Human lymphocyte motility: normal characteristics and anomalous behavior of chronic lymphocytic leukemia cells.
The characteristics of human lymphocyte motility and its relationship to the redistribution of surface membrane antigens (capping) are poorly defined. Since chronic lymphocytic leukemia (CLL) cells cap poorly when compared with normal human lymphocytes, this study was undertaken to compare the motility of these two cell types. A modification of the Boyden chamber system was employed to quantify lymphocyte motility by placing lymphocyte suspensions on 8-mum convoluted-pore nitrocellulose filters and measuring the depth of migration of the cells into the filter at 37 degrees C. After 3 hr of incubation, CLL cells migrated significantly less into the filter than normal cells. Incubation in the presence of sodium azide or at 4 degrees C abolished all motility, indicating the active nature of the process. The relative motility of individual CLL patients' cells correlated best with the proportion of abnormal cells present as determined by surface receptor assays. The possibility that decreased cell motility in CLL was a reflection of enrichment by a "bone marrow-derived" (B cell) population was eliminated by the finding that normal B cells purified by gradient separation of rosetted cells migrated faster than normal T cells and considerably faster than CLL cells. Motility of normal and CLL lymphocytes was decreased by cytochalasin B and increased by colchicine, vincristine, and vinblastine. Thus, human lymphocyte motility appears to be dependent on microfilament integrity but not to require the colchicine-sensitive cytoskeleton. The decreased motility of CLL cells is the result of an intrinsic cell abnormality, but this finding cannot fully explain the decreased capping, since in human lymphocytes the latter is not prevented by an inhibitor of motility. (+info)
Effect of a low protein diet during pregnancy of the rhesus monkey. II. Physiological adaptation of the infant.
Heart rate, respiratory rate, and ability to maintain body temperature were evaluated in six infants born to rhesus monkeys that had been fed a low protein diet throughout pregnancy. All infants were kept in incubations equipped with a "servo" control thermal unit that maintained the infants' skin temperature at 37.0 C (98.6 F). The thermal units were disconnected and the infants were exposed to room temperature (approximately 27 C) for 6-hr periods each day after 24 hr of age in order to determine the efficiency of thermal control mechanism. The thermal servo control units were "on" for a longer period of time in experimental animals than in control animals during the first 24 hr of life. Infants from mothers fed the low protein diets were also less able to maintain their own body temperatures after exposure to room temperature. This function was seriously compromised in two of the six experimental infants. The compromised temperature control mechanism seen in these infant monkeys is a serious and potentially lethal side-effect of protein-calorie malnutrition during pregnancy. The possible relationship of inadequate maternal nutrition to the inefficient thermal mechanism of certain "high risk" human newborns should be reevaluated. (+info)
Early thermal experience has different effects on growth and muscle fibre recruitment in spring- and autumn-running Atlantic salmon populations.
The consequence of early thermal experience for subsequent growth patterns was investigated in Atlantic salmon (Salmo salar L.). Spring- and autumn-running salmon were caught in upland (Baddoch) and lowland (Sheeoch) tributaries of the River Dee, Aberdeenshire, Scotland, respectively, on the final stages of their spawning migrations. The eggs were incubated at the simulated natural temperature regime of each stream, which was on average 2.8 degrees C lower for the Baddoch. The offspring, representing 11 families per population, were transferred at first feeding to constant environmental conditions (12-14 degrees C; 16h:8h light:dark photoperiod) and reared in replicate tanks. Salmon of both populations were longer and heavier at 6 and 12 weeks in fish initially reared under the cooler Baddoch regime. Length frequency distributions became bimodal after 18 weeks, and only the upper growth mode was studied. Modelling of length distributions at 40 weeks revealed significantly different patterns of muscle growth according to initial temperature regime, but only for the Sheeoch salmon. In fish of Sheeoch origin, significantly more white muscle fibres were recruited per mm(2) increase in myotomal cross-sectional area at Sheeoch than at Baddoch temperatures (P<0.01). After 40 weeks, the density of white fibres was 10.4 % higher in fish initially reared at the Sheeoch (533+/-6 mm(-2)) than at the Baddoch (483+/-5 mm(-2)) thermal regimes (means +/- s.e.m., 16 fish per group; P<0.001). Muscle satellite cells were identified using an antibody to c-met. At 24 weeks, the density of muscle satellite cells was 29 % higher in Sheeoch salmon reared to first feeding at the temperature of their natal stream than at cooler Baddoch temperatures (P<0.01). In contrast, the number and size distributions of white muscle fibres in the myotomes of Baddoch salmon were independent of early thermal experience. (+info)
Antimicrobial susceptibilities of Helicobacter pylori isolates under microaerophilic atmospheres established by two different methods.
The MICs of clarithromycin, amoxicillin, and metronidazole for 150 Helicobacter pylori isolates were determined using the AnaeroPack system and were compared with those determined using a microaerophilic incubator. The MICs of clarithromycin, amoxicillin, and metronidazole determined under both microaerophilic atmospheres were mostly within one twofold dilution for 146 (97.3%), 150 (100%), and 149 (99.3%) of the isolates, respectively. (+info)
Objective assessments of temperature maintenance using in vitro culture techniques.
