Cloning of a harvest-induced beta-galactosidase from tips of harvested asparagus spears.
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Cloning, Molecular↗
Biomedical subjects
G A King
Publications and source records attributed to G A King.
Radiotherapy for nasolacrimal tract epithelial cancer.
PURPOSE: Tumors of the lacrimal sac are rare and have traditionally been treated surgically. We investigated the use of irradiation for treatment. METHODS AND MATERIALS: Three consecutive patients with primary epithelial cancer of the nasolacrimal apparatus were treated with irradiation. A tumor dose of 52-66 Gy was delivered with conventional fractionation to fields limited to the primary site and immediately surrounding tissues. RESULTS: Local tumor control was achieved in all three patients. Two patients subsequently developed metastatic cervical adenopathy; both were controlled with irradiation to the neck. One of these two died of distant metastases. Two patients are alive and well at 13 years and at 26 months. CONCLUSION: We conclude that epithelial lacrimal sac tumors are controllable by radiation therapy and with a good cosmetic result. Poorly differentiated lesions require elective cervical nodal irradiation.
Carcinoma↗
Parenting behavior rating scales: preliminary validation with intrusive, abusive mothers.
This study provided preliminary evidence of the reliability and validity of a set of scales fro rating the parenting behaviors of physically abusive mothers. Thirty-two male and 32 female undergraduates viewed a video-tape of either an abusive or matched control mother interacting with her son. Two tapes of abusive mothers, representative of the Intrusive mother type, and two matched control tapes were used. Subjects rated the mother's behavior on rating scales matching an observational coding system developed by Oldershaw, Walters, and Hall (1989). Mean ratings of between six to eight raters displayed sufficiently high reliability. As expected, ratings correlated with behavior frequency data. Observers' ratings discriminated between abusive mothers and nonabusive controls, providing evidence of construct validity of the scales. The potential utility of a rating scale instrument for the clinical assessment of parenting behaviors is discussed.
Adult↗
Stereotactic linear radiosurgery for cavernous angiomas.
Optimal management of symptomatic cavernous angiomas (CA) located in the thalamus and the brainstem is problematic. Clinical and radiological (MRI) follow-up series suggest that having hemorrhaged once, recurrent hemorrhage with progressive neurologic dysfunction may commonly occur. We have therefore chosen to treat these lesions when first symptomatic with stereotactic linear radiosurgery (SLR). We now report, after a median follow-up of 27 months, 12 patients with CAs (9 women, 3 men, mean age 40 years) treated in this fashion. Ten patients presented with hemorrhage (3 had more than one hemorrhage): two patients had new onset seizures. All patients had enhanced MRI/MRAs characteristic of CA. There were five brainstem and five thalamic CAs, and one each in the temporal lobe and insula. Cerebral angiograms were done in 8 patients for comparison with their respective MRAs. Only one CA was visualized in the late venous phase on cerebral angiogram and identical vascular features were appreciated on the MRA. The diameter of the CAs ranged from 1.0 to 3.0 cm with a mean of 1.6 cm. Dosimetry planning was based on MRI/CT features and the mean dose at the isocenter was 2.167 cGy (range = 2,000-2,500 cGy) delivered with a mean collimation diameter of 1.46 cm (range = 1.0-2.0 cm). All 12 patients continued to improve neurologically after SLR and had MRI-documented changes in their lesions: in general, the lesions became smaller and signal characteristics converted to methemaglobin. However, 1 patient had an early post-SLR hemorrhage, documented by MRI, at 4.5 months.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
Protection of rabbit lungs from endotoxin injury by in vivo hyperexpression of the prostaglandin G/H synthase gene.
