Absence of parvovirus and cytomegalovirus in red-cell aplastic crises in glucose-6-phosphate dehydrogenase deficiency.
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Adolescent↗
Biomedical subjects
R Reynolds
Publications and source records attributed to R Reynolds.
Children, bicycles and head injuries.
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Australia↗
High-affinity uptake of gamma-[3H]aminobutyric acid by isolated mouse oligodendrocytes in culture.
Oligodendrocytes were isolated from mixed glial cultures of neonatal mouse forebrain and further grown in serum-free hormone supplemented culture medium. Cell populations were identified by indirect immunofluorescence using a range of specific antibodies, revealing a predominantly immature population of oligodendrocytes, the majority expressing the myelin glycolipids galactocerebroside and sulfatide on their plasma membrane. Astroglial contamination was found to be minimal. Simultaneous autoradiography and immunofluorescence demonstrated the presence of a transport system for the major inhibitory neurotransmitter GABA in the oligodendrocytes. The transport system was found to be energy, sodium and temperature dependent. Kinetic analysis revealed a high affinity system, with a Km of 6.27 microM and Vmax of 0.714 nmol/min/mg protein, which is comparable to that found previously for CNS neurons and astrocytes.
Animals↗
Simultaneous immunofluorescence and autoradiography: a useful technique for investigating neurotransmitter uptake by neurons and glia in primary central nervous system culture.
Previous studies on the localization of radiolabelled neurotransmitters in cultured cells of neural origin have relied on the comparison of cell morphology, as determined by immunocytochemistry, with the patterns of labelling on autoradiograms. We present here a method combining simultaneously autoradiography, following the uptake of tritium-labelled amino acid transmitters, with indirect immunofluorescence using antibodies against both surface and intracellular antigens. Using a fixative containing only a low concentration of glutaraldehyde (4% paraformaldehyde, 0.1% glutaraldehyde), a similar retention of gamma-[3H]aminobutyric acid (GABA) and D-[3H]aspartate was achieved as with the higher concentrations commonly used, with the advantage that the autofluorescence associated with glutaraldehyde fixed tissue was eliminated, and the immunoreactivity of the antigens to be localized was not destroyed. Using this method GABA and D-aspartate accumulating cells, in dissociated mouse central nervous system (CNS) cultures, could be reliably identified as oligodendrocytes, and some multiprocessed astrocytes, by anti-galactocerebroside (GC) and anti-glial fibrillary acidic protein (GFAP) immunofluorescence respectively. GABA-accumulating neuron-specific enolase (NSE) positive neurons could be clearly identified but no D-aspartate accumulating neurons were found. This technique should have a wide application in the investigation of whether selective transport mechanisms coexist with antigens characteristic of a certain cell type or sub-type.
Animals↗
Oligodendroglial and astroglial heterogeneity in mouse primary central nervous system culture as demonstrated by differences in GABA and D-aspartate transport and immunocytochemistry.
Using simultaneous autoradiography and immunofluorescence we have investigated the functional heterogeneity amongst oligodendrocytes and astrocytes in primary mouse central nervous system (CNS) culture as expressed by differences in their ability to accumulate gamma-[3H]aminobutyric acid [( 3H]GABA) and D-[3H]aspartate. We have used a range of specific antibodies that identify oligodendrocytes and astrocytes, from precursor to fully mature cells, to address the question of whether all neuroglial cells are capable of expressing this function. Our results showing that A2B5-, 03-, and galactocerebroside-positive cells became heavily labelled with these two neuroactive amino acids, whereas cells expressing the myelin proteins 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) and myelin basic protein (MBP) did not, demonstrate that this capacity is already present in oligodendrocytes at early developmental stages but may not extend to fully mature cells. Astrocytes in culture exhibited a large degree of variability with respect to their ability to transport GABA and D-aspartate. When grown in either serum-containing or serum-free hormone supplemented culture medium two morphologically distinct of glial fibrillary acidic protein (GFAP)-positive astrocyte were identified, process-bearing and epithelioid. Process-bearing cells became heavily labelled with the amino acids under both growth conditions, whereas, data showed that although epithelioid astrocytes were not, or only lightly, labelled with either amino acid in serum-containing cultures, when grown in serum-free culture medium they became more heavily labelled. Thus the expression, in culture, by epithelioid astrocytes, of one of the functions attributed to these cells is largely dependent on growth conditions.
Animals↗
Selective uptake of neuroactive amino acids by both oligodendrocytes and astrocytes in primary dissociated culture: a possible role for oligodendrocytes in neurotransmitter metabolism.
