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Biomedical subjects

J Nelson

Publications and source records attributed to J Nelson.

At least 19 recordsLinked to original sources
PubMed1992

Progressive disease after high-dose therapy and autologous transplantation for lymphoid malignancy: clinical course and patient follow-up.

Of 364 patients with lymphoid malignancy who underwent high-dose therapy with autologous bone marrow transplantation (ABMT) or peripheral stem cell transplantation (PSCT), 169 patients have had progressive disease after the procedure. The median survival from the time of relapse for patients with Hodgkin's disease (HD) who progressed after the transplant was 10.5 months. This compares with a median survival of 3 months for relapsed non-Hodgkin's lymphoma (NHL) patients (P = .0036). After failing transplantation, 56 patients were treated with further chemotherapy, 35 with involved field irradiation therapy, and 18 patients were treated with combination chemotherapy and irradiation. Seven patients received biologic therapy and seven patients underwent a second bone marrow transplant. The remainder of the patients were believed to be too ill for further therapy or chose not to receive further treatment for their recurrent lymphoid malignancy. Sixty of the 169 patients with progressive disease after the transplant are still alive; however, only 18 patients are alive off therapy without evidence of active disease after their relapse. Ten of the 18 patients are still less than 12 months past their posttransplant salvage therapy and are at high-risk for relapse. Five patients are progression free at 15 to 36 months after their posttransplant relapse. Only three patients (two NHL and one HD) treated with other modalities after autologous transplant failure are alive without evidence of disease and have been observed at least 4 years postrelapse. Although a few patients will have a durable response to subsequent therapy, the majority of patients who have progressive disease after an autologous transplant for lymphoid malignancy will succumb to recurrent disease within a short period of time.

Adult↗
PubMed1992

The synthesis, kinetic characterization and application of a novel biotinylated affinity label for cathepsin B.

In this study we report on the synthesis, kinetic characterization and application of a novel biotinylated and active-site-directed inactivator of cathepsin B. Thus the peptidyldiazomethane biotinyl-Phe-Ala-diazomethane has been synthesized by a combination of solid-phase and solution methodologies and has been shown to be a very efficient inactivator of bovine and human cathepsin B. The respective apparent second-order rate constants (k0bs./[I]) for the inactivation of the human and bovine enzymes by this reagent, namely approximately 5.4 x 10(4) M-1.min-1 and approximately 7.8 x 10(4) M-1.min-1, compare very favourably with those values determined for the urethane-protected analogue benzyloxycarbonyl-Phe-Ala-chloromethane first described by Green & Shaw [(1981) J. Biol. Chem. 256, 1923-1928], thus demonstrating that the presence of the biotin moiety at the P3 position is compatible with inhibitor effectiveness. The utilization of this reagent for the detection of cathepsin B in electrophoretic gels, using Western blotting and in combination with a streptavidin/alkaline phosphatase detection system, is also demonstrated. Given that the peptidyldiazomethanes exhibit a pronounced reactivity towards cysteine proteinases, we feel that the present label may well constitute the archetypal example of a wide range of reagents for the selective labelling of this class of proteinase, even in a complex biological milieu containing additional classes of proteinases.

Affinity Labels↗
PubMed1992

The synthesis, kinetic characterization and application of biotinylated aminoacylchloromethanes for the detection of chymotrypsin and trypsin-like serine proteinases.

The synthesis of two biotinylated affinity labels for chymotrypsin and trypsin-like serine proteinases is described, along with their kinetic characterization and application to the detection of these proteinases after PAGE and Western blotting. Thus the chloromethane analogues biotinylphenylalanylchloromethane (Bio-Phe-CH2Cl; reagent 1) and biotinylarginylchloromethane (Bio-Arg-CH2Cl, reagent 2), have been shown to be potent active-site-directed inactivators of chymotrypsin and trypsin respectively. The apparent overall second-order rate constants (kobs./[I]) for the inactivation of chymotrypsin and trypsin by reagent 1 (approximately 4.9 x 10(3) M-1.min-1) and reagent 2 (approximately 1.0 x 10(5) M-1.min-1) respectively are comparable with those obtained by other workers with simple urethane-protected analogues and demonstrates that the presence of the bulky biotinyl moiety is compatible with inhibitor effectiveness. Samples of chymotrypsin and trypsin that have been inactivated by reagents 1 and 2 respectively and which have been subjected to SDS/PAGE and Western blotting can be revealed with a streptavidin/alkaline phosphatase label. We can presently detect down to 20 ng of inactivated proteinase by using this system. The utility of the arginine derivative for the detection of the plasma trypsin-like proteinases plasmin and thrombin has also been demonstrated, thus holding out the possibility that this reagent may find general application as an active-site-directed label for this class of proteinase.

