromosomal Inheritance , Genes, Plant/genetics , Mitochondria/genetics , Mitochondrial Proteins , Plant Proteins/genetics , Vegetables/genetics , Amino Acid Sequence , Base Sequence , Fertility/genetics , Mitochondria/metabolism , Molecular Sequence Data , Open Reading Frames/genetics , Plant Proteins/biosynthesis , Plant Proteins/isolation & purification , Plant Roots/metabolism , Plant Shoots/metabolism , RNA Editing , Reproduction/genetics , Sequence Homology, Amino Acid , Tissue Distribution , Transcription, Genetic , Vegetables/physiology
16.
Plant Mol Biol ; 24(1): 129-41, 1994 Jan.
Article in English | MEDLINE | ID: mdl-8111012

ABSTRACT

RNA editing and NH2-terminal processing of subunit 6 (atp6) of the mitochondrial Fo-ATPase complex has been investigated for the normal (fertile) and Ogura (male-sterile) radish cytoplasms to determine if previously identified differences between the Ogura atp6 locus and its normal radish counterpart are associated with cytoplasmic male sterility. Analysis of cDNA clones from five different sterile and fertile radish lines identified one C-to-U transition, which results in the replacement of a proline with a serine, in several of the lines. No editing of atp6 transcripts was observed in two lines, Scarlet Knight (normal radish) and sterile CrGC15 (Ogura radish). This is the first example of a naturally occurring plant mitochondrial gene that is not edited. The Ogura atp6 polypeptide is synthesized with a predicted NH2-terminal extension of 174 amino acids in contrast to the nine amino acid extension found in normal radish. In spite of the lack of similarity between the two extensions, NH2-terminal sequence analysis indicates that both polypeptides are processed to yield identical core proteins with a serine as the NH2-terminal residue. These results indicate that ATPase subunit 6 is synthesized normally in Ogura radish, and that it is unlikely that the atp6 locus is associated with Ogura cytoplasmic male sterility.


Subject(s)
Cytoplasm/enzymology , Proton-Translocating ATPases/genetics , RNA Editing , Sequence Analysis/methods , Vegetables/genetics , Amino Acid Sequence , Base Sequence , DNA , Fertility , Molecular Sequence Data , Protein Biosynthesis , Sequence Homology, Amino Acid
17.
Int J Radiat Oncol Biol Phys ; 27(1): 137-43, 1993 Sep 01.
Article in English | MEDLINE | ID: mdl-8365934

ABSTRACT

PURPOSE: With on-line portal imaging devices and image registration tools, the verification of radiation field position prior to each treatment becomes technically feasible. In this paper, we analyze the impact of pre-treatment verification and field position adjustment on target coverage and normal tissue sparing. METHODS AND MATERIALS: Port films were compared with corresponding simulation films to determine the magnitude of setup variations in patients treated for prostate cancer. From these data, an analytic function was determined between geometric coverage of the target and field margin size. A paradigm for on-line patient repositioning was employed to generate a new relationship between margin and target coverage. Margins were selected for the situations of normal treatment and on-line repositioning to ensure target coverage. Dose-volume histograms were generated for a typical prostate treatment using these margins. RESULTS: On-line repositioning, when setup errors exceed 1 cm, results in a 6 mm reduction in margin, suggesting that 10% of the volume of bladder and rectum may be spared of high dose. CONCLUSION: The use of on-line imaging and image registration to guide adjustment of patient setup may lead to a reduction in the volume of normal tissues irradiated, and possibly improve the probability of complication-free survival in future treatments.


Subject(s)
Posture , Prostatic Neoplasms/radiotherapy , Radiotherapy, Computer-Assisted/methods , Computer Simulation , Humans , Male , Prostatic Neoplasms/diagnostic imaging , Radiography
18.
Ann Oncol ; 4(7): 591-5, 1993 Aug.
Article in English | MEDLINE | ID: mdl-8395874

