Abnormal activation of lipoprotein lipase by non-equilibrating apoC-II: further evidence for the presence of non-equilibrating pools of apolipoproteins C-II and C-III in plasma lipoproteins

Using artificial triglyceride emulsions, we have demonstrated the presence of non-equilibrating pools of apolipoproteins C-II and C-III in human plasma lipoproteins. As the concentrations of acceptor triglycerides were increased, a greater fraction of both apoC-II and apoC-III shifted away from the native plasma lipoproteins to the artificial lipid emulsions. All of the apoC-II and apoC-III in very low density and high density lipoproteins (VLDL and HDL), however, could not be removed from native plasma lipoproteins. The percent of total plasma apoC-II and apoC-III that could be recovered in the VLDL and HDL density fractions varied when plasma from different individuals was used. When plasma samples from normotriglyceridemic subjects were used, HDL was the primary donor of apoCs. The percent of total plasma apoCs associated with HDL decreased from 60% to 25% for apoC-II and from 65% to 15% for apoC-III. When plasma samples from hypertriglyceridemic subjects were incubated with artificial lipid emulsions, VLDL was the primary donor of apoCs. HDL from hypertriglyceridemic subjects only accounted for 5-10% of total fasting plasma apoCs and did not contribute significantly to the final apoC contents of the artificial triglyceride emulsions. To evaluate the significance of the depletion of exchangeable apoCs from plasma HDL, we also examined the ability of control and apoC-depleted HDL to serve as activator for bovine milk lipoprotein lipase (LPL) in vitro. When HDL depleted of exchangeable apoCs were used as the source of plasma apolipoproteins for the activation of LPL in vitro, only 5-10% of the maximal activity obtained with native HDL was demonstrated. In fact, in the presence of comparable concentrations of HDL apoC-II, activation of LPL was the least with HDL which lacked exchangeable apoCs. Our data thus indicated that the presence of exchangeable apoC-II on HDL is necessary for the activation of LPL in vitro. This finding is consistent with our data that suggest that HDL from hypertriglyceridemic subjects do not stimulate LPL as well as HDL from normolipidemic subjects.     

Improvement of plasma lipoprotein profiles during high-flux dialysis

Patients undergoing maintenance hemodialysis therapy have increased mortality due to cardiovascular disease. One possible etiologic factor for this increased mortality is the lipid abnormalities associated with chronic renal failure. These include elevated triglyceride (TG) and decreased high-density lipoprotein (HDL) concentrations. Lipoprotein profiles of patients undergoing chronic hemodialysis with either saponified cellulose ester (CE) (N = 9) or polysulfone (PS) high-flux dialysis membranes (N = 10) were compared. Patients in each group received similar amounts of heparin during the dialysis. CE-dialyzed patients showed no alteration in serum TG, HDL, low-density lipoprotein, or total cholesterol when predialysis and postdialysis values were compared. PS patients, on the other hand, had a significant decrease in TG concentrations (P < 0.01) as well as a significant rise in HDL (P < 0.01). These changes might signify activation of lipoprotein lipase (LPL) during dialysis. LPL activity in PS sera was significantly greater than LPL in CE sera. Moreover, sera from PS patients inhibited LPL much less than did sera from CE patients. These findings suggest that a circulating substance not dialyzable with cellulosic membranes inhibits LPL in uremic subjects and is removed during dialysis with a PS membrane. Alternatively, the greater biocompatibility of PS may produce less LPL inhibitory cytokines during dialysis. The improvement of lipoprotein profiles in patients receiving dialysis with PS membranes may, in the long term, lead to less morbidity and mortality from atherosclerotic disease.     

In vitro cross-resistance and collateral sensitivity in seven resistant small-cell lung cancer cell lines: preclinical identification of suitable drug partners to taxotere, taxol, topotecan and gemcitabin

The thiazole orange dye 1,1'-(4,4,8,8-tetramethyl-4,8-diazaundecamethylene)-bis[4-[3-methy l-2, 3-dihydro(benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO) binds to double-stranded DNA (dsDNA) in a sequence selective bisintercalation. Each chromophore is sandwiched between two base pairs in a (5'-CpT-3'):(5'-ApG-3') site, and the linker spans over two base pairs in the minor groove. The binding of analogs of TOTO in which the linker has been modified is examined. The aim of the study is to utilize the sequence selectivity of the TOTO chromophores to enhance and/or alter the overall selectivity of the binding. One- and two-dimensional 1H-NMR investigations of complexes between TOTO analogs and various dsDNA oligonucleotides are reported. The following analogs were synthesized and used: 1,1'-(4,4,8,8-tetramethyl-4,8-diazadodecamethylene) -bis[4-[3-methyl-2,3-dihydro- (benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO10), 1,1'-(5,5,9,9-tetramethyl-5,9-diazatridecamethylene)-bis[4-[3-meth yl-2, 3-dihydro(benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO11), and 1,1'-(6,6,10,10-tetramethyl-6,10-diazapentadecamethylene)-bis[4-[3 -methyl-2, 3-dihydro(benzo-1,3-thiazole)-2-methylidene]]quinolinium tetraiodide (TOTO13). The results show that with a longer linker the dyes can bisintercalate into two (5'-CpT-3'):(5'-ApG-3') sites separated by one or two base pairs. Bisintercalation in two such "isolated" binding sites yields non-nearest-neighbor bisintercalation in which the linker spans over more than two base pairs. The investigations also showed that an exact length of the linker is not crucial for the site selectivity since TOTO, TOTO10, and TOTO11 are almost equally suitable in binding selectively to the (5'CTAG-3')2 sequence. Fluorescence measurements show that TOTO10, TOTO11, and TOTO13 have higher fluorescence quantum yields than TOTO when bound to d(CGCTAGCG)2. This indicates that the length of the linker in TOTO may not be the optimum one in terms of using the dye as a fluorescence marker.     