A recombinant prostaglandin G/H (PGH) synthase gene has been expressed in vitro in bovine pulmonary artery endothelial cells and in vivo in rabbits by transfection with a plasmid using cationic liposomes. Transfection of bovine pulmonary artery endothelial cells with the PGH synthase cDNA resulted in increased intracellular PGH synthase protein (determined by Western blot analysis) and increased release of prostacyclin. Rabbits intravenously transfected with the PGH synthase gene had increased plasma levels of prostacyclin and PGE2, and their lungs produced increased amounts of the same eicosanoids. In an in situ, perfused preparation of PGH synthase transfected rabbit lungs, the pressor response to endotoxin was markedly attenuated. In addition, pulmonary edema and release of thromboxane B2 into the perfusate after endotoxin infusion were markedly decreased in transfected lungs compared to controls (animals transfected with a pCMV4 construct that did not contain a cDNA insert). The data suggest that augmented endogenous production of prostacyclin and PGE2, achieved by liposome-mediated gene transfer, protects the lungs from endotoxin. This may be caused in part by suppression of endotoxin-stimulated thromboxane B2 production. Modification of lipid mediator responses by in vivo transfection is a potential approach to the therapy of acute lung injury.
Animals↗
Induction and repair of DNA double-strand breaks in the same dose range as the shoulder of the survival curve.
We have used pulsed-field gel electrophoresis (PFGE) to test two hypotheses that have been proposed to explain the survival curves with shoulders which are characteristic of low-LET ionizing radiation: (1) Neutral elution studies of the induction of double-strand breaks (DSBs) have suggested that ionizing radiation might induce DSBs in a nonlinear fashion at low doses. (2) Based on analogies to enzyme kinetics, DSB repair might be saturating in the shoulder region. We quantified DSB induction and survival resulting from doses between 0 and 5 Gy spanning the shoulder region of the survival curve. We found that DSB induction was linear at all doses tested down to 0.5 Gy, the limits of sensitivity. Therefore, nonlinear DSB induction cannot account for the shape of the survival curve. To determine whether the DSB repair system was saturated in the shoulder region, we quantified the rate of DSB repair as a function of dose of X rays between 1.25 and 20 Gy. The repair of DSBs was exponential with half-times of repair constant for doses below 10 Gy, and averaged 28 min. We determined the initial rate of repair from the exponential repair kinetics for each dose. The initial rate of repair after radiation treatment increased linearly with dose up to at least 10 Gy. Therefore, saturating DSB repair cannot explain the shoulder of the survival curve.
Cell Survival↗
Isolation and characterization of a cDNA clone for a harvest-induced asparagine synthetase from Asparagus officinalis L.
A full-length cDNA clone (pTIP27) encoding asparagine synthetase (AS; EC 6.3.5.4) was isolated from a cDNA library prepared from the tip section (apex to 30 mm) of Asparagus officinalis L. spears. The cDNA clone encodes an mRNA of 1978 bp, giving a derived protein of 66.5 kD molecular mass. The derived amino acid sequence is 81% homologous to AS from Pisum sativum. Only low levels of transcript for AS could be detected in growing spears, roots, or ferns. However, AS mRNA levels began to increase in the tips of harvested spears after 2 h at 20 degrees C, and in the other sections of the spear after 4 h, suggesting that all sections of the spear were responding to the same postharvest signal. The results are discussed in relation to metabolic changes occurring in harvested spears.
Amino Acid Sequence↗
An imbalance between the excretion of thromboxane and prostacyclin metabolites in pulmonary hypertension.
