Two new variants of RAG-1 protein predicted by SSCP.
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Homeodomain Proteins↗
Biomedical subjects
J Soler
Publications and source records attributed to J Soler.
Purification and characterization of an extracellular aspartate protease from Phycomyces blakesleeanus.
An acid protease has been found in the culture broth of Phycomyces blakesleeanus growing under standard conditions. It has been induced up to 70-fold with several complex growth media and the enzyme has been purified to homogeneity and characterized. The molecular mass of the native enzyme was estimated by gel filtration to be 40 kDa. The acid protease of Phycomyces migrated as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, corresponding to a molecular mass of 35 kDa. The glycoprotein nature for the acid protease was deduced from its binding to a concanavalin A-Sepharose 4B column. The carbohydrate moiety is composed of mannose and rhamnose. Its amino acid composition was determined, and its isoelectric point was estimated to be 4.2, the optimum pH was 2.5 to 3, and the optimum temperature was 70 degrees C, using hemoglobin as a substrate. The enzyme showed thermal stability between 37 and 50 degrees C. The thermodynamic parameters for hemoglobin hydrolysis and thermal inactivation were calculated. With Lys-Pro-Ile-Glu-Phe-Phe(4-N02)-Arg-Leu as the substrate, the Km, kcat, and Vmax values were 8.78 microM, 1.25 s(-1), and 2.12 mumol min(-1) mg(-1), respectively. The protease was insensitive to phenylmethylsulfonyl fluoride, O-phenanthroline, N-ethylmaleimide, iodoacetamide, ethylenediaminetetraacetate, [ethyl-enebis(oxyethylenenitrilo)]tetraacetic acid, and trypsin inhibitor. However, pepstatin A established a strong competitive inhibition against it, with a K(i) value of 1.33 nM. The data suggest that this protease has properties of an aspartate-type proteinase.
Amino Acid Sequence↗
Expression of ICAM-1 in distant metastatic thyroid carcinoma.
Distant metastasis from follicular thyroid carcinoma developed in a 48-year-old woman 11 yr after the resection of the primary tumor. Distant metastasis consisted in invasion of the left infraspinatus muscle by malignant thyroid cells. After its surgical removal, in the following 6-8 months multiple metastasis to distant skeletal muscle and brain appeared, and despite chemotherapy and local radiotherapy, the patient finally died. Intercellular adhesion molecule-1 (ICAM-1), a single chain transmembrane glycoprotein, was detected on the surface of cells of the metastatic tissue. Although ICAM-1-positive staining has been recently described in primary tumors such as papillary adenocarcinoma, and metastatic tumors from skin, brain, thymus, liver, adrenal gland and prostate, to our knowledge its expression on distant metastasis from thyroid carcinoma has not been previously reported.
Adenoma↗
Rapid sequencing protocol using microconcentrators.
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Genes, RAG-1↗
Adhesion of K88ab to guinea pig erythrocytes: effect on membrane enzyme activities.
Nontoxigenic Escherichia coli strains bearing K88 fimbriae have been associated with diarrhea in piglets. We have used guinea pig erythrocytes as a model of the host cell to study the cellular alterations after adherence of purified K88ab fimbriae. Although Mg2+-dependent ATPase was inhibited (up to 61%), Na/K ATPase was not. Metabolic enzymes were not significantly affected.
Adenosine Triphosphatases↗
Beta-cell growth and mass are preserved in long-term syngeneic islet transplantation in streptozocin-induced diabetic Lewis rats.
We determined beta-cell replication and mass in basal and stimulated conditions in long-term transplanted islets. Three groups of streptozocin-induced diabetic Lewis rats were transplanted with 1,000 islets (500 islets under left and right kidney capsules). At 2 (Tx-2), 5 (Tx-5), or 9 (Tx-9) months after transplantation, one of the two grafts (basal) was harvested; 14 days later, the contralateral graft (stimulated) was also harvested. Normoglycemia was achieved and maintained in all transplanted rats, although the capacity to respond to a glucose challenge deteriorated slightly 9 months after transplantation. Beta-cell replication remained stable in Tx-2, Tx-5, and Tx-9 basal grafts and was similar to replication in a control group of nontransplanted rats (0.28 +/- 0.06%); replication increased in Tx-2 (0.90 +/- 0.23%, P < 0.05) and Tx-9 (0.72 +/- 0.09%, P < 0.05) stimulated grafts. Beta-cell mass in basal grafts was similar to the initially transplanted mass (1.24 +/- 0.06 mg) and increased in stimulated grafts in Tx-2 (1.91 +/- 0.38 mg, P < 0.05) and Tx-5 (1.73 +/- 0.27 mg, P = 0.01) groups, compared with basal grafts, and in Tx-2 and Tx-9 groups (1.92 +/- 0.30 mg, P < 0.05), compared with initially transplanted mass. Therefore, beta-cell replication and mass were preserved up to 9 months after syngeneic transplantation, and beta-cells maintained the capacity to respond to increased metabolic demand, suggesting that replication is not a limiting factor in the survival of transplanted islets.
