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Biomedical subjects

M H Fox

Publications and source records attributed to M H Fox.

At least 55 records · Page 3Linked to original sources
PubMed1990

The concentration of calcium in neonatal cardioplegia.

The optimal calcium concentration in cardioplegia for the newborn has not been determined. Therefore, the effect of 0, 0.6, 1.2, 1.8, and 2.4 mmol/L calcium in modified St. Thomas cardioplegia was evaluated in isolated working hearts of 7- to 10-day-old rabbits. Functional recovery was determined by comparing aortic flow, developed pressure, and first derivative of left ventricular pressure (dP/dt) before and after 1 hour of normothermic (37 degrees C) ischemia. As percentages of baseline values, recovery of developed pressure and dP/dt averaged 10% +/- 1% (mean +/- standard error of the mean) and 10% +/- 1% with 0 mmol/L, 46% +/- 7% and 44% +/- 8% with 0.6 mmol/L, 79% +/- 2% and 76% +/- 2% with 1.2 mmol/L, 67% +/- 2% and 61% +/- 5% with 1.8 mmol/L, and 65% +/- 5% and 65% +/- 7% with 2.4 mmol/L calcium, respectively. Significant improvement in recovery of developed pressure and dP/dt was detected when the calcium concentration was increased from 0 to 0.6 mmol/L and from 0.6 to 1.2 mmol/L, but the groups with 1.2, 1.8, and 2.4 mmol/L did not differ from one another significantly in terms of developed pressure and dP/dt recovery. There was no recovery of aortic flow when 0 mmol/L calcium was used; at calcium concentrations of 0.6, 1.2, 1.8, and 2.4 mmol/L, recovery of aortic flow averaged 16% +/- 7%, 63% +/- 10%, 23% +/- 10%, and 36% +/- 11% of baseline values, respectively. Recovery of aortic flow with 1.2 mmol/L calcium was significantly higher than at concentrations of 0.6 and 1.8 mmol/L.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

In vitro expression of bovine leukemia virus in isolated B-lymphocytes of cattle and sheep.

The purpose of this study was to determine the effect of T-lymphocytes and phytohemagglutinin (PHA), a T-cell mitogen, on the expression of bovine leukemia virus (BLV) in cultured B-lymphocytes from BLV-infected cattle and sheep. Bovine B-lymphocytes were isolated by negative selection via complement-mediated lysis of T-lymphocytes. Additionally, bovine and ovine B-lymphocytes were positively selected using fluorescence activated cell sorting. Expression of BLV in cultured bovine and ovine B-lymphocytes occurred in the absence of T-lymphocytes and without PHA stimulation. The results of this study demonstrate that BLV replication in cultured B-lymphocytes is T-cell independent. This finding may have implications for the mechanism of viral latency within infected B-lymphocytes.

Animals↗
PubMed1988

Effects of chronic pH 6.6 on growth, intracellular pH, and response to 42.0 degrees C hyperthermia of Chinese hamster ovary cells.

Culturing Chinese hamster ovary cells in low pH (6.6) medium for several months altered the reproductive survival of these cells to combined low pH treatments and 42.0 degrees C heating. We isolated new pH-resistant cells (identified as pHV-2) with enhanced ability to grow and divide under a low pH (6.6) environment. Their growth characteristics include (a) a plating efficiency of 70%, (b) a doubling time of 16 to 17 h, and (c) a steady state intracellular pH 0.12 pH units higher than for cells grown at a normal pH of 7.3. The pHV-2 cells had 100- to 200-fold increases in survival after 5 h of heating compared to cells incubated at low pH (6.6) for 4 h prior to and during the heat treatments. In addition, they developed a significant degree of thermotolerance. We measured a progressive decline in the intracellular pH as a function of time at 42.0 degrees C. However, the decrease in the intracellular pH did not seem to be correlated with the increased heat sensitivity. The ability to select for low pH variants may have important implications in the extrapolation of in vitro hyperthermic data to the in vivo situation.

Animals↗
PubMed1988

Effects of acute pH 6.6 and 42.0 degrees C heating on the intracellular pH of Chinese hamster ovary cells.

Incubation of Chinese hamster ovary cells in pH 6.6 medium for 4 h prior to and during 42.0 degrees C heating enhanced thermal cell killing compared to cells heated under normal pH 7.3 conditions. We examined the relationship between the extracellular pH and intracellular pH (pHi) of Chinese hamster ovary cells using a flow cytometer with the pH-sensitive fluorescent molecule 2,3-dicyanohydroquinone. Using either normal (7.3) or low (6.6) pH conditions, the mean pHi and population pHi heterogeneity was studied as a function of time at 42.0 degrees C. Cells incubated at pH 6.6 for 4 h had a resting pHi 0.14 to 0.19 pH units lower than cells at normal pH 7.3, indicating the presence of an active pHi regulatory system. Heating 1 h at 42.0 degrees C at normal pH caused an increase in the pHi of 0.14 pH units. With further heating the cells gradually returned to the unheated (7.3) control levels. Similar pHi changes were observed with the cells incubated and heated at pH 6.6. However, the mean pHi was always more acidic than cells heated at normal pH. Active pHi regulation was still possible for a substantial (greater than 30%) number of cells even after 10 h of heating under low pH conditions. These results suggest that a breakdown in pHi regulation is not the mechanism of low pH-induced heat sensitization.