BACKGROUND: Constriction of small pulmonary arteries and arterioles and focal vascular injury are features of pulmonary hypertension. Because thromboxane A2 is both a vasoconstrictor and a potent stimulus for platelet aggregation, it may be an important mediator of pulmonary hypertension. Its effects are antagonized by prostacyclin, which is released by vascular endothelial cells. We tested the hypothesis that there may be an imbalance between the release of thromboxane A2 and prostacyclin in pulmonary hypertension, reflecting platelet activation and an abnormal response of the pulmonary vascular endothelium. METHODS: We used radioimmunoassays to measure the 24-hour urinary excretion of two stable metabolites of thromboxane A2 and a metabolite of prostacyclin in 20 patients with primary pulmonary hypertension, 14 with secondary pulmonary hypertension, 9 with severe chronic obstructive pulmonary disease (COPD) but no clinical evidence of pulmonary hypertension, and 23 normal controls. RESULTS: The 24-hour excretion of 11-dehydro-thromboxane B2 (a stable metabolite of thromboxane A2) was increased in patients with primary pulmonary hypertension and patients with secondary pulmonary hypertension, as compared with normal controls (3224 +/- 482, 5392 +/- 1640, and 1145 +/- 221 pg per milligram of creatinine, respectively; P less than 0.05), whereas the 24-hour excretion of 2,3-dinor-6-keto-prostaglandin F1 alpha (a stable metabolite of prostacyclin) was decreased (369 +/- 106, 304 +/- 76, and 644 +/- 124 pg per milligram of creatinine, respectively; P less than 0.05). The rate of excretion of all metabolites in the patients with COPD but no clinical evidence of pulmonary hypertension was similar to that in the normal controls. CONCLUSIONS: An increase in the release of the vasoconstrictor thromboxane A2, suggesting the activation of platelets, occurs in both the primary and secondary forms of pulmonary hypertension. By contrast, the release of prostacyclin is depressed in these patients. Whether the imbalance in the release of these mediators is a cause or a result of pulmonary hypertension is unknown, but it may play a part in the development and maintenance of both forms of the disorder.
6-Ketoprostaglandin F1 alpha↗
Recombinant protein production in insect cell cultures infected with a temperature-sensitive baculovirus.
Spodoptera frugiperda (IPLB-SF-21) insect cells were grown in shake-flasks and infected with a temperature-sensitive baculovirus to express the gene of chloramphenicol acetyl transferase (CAT) in serum-free medium (SF-900) and two serum-supplemented media (IPL-41 and Grace's). In temperature-shift experiments (cell growth at 33 degrees C followed by virus replication at 27 degrees C 3-4 days later), virus and CAT production were much poorer in the serum-free medium than in serum-supplemented media, though cell growth was virtually the same in the different media tested. In all the three media, highest virus and CAT titers were obtained at the lowest MOI (multiplicity of infection 0.02). This result is contrary to that obtained in constant-temperature culture (27 degrees C for both cell growth and virus replication). Virus and CAT production was greatly improved when the entire culture was run at constant temperature. It appeared that infected cells were severely damaged at 33 degrees C (6 degrees C above the optimal 27 degrees C), resulting in little or no virus and protein production. As a result of these temperature-shift experiments, a larger-scale (14 1 air-lift bioreactor) serum-free culture of Sf-9 insect cells was conducted at constant temperature (27 degrees C) to produce recombinant protein (beta-galactosidase). A cell density as high as 1 x 10(7) cells.ml-1, and a beta-gal concentration of up to 104,000 unit.ml-1 were achieved.
Animals↗
Effects of thromboxane synthase and cyclooxygenase inhibition on PAF-induced changes in lung function and arachidonic acid metabolism.
PAF was administered as an intravenous bolus (0.1 micrograms/kg) to eight chronically instrumented awake sheep. The effects of pretreatment with an inhibitor of cyclooxygenase (meclofenamate) on PAF-induced changes in lung function were compared to those observed with a specific inhibitor of thromboxane synthase (DP1904). Each animal was studied four times in varied order: PAF alone, PAF + DP1904, PAF + meclofenamate, and DP1904 alone. Saline alone (control), DP1904 alone, and meclofenamate alone did not cause changes in any of the measured variables. DP1904 and meclofenamate significantly attenuated the PAF-induced fall in lung compliance, elevation in peak pulmonary artery pressure, and increased lung lymph flow. Both drugs abolished the PAF-induced increases in lung lymph thromboxane B2 concentrations. Meclofenamate, but not DP1904, blocked the rise in lymph 6-keto-PGF1 alpha. Although meclofenamate blocked the rise in lymph PGE2, DP1904 resulted in levels 2.7 times higher than PAF alone. We conclude that: (1) inhibition of thromboxane synthase is as effective as inhibition of cyclooxygenase in attenuating PAF-induced changes in lung function, and (2) thromboxane synthase inhibition results in augmented production of PGE2 following PAF administration in vivo.
Animals↗
External beam radiotherapy for primary spinal cord tumors.