CNS glia may be involved in the modulation of neuronal excitability through their capacity to accumulate and metabolize neuroactive amino acids. To investigate the possible role of oligodendrocytes in amino acid neurotransmitter metabolism, we have used light microscopic autoradiography, following the uptake of 3H-labelled amino acids by dissociated cultures of neonatal mouse brain, characterized immunocytochemically using cell-type specific markers. Oligodendrocytes, recognized by their characteristic galactocerebroside membrane staining, rapidly accumulated [3H] gamma-aminobutyric acid (GABA), becoming intensely labelled over cell body and processes after short incubations. In contrast, oligodendrocytes became only lightly labelled with [3H]L-glutamate and aspartate, which preferentially labelled astrocytes. [3H]D-aspartate, a non-metabolized analogue of L-glutamate, was avidly accumulated by oligodendrocytes, labelling cell bodies and processes after short incubations, to a similar extent as GABA. Thus, oligodendrocytes possess a transport mechanism for these excitatory amino acids, but rapidly metabolize them and release the metabolites. Not only the GC-positive cells but also the GC-negative undifferentiated oligodendrocyte precursors accumulated both GABA and D-aspartate, suggesting that this may be a function expressed early in the differentiation of oligodendrocytes. Net uptake of [3H] beta-alanine and [3H]glycine by oligodendrocytes was not observed under any conditions tested. A small number of oligodendrocytes were labelled with [3H]taurine after longer incubations. The uptake of certain neuroactive amino acids is thus a property shared by astrocytes and oligodendrocytes, the latter acting in a protective fashion around neuronal perikarya and axons.
Alanine↗
Long-term survival following relapse after 5-FU but not CMF adjuvant breast cancer therapy.
Beginning in 1974, patients with greater than or equal to 4 nodes positive following mastectomy were randomized to receive either 5-FU i.v. weekly or CMF i.v. every 2 weeks, both given for 12 months. Median follow-up now exceeds 112 months with nine year results below: (table; see text) Early results based on relapse-free survival favored CMF, but more patients currently are alive on the 5-FU arm. As the survival curves cross at 40 months, the 20% survival advantage for 5-FU did not achieve statistical significance. For 34% of patients failing adjuvant 5-FU, use of combination chemotherapy after relapse (commonly with CMFVP or CMF) resulted in long term survival. In contrast, long-term survival for patients failing adjuvant CMF was unusual. Relapse was detected while under weekly observation in a greater proportion of patients on 5-FU (36%) compared to CMF (6%) adjuvant treatment (p less than 0.05), potentially influencing tumor burden at recurrence. Hormonal therapy or radiation therapy as initial therapy after relapse was ineffective, with no long term survivors resulting on either arm. Weight increase on adjuvant chemotherapy was commonly seen, with weight increase greater than 10 kg associated with a poor prognosis. We conclude that initial improvement in relapse-free survival may not predict long term survival in adjuvant breast cancer trials since both the specific adjuvant therapy given pre-relapse as well as the type of salvage therapy given post-relapse may influence ultimate patient outcome.
Antineoplastic Combined Chemotherapy Protocols↗
Health promotion for educators: impact on absenteeism.
The impact of a comprehensive worksite health promotion program on employee absenteeism in a large metropolitan school district was evaluated. Participants in the program (n = 3,846) did not differ from nonparticipants (n = 8,290) in age or sex distributions. Program participants improved their physical fitness, body composition, coronary risk factors, and overall feeling of well-being. Absenteeism was determined for the study year and for the previous year from records in the district personnel office. Absenteeism differences were evaluated by analysis of covariance by using age, sex, ethnic group, and the previous year's absenteeism as covariates. Participants who completed the health promotion program had an average of 1.25 days less absenteeism (P less than 0.0001) during the study year than nonparticipants. Regression analyses indicated that improvement in physical fitness was associated with less absenteeism. The results suggest that a reduction in absenteeism due to the health promotion program was possibly associated with an improvement in physical fitness.
Absenteeism↗
Biodegradable resurfacing of the hip in dogs.
Arthroplasties of the hips in dogs using a biodegradable poly-L-lactide cup to protect the regenerating cartilage following removal of normal articular cartilage and the drilling of multiple full thickness defects into subchondral bone were performed to determine whether or not the repair response could be improved. The poly-L-lactide cup degraded in approximately 60 days. The regenerating cartilage was much better in the experimental than the control dogs initially, but ultimately failed, perhaps due to inadequate duration of protection of the immature cartilage by the arthroplasty material.
Animals↗
Prednimustine in refractory non-Hodgkin's lymphoma: a phase II study of the Northern California Oncology Group.