Affinity Labels↗
PubMed1992

The application of a novel biotinylated affinity label for the detection of a cathepsin B-like precursor produced by breast-tumour cells in culture.

In this report we demonstrate how the recently developed biotinylated affinity label biotinyl-Phe-Ala-diazomethane (Bio-Phe-Ala-CHN2) [Cullen, McGinty, Walker, Nelson, Halliday, Bailie & Kay (1990) Biochem. Soc. Trans. 18, 315-316; Walker, Cullen, Kay, Halliday, McGinty & Nelson (1992) Biochem. J. 283, 449-453] can be used for the detection of a precursor form of a cathepsin B-like enzyme produced by breast-tumour cells in culture. Thus the cell lines MDA-MB-436, ZR-75-1 and T47-D produce a soluble protein that can be allowed to react with the biotinylated affinity label to yield an SDS-resistant complex; this can be revealed with a streptavidin/alkaline phosphatase label after PAGE and Western blotting. This protein (molecular mass 47 kDa) can also be detected by immunoblotting using sheep anti-(cathepsin B) antibodies in conjunction with a donkey anti-sheep IgG label. None of the cell lines studied produced any mature cathepsin B-like activity, as gauged by the lack of turnover of the fluorogenic substrate benzyloxycarbonyl-Arg-Arg-4-methylcoumarin-7-ylamide (Cbz-Arg-Arg-NH-Mec). However, treatment of medium samples with pepsin resulted in the generation of such activity. When the pepsin-catalysed activation step was analysed by SDS/PAGE, the protein of 47 kDa was completely converted into two species of very similar molecular masses of 30.5 kDa and 29 kDa. Both these proteins can incorporate the biotinylated probe and, in common with the 47 kD species, they can be detected with the streptavidin/alkaline phosphatase label and immunoblotting. We propose that the 47 kD form is the pepsin-activable proform of these lower-molecular-mass species. The release of the proform from the oestrogen-receptor (ER)-positive breast-tumour cell lines ZR-75-1 and T47-D is stimulated 5-10-fold when these cells are grown in medium containing epidermal growth factor (EGF) at a concentration of 10 ng/ml. In contrast, there is no modulation in the amount of proform released by the ER-negative cell line MDA-MB-436, over a range of EGF concentrations from 0 to 100 ng/ml.

Affinity Labels↗
PubMed1992

Development of the vestibular and auditory system of the northern native cat, Dasyurus hallucatus.

The developing vestibular and auditory system of the native cat Dasyurus hallucatus was examined from birth to day 55 postpartum to determine when the six sensory regions had an adult structure. The utricle, a sensory epithelium with an overlying discrete population of otoliths, was present in the newborn native cat. The saccule, which has a similar structure to the utricle, and the three crista ampullaris of the semicircular canals, were present by day 21 postpartum. The organ of Corti was formed by day 50 and the external ear duct was patent by day 55 postpartum. Hair is first seen on the native cat by day 45 when the young first leave the pouch. They are left in the nest and travel on the back of the lactating mother until about day 80. The eyes open on approximately day 75. Thus, the native cat has fully functional visual, auditory, and vestibular systems when the juvenile leaves the nest.

Age Factors↗
PubMed1992

Measurement of blood-brain barrier permeability in a tumor model using magnetic resonance imaging with gadolinium-DTPA.

Sequential MR imaging with gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA) and sequential measurements of plasma Gd-DTPA concentration by inductively coupled plasma atomic emission spectroscopy (ICP-AES) were used to estimate the blood-to-tissue transport coefficient (Ki) in the 36B-10 rat glioma model. For these measurements, tissue Gd-DTPA concentration was estimated from tumor enhancement by correlation with calibration measurements obtained by ICP-AES analysis of tumor tissue. The 14 animals for which Ki was calculated can be grouped into those imaged at 11 days following tumor implantation, at 13-18 days, and at 20 days. The mean (+SEM) Ki values for these groups were 1.1 + 0.24, 9.2 + 0.8, and 13.4 + 1.7 ml/kg-min, respectively. These results correspond well with published data obtained by quantitative autoradiography. It is concluded that frequent sequential imaging and a graphical approach to Ki calculation are promising methods for determining the blood-to-tissue transport coefficient noninvasively by contrast-enhanced MRI.

