Implications of p75NTR for dentate gyrus morphology and hippocampus-related behavior revisited.

The pan-neurotrophin receptor p75NTR is expressed in the adult brain in a discrete pattern. Although numerous studies have addressed its implications for hippocampal functions, the generated sets of data are surprisingly conflicting. We have therefore set out to re-investigate the impact of a deletion of the full-length p75NTR receptor on several parameters of the dentate gyrus (DG), including neurogenesis and hippocampus-related behavior by using p75NTR(ExIII) knockout mice. Moreover, we investigated further parameters of the DG (cholinergic innervation, dendritic spines). In addition, we analyzed on the morphological level the impact of aging by comparing adult and aged p75NTR(ExIII) mice and their age-matched littermates. Adult (4-6 months old), but not aged (20 months old), p75NTR(ExIII) knockout mice display an enhanced volume of the DG. However, adult neurogenesis within the adult DG was unaffected in both adult and aged p75NTR(ExIII) knockout mice. We could further demonstrate that the change in the volume of the DG was accompanied by an increased cholinergic innervation and increased spine densities of granule cells in adult, but not aged p75NTR deficient mice. These morphological changes in the adult p75NTR deficient mice were accompanied by specific alterations in their behavior, including altered behavior in the Morris water maze test, indicating impairments in spatial memory retention.

Biomechanical comparison of conventional open reduction and internal fixation versus calcium phosphate cement fixation of a central depressed tibial plateau fracture.

OBJECTIVE: To evaluate the effect of calcium phosphate bone cement on stability and strength of the fracture repair in a central depressed tibial plateau fracture cadaveric model. DESIGN: Paired human cadaveric tibial specimens. SETTING: Biomechanics laboratory. PATIENTS: Uniform pure depression fractures of lateral tibial plateau were created in twenty human cadaveric tibial specimens. INTERVENTION: The first part of the study used thirteen pairs of tibiae in two groups: a control group receiving the conventional treatment of morselized bone graft and two cancellous screws and an experimental group receiving calcium phosphate bone cement only. The second part of the study used seven pairs of tibiae in two experimental groups: one receiving calcium phosphate bone cement with a more extensive void preparation and the other group receiving calcium phosphate bone cement with a more extensive void preparation and two screws. MAIN OUTCOME MEASUREMENTS: Each tibia was loaded on a Material Testing Systems machine from twenty newtons to 250 newtons for 10,000 cycles to simulate immediate postoperative load transmission to the tibial plateau. Specimens were then loaded to failure to determine the ultimate strength of the reconstruction. Displacement of the articular fragment and stiffness at each cycle were measured during dynamic loading. Peak load, deformation at peak load, and resistance to depression were measured during the load to failure. RESULTS: The treatment of depressed tibia plateau fractures with a calcium phosphate cement provides equivalent or better stability than conventional open reduction and internal fixation in pure depression tibial plateau fractures. If the fracture void is prepared by eliminating the cancellous bone under the subchondral plate, the results are further improved. CONCLUSIONS: This study suggests that the non-weight-bearing postoperative period may be significantly reduced without clinically significant articular collapse.

Biomechanical evaluation of calcium phosphate cement-augmented fixation of unstable intertrochanteric fractures.

OBJECTIVE: The purpose of this study was to evaluate the mechanical effects of using an injectable calcium phosphate cement, Norian Skeletal Replacement System (SRS), which is replaced by the native remodeling process, to augment sliding hip screw fixation of unstable intertrochanteric fractures in a cadaver model. DESIGN: Ten matched pairs of human cadaver femora were used. One randomly selected femur from each pair was designated as experimental, whereas the contralateral femur served as the control. SETTING: Testing was performed in the Orthopaedic Research Laboratories, The University of Michigan, Ann Arbor, Michigan, U.S.A. PATIENTS/PARTICIPANTS: Femora with no radiographic evidence of pathology and with below-normal bone mineral density in the neck region were accepted into the study. INTERVENTION: Three-part, intertrochanteric fractures were repaired by using a sliding hip screw and sideplate, without replacement of the posteromedial fragment. In the experimental side of each pair, SRS was used to grout the hip screw and to fill the posteromedial defect. MAIN OUTCOME MEASUREMENTS: Femora were subjected to simulated single-leg stance loading to approximately one body weight in a servohydraulic testing machine. Measurements of stiffness, medial bone surface strain, hip screw displacement, and sideplate strain were made in fractured femora. Stiffness and medial bone surface strain baseline data were obtained in the intact bone before fracture creation as well. RESULTS: Augmenting fixation with Norian SRS increased the fracture construct stiffness and minimized sliding hip screw displacement. It also maintained medial bone surface strain closer to the intact state and lowered sideplate strain relative to controls. However, there was no difference in the load to failure between SRS-augmented and control femora. CONCLUSIONS: SRS augmentation of unstable, intertrochanteric fractures significantly improved overall stability, facilitated load transfer across the fracture, and decreased both shortening of the proximal femur and stress on the sliding hip screw.