Representing primary electrophoretic data in the 1/time domain: comparison to representations in the time domain

A plot of absorbance vs 1/time (the "1/time domain") is a more useful representation of the primary data in capillary electrophoresis than traditional plots of absorbance vs time (the "time domain") in a wide set of circumstances, especially when comparing electropherograms in which the rate of electroosmotic flow is not precisely the same. The quantity that is of fundamental interest in capillary electrophoresis (CE) is the electrophoretic mobility of an analyte. The electrophoretic mobility of a species is nonlinearly proportional to time and, therefore, not linearly represented in the time domain: that is, the distance between two peaks along the time axis is not linearly related to the difference in their electrophoretic mobilities. In contrast, the electrophoretic mobility is linearly proportional to 1/time, and the distance between two peaks along the 1/time axis is linearly related to the difference in electrophoretic mobilities. Plots in the 1/time domain are similar to the familiar plots in the time domain (each analyte is represented by a peak, and the order of peaks corresponds to the order in which these analytes reach the detector), but the spacing between the peaks corresponds linearly to differences in mobility. This article derives this useful, visually appealing, and broadly applicable plotting strategy and illustrates common situations in which these plots are more useful than plots in the time domain.     

Prognostic relevance of urokinase plasminogen activator detection in micrometastatic cells in the bone marrow of patients with primary breast cancer

The "attentional model" of time estimation assumes that temporal judgments depend on the amount of attention allocated to the temporal processor (the timer). One of the main predictions of this model is that an interval will be judged shorter when attention is not allocated to the temporal parameters of the task. Previous studies combining temporal and nontemporal tasks (dual-task method) have suggested that the time spent processing the target duration might be a key factor. The less time devoted by the subject to the temporal task, the shorter the judged duration. In the two experiments presented here, subjects were asked to judge both the duration of a visual stimulus and an increment in intensity occurring at any time during this stimulus. In the second experiment, trials without intensity increments were added. The main result is that the judged duration was shorter when the increment occurred later in the stimulus or did not occur. In those cases, subjects had been expecting increment occurrence during most part of the stimulus and thus had focused for a shorter time on stimulus duration. We propose that attention shifts related to expectancy and to detection of the increment reduce subjective duration.     

Behavior and behavioral needs

An understanding of behavior is important in any consideration of poultry welfare. Behavior is a good indicator of states of suffering such as fear, frustration, and pain. It might also be possible to use social interactions as indicators of welfare. The possibility of using "luxury" behavior, such as play and exploratory behavior, as an indicator of positive emotional states, requires investigation. Important welfare consequences arise from the fact that some behavior may be so strongly motivated as to constitute a "need". A behavioral need will arise in the case of behavior, such as nesting, that is controlled largely by internal factors, because these factors will be present no matter what type of environment is provided. Behavior triggered largely by external stimuli, such as responses to predators, will not give rise to a need if the external factors can be removed from the environment. Dustbathing is an example of behavior controlled by complex interactions between internal and external factors; the extent to which this constitutes a need is still being debated. If a behavioral need arises, then it is important that the environment provided allows it to be performed without damage to the performer or other birds. It should also be remembered that birds may need to perform behavior, including appetitive as well as consummatory elements, although the functional consequences of these are no longer required for survival. Finally, the performance of certain behavior leads to an increase in health or physical condition that improves welfare later in life.     