Animals↗
Hematopoietic growth factor (G-CSF or GM-CSF) after salvage chemotherapy in refractory or relapsed adult de novo acute leukemia.
Nineteen adults with primary refractory or relapsed acute leukemia (12 ALL and 7 ANLL) were treated with an intensive salvage chemotherapy (intermediate-dose ara-C, intermediate-dose methotrexate, vindesine, cyclophosphamide, mitoxantrone and prednisolone) followed by a hematopoietic growth factor (HGF), either granulocyte colony-stimulating factor (5 micrograms/kg) or granulocyte-macrophage colony-stimulating factor (10 micrograms/kg). Both were given from the day after chemotherapy ended and until the neutrophil count rose above 1 X 10(9)/l for three consecutive days. Eleven patients (58%, 95% CI 33% to 82%) achieved complete remission, and 15 courses of salvage therapy were given to these complete responders. In a historical control group that did not receive HGF, 23 out of 38 patients (60%, 95% CI 44% to 77%) achieved complete remission, and 27 courses of therapy were delivered to complete responders. Treatment with a HGF accelerated the recovery of neutrophils to 0.5 X 10(9)/l significantly, shortening it from a mean of 28 to 22 days (p = .0002), with no effect on platelet recovery. There were no differences in the rates of documented and fatal infections, which were relatively high in both groups. In the patients with ANLL, there was no evidence that HGF accelerated leukemic regrowth. We conclude that HGF accelerates neutrophilic recovery following intensive salvage chemotherapy for acute leukemia, although no reduction in documented infections was found. Many factors, including the small patient sample, may have contributed to this latter finding.
Acute Disease↗
Nitrendipine and enalapril in the treatment of diabetic hypertensive patients with microalbuminuria.
A trial to study the efficacy, safety and tolerability of nitrendipine and enalapril in the treatment of diabetic hypertensive patients with microalbuminuria (MA) was performed to compare the effects of both drugs in the prevention of the renal impairment. Twenty-eight valid patients [13 with nitrendipine (N) and 15 with enalapril (E) with NIDDM, hypertension (diastolic blood pressure between 90 to 114 mm Hg) and MA (urinary albumin between 30 to 300 mg/24 hr) were recruited in a double blind, randomized trial. Following a placebo run-in period of two to four weeks, all eligible patients were randomly allocated to either N or E treatment. Treatment lasted six months, with two different visits at three and six months in which blood pressure (BP), heart rate (HR), renal function and MA were measured. No statistically significant differences on BP and metabolic parameters were found between both treatment groups. The geometric mean of final glomerular filtration rate (GFR) in the N group was 34.5% higher than in the E group, while the reduction on MA was most important in the E group. Eleven patients reported adverse events (AEs) and there were four dropouts, three of them due to AEs. We conclude that both treatments are a good choice for treating diabetic hypertensive patients with early altered renal function, as they reduce BP without altering metabolic parameters, increase GFR and reduce MA with a low frequency of AEs.
Adult↗
GH response to GH-releasing hormone (GHRH) in Alzheimer and vascular dementia. Relation with somatostatin cerebrospinal levels.