Biomechanical evaluation of fixation of intra-articular fractures of the distal part of the radius in cadavera: Kirschner wires compared with calcium-phosphate bone cement.

BACKGROUND: The purpose of this study was to compare the biomechanical efficacy of an injectable calcium-phosphate bone cement (Skeletal Repair System [SRS]) with that of Kirschner wires for the fixation of intraarticular fractures of the distal part of the radius. METHODS: Colles fractures (AO pattern, C2.1) were produced in ten pairs of fresh-frozen human cadaveric radii. One radius from each pair was randomly chosen for stabilization with SRS bone cement. These ten radii were treated with open incision, impaction of loose cancellous bone with use of a Freer elevator, and placement of the SRS bone cement by injection. In the ten control specimens, the fracture was stabilized with use of two horizontal and two oblique Kirschner wires. The specimens were cyclically loaded to a peak load of 200 newtons for 2000 cycles to evaluate the amount of settling, or radial shortening, under conditions simulating postoperative loading with the limb in a cast. Each specimen then was loaded to failure to determine its ultimate strength. RESULTS: The amount of radial shortening was highly variable among the specimens, but it was consistently higher in the Kirschner-wire constructs than in the bone fixed with SRS bone cement within each pair of radii. The range of shortening for all twenty specimens was 0.18 to 4.51 millimeters. The average amount of shortening in the SRS constructs was 50 percent of that in the Kirschner-wire constructs (0.51+/-0.34 compared with 1.01+/-1.23 millimeters; p = 0.015). With the numbers available, no significant difference in ultimate strength was detected between the two fixation groups. CONCLUSIONS: This study showed that fixation of an intra-articular fracture of the distal part of a cadaveric radius with biocompatible calcium-phosphate bone cement produced results that were biomechanically comparable with those produced by fixation with Kirschner wires. However, the constructs that were fixed with calcium-phosphate bone cement demonstrated less shortening under simulated cyclic load-bearing.

Superior compressive strength of a calcaneal fracture construct augmented with remodelable cancellous bone cement.

Twenty-six paired, fresh-frozen cadaveric feet were disarticulated at the ankle joint, and the dome of the talus was potted. Stress-risers were placed along the medial, lateral, and posterior aspects of the calcaneus, and the specimen was loaded rapidly to failure in a testing machine to produce a type-IIB displaced intra-articular fracture according to the classification system of Sanders et al. One specimen of each pair was treated with standard internal fixation with bone-grafting (the control group), and the other was treated with similar fixation but with SRS (Skeletal Repair System) calcium phosphate bone cement placed in any osseous defect. All of the specimens were cured for twenty-four hours in a bath of saline solution at 37 degrees Celsius. The specimens were tested cyclically for ten cycles from zero to 100 newtons at one hertz and for 1010 cycles from zero to 350 newtons at one hertz. The deformation per cycle (millimeters per cycle), first-cycle deformation (millimeters), number of cycles to failure, and number of specimens withstanding the cyclical testing were calculated. The specimens were examined radiographically before and after fracture and after reconstruction and testing. A large difference in the results of the cyclical testing was noted. The specimens that had been augmented with the SRS bone cement had an average deformation of 0.00195 millimeter per cycle compared with 1.013 millimeters per cycle in the control group (p < 0.005). A similar magnitude of difference was noted when the results were stratified for good and poor-quality bone. Visual examination and radiographs demonstrated that a type-IIB displaced intra-articular fracture had been created reproducibly, and computed tomographic scans showed that nearly anatomical reconstruction had been achieved in all of the specimens. The computerized tomographic scans revealed good filling of the osseous voids and no evidence of failure of the cement after cyclical loading.