Of 34 evaluated patients with primary spinal cord tumors, 32 were irradiated at our institutions between 1969 and 1983. The results are reported of 32 patients, 16 with ependymoma and 16 with astrocytoma, who were treated with postoperative external beam radiotherapy following biopsy or subtotal resection. Twenty-nine patients received 45-55 Gy megavoltage beam irradiation in five to six weeks, and the remaining three patients received less than 40 Gy. The minimum follow-up was five years. Five- and ten-year actuarial survival rates for the entire group of patients were 73% (22/30) and 50% (8/16), respectively, including three patients who were salvaged by surgery after radiotherapy failures. Corresponding five- and ten-year relapse-free survival rates were 60% (18/30) and 32% (6/19), respectively. Of the 29 patients who received more than 45 Gy, relapse-free survival at five years was 63% (17/27). Treatment failed in 13 patients, and all of those failures were within the irradiated portals. Patients with ependymomas have significantly better relapse free survival than those with astrocytomas, 80% vs. 40% (p less than 0.05). There was a significant difference in survival between patients with tumors involving the cervical spine and those with tumors in the other locations, 45% vs. 89% (p less than 0.05). There was no significant difference in survival between patients with cauda equina tumors and those with tumors of the spinal cord, 100% vs. 68% (p less than 0.05). No radiotherapy-related neurological deficit was noted with a maximum 20 year follow-up. This study confirms that external beam radiotherapy is a safe and effective treatment modality for primary spinal cord tumors.
Adult↗
The influence of maternal labels on dietary preferences of spiny mouse weanlings.
To assess the influence of conspecific chemical cues on the development of dietary preferences, spiny mouse (Acomys cahirinus) weanling were tested for their responses to food items that were partially eaten (thus, presumably labeled) by various categories of conspecifics. Consumption of a highly preferred novel food (fresh carrot) labeled by the pups' parents, or mother alone, was greater than that of a clean sample of the same food. The presence of labels emanating from the father or an alien mother had no effect on food consumption. Salient maternal labels that elicit preferential feeding by young appear to be individually unique, therefore allowing pups to recognize such cues emanating from their own mother. The attractive substance that mothers transfer to food items while eating (the maternal label per se) has not been identified, but saliva is a likely candidate.
Animals↗
Dosimetry of I-125 seeds implanted on the surface of a cavity.
Dosimetry of a new implant technique to treat brain tumors is presented. High grade gliomas or astrocytomas are surgically removed, and radioactive I-125 seeds are implanted on the surface of the cavity. A computational model is presented to determine the number of seeds and the activity of the seeds for a given dose and cavity size.
Astrocytoma↗
External beam radiotherapy for carcinoma of the prostate.
Five hundred nineteen patients with prostate cancer were seen in the Radiation Oncology Division of the State University of New York (SUNY) Health Science Center, Syracuse, New York, between 1969 and 1981. The results for the 239 patients treated with radical intent are reported here. All patients received 60 to 70 Gy to the prostate with megavoltage beam irradiation; 142 with a small field (10 X 10 cm) 360 degrees rotational technique for Stage A, B, or C disease and 69 with a four-field pelvic brick technique (followed by a boost to the prostate) for Stage A through C and D1 disease. Twenty-eight patients were treated postoperatively for residual disease after radical prostatectomy or for recurrent tumor. The minimum follow-up time was 5 years. Actuarial 5-year and 7-year survival rates for Stage A (n = 34), B (n = 100), C (n = 63), and D1 (n = 14) were 91% and 76%, 86% and 75%, 67% and 40%, and 46% and 36%, respectively. The corresponding 5-year and 7-year relapse-free survival rates were 72% and 65%, 77% and 60%, 46% and 28%, and 38% and 25%. The local tumor control rates at 5 years were 91%, 85%, 77%, and 62% for Stage A, B, C, and D1, respectively. In our experience, there was no significant difference in relapse-free survival rates for patients who underwent transurethral resection (TURP) versus those who did not (67% versus 78% for Stage B [P greater than 0.25] and 38% versus 47% for Stage C [P greater than 0.25], respectively). Also there was no significant difference in relapse-free survival rates between large and small field techniques (64% versus 77% for Stage B [P greater than 0.25] and 56% versus 41% for Stage C [P greater than 0.25], respectively). The 5-year and 7-year actuarial survival rates were 90% and 71%, respectively, for the 15 patients with residual tumor and 58% and 33%, respectively, for the 13 patients treated for postprostatectomy recurrence. Severe complications were documented in only nine patients (3.7%) and mild to moderate complications in 53 patients (22%). Larger fields did not cause a higher rate of complications, although small fields were tolerated better than large fields; the significant acute reaction rate was 27% for large field techniques versus 11% for small field techniques (P greater than 0.01). These results confirm that external beam irradiation is an effective treatment for prostate cancer.
