Kinetic correlates of methotrexate transport and therapeutic responsiveness in murine tumors

Net accumulation of methotrexate by carrier-mediated transport in different murine tumor cells in vitro exhibits a positive correlation with the relative drug pharmacokinetics and therapeutic responsiveness in these tumors in vivo. The transport of methotrexate by Sarcoma 180, Ehrlich carcinoma, P388, P288, and L1210 leukemia cells is qualitatively similar. Influx of drug exhibits saturation kinetics and is highly temperature dependent (Q10, 6.1 to 9.4). Efflux of exchangeable methotrexate from all of the different tumor cells exhibited first-order kinetics and the same high temperature dependence seen for influx (Q10, 6.1 to 8.0). The major kinetic determinant of responsiveness is the Km for influx. Values vary from 3.1 to 11.2 X 10(-6) M and are highest in cells from a nonresponsive Sarcoma 180 tumor, somewhat lower in the poorly responsive Ehrlich tumor, lower in moderately responsive P388 and P288 leukemias, and lowest in the highly responsive L1210 leukemia. Values for the influx Vmax differ to some extent, but in a manner not correlatable with responsiveness. The level of responsiveness of the P388 leukemia in vivo can also be partially attributed to an efflux rate that is lower than that measured for the other tumor cells. Steady-state levels of drug accumulation in vitro reflected influx and efflux rates and were consistently correlatable with therapeutic responsiveness. There was no significant difference in the extent to which folate and reduced 5-substituted folate derivatives compete with methotrexate for uptake in cells from all five tumors. The average value for Ki measured with folate for each tumor cell type was 50- and 80-fold higher than for 5-formyltetrahydrofolate and 5-methyltetrahydrofolate.     

Melatonin induction of testicular recrudescence in hamsters and its subsequent inhibitory action on the antigonadotrophic influence of darkness on the pituitary-gonadal axis

The results of this animal study confirm the induction of delayed hypersensitivity to a halothane metabolite, but the sensitization does not cause hepatic damage on subsequent halothane exposures. Before the hypothesis that hepatic necrosis following halothane has an immunologic basis can be accepted or rejected, more critical studies with different approaches must be made.     

Frontal cortex and response suppression in the rat

Parts of the rat's neocortex were mapped for sites where electrical stimulation, square waves at 10/sec, will suppress a bar-press response for food. Effective sites were found in the frontal pole and over most of the frontal dorsolateral cortex. Within these regions the strongest inhibitory influences were at sites in the frontal pole and adjacent frontal cortex, and at sites along the midline. These data generally agree with results of previous cortical ablation studies, but they suggest that inhibitory processes are more widespread in the dorsolateral cortex than these studies indicated.     

Force and motion analysis of the normal, diseased, and prosthetic ankle joint

A 2-dimensional motion and force study of the ankle joint during gait has been carried out on normal subjects and patients with ankle joint disease, before and 1 year following total ankle replacemetn. The methods employed involved the use of high-speed motion picture film, force plate and foot-switch data. The Achilles and anterior tibial tendon forces, the compressive and tangential (shear) forces across the ankle during stance phase of gait were determined, based on a quasi-static analysis. During stance phase of gait normal subjects used a mean of 24.4 degree of sagittal plane ankle motion. Patients with ankle joint disease showed reduced motion which returned to near normal values 1 year following total ankle replacement. Compressive force across the ankle joint rose to about 5 times body weight during the latter part of stance phase. Backward, or aft, shear forces or nearly full body weight were demonstrated during all but the last 20% of stance phase. Patients with ankle joint disease apparently altered their gait to markedly reduce these forces. Following total ankle replacement, shear forces returned toward more normal values, but compressive forces were not significantly changed.     

Effect of histamine on technetium-99m excretion by gastric mucosa

The ability of histamine to increase excretion of 99mtechnetium by gastric mucosa was investigated in dogs with Heidenhain pouches and denervated antral pouches. Histamine increased Heidenhain pouch 99mtechnetium output in a dose-related manner, and 99mtechnetium output was related linearly to acid output. Antral pouch 99mtechnetium output was unchanged by increasing doses of histamine. The study suggests that concomitant use of histamine may improve the accuracy of 99mtechnetium scanning in the clinical diagnosis of conditions caused by ectopic gastric mucosa.     

Intestinal ascorbic acid transport following diets of high or low ascorbic acid content

Active transport of ascorbic acid in ileum is mediated by a carrier mechanism at the brush border membrane. This mechanism may show compensatory changes in activity in response to alterations of dietary ascorbic acid content. The unidirectional influx of ascorbic acid across the brush border into epithelial cells of guinea pig ileum was determined in vitro. Influx was significantly reduced in scorbutic animals and following 14 or 28 days of high doses (5 or 25 times normal) of ascorbic acid. The transport rate was reduced by intramuscular administration of ascorbic acid, suggesting that the transport mechanism may respond to circulating levels of the vitamin.     

Flow-microfluorometric analysis of nuclei isolated from various normal and malignant human epithelial tissues. A preliminary report

In order to obviate some of the technical problems associated with preparation of monocellular cell suspensions required for flow fluorometry, isolation of nuclei from several types of benign and malignant human tissues was undertaken. Satisfactory preparations of nuclei were obtained from epithelia of the uterine cervix and colon and from lung tissue using the citric acid method. The sucrose method was effective with colonic epithelium only. Distribution of deoxyribonucleic acid content in these nuclei was measured based on green fluorescence of acridine orange and red fluorescence of propidium iodide in a Bio-Physics Cytofluorograph. Essentially diploid patterns of deoxyribonucleic acid distribution were observed for all benign samples regardless of tissue origin whereas the malignant samples gave histograms suggestive of abnormal deoxyribonucleic acid distribution. Preliminary observations on distribution of single-stranded nucleic acids using acridine orange red fluorescence showed marked differences between populations of benign and malignant nuclei. Isolated nuclei appear to be suitable for flow-through microfluorometric analysis and offer some significant advantages over intact cells.     

Symmetrical peripheral gangrene

Two cases of symmetrical peripheral gangrene associated with intracardiac lesions are described. One had a secondary deposit from a uterine leiomyosarcoma in the wall of the right ventricle, the other had a free ball thrombus in the right atrium; there were no valvular lesions in either case. The published reports of this type of gangrene indicate that it may result from (1) va sospastic conditions, (2) small vessel obstruction, or (3) conditions producing a very low cardiac output. The two patients reported are unusual examples of the last.