Histological, chemical, and crystallographic analysis of four calcium phosphate cements in different rabbit osseous sites.

Four calcium phosphate cement formulations were implanted in the rabbit distal femoral metaphysis and middiaphysis. Chemical, crystallographic, and histological analyses were made at 2, 4, and 8 weeks after implantation. When implanted into the metaphysis, part of the brushite cement was converted into carbonated apatite by 2 weeks. Some of the brushite cement was removed by mononuclear macrophages prior to its conversion into apatite. Osteoclastlike cell mediated remodeling was predominant at 8 weeks after brushite had converted to apatite. The same histological results were seen for brushite plus calcite aggregate cement, except with calcite aggregates still present at 8 weeks. However, when implanted in the diaphysis, brushite and brushite plus calcite aggregate did not convert to another calcium phosphate phase by 4 weeks. Carbonated apatite cement implanted in the metaphysis did not transform to another calcium phosphate phase. There was no evidence of adverse foreign body reaction. Osteoclastlike cell mediated remodeling was predominant at 8 weeks. The apatite plus calcite aggregate cement implanted in the metaphysis that was not remodeled remained as poorly crystalline apatite. Calcite aggregates were still present at 8 weeks. There was no evidence of foreign body reaction. Osteoclastlike cell remodeling was predominant at 8 weeks. Response to brushite cements prior to conversion to apatite was macrophage dominated, and response to apatite cements was osteoclast dominated. Mineralogy, chemical composition, and osseous implantation site of these calcium phosphates significantly affected their in vivo host response.

Biomechanical and histological evaluation of a calcium phosphate cement.

It is often difficult to achieve stable fixation of a comminuted fracture associated with a metaphyseal defect. The injection of a resorbable cement into an osseous defect may help to stabilize the fracture and to maintain osseous integrity as the cement is resorbed and replaced by bone. The purpose of the present study was to evaluate the repair of a metaphyseal defect after treatment with an injectable calcium-phosphate cement. The injectable cement undergoes isothermic curing in vivo to form a carbonated apatite (dahllite) with a compressive strength of twenty-five megapascals. Either the cement or allograft bone was placed in proximal tibial metaphyseal and distal femoral metaphyseal defects in seventy-two dogs and was evaluated from twenty-four hours to seventy-eight weeks postoperatively. Histological examination showed that the cement was osteoconductive; nearly the entire surface area was covered with bone two weeks after the injection. The resulting bone-cement composite underwent gradual remodeling over time in a pattern that was qualitatively similar to the remodeling of normal cortical and cancellous bone. Osteoclasts were found to resorb the cement and were usually associated with adjacent new-bone formation. With increasing time in vivo, the cement was penetrated by small blood vessels that became surrounded by circumferential lamellae of bone and that closely resembled evolving haversian systems. This process occurred more rapidly in the cortex than in the medulla. Mechanical testing showed that, by eight weeks, the tibiae that had been treated with cement had reached nearly 100 per cent of the torsional strength of the contralateral, control (intact) tibiae; this finding paralleled the histological observations of bone apposition to the cement and rapid restoration of the cortex. At no time was fibrous tissue present between the cement and the bone, and there was no evidence of acute inflammation. Small particles of cement were present within occasional macrophages during the process of cement resorption, but the macrophages disappeared over time and were not associated with fibrosis or unexpected resorption of bone. Resorption of the cement was incomplete in the medullary area at seventy-eight weeks, but the pattern of cement resorption and bone-remodeling suggested gradual restoration of a physiological proportion of bone and marrow in both the cortical and the medullary region with maintenance of mechanical function.

Norian SRS cement augmentation in hip fracture treatment. Laboratory and initial clinical results.