Acid-Base Equilibrium↗
PubMed1988

Intracellular pH measurements using flow cytometry with 1,4-diacetoxy-2,3-dicyanobenzene.

1,4-Diacetoxy-2,3-dicyanobenzene (ADB) has been increasingly used for measurement of intracellular pH by flow cytometry. ADB rapidly enters cells and is cleaved to the fluorescent pH indicator 2,3-dicyano-hydroquinone (DCH). We have analyzed several potential problems that can affect its usefulness as a pH indicator. Hydrolysis of ADB in aqueous solutions reveals the temporary presence of a fluorescent species blue-shifted from DCH at the same pH. The presence of this species with DCH can lead to erroneous pH measurements. Stable pH measurements with ADB depend on the incubation conditions and esterase activity. Heated cells required 20 min for stable measurements, whereas control cells required 5 to 10 min. The reproducibility of pH measurements was excellent, with a resolution of less than or equal to 0.05 pH units in the range of 6.4 to 8.0. Absolute calibration curves of intracellular pH using the ionophore nigericin depended on matching the intracellular K+ concentration with the buffer, but relative measurements of intracellular pH were insensitive to K+. ADB was nontoxic to Chinese hamster ovary cells at up to 20 micrograms/ml. However, when cells loaded with dye were passed through a UV laser beam, concentrations of dye greater than 5 micrograms/ml were highly toxic. Viable cells could be sorted on the basis of intracellular pH if ADB were used at low concentrations.

Animals↗
PubMed1988

Protection of the neonatal heart following normothermic ischemia: a comparison of oxygenated saline and oxygenated versus nonoxygenated cardioplegia.

Optimal methods of myocardial preservation in the neonate remain unknown. Hypothermia and cardioplegia have been shown to protect neonatal hearts, but few studies have examined the effects of cardioplegia when administered at normothermia. Accordingly, the role of 37 degrees C St. Thomas' cardioplegic solution in protecting the neonatal heart during 1 hour of ischemia in an isolated working rabbit heart model was examined. Both oxygenated and nonoxygenated cardioplegic solutions (CSs) were evaluated and compared with an oxygenated physiological saline solution (PSS). Following ischemia, control hearts were characterized by severely impaired left ventricular function, whereas all three treatment groups recovered well, indicating that the treatments provided substantial protection. Aortic flow recovered to 62, 63, and 57% of preischemic values for the oxygenated CS, nonoxygenated CS, and oxygenated PSS groups, respectively. Similarly, rate of change of pressure recovered to 76, 80, and 76% of preischemic values for oxygenated CS, nonoxygenated CS, and oxygenated PSS groups. All values were significantly greater than those for the control group. Recovery of developed pressure was significantly improved in all three groups. End-diastolic pressure rose markedly following ischemia in control hearts, was not increased after ischemia in hearts receiving oxygenated and nonoxygenated CS, but was increased in the oxygenated PSS group. These data indicate that crystalloid cardioplegia and oxygenated PSS provide substantial protection in neonatal rabbit hearts, even when delivered at 37 degrees C. No additional benefit was seen when the cardioplegic solution was oxygenated. Therefore, either method of balancing the oxygen supply/demand ratio appears to be beneficial: supplying oxygen intermittently during ischemia (oxygenated PSS group) or decreasing oxygen demand during the ischemic period (cardioplegia groups).

Animals↗
PubMed1988

Intracellular pH of Chinese hamster ovary cells heated at 45.0 degrees C at pH 6.6.

The mechanism by which low pH sensitizes cells to hyperthermic treatments is unknown. In this report we have examined the relationship between the extracellular and intracellular pH of Chinese hamster ovary (CHO) cells after 45.0 degrees C heating. The intracellular pH (pHi) was measured with a fluorescence technique developed for flow cytometric instrumentation using the dye 1,4-diacetoxy-2,3-dicyanobenzene. Cellular esterase activity was modified by heating, requiring 20 min incubation time with the dye to achieve stable intracellular pH measurements. The intracellular pH of CHO cells incubated under normal pH (7.3) conditions was not perturbed significantly with up to 20 min of heating. Heating from 5 to 15 min induced increases in the pHi of CHO cells incubated under low pH (6.6) conditions. Cells heated in either normal pH or low pH medium became acidified with prolonged heating times. However, the distributions of intracellular pH showed that 20 to 30% of the population heated for up to 60 min at either pH 7.3 or 6.6 overlapped with the pHi distributions of normal cells. A decrease in intracellular pH does not seem to be the principal reason for the heat sensitization caused by low extracellular pH.

Animals↗
PubMed1988

Development of thermotolerance and changes in intracellular pH in CHO cells heated at 45.0 degrees C at pH 6.6.