PURPOSE: To assess the ability of various facets of embryo culture (microscope stage warmers, volumes of culture media, culture vessel lids, and type of culture incubator) to maintain a constant temperature in vitro. METHODS: Ability to maintain 37.0 degrees C in the microenvironment of gametes was recorded by digital thermocouple in the chosen facets of in vitro culture. RESULTS: Stage warmers are highly variable in their ability to maintain the set temperature (range 33.8 degrees C-37.0 degrees C after 60 s). Temperature loss in culture media is both volume and vessel dependent, and the direct heat transfer culture incubator (MINC) has superior temperature maintenance compared with a large volume air convection incubator (FORMA), where temperature regain from 35.0 degrees C to 37.0 degrees C took 5.5 min compared to >20 min. CONCLUSIONS: There are large measurable differences in the ability to maintain set temperature that depend on the stage warmer used, volume of media, use of vessel lids, and the type of incubator chosen for IVF culture. (+info)
Heart period variability of intubated very-low-birth-weight infants during incubator care and maternal holding.
BACKGROUND: Heart rate has been used to measure infants' physiological stability during skin-to-skin holding. Variability in heart period (interbeat interval), a more sensitive measure of autonomic nervous system tone, has not. OBJECTIVE: To describe heart period variability in intubated very-low-birth-weight infants during incubator care and during maternal skin-to-skin holding. DESIGN/METHODS: An experimental, interrupted time series, crossover design was used; infants served as their own controls. Infants were randomly assigned to treatment order: 2 hours of intermittent skin-to-skin holding for 2 consecutive days followed by 2 days of incubator care or vice versa. The analog signal representing heart period was sampled and quantized at 5 Hz via a dedicated computer system in multiple 300-second epochs each day. RESULTS: Fourteen infants with similar characteristics completed the protocol. The mean interbeat interval was 332 ms during skin-to-skin care and 368 ms during incubator care. Power within the low- and high-frequency regions of heart period was not significantly different between skin-to-skin holding and incubator care. Mean low-frequency power was 124.6 ms2 during skin-to-skin holding and ranged from 51.9 ms2 to 71.4 ms2 during all periods of incubator care. Mean high-frequency power was similar during skin-to-skin holding and incubator care (8.8 ms2 and 6.1 ms2). Infants of 32 to 34 weeks' corrected gestational age had increased power in the low- and high-frequency regions. CONCLUSIONS: Heart period variability did not improve during skin-to-skin holding. Gestationally older infants had increased power in the low- and high-frequency regions, suggesting a maturing autonomic nervous system. (+info)
Accumulation of genistein and lipophilic genistein derivatives in lipoproteins during incubation with human plasma in vitro.
Atherosclerosis is initiated by the uptake and retention of oxidized low-density lipoprotein (LDL) into the arterial intima. We have previously shown that dietary isoflavone phytoestrogens inhibit LDL oxidation in vitro. The inhibition could have been caused by undetected isoflavone metabolites associated with lipoproteins. In the present study, we incubated human plasma with [3H]genistein, both with and without the lecithin:cholesterol acyltransferase (LCAT) inhibitor dithionitrobenzoic acid (DTNB). Our results indicated that the 3H-label was attached to both high-density lipoprotein (HDL) and LDL, and that it represented both underivatized genistein and lipophilic derivatives of genistein, part of which were identified as fatty acid monoesters. The latter was demonstrated by the findings that DTNB decreased the HDL and LDL associated radioactivity in the lipophilic fraction isolated by hydrophobic chromatography and that saponification hydrolysis liberated a corresponding part of the 3H-label. Two-dimensional reversed-phase thin-layer chromatography (TLC) demonstrated that a corresponding part of the radioactivity comigrated with genistein monoester standards in the absence of DTNB but was abolished if DTNB had been present in the incubation. In summary, incubation of plasma with [3H]genistein resulted in accumulation of underivatized genistein as well as lipophilic genistein derivatives in lipoproteins. A smaller part of the latter were genistein monoesters, while part remained unidentified. Our results suggest an explanation for the increased oxidation resistance of isolated LDL during intake of soybean isoflavones. (+info)
Differences in troglitazone action on glucose metabolism in freshly isolated vs long-term incubated rat skeletal muscle.
1. Exposure of isolated skeletal muscle to troglitazone has resulted in inconsistent findings ranging from inhibition to stimulation of fuel oxidation and the glycogenic pathway. To better understand such variation in outcome, the present study used isolated rat soleus muscle strips to examine the interdependent influences of prolonged maintenance in vitro and of troglitazone exposure. 2. If freshly isolated muscle strips were exposed to troglitazone (1 micro mol l(-1)) for 24 h, glucose oxidation was markedly reduced (-26+/-1%, P<0.0001), whereas glycogen synthesis remained unaffected (+9+/-7%, n.s.). 3. In contrast, extended exposure to troglitazone for 72 h increased both glucose oxidation (+65+/-28%, P<0.05) and glycogen synthesis (+46+/-11%, P<0.005), and a similar stimulatory effect was also observed in muscles exposed to troglitazone only during the last 24 h of their 72 h preincubation period (glucose oxidation: +61+/-15%, P<0.001; glycogen synthesis: +43+/-15%, P<0.01). 4. Troglitazone thus stimulated glucose utilization in long-term incubated muscle independent of the duration of exposure (24 or 72 h), whereas it inhibited glucose utilization in freshly isolated muscle. 5. The observed differences in troglitazone action on freshly isolated vs long-term incubated muscle suggest that findings on muscle tissue subject to prolonged maintenance in vitro cannot be extrapolated to native muscle in vivo. (+info)