Bone quality, initial fracture displacement, severity of fracture comminution, accuracy of fracture reduction, and the placement of the internal fixation device are important factors that affect fixation stability. New high strength cements that are susceptible to remodeling and replacement for fracture fixation may lead to improved clinical outcome in the treatment of hip fractures. Norian SRS is an injectable, fast setting cement that cures in vivo to form an osteoconductive carbonated apatite of high compressive strength (55 MPa) with chemical and physical characteristics similar to the mineral phase of bone. It can be used as a space filling internal fixation device to facilitate the geometric reconstruction, load transfer, and healing of bone with defects and/or fractures in regions of cancellous bone. Furthermore, this cement can improve the mechanical holding strength of conventional fixation devices. Use of this material potentially could improve fracture stability, retain anatomy during fracture healing and improve hip function, thus achieving better clinical outcomes. In vivo animal studies have shown the material's biocompatibility, and cadaveric studies have shown the biomechanical advantage of its use in hip fractures. Initial clinical experience (in 52 femoral neck fractures and 39 intertrochanteric fractures) showed the potential clinical use of this innovative cement in the treatment of hip fractures.

Carbonated apatite cement augmentation of pedicle screw fixation in the lumbar spine.

STUDY DESIGN: Biomechanical testing with human cadaveric lumbar vertebral bodies was used to determine the utility of an injectable carbonated apatite cancellous bone cement for improving the structural performance of pedicle screws subjected to axial pull-out or transverse cyclic loading. OBJECTIVES: To ascertain whether augmentation with a carbonated apatite cement can enhance pedicle screw fixation in the lumbar spine. SUMMARY OF BACKGROUND DATA: The beneficial effects of polymethylmethacrylate augmentation on pedicle screw pull-out strength have been demonstrated. Cancellous bone cement, however, may provide an attractive alternative in this application, as it is remodelable, biocompatible, and nonexothermic. METHODS: Forty-three cadaveric lumbar vertebral bodies were instrumented with pedicle screws. In 20 of these specimens, axial pull-out strength was compared between the control screws and those augmented with cancellous bone cement. In the remaining 23 specimens, the screws were loaded in the superior-inferior direction with a peak displacement of +/- 1 mm at a frequency of 3 Hz for 5000 cycles. Three parameters were calculated from the force-versus-time data: 1) the energy dissipated, 2) the peak force at the start of the test, and 3) the peak force at the end of 5000 cycles. RESULTS: The pull-out strength of the augmented pedicles averaged 68% greater than that of the control side. In response to cyclic loading, all measures of bio-mechanical performance improved 30-63%. CONCLUSIONS: The data suggest that augmentation with this carbonated apatite cancellous bone cement can enhance immediate screw fixation.

Augmentation of femoral neck fracture fixation with an injectable calcium-phosphate bone mineral cement.

The first goal of this study was to determine if augmentation with an injectable, in situ setting, calcium-phosphate cement that is capable of being remodeled and was designed to mimic bone mineral significantly improved the strength and stiffness of fixation in a cadaveric femoral neck fracture model. The second goal was to determine if greater increases in fixation strength were achieved as the bone density of the specimen decreased. Sixteen pairs of fresh cadaveric human femora with a mean age of 70.9 years (SD = 17.2 years) were utilized. The bone density of the femoral neck was measured with dual-energy x-ray absorptiometry. The femoral head was impacted vertically with the femoral shaft fixed in 12 degrees of adduction using a materials testing machine to create a fully displaced fracture. Following fracture, 30% inferior comminution was created in each specimen. One randomly chosen femur from each pair underwent anatomic reduction and fixation with three cannulated cancellous bone screws, 7 mm in diameter, in an inverted triangle configuration. The contralateral femur underwent the same fixation augmented with calcium-phosphate cement. Specimens were preconditioned followed by 1.000 cycles to one body weight (611.6 N) at 0.5 Hz to simulate single-limb stance loading. The stiffness in the first cycle was observed to be significantly greater in cement-augmented specimens compared with unaugmented controls (p < 0.05). After cycling, each specimen was loaded at 10 mm/min until complete displacement of the fracture surface and failure of fixation occurred. Specimens augmented with bone mineral cement failed at a mean of 4,573 N (SD = 1,243 N); this was significantly greater (p < 0.01) than the mean for controls (3,092 N, SD = 1,258 N). The relative improvement in fixation strength (augmented/control x 100%) was not inversely correlated to femoral neck bone density (p = 0.25, R2 = 0.09), was weakly correlated to the volume of cement injected (p = 0.07, R2 = 0.22), and was inversely related to the fixation failure load of the control specimen (p = 0.001, R2 = 0.54). There was a mean relative improvement in fixation strength of 169.6% (SD = 77.5). These findings suggest that calcium-phosphate cement provides initial beneficial augmentation to fixation of femoral neck fractures.