Animals↗
PubMed1992

Glutamate production in islets of Langerhans: properties of phosphate-activated glutaminase.

Homogenates of rat pancreas, pancreatic islets, and HIT-T15 cells (a clonal line derived from B cells) catalyzed the breakdown of glutamine to glutamate. This activity was markedly stimulated by the addition of orthophosphate and was much greater in homogenates from islets and the B-cell-derived clonal cell line than in those from whole pancreas. Islet glutaminase was half-maximally stimulated with 40 mmol/L phosphate. Kinetic analyses of the rates of glutamine hydrolysis showed that the Vmax for the reaction increased with the increase in phosphate concentration, whereas the Km for glutamine (2.6 +/- 0.2 mmol/L) was unaltered. The pH optimum for enzyme activity was 8.0 to 8.5 at all phosphate concentrations studied. Glutamine breakdown was enhanced by adenosine triphosphate ([ATP] approximately 100% at 10 mmol/L) and citrate (approximately 30% at 10 mmol/L), but it was unaffected by malate, 2-oxoglutarate, lactate, and ammonia. Glutamate significantly inhibited glutamine hydrolysis. Freshly isolated islets had a low content of both glutamate and glutamine. After culturing for 1 hour in an amino acid-containing medium, the concentrations of glutamine and glutamate increased. Subsequent perifusion without amino acids caused a loss of glutamine and a concomitant increase in glutamate level. Perifusion with 1 mmol/L glutamine led to an increase in both internal glutamine and glutamate. The addition to the perifusion medium of either 10 mmol/L glutamine, 10 mmol/L orthophosphate, or both substantially enhanced insulin release evoked by 10 mmol/L leucine.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1992

Grading nephroureteral dilatation detected in the first year of life: correlation with obstruction.

To understand better the significance of pediatric idiopathic nephroureteral dilatation the renal ultrasound images of patients less than 1 year old with hydronephrosis or hydroureteronephrosis were graded and compared to the radiological diagnosis of obstruction as determined by diuresis renography and/or urography. The study included 73 boys and 30 girls with hydronephrosis (76 patients) or hydroureteronephrosis (27). For hydronephrosis obstruction was diagnosed in 56 children (74%) and involved 61 of 97 kidneys (63%). For obstructed kidneys the mean grade of hydronephrosis (3.4 +/- 0.7 standard deviation) was statistically different from that of nonobstructed kidneys (1.6 +/- 0.8 standard deviation) (p less than 0.05). When the value to predict obstruction was set at grade 3 hydronephrosis or greater there was an 88% sensitivity and 95% specificity. For hydroureteronephrosis obstruction was diagnosed in 15 of 27 children (56%) and involved 17 of 34 kidneys (50%). The degree of dilatation was weighted as a score to assess the grades of hydronephrosis and ureteral dilatation, namely hydroureteronephrosis score equals grade of hydronephrosis plus grade of ureteral dilatation. In obstructed megaureters the mean hydroureteronephrosis score (5.8 +/- 1.0) was statistically different from that for nonobstructed megaureters (mean hydroureteronephrosis score 2.7 +/- 1.9) (p less than 0.001). When the value to predict obstruction was set at hydroureteronephrosis score of 5 or greater there was a 94% sensitivity and 80% specificity. Although ultrasound examination alone cannot be used to diagnose urinary obstruction, the radiological diagnosis of obstruction is linked with the grade of hydronephrosis or score of hydroureteronephrosis.

Age Factors↗
PubMed1992

Report of the Conference on Low Blood Cholesterol: Mortality Associations.