OBJECTIVE: To clarify the functional state of the somatotropinergic system in patients with Alzheimer's disease (AD) and in patients with vascular dementia (VD) in relation with somatostatin cerebrospinal levels. DESIGN We studied the GH response to GHRH in controls and in AD and VD patients and correlated their responses with somatostatin cerebrospinal levels. PATIENTS: Twelve control subjects and 56 patients, 28 with AD and 28 with VD, were studied. A GHRH test was performed in every case. One hundred mcg of GHRH was administered as a intravenous bolus and blood samples were drawn at -15, 0, 30, 60, 90 and 120 minutes. Cerebrospinal fluid was obtained after lumbar puncture. Measurements of GH and somatostatin were determined by specific radioimmunoassay. RESULTS: The GH response to GHRH exhibited no difference between AD and VD patients (12.2 +/- 2 mcg/l vs 9.9 +/- 1.8). There were 17 AD patients and 16 VD patients with exaggerated GH response to GHRH. There was no difference between cerebrospinal levels of somatostatin (30 +/- 2.1 pg/mL in AD patients vs 42 +/- 2.4 in VD patients), and there was no correlation among GH response to GHRH and cerebrospinal levels. CONCLUSION: In our study we observed no difference between the AD and VD patients in GH response to GHRH and no difference in somatostatin cerebrospinal levels in these patients.
Aged↗
Mutational analysis of RAG-1 in lymphoid malignancies.
Illegitimate recombinase activity promoted by the recombination activating RAG-1 and RAG-2 is assumed to be involved in the pathogenesis of the chromosomal translocations observed in lymphoid neoplasias. We analyzed the complete coding region of the RAG-1 gene in patients with lymphoid neoplasis using a multiple PCR-SSCP (single strand conformation polymorphism) strategy. Nine multiple myelomas, 17 non-Hodgkin's lymphomas, 18 acute lymphocytic leukemias, 37 chronic lymphocytic leukemias and 33 non-neoplastic controls were studied. To screen the entire RAG-1 gene we used primers overlapping genomic segments of the RAG-1 coding sequence (nucleotides 87 to 3311). Samples with an abnormal band pattern in the SSCP were cloned and sequenced. Successful amplification was achieved with our protocol. The multiple PCR-SSCP analysis proved to be a feasible and sensitive strategy for studying variations in the sequence of the RAG-1 gene. No mutations other than the three previously reported sequence variations were detected. Although mutations in this gene do not appear to be common in lymphoid neoplasias, it would be interesting to ascertain whether the different variant forms of RAG-1 protein have an abnormal recombinase activity.
DNA Mutational Analysis↗
Ketoacidosis at diagnosis is predictive of lower residual beta-cell function and poor metabolic control in type 1 diabetes.
To determine the factors at diagnosis predictive of changes in residual beta-cell function and metabolic control in Type 1 diabetes, 125 patients older than 7 years of age consecutively diagnosed between March 1986 and June 1991 were followed prospectively for two years. The effect of age, gender and the presence of ketoacidosis (DKA) and islet-cell antibodies (ICA) on beta-cell function, metabolic control and insulin requirements were studied by multivariate analysis of variance (repeated measurements over time) in 90 patients who completed follow-up. DKA had an independent negative effect on residual beta-cell function over time (p = 0.001). ICA-positive patients had lower residual beta-cell function at the end of follow-up (p < 0.05), but overall differences were not significant. DKA and younger age had an independent negative influence on metabolic control (p < 0.05) and insulin requirements (p < 0.001) over time. It is concluded that residual beta-cell function in Type 1 diabetic patients two years after diagnosis was independently influenced by DKA and ICA at diagnosis. Moreover, DKA and age influenced metabolic control and could thus be used to predict those patients with rapidly deteriorating metabolic control who might benefit from a more intensive therapeutic approach.
Adolescent↗
A multicenter, parallel comparative study of the antihypertensive efficacy of once-daily lisinopril vs enalapril with 24-h ambulatory blood pressure monitoring in essential hypertension.
The aim of the study was to compare the antihypertensive efficacy of once-daily lisinopril vs enalapril both during normal daily activity and sleep, in mild-to-moderate essential hypertension. After a 4-week wash-out period, 34 patients (17 M, 17 F) aged 22 to 67 years were randomized in a multicenter, open, parallel fashion: 17 received lisinopril (10-20 mg) and 17 enalapril (10-20 mg) for a 12-week period. Twenty-four hour ambulatory blood pressure monitoring (ABPM) was performed using an oscillometric non-invasive automated device at both the end of the 4-week drug-free baseline period and during the last week of treatment. With no differences in initial blood pressure (BP) between groups, both drugs significantly reduced office and ABPM values. Lisinopril tended to reduce BP in a greater extension than enalapril, but only the reduction of office systolic BP (SBP) (p = 0.0062), 24-h SBP load (P = 0.0182) and night time SBP load (P = 0.0316) reached statistical significance. We conclude that, in spite of a more prominent reduction of SBP by lisinopril, both drugs have a similar efficacy in reducing BP, assessed by both office and ABPM measurements.