Skeletal repair by in situ formation of the mineral phase of bone.

A process has been developed for the in situ formation of the mineral phase of bone. Inorganic calcium and phosphate sources are combined to form a paste that is surgically implanted by injection. Under physiological conditions, the material hardens in minutes concurrent with the formation of dahllite. After 12 hours, dahllite formation was nearly complete, and an ultimate compressive strength of 55 megapascals was achieved. The composition and crystal morphology of the dahllite formed are similar to those of bone. Animal studies provide evidence that the material is remodeled in vivo. A novel approach to skeletal repair is being tested in human trials for various applications; in one of the trials the new biomaterial is being percutaneously placed into acute fractures. After hardening, it serves as internal fixation to maintain proper alignment while healing occurs.

Bone remodeling and in vivo strain analysis of intact and implanted greyhound proximal femora.

Pre- and poststudy motion and gait analyses of eight size-matched male greyhounds confirmed uniform loading of their femora. Subminiature strain gages implanted on the intact inferior and anterior aspects of the femoral neck in six greyhounds indicated in vivo strain variations among test animals. Motion and gait analyses confirmed uniform loading of femora following unilateral hemiarthroplasty with cobalt-chromium hip implants. In vivo strain measurements adjacent to the implants indicated large variations among test animals. A consistent direction of strain change relative to the intact femur was noted, even though strain changes varied in magnitude. Image analysis of microradiographs indicated insignificant differences in the cortical areas of implanted and intact femora. Extensive new trabecular bone formation was noted along the implant in the endosteal cavity and correlated with a combination of implant placement and exercise level. Most of the bone was formed with centrally placed implants in exercised dogs, and the least with stems on the medial neck surface in rested dogs. Iliac crest biopsies indicated that bone formation rates slowed in rested animals and remained constant throughout the study in exercised animals. All implanted femora had a thin (< 1 mm thick) aligned fibrous tissue layer separating the implant from bone. It varied in thickness as a function of the aspect of the implant. Exercised dogs had a larger proportion of fibrous tissue on the anterior and posterior aspects, while rested dogs had a larger proportion of fibrous tissue on the medial and lateral aspects.

[Randomized double-blind study of therapy of sudden deafness. Low molecular weight dextran + naftidrofuryl vs. low molecular weight dextran + placebo]. / Randomisierte Doppelblindstudie zur Hörsturztherapie. Niedermolekulares Dextran + Naftidrofuryl vs. niedermolekulares Dextran + Plazebo.

Eighty patients with idiopathic sudden deafness existing no longer than 10 days were included in a prospective randomized double-blind study. Patients were treated for 10 days with infusions of either 10% low-molecular weight dextran or the combination of low-molecular weight dextran with naftidrofuryl. Before treatment and after 10 days hearing loss in the affected ear was determined at 0.5, 1, 2, 3, 4 and 6 kHz. The mean hearing loss was then calculated as the average from these values. During monotherapy with low-molecular weight dextran the mean hearing loss decreased from 40 to 27 dB compared to 38 to 17 dB when naftidrofuryl treatment was added (p < 0.01 between groups). A significant benefit of naftidrofuryl on hearing loss was also found at frequencies between 0.5 and 3 kHz. Furthermore, patients reported better improvement of tinnitus when naftidrofuryl was combined with dextran. Two patients receiving dextran alone developed side effects: one had an allergic reaction causing withdrawal of treatment, which the other case had vertigo, nausea and headache with spontaneous recovery. The results of the study showed that treatment with naftidrofuryl in addition to hemodilution with low-molecular weight dextran was of therapeutic benefit in the therapy of sudden deafness without increasing the rate of side effects.

[Frontal lobe abscess as a complication of frontal sinus osteoma]. / Frontalhirnabszess als Komplikation eines Stirnhöhlenosteoms.

A frontal brain abscess was caused by an osteoma of the frontal sinus. This osteoma had destroyed the posterior wall and was blocking the infundibulum. Under these preconditions a sinusitis frontalis was followed by an intracranial infection and a brain abscess.