BACKGROUND: A National Heart, Lung, and Blood Institute (NHLBI) Conference was held October 9-10, 1990, to review and discuss existing data on U-shaped relations found between mortality rates and blood total cholesterol levels (TC) in some but not other studies. Presentations were given from 19 cohort studies from the United States, Europe, Israel, and Japan. A representative of each study presented its findings and also submitted tables of proportional hazards regression coefficients for entry TC levels in regard to death, and these were incorporated into a formal statistical overview adjusted for age, diastolic blood pressure, cigarette smoking, body mass index, and alcohol intake, as available. METHODS AND RESULTS: The U-shape for total mortality in men and the flat relation in women resulted largely from a positive relation of TC with coronary heart disease death and an inverse relation with deaths caused by some cancers (e.g., lung but not colon), respiratory disease, digestive disease, trauma, and residual deaths. Risk for combined noncardiovascular, noncancer causes of death decreased steadily across the range of TC. The conference considered possible explanations for the statistical associations found between low TC levels or active TC lowering and certain causes of death. One is that TC is lowered by some disease conditions themselves, such as wasting in chronic pulmonary disease or reduced production and secretion of cholesterol-bearing lipoproteins with liver disease. In this sort of situation, the TC:mortality association found in observational studies may be due to preexisting disease. This was addressed by excluding early deaths from the analysis, which did not change the results. The conference considered as well the biological function of cholesterol, which, if seriously deranged, might hypothetically cause a wide variety of diseases and dysfunction. The conference also considered the biological functions that might provide plausible mechanisms for the associations found. CONCLUSIONS: Definitive interpretation of the associations observed was not possible, although most participants considered it likely that many of the statistical associations of low or lowered TC level are explainable by confounding in one form or another. The conference focused on the apparent existence and nature of these associations and on the need to understand their source rather than on any pertinence of the findings for public health policy. Further research is recommended to explain the observed associations of low TC levels (and TC lowering) with certain noncardiovascular diseases. This includes studies of the time course of TC change in disease, the relation of TC to morbidity, further studies of possible epidemiological confounding, monitoring of population trends in TC and mortality, further studies of the relations in women, auditing of noncardiovascular events in trials, studies of cell membrane, genetic and molecular links to cholesterol metabolism, TC level and disease, studies of disease manifestations in specific lipid disorders, and further study of the proposed causal mechanisms linking low TC and hemorrhagic stroke.

Alcohol Drinking↗
PubMed1992

Simple surgical treatment of nonparasitic hepatic cysts.

Benign nonparasitic liver cysts are uncommon lesions. Incidental diagnosis is increasing with the advent of routine abdominal computed tomography and ultrasound scanning. Cysts that attain massive proportions often become symptomatic and require therapeutic intervention. Surgical resection and Roux-en-Y cystojejunostomy drainage have been the treatments of choice, but simpler unroofing techniques without drainage have recently been employed with success. Three patients with symptomatic, large, nonparasitic cysts were surgically treated in such a fashion without complication and form the basis of this report. The technique of wide unroofing involves excision of the nonhepatic cyst wall with oversewing of communicating biliary radicals. No recurrences have been detected in follow-up screening. Wide unroofing is a simple and yet reliable surgical option for the treatment of symptomatic hepatic cysts.

Aged↗
PubMed1992

[Mechanism of the effect of serine and threonine on ammoniagenesis and biosynthesis of orotate in the mouse].

We have evaluated the effects of serine and threonine on orotate metabolism in mice, by injecting i.p. 0.5 to 1.5 mmol/100 g of body weight, during 4 weeks of experimentation. The results indicate that serine as well as threonine cause a significant increase (p less than 0.01) of plasma ammonia and urinary orotate. We have also studied the effects of various inhibitors, adenine (0.3% diet), N-(phosphonoacetyl)-L-aspartate (PALA) (10 mg/100 g, i.p.), acivicin (1 mg/100 g, i.p.) and cycloheximide (1.5 mg/kg, i.p.) on orotate synthesis in mice injected with serine (2.5 mmol/100 g, i.p.) or threonine (1.25 mmol/100 g, i.p.). The results show an increase (p less than 0.05) of urinary orotate following an adenine-rich diet, and a reduction of orotate following PALA (p less than 0.01). The orotic aciduria remained insensitive to acivicin while it was inhibited by cycloheximide. These results suggest that adenine blocks the utilisation of orotate and consequently could affect the biosynthesis of pyrimidines; the induction of orotate by serine and threonine is controlled by the translation of a specific protein synthesis, and necessarily implicates the mitochondrial carbamyl phosphate synthetase-I (CPS-I). The regulating step in this synthesis of orotate could also be the transport of carbamyl-phosphate from the mitochondria to cytosol.

Adenine↗
PubMed1991

Relationships between energy level and insulin secretion in isolated rat islets of Langerhans. A study at various pH values.