ABSTRACT

BACKGROUND: Iododeoxyuridine (IUdR) is a known radiation enhancer, and interacts biochemically with 5-fluorouracil (5-FU) and hydroxyurea (HU). PATIENTS AND METHODS: IUdR was added to the previously studied regimen of continuous infusion 5-FU at 300 mg/m2/day for 5 days, HU 500 mg every 12 hours for 11 doses and radiotherapy 200 cGy/day for 5 days, all administered for 7 consecutive weeks to patients with malignant glioma. IUdR was administered as 5-day continuous intravenous infusion during weeks 1 and 4. The IUdR dose was changed in cohorts of patients. IUdR plasma concentrations were determined during weeks 1 and 4, and IUdR incorporation into the DNA of granulocytes was measured on weeks 2 and 5. RESULTS: Two patients treated at the initial IUdR dose of 500 mg/m2/day developed grade 3 or 4 myelosuppression and mucositis. Additional dose levels of IUdR tested were 250 mg/m2/day and 125 mg/m2/day; at the latter dose, severe or life-threatening toxicity was seen in only 3 of 8 patients treated. IUdR incorporation into DNA of granulocytes was 10.5(+/- 2.3)% at an IUdR dose of 500 mg/m2/day but decreased to 0.76(+/- 0.3)% at 125 mg/m2/day. Similarly, IUdR plasma concentrations decreased from 436 (+/- 114) ng/ml to 99 (+/- 29) ng/ml. CONCLUSIONS: The addition of IUdR to 5-FU and HU results in significant systemic toxicity necessitating limitation of the IUdR dose to 125 mg/m2/day. There is a significant biochemical interaction between IUdR, 5-FU and HU leading to increased IUdR incorporation into DNA and to substantial clinical toxicity. Further clinical studies to exploit this interaction at more feasible schedules may be useful.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Astrocytoma/drug therapy , Glioblastoma/drug therapy , Idoxuridine/administration & dosage , Adult , Aged , Astrocytoma/metabolism , Astrocytoma/radiotherapy , Chemotherapy, Adjuvant , Drug Administration Schedule , Female , Fluorouracil/administration & dosage , Glioblastoma/metabolism , Glioblastoma/radiotherapy , Humans , Hydroxyurea/administration & dosage , Idoxuridine/metabolism , Male , Middle Aged , Remission Induction
19.
Curr Genet ; 24(1-2): 156-63, 1993.
Article in English | MEDLINE | ID: mdl-8358822

ABSTRACT

The orfB locus of the normal (fertile) and Ogura (male-sterile) radish mitochondrial genomes has been characterized in order to determine if this region, which has previously been correlated with cytoplasmic male sterility (CMS) in Brassica napus cybrids (Bonhomme et al. 1991; Temple et al. 1992), could also be involved in radish CMS. In normal radish, orfB is expressed as a 600-nucleotide (nt) transcript. In Ogura radish, orfB is present as the second gene of a 1200-nt transcript that also contains a 138-codon open reading frame (orf138). Sequences showing similarity to orf138 are present in normal radish, but are not expressed.


Subject(s)
DNA, Mitochondrial/genetics , Plants/genetics , Amino Acid Sequence , Base Sequence , Blotting, Northern , Blotting, Southern , Fertility/genetics , Molecular Sequence Data , Open Reading Frames , Restriction Mapping , Sequence Homology, Amino Acid , Transcription, Genetic
20.
Am J Clin Oncol ; 15(5): 405-11, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1326225

ABSTRACT

Forty-nine consecutive patients with pathologic Stage II non-small-cell lung cancer treated over a 15-year period were retrospectively reviewed. The treatment strategy evolved during the period of review. Early patients were treated with surgery alone (S); subsequent patients were treated with adjuvant radiation therapy (SR); and more recent patients were treated with postoperative chemotherapy and radiation therapy (SRC). Fifteen patients received S alone, 10 patients received SR, and 24 patients received SRC. The median survival time (MST) of all 49 patients was 20 months, and the estimated 5-year survival was 25%. The MST of patients in each of the three treatment arms was S-6 months; SR-19 months; and SRC-25 months. The majority of patients died from systemic relapses or second primary lung cancers. The addition of adjuvant therapy (SR, SRC) significantly improved the MST of patients compared to surgery alone (S). The overall survival of patients did not change between treatment arms.


Subject(s)
Carcinoma, Non-Small-Cell Lung/therapy , Lung Neoplasms/therapy , Adult , Aged , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/pathology , Carcinoma, Non-Small-Cell Lung/radiotherapy , Carcinoma, Non-Small-Cell Lung/surgery , Chemotherapy, Adjuvant , Combined Modality Therapy , Cyclophosphamide/administration & dosage , Doxorubicin/administration & dosage , Female , Humans , Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Lung Neoplasms/radiotherapy , Lung Neoplasms/surgery , Male , Methotrexate/administration & dosage , Middle Aged , Neoplasm Staging , Procarbazine/administration & dosage , Retrospective Studies , Survival Analysis
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