Adult↗
[The value of the parathyrin-related protein (PTH-RP) in the diagnosis of cancer-associated hypercalcemia].
BACKGROUND: The parathyrine related protein (PTH-RP) is very similar, both in structure and in function, to the PTH and is considered as a mediator in humoral hypercalcemia in cancer. The aim of this study was to know the clinical value of PTH-RP measurement. METHODS: Serum PTH-RP concentrations were studied in 22 healthy subjects, 13 patients with primary hyperparathyroidism, 9 patients with solid neoplasms and normocalcemia, 26 patients with solid neoplasms and hypercalcemia and 4 patients with hematologic neoplasms and hypercalcemia. The PTH-RP was quantified by a competitive radioimmunoassay technique using a specific antibody of the PTH-RP 1-40 fragment. Intact parathyrine (i-PTH) was quantified by an IRMA method using 2 polyclonal antibodies (INCSTAR). RESULTS: Fifteen (68%) of the healthy controls presented undetectable serum PTH-RP concentrations. The serum PTH-RP concentration was normal in all those patients with hyperparathyroidism. Elevated serum PTH-RP values were not found in patients with solid neoplasms and normocalcemia or in those with hematologic neoplasms and hypercalcemia. High values of PTH-RP were observed in 8 out of 9 (88%) of the patients with solid neoplasms and hypercalcemia with bone metastasis and in 7 out of 11 (63%) of the patients with bone involvement. CONCLUSIONS: Serum parathyrine-related protein was found to be high in a large proportion of patients with solid neoplasms and hypercalcemia. Serum PTH-RP determination is useful in the clinical investigation of patients with hypercalcemia. Even in patients with bone metastasis, hypercalcemia may have a humoral background.
Adult↗
The pH dependence and modification by diethyl pyrocarbonate of isocitrate lyase from Phycomyces blakesleeanus.
We determined the variation with pH of the kinetic parameters for the isocitrate cleavage reaction catalyzed by Phycomyces isocitrate lyase, with the aim of elucidating the role played by ionising amino acid residues in binding and catalysis. The log VmaxpH profile shows that the enzyme possesses two ionising groups with pK values of 6.1 and 8.3. The first group is also observed in the VmaxpH/KmpH and pKmpH profiles, so this group is involved in catalysis. The last two profiles exhibit a similar pK value of 16 on the basic side, which represents the sum of the pK values for two ionising groups with pK values that differ by less than two pH units. Diethyl pyrocarbonate inactivated isocitrate lyase from Phycomyces with a second-order rate constant of 18.58 M-1 s-1 (at pH 6.0 and 20 degrees C). The difference spectra of the modified enzyme revealed an absorption maximum at 242 nm, characteristic of N-carbethoxyhistidine isocitrate lyase. No trough at around 280 nm due to O-carbethoxytyrosine is observed. Quantification of the increase in absorbance to 242 nm due to N-carbethoxyhistidine showed that ten histidine residues/active site were modified during total inactivation. However, only one of them was essential for catalysis. Treatment of the partially inactivated enzyme with hydroxylamine led to recovery of a substantial part of the original activity. The reactivity of isocitrate lyase towards diethyl pyrocarbonate declined with pH, following a titration curve for a group of pK 6.1. The presence of substrate decreased the rate of inactivation. Data-protection analyses indicate that the reactive histidine residues are within the active site of the enzyme.
Binding Sites↗
Isocitrate lyase from Cephalosporium acremonium. Role of Mg2+ ions, kinetics, and evidence for a histidine residue in the active site of the enzyme.