Seasonal changes in the negative feedback regulation of the secretion of the gonadotrophins by testosterone and inhibin in rams

Three experiments were conducted with castrated Romney Marsh rams (wethers) to investigate the ability of testosterone and inhibin to suppress the secretion of LH and FSH during the breeding and the non-breeding seasons. In Experiment 1, wethers (n=5/group) were treated every 12 h for 7 days with oil or 16 mg testosterone propionate (i.m.) and were then given two i.v. injections either of vehicle or of 0.64 microg/kg human recombinant inhibin A (hr-inhibin) 6 h apart. Blood samples were collected for 4 h before inhibin or vehicle treatment and for 6 h afterwards for the assay of LH and FSH. In Experiments 2 and 3 wethers underwent hypothalamo-pituitary disconnection (HPD) and were given 125 ng GnRH i.v. every 2 h. In Experiment 2, HPD wethers (n=3/group) were injected (i.m.) every 12 h with oil or testosterone and blood samples were collected over 9 h before treatment and 7 days after treatment. In Experiment 3, HPD (n=5/group) wethers were treated with vehicle or hr-inhibin, as in Experiment 1, after treatment with oil, or 4, 8 or 16 mg testosterone twice daily for 7 days. Blood samples were collected over 4 h before treatment with vehicle or hr-inhibin and for 6 h afterwards. Treatment of wethers with testosterone (Experiment 1) resulted in a significant decrease in the plasma concentrations of LH and number of LH pulses per hour but the magnitude of these reductions did not differ between seasons. Testosterone treatment had no effect on LH secretion in GnRH-pulsed HPD wethers in either season and treatment with hr-inhibin did not affect LH secretion in wethers or HPD wethers in any instance. Plasma concentrations of FSH were significantly (P<0.05) reduced following treatment with testosterone alone during the breeding season but not during the non-breeding season. FSH levels were reduced to a greater extent by treatment with hr-inhibin but this effect was not influenced by season. During the non-breeding season, the effect of hr-inhibin to suppress FSH secretion was enhanced in the presence of testosterone. These experiments demonstrate that the negative feedback actions of testosterone on the secretion of LH in this breed of rams occurs at the hypothalamic level and is not influenced by season. In contrast, both testosterone and inhibin act on the pituitary gland to suppress the secretion of FSH and these responses are affected by season. Testosterone and inhibin synergize at the pituitary to regulate FSH secretion during the non-breeding season but not during the breeding season.     

NO- and NO2-carrying molecules potentiate photorelaxation in rat trachea and aorta

Photorelaxation elicited by ultraviolet light (366 nm) was investigated on isolated rat thoracic aorta and trachealis. Rat tracheal smooth muscle but not aorta did not show UV-induced photorelaxation. Both streptozotocin, NO-carrying molecule and N omega-nitro-L-arginine, NO2-carrying molecule significantly enhanced photorelaxation, concentration-dependently, in rat trachealis and aorta. Methylene blue (10 microM) inhibited the potentiation action of streptozotocin and N omega-nitro-L-arginine in both tissues. Superoxide dismutase (300 U/ml) enhanced streptozotocin- and N omega-nitro-L-arginine-potentiated photorelaxation in rat trachealis, while pyrogallol (0.1 mM), a potent O2- generating agent, inhibited streptozotocin-potentiated photorelaxation in trachealis. Streptozotocin was much more effective than N omega-nitro-L-Arginine in potentiating of photorelaxation elicited by UV light in both tissues. From these findings, we conclude that streptozotocin and N omega-nitro-L-arginine produce EDRF like labile substance(s) by UV irradiation.     

Computerized analysis of isometric tension studies provides important additional information about vasomotor activity

Concentration-response curves of isometric tension studies on isolated blood vessels are obtained traditionally. Although parameters such as Imax, EC50 and pA2 may be readily calculated, this method does not provide information on the temporal profile of the responses or the actual nature of the reaction curves. Computerized data acquisition systems can be used to obtain average data that represent a new source of otherwise inaccessible information, since early and late responses may be observed separately in detail.     

Systemic targeting of liposome-encapsulated immunomodulators to macrophages for treatment of cancer metastasis

The therapy of cancer is, in reality, the design of therapeutic strategies for therapy of metastatic disease. Metastases consist of unique subpopulations of tumor cells that are derived from the primary tumor, colonize distant target organs, and are able to subvert host immune responses, establish necessary angiogenesis, and obtain a sufficient nutrient supply while evolving to become autonomous from homeostatic mechanisms that function within normal, differentiated tissues. Attempts at eradication of metastases by conventional therapies have generally been unsuccessful due to genetic instability and heterogeneity of metastatic tumors; these properties lead to the emergence of tumor cells that are resistant to most conventional treatments. It may be possible to circumvent this heterogeneity by the activation of tissue macrophages to the tumoricidal state. Activated macrophages are able to kill tumor cells while sparing normal tissues, and efficient activation can be achieved by encapsulation of synthetic muramyl tripeptide analogues into multilamellar vesicles composed of phospholipids. Systemic administration of these liposome-encapsulated compounds leads to tumoricidal activation of alveolar and peritoneal macrophages and eradication of established tumor metastasis in numerous animal tumor models, and this form of therapy is enhanced by combination with parenteral administration of cytokines. Phase III clinical trials of recurrent osteosarcoma are currently in progress. Modulation of the tumor microenvironment by activated macrophages may prove to be an additional modality in treatment strategies that combine the use of biological response modifiers with conventional therapies.