Adult↗
Stage II seminoma: results of postorchiectomy irradiation.
One hundred forty patients with testicular seminoma were treated at the State University of New York Health Science Center at Syracuse between 1966 and 1985. Disease was classified as stage II in 32 patients (23%): stage IIA in 21 patients and stage IIB in 11 patients. All patients underwent irradiation below the diaphragm after a radical orchiectomy, and 28 underwent planned mediastinal and supraclavicular irradiation. The median follow-up was 8 1/2 years; 24 of 32 patients have been followed up for more than 5 years. Twenty-eight patients remain alive and well; four patients died of intercurrent disease. Two patients developed a recurrent seminoma in the mediastinum; a variant lymphangiographic pattern was shown in these patients, and they were cured. A third patient developed a non-seminomatous "recurrence" in the ipsilateral, unirradiated, inguinal nodes and is well after chemotherapy.
Adult↗
The (mis)measurement of restraint: an analysis of conceptual and psychometric issues.
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Diet, Reducing↗
Stage III thymoma: results of postoperative radiation therapy.
The results of postoperative radiation therapy in 12 patients with stage III thymoma treated during 1966-1986 were reviewed. Surgical therapy consisted of total resection in one, subtotal resection in seven, and biopsy only in four. Megavoltage irradiation in the dose range of 3,000-5,600 cGy was employed, with nine patients receiving a dose of at least 5,000 cGy. The local control rate was 67%. The actuarial observed and adjusted 5-year survival rates were 57% and 75%, respectively. These results indicate that postoperative radiation therapy is an effective therapeutic modality in the control of stage III thymoma.
Actuarial Analysis↗
Role of circulating platelets and granulocytes in PAF-induced pulmonary dysfunction in awake sheep.
The effects of a single intravascular bolus injection of platelet-activating factor (PAF) on pulmonary hemodynamics, lung mechanics, and lung fluid and solute exchange were studied in 13 chronically instrumented unanesthetized sheep. Since PAF has profound effects on both platelets and granulocytes, we investigated the effects of platelet and granulocyte depletion on the sheep's response to exogenous PAF. Sheep received PAF when granulocyte and platelets counts were normal and after platelet depletion with rabbit antisheep platelet antibodies (n = 5) or granulocyte depletion with hydroxyurea (n = 5). Sheep served as their own controls, and the order of experimentation was varied. Bolus injections of PAF had reproducible effects on pulmonary hemodynamics (pulmonary arterial pressure increased acutely to 85 +/- 3.7 cmH2O) and lung mechanics (dynamic compliance of the lungs decreased to 24.5 +/- 3.8% of base line and resistance to airflow across the lungs increased greater than 10-fold) and caused marked increases in lung lymph concentrations of thromboxane B2 and 6-ketoprostaglandin F1 alpha. The single bolus injection of PAF also caused marked prolonged elevations in lung lymph flow and increases in the lymph-to-plasma protein concentration ratio for 3 h after PAF. PAF had profound effects despite platelet and granulocyte depletion. Platelet depletion slightly attenuated the pulmonary hypertension observed after PAF injection. Platelet depletion also attenuated the increases in thromboxane B2 concentrations in lung lymph, and lung mechanics normalized more rapidly in platelet-depleted sheep. There were no statistically significant effects of granulocyte depletion to less than 200 granulocytes/mm3 on any of the measured variables.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