Fifty-six patients with advanced non-Hodgkin's lymphoma (NHL) were entered into a phase II study of prednimustine, an ester of chlorambucil and prednisolone. All patients were refractory to extensive prior combination chemotherapy. Therapy with prednimustine, 100 mg/m2/day orally, was given for three consecutive days every 2 weeks. The overall response rate in 43 evaluable patients was 30% (13/43), with 9% (4/43) achieving complete response (CR) and 21% (9/43) achieving partial response (PR). In the favorable histology subgroup (23 patients), the response rate was 39% (9/23), with 4% (1/23) achieving CR and 35% (8/23) achieving PR. In the unfavorable histology subgroup (20 patients), responses were seen in 20% (4/20) with 15% (3/20) achieving CR, all in heavily pretreated diffuse histiocytic lymphoma. Toxicity of this regimen was mild, with leukopenia below 3,000/mm3 in 22% and thrombocytopenia below 90,000/mm3 in 16% of patients. A positive correlation was observed between response and hematologic toxicity, indicating the potential for a dose-escalation schedule in future trials. These data confirm activity of prednimustine in NHL refractory to standard treatment. In view of its relatively mild toxicity, we conclude that prednimustine is an appropriate agent to test in combination chemotherapy regimens in this group of lymphomas.
Adult↗
Release of arachidonate from membrane phospholipids in cultured neonatal rat myocardial cells during adenosine triphosphate depletion. Correlation with the progression of cell injury.
The present study utilized a cultured myocardial cell model to evaluate the relationship between the release of arachidonate from membrane phospholipids, and the progression of cell injury during ATP depletion. High-energy phosphate depletion was induced by incubating cultured neonatal rat myocardial cells with various combinations of metabolic inhibitors (deoxyglucose, oligomycin, cyanide, and iodoacetate). Phospholipid degradation was assessed by the release of radiolabeled arachidonate from membrane phospholipids. In this model, the current study demonstrates that (a) cultured myocardial cells display a time-dependent progression of cell injury during ATP depletion; (b) the morphologic patterns of mild and severe cell injury in the cultured cells are similar to those found in intact ischemic canine myocardial models; (c) cultured myocardial cells release arachidonate from membrane phospholipids during ATP depletion; and (d) using two separate combinations of metabolic inhibitors, there is a correlation between the release of arachidonate, the development of severe cellular and sarcolemmal damage, the release of creatine kinase into the extracellular medium, and the loss of the ability of the myocardial cells to regenerate ATP when the metabolic inhibitors are removed. Thus, the present results suggest that during ATP depletion, in cultured neonatal rat myocardial cells, the release of arachidonate from myocardial membrane phospholipids is linked to the development of membrane defects and the associated loss of cell viability.
Adenosine Triphosphate↗
The use of topical cocaine to prevent laryngospasm after general anesthesia on endoscopy procedures.
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Adult↗
Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results.
We describe the development of a simple and highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) for measuring IgG and IgM anticardiolipin antibodies (ACA). Microtitre plates were coated with cardiolipin at a concentration of 45 micrograms/ml by evaporation under nitrogen. Non-specific binding of diluted sera was eliminated by blocking of plates with 10% fetal calf serum in phosphate buffered saline (PBS/FCS) for 2 h. Then sera (100 microliters) at a dilution of 1:100 were incubated in the wells for 1 h. Affinity purified goat anti-human IgG or IgM (100 microliters) at a concentration of 1 microgram/ml was subsequently added and allowed to incubate for 1 h; detection of ACA was achieved using an alkaline phosphatase conjugated rabbit anti-goat IgG reagent by reading the colorimetric yield at 405 nm after incubation with substrate. Reference serum pools were established to study reproducibility of the assay throughout its sensitivity range, and Standard curves were established. The quantitative normal range was 0-9.0 Anticardiolipin ELISA Units (AEU) for IgG and 0-8.0 (AEU) for IgM-ACA. A strong correlation was found between the ELISA and radioimmunoassay methods for measuring ACA of both IgG and IgM classes. Results from 65 patients with systemic lupus erythematosus (SLE) and 45 patients with seropositive rheumatoid arthritis are also reported. The advantages of the ELISA method for quantitative determination of ACA levels, should make it a useful and reliable method for clinical and experimental monitoring of patients with SLE and associated autoimmune disorders.
Arthritis, Rheumatoid↗
Uptake of [3H]GABA by oligodendrocytes in dissociated brain cell culture: a combined autoradiographic and immunocytochemical study.
