Regulation of protein tyrosine kinase activation by the T-cell antigen receptor zeta chain.
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Amino Acid Sequence↗
Biomedical subjects
D Straus
Publications and source records attributed to D Straus.
Genomic subtraction for cloning DNA corresponding to deletion mutations.
We have developed a technique, called genomic subtraction, for isolating the DNA that is absent in deletion mutants. The method removes from wild-type DNA the sequences that are present in both the wild-type and the deletion mutant genomes. The DNA that corresponds to the deleted region remains. Enrichment for the deleted sequences is achieved by allowing a mixture of denatured wild-type and biotinylated mutant DNA to reassociate. After reassociation, the biotinylated sequences are removed by binding to avidin-coated beads. This subtraction process is then repeated several times. In each cycle we hybridize the unbound wild-type DNA from the previous round with fresh biotinylated deletion mutant DNA. The unbound DNA from the final cycle is ligated to adaptors and amplified by using one strand of the adaptor as a primer in the polymerase chain reaction. The amplified sequences can then be used to probe a genomic library. We applied genomic subtraction to a yeast strain that has a 5-kilobase deletion, corresponding to 1/4000th of the genome. In the experiment reported here, three rounds of subtraction were sufficient to accurately identify genomic clones containing sequences that are missing in the deletion mutant. We discuss the limitations and some potential applications of the method.
Affinity Labels↗
DnaK, DnaJ, and GrpE heat shock proteins negatively regulate heat shock gene expression by controlling the synthesis and stability of sigma 32.
The Escherichia coli DnaK heat shock protein has been identified previously as a negative regulator of E. coli heat shock gene expression. We report that two other heat shock proteins, DnaJ and GrpE, are also involved in the negative regulation of heat shock gene expression. Strains carrying defective dnaK, dnaJ, or grpE alleles have enhanced synthesis of heat shock proteins at low temperature and fail to shut off the heat shock response after shift to high temperature. These regulatory defects are due to the loss of normal control over the synthesis and stability of sigma 32, the alternate RNA polymerase sigma-factor required for heat shock gene expression. We conclude that DnaK, DnaJ, and GrpE regulate the concentration of sigma 32. We suggest that the synthesis of heat shock proteins is controlled by a homeostatic mechanism linking the function of heat shock proteins to the concentration of sigma 32.
Alleles↗
Definitive irradiation for localized non-Hodgkin's lymphoma of breast.
A diagnosis of primary non-Hodgkin's lymphoma (NHL) confined to the breast or breast and ipsilateral lymph nodes was biopsy proven in 19 patients who presented to our institution between 1970 and 1984. Four patients were treated with a modified radical mastectomy, and one with chemotherapy only. The remaining 14 patients comprise the basis of this study. All patients were clinically staged. There were 12 patients with 1A-E disease and two with IIA-E disease. All patients were treated with breast radiation. The axillary nodes were included in seven of the patients and two, in addition, received "mantle" irradiation. The majority of the patients received between 3500-4400 cGy, but four received lower doses. One patient, with Stage IIA-E disease, received 6 months of systemic chemotherapy, followed by 3500 cGy to the breast and axilla. Three of the 14 patients relapsed within the treated area, with a local control rate of 78%. One was salvaged with a mastectomy and remains alive and well 15 years after diagnosis. Seven of the 14 patients relapsed distantly, including two with local failures, but only three have died of disease. An additional two patients have died of other causes without evidence of disease, for a survival of 66% at 48 months (Kaplan-Meier Calculations).
Aged↗
Use of topical antiseptic in prophylaxis of catheter-related septic complications.
Pretreatment of catheters by a simple new procedure designed to reduce the incidence of septic complications was tested in both in vitro and in vivo studies. In the former experiments, the high sensitivity of gram-negative bacteria and fungi to Silastic (silicone rubber) catheters pretreated with silver nitrate solution was determined. The antimicrobrial activity remained unchanged after both sterilization and up to six weeks of storage. Furthermore, prolonged incubation of treated catheter segments in Escherichia coli inoculated plasma resulted in a significant reduction of organisms in the media and those adherent to the surface of the catheters. The in vivo experiments were performed upon two groups of rabbits. In the first group, catheters (one treated and one untreated) were implanted into contralateral jugular veins. In the second group, alternating treated and control catheter segments were threaded onto polyethylene core tubing; the resulting string of segments was positioned in the inferior vena cava. Five to 11 days after implantation of the catheter, all rabbits were intravenously injected with live E. coli (approximately 10(8) to 10(9) organisms per kilogram of weight); 18 to 24 hours later, the catheters and blood samples were removed for cultivation. Significant reductions in both incidence and magnitude of colonization in treated catheters by E. coli were observed in both rabbit groups. Additionally, histologic examination did not reveal any significant differences between contralateral jugular veins (previously in prolonged and intimate contact with the indwelling catheters), confirming the absence of any local adverse effects of silver nitrate.