To define better the role of [ATP]/[ADP] in insulin release from pancreatic islets, changes in the adenine nucleotide ratios elicited by alterations in external pH were correlated with the secretion profiles produced by administration of two metabolic secretagogues, 16 mM glucose and 10 mM alpha-ketoisocaproic acid. Experiments were carried out in buffers with and without bicarbonate, in the pH range 6.5-7.7. Insulin release was dependent on pHe irrespective of the secretagogue used. Secretion profiles for alpha-ketoisocaproic acid were the same both with and without bicarbonate; the release was decreased below pH 7.1 but maintained at 7.4-7.7. The same pattern was seen with glucose in media buffered with Hepes. With bicarbonate present, secretion caused by high glucose showed a bell-shaped dependence on [H+], with reductions at the acid and alkaline sides of pH 7.1-7.4. [ATP] and [ADP] were higher when Hepes was the buffer, at all pH values studied. The [ATP]/[ADP] declined with increasing pH under both basal and stimulated conditions; the values were always larger after stimulation although at pH 7.7 with bicarbonate present and glucose as the stimulant the difference was very small. It is concluded that: (i) the [ATP]/[ADP] in pancreatic islets is markedly dependent on pHe; (ii) there is no straight-forward correlation between either [ATP] or the absolute value for [ATP]/[ADP] and insulin secretion; and (iii) a rise in [ATP]/[ADP] is necessary for glucose-stimulated insulin release although it is not always the rate-determining event.

Adenosine Diphosphate↗
PubMed1991

Neurogenesis in the brain auditory pathway of a marsupial, the northern native cat (Dasyurus hallucatus).

Neurogenesis in the auditory pathway of the marsupial Dasyurus hallucatus was studied. Intraperitoneal injections of tritiated thymidine (20-40 microCi) were made into pouch-young varying from 1 to 56 days pouch-life. Animals were killed as adults and brain sections were prepared for autoradiography and counterstained with a Nissl stain. Neurons in the ventral cochlear nucleus were generated prior to 3 days pouch-life, in the superior olive at 5-7 days, and in the dorsal cochlear nucleus over a prolonged period. Inferior collicular neurogenesis lagged behind that in the medial geniculate, the latter taking place between days 3 and 9 and the former between days 7 and 22. Neurogenesis began in the auditory cortex on day 9 and was completed by about day 42. Thus neurogenesis was complete in the medullary auditory nuclei before that in the midbrain commenced, and in the medial geniculate before that in the auditory cortex commenced. The time course of neurogenesis in the auditory pathway of the native cat was very similar to that in another marsupial, the brushtail possum. For both, neurogenesis occurred earlier than in eutherian mammals of a similar size but was more protracted.

Aging↗
PubMed1991

The effect of bicarbonate on resuscitation from cardiac arrest.

STUDY OBJECTIVES: This study attempted to determine the effect of bicarbonate administration on resuscitation in a porcine model of prolonged cardiac arrest. DESIGN: After instrumentation, 26 swine were subjected to ventricular fibrillation for 15 minutes (16 animals) or 20 minutes (ten animals) with no resuscitative efforts. INTERVENTIONS: Resuscitation attempts with open-chest cardiac massage and epinephrine were used in all animals after the arrest period. The experimental group was given sodium bicarbonate (3 mEq/kg), and the control group received 3% saline (5 mL/kg) at the initiation of cardiac massage. MEASUREMENTS: Resuscitation success, hemodynamics, and arterial and mixed venous gases were compared in the bicarbonate and hypertonic saline-treated groups. RESULTS: There was no difference in resuscitation rates between bicarbonate and nonbicarbonate-treated swine. After 15 minutes of ventricular fibrillation, six of eight bicarbonate-treated swine were resuscitated successfully compared with five of eight hypertonic saline-treated animals. None of the five bicarbonate-treated or five hypertonic saline-treated swine that underwent 20 minutes of ventricular fibrillation were resuscitated. The arterial and mixed venous pH values were significantly different in the bicarbonate-treated animals from values in the control group. There was no difference in systolic or diastolic blood pressures or myocardial perfusion pressure between the bicarbonate and hypertonic saline-treated animals. CONCLUSION: Despite correlation of arterial and venous acidemia, the use of sodium bicarbonate did not improve resuscitation from prolonged cardiac arrest.

Animals↗