Enhancement of folate analogue transport inward in L1210 cells during methotrexate therapy of leukemic mice: evidence of the nature of the effect, possible host mediation, and pharmacokinetic significance.

Studies are described that sought the basis for a discrepancy in values for a key kinetic parameter of methotrexate transport (influx Vmax) in L1210 cells derived alternately from biochemical or pharmacokinetic measurements. Our results show that, within a short period of time following administration of a therapeutic dose of methotrexate to leukemic mice, influx of this folate analogue measured in L1210 cells removed from these mice was markedly stimulated. Enhancement of [3H]methotrexate influx in these cells was observed within 15 min of drug administration, was maximum (up to 3-fold) within 2 to 3 h, then decreased with time until 24 h when influx was at the control level. Measurements of [3H]methotrexate influx in cells removed from drug-treated mice were made after a period of incubation in drug-free medium to allow for efflux of exchangeable drug. Enhanced influx of [3H]methotrexate was accounted for by an increase in influx Vmax (influx Km was unchanged) and was further enhanced (to a total of 5-fold) by coadministration of leucovorin. Also, enhancement of influx of [3H]methotrexate in L1210 cells did not occur following administration of 1-beta-D-arabinofuranosylcytidine at a therapeutically equivalent dose to leukemic mice or following exposure of these cells to methotrexate or methotrexate with leucovorin during growth in culture. Methotrexate therapy did not affect all transport systems, since the same therapy of leukemic mice had no effect on influx of the purine nucleoside analogue, 9-beta-D-arabinofuranosyl-2-fluoroadenine, in these same L1210 cells. These findings suggest that stimulation of [3H]methotrexate influx in L1210 cells during therapy with this folate analogue was not due to transstimulation during exchange between folate compounds and was not related to the antiproliferative effect of methotrexate on these tumor cells. The coadministration of cycloheximide with methotrexate to leukemic mice at a dose which markedly inhibited 3H-leucine incorporation into L1210 cell protein severely diminished the stimulation of [3H]methotrexate influx. However, in L1210 cells removed from leukemic mice treated with methotrexate, there was no increase compared to control cells in affinity labeling with the N-hydroxysuccinimide ester of [3H]methotrexate. This suggested that the effect of cycloheximide was not on increased synthesis of folate transporter and that increased rate of translocation of folate transporter, rather than increased amount of transporter, accounted for the increase in [3H]methotrexate influx.(ABSTRACT TRUNCATED AT 400 WORDS)

[Therapy of sudden deafness--naftidrofuryl (Dusodril) and pentoxifylline (Trental) compared]. / Zur Therapie des Hörsturzes--Naftidrofuryl (Dusodril) und Pentoxifyllin (Trental) im Vergleich.

In a prospectively randomized trial, conduced jointly by the University ENT Department, Göttingen, and the ENT Department of Zentralkrankenhaus St.-Juergen-Strasse, Bremen, the therapeutic efficiencies of naftidrofuryl and pentoxifyllin, both in combination with a basic medication (cortison, dextran), were compared in 151 patients with sudden deafness. Evaluation of the results revealed a decisive influence of cardiovascular risk factors (particularly vascular diseases) on the outcome of therapy. There was no difference between naftidrofuryl and pentoxifyllin with respect to therapeutic success. Since further improvement was evident 6 months later, long-term treatment is recommended for either substance. The therapeutic success rate of our own treatment schedule is not higher than that of other therapeutic schedules given in the literature.

Aggravation of arrhythmia induced with antiarrhythmic drugs during electrophysiologic testing.