Uptake of [3H]GABA by dissociated mixed cell cultures of fetal mouse brain was studied using light microscopic autoradiography. Major cell types in the cultures were identified and quantified by immunocytochemical localization of reliable cell type-specific antigenic markers. In 12 days in vitro (DIV) cultures [3H]GABA uptake was predominantly into neurons and oligodendrocytes, whilst at 28 DIV the only surface cells labeled were oligodendrocytes. This was confirmed by complement-dependent antibody-mediated cytotoxicity against galactocerebroside-positive oligodendrocytes. There was a moderate labeling of almost all flat cells, the majority of which were glial fibrillary acidic protein (GFAP)-positive astrocytes. Heavily labeled astrocytes were only occasionally observed. Oligodendrocytes accumulated [3H]GABA more rapidly than astrocytes but slower than neurons. Oligodendroglial labeling was predominantly over the cell body, whereas neuronal labeling was more uniformly distributed over cell body and processes. The uptake was inhibited by diaminobutyric acid (DABA) and nipecotic acid, but not by beta-alanine, and thus had similar characteristics to neuronal rather than astroglial uptake. Oligodendrocytes did not accumulate [3H]beta-alanine, which labeled only astrocytes. Oligodendroglial [3H]GABA uptake was Na+-dependent and sensitive to ouabain, but was only slightly enhanced by aminooxyacetic acid (AOAA), whereas astroglial uptake was not sensitive to ouabain but was markedly enhanced by AOAA. The results indicate that oligodendrocytes, in addition to astrocytes, may also be involved in the modification of neuronal function by the uptake and inactivation of neuroactive substances.
Animals↗
Chemotherapy of small-cell carcinoma of lung: a randomized comparison of alternating and sequential combination chemotherapy programs.
One hundred forty-seven eligible patients with small-cell carcinoma of the lung (SCCL) have been randomized to receive alternating (A) or sequential (S) combination chemotherapy. Initial treatment was with three cycles of VAM (A) or two cycles of POCC (S). VAM consists of VP16-213 200 mg/m2 intravenously (IV) day 1, Adriamycin (Adria Laboratories, Columbus, Ohio) 50 mg/m2 IV day 1, and methotrexate 30 mg/m2 IV day 1 repeated at 21-day intervals. POCC consists of cyclophosphamide 600 mg/m2 IV days 1 and 8, vincristine 1.5 mg/m2 (maximum, 2 mg) IV days 1 and 8, CCNU 60 mg/m2 po day 1, and procarbazine 100 mg/m2 po days 2 through 15. After initial treatment, all patients received whole brain radiation therapy (3,000 rad/10 fractions/2 wk). Patients with limited disease in addition received irradiation encompassing the tumor, hilar, mediastinal, and supraclavicular regions (5,000 rad/25 fractions/5 wk). After radiation, patients on arm A received POCC alternating with VAM; patients on arm S received POCC until progression when they were to be treated with VAM. The alternating arm was superior with respect to rate of complete remission (CR), median disease-free survival (MDFS), and median survival (MS). The advantage of alternating therapy was not as clearly demonstrated in the limited disease groups when interposition of involved field radiation delayed the initiation of the alternating schedule. In limited disease alone, comparing arm A with arm S, no statistically significant differences were noted. The CR rate was 42% v 54%, MDFS was 14 v 10 months, and MS was 16 v 10 months. In extensive disease, the CR rate was 44% v 20% (P = .03), MDFS was 6 v 4 months (P = .003), and MS was 10 v 7 months (P = .001). Improved treatment outcome in SCCL is achieved when combination chemotherapy regimens of similar effectiveness are administered in an alternating rather than sequential schedule.
Adult↗
Health promotion for educators: impact on health behaviors, satisfaction, and general well-being.
A random sample of 117 teachers in three treatment schools and one control school participated in a health survey at the beginning and end of the spring semester. Teachers in the treatment schools participated in a 10-week health promotion program which emphasized exercise, stress management, and nutrition. Comparison of pre- and post-survey data indicated that teachers in the treatment schools increased their participation in vigorous exercise, improved their physical fitness, lost weight, lowered their blood pressure, reported a higher level of general well-being, and were better able to handle job stress.
Adult↗
5-Fluorouracil + Oncovin + Adriamycin + mitomycin C (FOAM): an effective program for breast cancer, even for disease refractory to previous chemotherapy. A Northern California Oncology Group (NCOG) Study.
One hundred and thirty-one patients (118 evaluable) with disseminated breast cancer were treated with a combination of 5-fluorouracil + Oncovin + Adriamycin + mitomycin C (FOAM). The objective response rate for 82 evaluable patients whose disease was refractory to previous CMF or L-PAM chemotherapy was 35%; that for 36 evaluable patients who had not previously received chemotherapy, 56%. The hematologic toxicity of this therapy was generally mild and acceptable. It is believed that FOAM is an effective therapy for patients whose tumors are resistant to CMF.
Adult↗