Animals↗
Autologous bone marrow transplantation for patients with poor-prognosis lymphoma.
Review of prognostic factors at Memorial Hospital in New York City has shown that adult patients with large-cell lymphoma (diffuse histiocytic lymphoma by Rappaport classification) who have high lactic dehydrogenase (LDH) and/or bulky mediastinal or abdominal disease are destined to do poorly with conventional combination chemotherapy, with a 2-year disease-free survival of about 20%. Patients who relapse after conventional combination chemotherapy have a similar poor prognosis. Thirty-one such patients with lymphoma were studied to evaluate the efficacy of intensive radiotherapy (hyperfractionated total body irradiation [TBI] [1,320 rad]), and cyclophosphamide (60 mg/kg/d for two days) followed by autologous bone marrow transplantation (ABMT). Our results show a disease-free survival advantage (P = .002) for 14 patients who underwent ABMT immediately after induction of remission with 79% surviving at a median follow-up 49.2+ months, compared with a median survival of 5.2 months for 17 patients administered ABMT while in relapse and/or after failing conventional treatment. Our results support the use of aggressive therapy as early treatment for patients with poor prognostic features.
Adolescent↗
Lymphoblastic lymphoma in adults.
Fifty-one patients with lymphoblastic lymphoma (LBL) treated with one of five successive intensive chemotherapy protocols for acute lymphoblastic leukemia (ALL) since 1971 were reviewed. The patients were divided into leukemic and nonleukemic groups, and their clinical and laboratory parameters compared. The projected 5-year survival rate for all patients treated with the L10/17 protocols was 45% for both leukemic and nonleukemic LBL. The response to treatment was compared with that of 111 patients with ALL and was nearly identical. Poor prognostic factors were age beyond 30, WBC greater than 50,000/microL, failure to achieve a complete response (CR), and a late CR during induction. Leukemia at presentation, T cell surface markers, and the presence of a mediastinal mass did not adversely affect survival. The use of intensive chemotherapy protocols has proven to be a significant advance in the treatment of LBL.
Adolescent↗
Chicken triosephosphate isomerase complements an Escherichia coli deficiency.
We present the sequence of full-length chicken triosephosphate isomerase (D-glyceraldehyde 3-phosphate ketol-isomerase, EC 5.3.1.1) mRNA based on the analysis of cDNA and genomic clones. To isolate cDNA clones encoding the enzyme, we screened a muscle cDNA library with radioactively labeled cDNA made from RNA that had been enriched by immunoselection of polysomes. We blocked the signal caused by contaminating species in the probe with cloned DNA corresponding to the contaminants. Screening a chicken genomic library with cDNA coding for triosephosphate isomerase led to the isolation of phage containing the entire gene, which we used to map the transcriptional start. When placed downstream from a hybrid trp-lac promoter, the cDNA encoding the chicken enzyme programs the synthesis of functional protein, as judged by enzymatic criteria and by complementation of an Escherichia coli mutant that is deficient in bacterial triosephosphate isomerase.
Animals↗
Active site of triosephosphate isomerase: in vitro mutagenesis and characterization of an altered enzyme.
We have replaced the glutamic acid-165 at the active site of chicken triosephosphate isomerase with an aspartic acid residue using site-directed mutagenesis. Expression of the mutant protein in a strain of Escherichia coli that lacks the bacterial isomerase results in a complementation phenotype that is intermediate between strains that have no isomerase and strains that produce either the wild-type chicken enzyme or the native E. coli isomerase. The value of kcat for the purified mutant enzyme when glyceraldehyde 3-phosphate is the substrate is 1/1500th that of the wild-type enzyme, and the Km is decreased by a factor of 3.6. With dihydroxyacetone phosphate as substrate, the kcat value is 1/240th that of the wild-type enzyme, and Km is 2 times higher. The value of Ki for a competitive inhibitor, phosphoglycolate, is the same for the mutant and wild-type enzymes, at 2 X 10(-5) M. By treating the enzyme-catalyzed isomerization as a simple three step process and assuming that substrate binding is diffusion limited, it is evident that the mutation of glutamic acid-165 to aspartic acid principally affects the free energy of the transition state(s) for the catalytic reaction itself.
Animals↗
Genetic engineering in the Precambrian: structure of the chicken triosephosphate isomerase gene.