There is evidence that antiarrhythmic drugs can worsen ventricular arrhythmias in patients. In a previous study ventricular arrhythmias worsened 11% when noninvasive monitoring and exercise tests were performed to evaluate drug effect. How frequently this complication occurs when patients undergo electrophysiologic studies is not known. Electrophysiologic (EP) tests were carried out in 63 patients who had a history of malignant, sustained ventricular tachyarrhythmias. Monitoring and exercise tests showed low-frequency or nonreproducible ventricular arrhythmia. Criteria for definite drug-induced aggravation of arrhythmia included conversion of nonsustained ventricular tachycardia to a sustained ventricular arrhythmia and provocation of the end point with one extrastimulus when three were required during control. Aggravation was deemed possible when, as compared to a control group, the end point resulted with the use of one less extrastimulus and sustained tachycardia with a more rapid rate was provoked. A total of 216 single drug studies were performed (3.4/patient). In general, definite or possible aggravation occurred in 35 tests (16%). In 28 cases (12.9%) aggravation was categorized as definite, while in 7 cases (3.2%) the induced arrhythmia was deemed as possibly related to the use of the antiarrhythmic drugs. Drug tests with multiple agents caused aggravation of arrhythmia in 19 patients (30%). Therefore, exacerbation of arrhythmia by antiarrhythmic drugs also occurs during electrophysiologic study. The incidence approximates that reported when monitoring and exercise tests are used for evaluating drug efficacy.

Studies on the in vivo synthesis of methotrexate polyglutamates and their efflux properties in normal, proliferative, and neoplastic mouse tissues.

Synthesis of poly-gamma-glutamyl metabolites of methotrexate was demonstrated in mouse small intestine, liver and bone marrow, and in L1210 leukemia, Sarcoma 180 and Ehrlich tumor cells after sc injections of [3H]methotrexate to tumor bearing mice. Ion exchange chromatography of tissue extracts resolved six peaks of radioactivity believed to represent methotrexate and metabolites with up to 4 additional glutamyl residues. Polyglutamate formation in L1210 cells and small intestine was shown to be independent of dose at least to 400 mg/kg as long as intracellular levels of drug in excess of the dihydrofolate reductase binding capacity (exchangeable) were maintained. Both the total amount of polyglutamates and the average length of the polyglutamyl chain increased with time as long as exchangeable level of drug were present intracellularily. The results also showed differences in the extent of metabolism of methotrexate polyglutamates among the tissues examined. Although, these differences were at times very large, there was no consistent correlation between these differences and other pharmacologic parameters or cytoxicity. Tumor cells appeared to synthesize more polyglutamates than the normal tissues examined. However, differences in total drug persistence and sensitivity to drug among tumor cells and among normal tissues did not reflect the relative extent of polyglutamate synthesis in each group. We observed no selective retention of polyglutamates as compared to methotrexate by L1210 cells in vitro as indicated by the extracellular accumulation during efflux of methotrexate and the polyglutamates. This could only be demonstrated by allowing efflux of intracellular drug in the presence of extracellular dihydrofolate reductase, which averted hydrolysis of the polyglutamates. It is concluded that the extent of polyglutamate synthesis per se may not be a determinant of drug sensitivity in murine tissues. However, the accumulation of these metabolites may contribute in some way to overall therapeutic response or relative cytotoxicity.

Differential synthesis of methotrexate polyglutamates in normal proliferative and neoplastic mouse tissues in vivo.

Synthesis of poly-gamma-glutamyl metabolites of methotrexate was demonstrated in mouse small intestine, liver, and bone marrow and in L1210 leukemia, Sarcoma 180, and Ehrlich tumor cells after s.c. injections of [3H]methotrexate to tumor-bearing mice. Ion-exchange chromatography of tissue extracts resolved six peaks of radioactivity believed to represent methotrexate and metabolites with up to five additional glutamyl residues. Polyglutamate formation in L1210 cells and small intestine was shown to be independent of dose at least to 400 mg/kg as long as intracellular levels of drug in excess of the dihydrofolate reductase-binding capacity (exchangeable) were maintained. Both the total amount of polyglutamates and the average length of the polyglutamyl chain increased with time as long as exchangeable level of drug was present intracellularly. The results also showed differences in the extent of metabolism of methotrexate polyglutamates among the tissues examined. Although these differences were at times very large, there was no consistent correlation between these differences and other pharmacological parameters or cytotoxicity. Tumor cells appeared to synthesize more polyglutamates than did the normal tissues examined. However, differences in total drug persistence and sensitivity to drug among tumor cells and among normal tissues did not reflect the relative extent of polyglutamate synthesis in each group. It is concluded that the extent of polyglutamate synthesis per se may not be a determinant of drug sensitivity in murine tissues. However, the accumulation of these metabolites may contribute in some way to overall therapeutic response or relative cytotoxicity.