Isocitrate lyase was purified from Cephalosporium acremonium CW-19 from cultures growing with poly(oxyethylene)sorbitan monopalmitate as the carbon source. Its subunit M(r) and native M(r) were 63,000 +/- 2000 and 250,000 +/- 5000, respectively. We found the Mg(2+)-isocitrate complex to be the true substrate and that Mg2+ ions act as a nonessential activator, according to the model reported by Giachetti et al. (1988) [Giachetti, E., Pinzauti, G., Bonaccorsi, R., & Vanni, P. (1988) Eur. J. Biochem. 172, 85-91], from which the kinetic parameters were calculated. The kinetic study is consistent with an ordered Uni-Bi mechanism, and the kinetic and rate constants of the model were calculated. pH dependence of the cleavage reaction indicated that the catalysis was dependent on two dissociable groups on the enzyme-substrate complex. The enzyme was inactivated by diethyl pyrocarbonate following first-order kinetics at all reagent concentrations used. The pseudo-first-order rate constant of inactivation increases with pH, suggesting participation of an amino acid residue with pK 6.0. Hydroxylamine added to the inactivated enzyme quickly restored the incremental absorption at 240 nm and most of the activity. Data analyses indicated that diethyl pyrocarbonate inactivation is a consequence of modification of 11 histidine residues per enzyme subunit, and from statistical analysis, we concluded that one is catalytically important. Mg(2+)-isocitrate protects the enzyme against diethyl pyrocarbonate inactivation with a Ks value of 26.8 +/- 2.1 microM, close to the Km value. Isocitrate protects the enzyme but a high concentration, suggesting its binding to the catalytic site of the nonactivated enzyme. Mg2+ ions also produced total competitive protection.
Acremonium↗
Pituitary apoplexy into nonadenomatous tissue: case report and review.
Pituitary apoplexy into nonadenomatous tissue is extremely rare. The authors describe a 20-year-old woman with symptomatic pituitary hemorrhage into an apparently intrasellar malignant teratoma, which caused headache, visual impairment, involvement of III, IV, VI, and 1st division of the V cranial nerves, and hypopituitarism. Diabetes insipidus had developed previously. Magnetic resonance scans had a high-intensity signal in the pituitary on T1- and T2-weighted images, and lack of the signal of the posterior pituitary. Transsphenoidal approach, radiotherapy, and chemotherapy management did not preclude a fatal outcome.
Adult↗
Ogilvie's syndrome from disseminated varicella-zoster infection and infarcted celiac ganglia.
We report a patient who had refractory Hodgkin's disease and who received an autologous bone marrow transplantation and 8 months later developed abdominal pain associated with acute colonic dilation. The course of the patient was rapidly fatal due to a lobar pneumonia. Autopsy revealed signs of disseminated herpesvirus infection with marked hemorrhagic infarction of celiac sympathetic ganglia. This finding supports the hypothesis that denervation caused by virus reactivation and secondary hemorrhage is a main mechanism of acute colonic pseudoobstruction.
Adult↗
Kinetics and Thermostability of NADP-Isocitrate Dehydrogenase from Cephalosporium acremonium.
NADP-isocitrate dehydrogenase [isocitrate:NADP(sup+) oxidoreductase (decarboxylating); EC 1.1.1.42] was purified from Cephalosporium acremonium as a single species. The enzyme is a dimer of 140 kDa with identical subunits of 75 kDa. The existence of a monomer-dimer equilibrium is apparent as revealed by an enzyme dilution approach. The chelate complex of the tribasic form of isocitrate and Mg(sup2+) is the true substrate. The V(infmax) depends on a basic form of an ionizable group of the enzyme-substrate complex with a pK(infes) (pK of the enzyme-substrate complex) of 6.9 and a (Delta)H(infion) (activation enthalpy) of -2 (plusmn) 0.4 kcal mol(sup-1) (ca. 8 (plusmn) 2 kJ mol(sup-1)). The enzyme showed maximum activity at 60(deg)C, an unusually high temperature for a nonthermophilic fungus. The thermodynamic parameters for isocitrate oxidative decarboxylation and for the binding of isocitrate and NADP(sup+) were calculated. We analyzed the kinetic thermal stability of the enzyme at pH 6.5 and 7.6. It was inactivated above 40(deg)C following a first-order kinetics. The presence of 12 mM Mg(sup2+) plus 10 mM dl-isocitrate led to 100% protection of enzyme activity against inactivation at 60(deg)C for 120 min. Removal of either or both compounds led to activity loss. A greater stabilizing role for Mg(sup2+) was seen at pH 6.5 than at pH 7.6, whereas the stabilizing effect of isocitrate was not dependent on pH.
Journal Article↗