We report the sequence of the single chicken triosephosphate isomerase gene and its flanking regions. The 3-kilobase-long gene is composed of seven similarly sized exons and six introns. By using crystallographic and sequence data, we argue that this ancient gene was originally assembled from the genetic antecedents of exons.
Amino Acid Sequence↗
Treatment of diffuse poorly differentiated lymphocytic lymphoma. An analysis of prognostic variables.
Forty patients with diffuse poorly differentiated lymphocytic lymphoma (DPDL) Stages II-IV were treated with three different and successive combination chemotherapy protocols. Seventeen patients were treated with the cyclophosphamide (CTX) L2 protocol which included maintenance chemotherapy for 3 years. Only three patients were treated with the NHL-3 (non-Hodgkin's lymphoma) protocol, and 20 patients received the NHL-5 program. All protocols included radiotherapy (1500-4800 rad) to areas of initial bulky disease or persistent tumor, as well as central nervous system (CNS) prophylaxis with intrathecal methotrexate in patients with bone marrow involvement. Seventy-eight percent of local recurrences occurred in previously irradiated areas. Two-year survival rates were 55% and 70% for the CTX-L2 and NHL-5 protocols, respectively. Median disease-free survival for 24 complete response (CR) patients was 16.5 months. Of the 40 patients, 37 were evaluable for response to therapy. The CTX-L2 produced an 80% total response (TR) rate, a 60% CR, and a 20% partial response (PR). The patients on the NHL-5 achieved a TR rate of 95%, 74% CR, and 21% PR. Differences in TR and CR between the two protocols were not significant. Only 1 of 3 patients on the NHL-3 protocol achieved a CR. There was a trend for age greater than 50 years to lessen the chances of CR (P = 0.091); however, sex, symptoms, stage of disease, and LDH level were not significantly related to CR rate. Response to treatment (CR versus PR versus failure) was the most important factor influencing survival (P less than 0.001); age (greater than 50 years) was also significant (P = 0.008). Lactate dehydrogenase (LDH) was of borderline significance (P = 0.06). Cox regression model showed age (greater than 50 versus less than 50 years, P = 0.001), LDH (greater than 500 versus less than or equal to 500 U/L, P = 0.019) and symptoms (A or B) to be the best predictors of survival.
Adult↗
Primary gastrointestinal lymphoma: a 30-year review.
The authors reviewed all cases of non-Hodgkin's lymphoma primarily involving the gastrointestinal tract treated at Memorial Hospital during the period from 1949-1978. Complete clinical records were available in 104 cases. Slides of original pathology specimens were available in 81 cases. Tumors were classified by Rappaport, Lukes-Collins and modified Kiel classifications. All patients were staged retrospectively, using modified Ann Arbor staging. The primary tumor was in the stomach in 76 patients, in the small bowel in 15 and in the large bowel in 13. The life-table survival for all patients at five years was 44% and for the 81 Stage I and II patients it was 53%. We found a trend toward improved survival for patients treated in the last decade (P = 0.05). Using Cox regression analysis, survival was found to be correlated with stage (P less than 0.0001) and involvement of adjacent structures (P = 0.007). For Stage I patients, resection and radiation therapy were equally effective alone in controlling local tumor even though factors responsible for the selection of either treatment could not be identified. For Stage II patients, resection combined with radiation therapy controlled local disease better than either treatment alone. For Stage II, patient survival was correlated with the pattern of nodal involvement (P less than 0.0001). Neither the choice of treatment (resection, radiation therapy, or resection with radiation therapy; P = 0.17) nor the involvement of resected margins (P = 0.22) affects survival. Among 81 Stage I and II patients, 68% had recurrences outside the primary field of treatment and 60% outside the abdomen. Systemic multiple modality therapy should be considered for patients at high risk for recurrence.
Adult↗
Primary gastric lymphoma. A review of 50 cases.
The records of 50 patients with localized primary gastric lymphoma were reviewed and clinical and prognostic factors characterized. Pathologic material was reclassified according to Rappaport's, Lukes-Collins, and Lennert's Kiel classifications. Factors with the greatest prognostic significance included initial stage as determined by surgery and pathology, absolute tumor size, degree of penetration through the stomach wall, and histologic grade of the lymphoma. After surgical resection for cure, the overall 5-year disease-free survival was 47%. For stage I disease, this was 78% vs 29% for stage II (P = 0.006). Patients with lymphomas less than 5 cm in diameter had 58% 5-year disease-free survival vs 32% for those with tumors greater than 10 cm (P = 0.06). Full-thickness penetration decreased 5-year survival from 75% to 38% (P = 0.06). Patients with histologically low-grade lymphomas had a better prognosis than those with high-grade lymphomas. The most significant correlation of histology to survival was seen with the Kiel classification with a 5-year survival of 39% for centroblastic polymorphous lymphoma vs 66% for LP immunocytoma. When lymphoma recurred it developed outside the abdomen in a majority of patients. The addition of abdominal radiation therapy to surgical resection made no significant impact on survival for either stage I or II disease.
Adult↗
Combination chemotherapy of advanced chronic lymphocytic leukemia: the M-2 protocol (vincristine, BCNU, cyclophosphamide, melphalan, and prednisone).
The M-2 protocol (vincristine, cyclophosphamide, BCNU, melphalan, and prednisone) was administered monthly to 63 evaluable patients with advanced chronic lymphocytic leukemia. Complete remission (absence of all clinical and bone marrow evidence of leukemia) and partial response (greater than 50% decrease in organ enlargement and reduction of WBC count to below 15,000 x 10(6)/liter) were achieved in 17% and 44%, respectively, for a total response rate of 61%. The median survivals from therapy of patients achieving a CR, RR, or no response were 73+, 40, and 14 mo respectively. The median survival time from onset of treatment for stages II, III, and IV disease were 47, 20 and 19 mo, respectively, which was not statistically different from historical controls. However, when untreated patients are compared to this latter group, a significant survival advantage from diagnosis was found (p = 0.01), stressing the importance of prior therapy as the only unfavorable prognostic factor. Although complete remissions in CLL, as reflected in apparently normal bone marrow B-lymphocyte markers, can be induced wih acceptable morbidity, the majority of patients relapse after cessation of therapy. An alternative approach to the M-2 protocol will be needed to eradicate the disease.
Antineoplastic Agents↗
Studies in acute leukemia. II. Chemotaxis of leukemic blasts from patients with acute nonlymphoid leukemia: its relationship with IgG Fc receptors and role of surface-bound immunoglobulin.
Leukemic blasts from patients with acute myeloblastic and acute monocytic leukemia were studied for their chemotaxis towards casein- or endotoxin-activated serum as well as for the presence of Fc receptors and surface immunoglobulins. Leukemic blasts from patients with acute myeloblastic leukemia lacked both Fc receptors and surface immunoglobulin and did not migrate towards chemoattractants. Leukemic blasts from patients with acute monocytic leukemia, however, demonstrated heterogeneity with regard to Fc receptors, surface immunoglobulins and chemotaxis, and could be divided into three groups: in group I, leukemic blasts which lacked both Fc receptors and surface immunoglobulin moved poorly; in group II, large proportions of leukemic blasts and Fc receptors but lacked surface immunoglobulins. These blasts migrated well towards the chemoattractants, and in group III, leukemic blasts which had both Fc receptors and surface immunoglobulins lacked chemotactic property. A correlation between the presence or absence of Fc receptors and chemotaxis was observed. In group III, lack of blast cell migration appears to be due to passive binding of IgG via the Fc receptors.
Acute Disease↗
Studies in acute leukemia. I. Antibody-dependent and spontaneous cellular cytotoxicity by leukemic blasts from patients with acute nonlymphoid leukemia.
Leukemic blasts from patients with acute nonlymphoid leukemia were examined for the presence of Ig, receptors for IgGFc, and for their capacity to mediate antibody-dependent cellular cytotoxicity (ADCC) against chicken red blood cells (RBC) coated with IgG and spontaneous cell-mediated cytotoxicity (SCMC) against cells of K562 cell line. Leukemic blasts from acute myeloblastic leukemia (AML) patients lacked both Fc receptors and Ig on their surface, had no SCMC activity and majority, but not all of them, lacked ADCC activity. Leukemic blasts from patients with acute monocytic leukemia (AMOL) had Fc receptors, and 50% had IgG on their surface. IgG was cytophilic and appeared not to be directed against cell-surface antigens. This antibody did not interfere with the ADCC activity of leukemic cells. Leukemic blasts from majority of patients with AMOL mediated ADCC, but had no SCMC activity. An association between ADCC and presence of Fc receptor was observed.
Adult↗
Phase II trial of vindesine in patients with malignant lymphoma.
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Adult↗
Making board meetings work: the Doyle/Straus interaction method.
The Doyle/Straus Interaction Method is just now beginning to be used at corporate board meetings on the West Coast. The methodology, based on ten years of experimentation with problem-solving and decisionmaking groups of all kinds, seems to hold a host of benefits not only for meetings at board level but for committee meetings of all kinds. Their step-by-step analysis of the problems that are caused by people and space promises much for the corporations that want to move the directorate into the 20th Century or even the 21st!
Decision Making↗