Chinese hamster ovary (CHO) cells were given short heat pulses (5 to 20 min) at 45.0 degrees C and incubated at 37 degrees C for up to 20 h under either pH 7.3 or 6.6 conditions. Thermotolerance developed under both pH conditions, but at a slower rate in the pH 6.6 medium. Intracellular pH (pHi) was measured with the dye, 1,4-diacetoxy-2,3-dicyanobenzene, combined with flow cytometry. Time-dependent changes in the intracellular pH occurred under either pH condition. CHO cells incubated under normal pH conditions had a transient increase in the pHi. This pHi elevation was followed by a rapid intracellular acidification of approximately 0.15 to 0.25 pH units. The timing of both the increases and decreases in the pHi was dependent on the magnitude of the initial heat dose. With heat doses less than or equal to 10 min, the pHi returned to normal unheated levels after the acidification phase. Although cells incubated under low pH (6.6) conditions showed similar pHi alterations, differences in the kinetics were measured. The intracellular pH increased immediately after heating. In addition, when intracellular acidification occurred, the rate of acidification was significantly reduced. With heat doses longer than 5 min under the low pH conditions, the pHi did not return to normal unheated levels.

Animals↗
PubMed1987

Membrane fluidity measured by fluorescence polarization using an EPICS V cell sorter.

We have characterized the measurement of fluorescence polarization on single cells using an EPICS V cell sorter. A critical analysis is made of the balancing and calibration of the system. The system is highly linear for polarization measurements. Cellular membranes were labeled with 1,6-diphenyl-1,3,5-hexatriene (DPH) to measure membrane fluidity. Fluorescence polarization histograms had coefficients of variation as low as 7%. Cells labeled with DPH after 24 hr incubation in medium lacking serum showed a significantly higher fluorescence polarization than cells in medium containing serum. The fluorescence polarization measured at 15 degrees C was 0.311 compared to 0.270 at 25 degrees C for cells labeled with DPH, verifying that temperature affects the membrane fluidity as measured by flow cytometry.

Animals↗
PubMed1987

Comparison of synchronized Chinese hamster ovary cells obtained by mitotic shake-off, hydroxyurea, aphidicolin, or methotrexate.

Synchronized cell populations are necessary to study many aspects of cell biology. We have developed a method to obtain highly synchronized Chinese hamster ovary cell populations in S phase or G2 phase by utilizing mitotic selection followed by incubation with either hydroxyurea, aphidicolin, or methotrexate for 12 h. Flow cytometry analysis shows that the coefficient of variation in the spread of the cell population in S phase is as low as 6%. Drug toxicity studies compare the effects of the various drugs on G1 and S phase cells. The use of aphidicolin or hydroxyurea results in the most highly synchronized cell populations, but methotrexate yields inadequate synchronization. These results demonstrate that both aphidicolin and hydroxyurea are useful drugs for obtaining highly synchronized cell populations after an initial synchrony in mitosis. Aphidicolin is perhaps the best choice because of less toxicity to S phase cells when used in low concentrations.

Animals↗
PubMed1987

Flow cytometric analysis of intracellular pH in 3T3 cells.

Techniques to determine intracellular pH generally report the average pH of population and do not indicate whether or not there is significant variance among cells within the population. Population variance is important to ascribe pH changes on a per cell basis. The magnitude of the pH change in individual cells is important to ascribe physiological function to changes in pH. To determine the variability of cell responses, we have used dual wavelength fluorescence emission spectroscopy of intracellular dicyanohydroquinone monitored with flow cytometry to determine the pH of normal and transformed 3T3 cells in response to serum or serum components. All cells were mechanically harvested from subconfluent cultures. Large differences in pH were observed between serum-deprived and serum-conditioned normal, but not transformed, cells. Addition of serum caused cytosolic alkalinization, with the serum-deprived cells responding more slowly. Titration of cells with submaximal doses of serum indicate that the response of pH is graded, that all cells respond in similar manner, and that the relative affinity of transformed cells for the serum components causing the pH effect is about twice that of normal cells.

Animals↗
PubMed1987

Risk factors among patients undergoing repeat aorta-coronary bypass procedures.

It is estimated that as many as 7% of patients who have an aorta-coronary bypass operation will require a second bypass procedure within 10 to 12 years. Using information from the Milwaukee Cardiovascular Data Registry, we matched 166 men who underwent two coronary bypass operations at least 6 months apart, between 1968 and 1981, with 428 patients who had a single procedure. Patients were matched according to date of operation and left ventricular wall motility function for analysis of risk factors for repeat operation. Elevated triglyceride levels were found to be the strongest risk factors associated with reoperation. In addition, both younger age and less complete revascularization during the first operation were significant predictive factors of repeat operation. The results suggest that efforts to reduce plasma triglyceride levels and ensure adequate revascularization may significantly reduce the need for repeat coronary bypass.

Age Factors↗
PubMed1985

The relationship between genome size and synaptonemal complex length in higher plants.

There appears to be only a weak correlation between genome size and the corresponding total length of a complete set of synaptonemal complexes (SCs) based on published evidence for several fungal, plant, and animal species. This result is unexpected, considering the strong positive correlations between genome size (DNA amount) and total chromosome length and volume and between relative lengths of chromosomes and SCs. Because the observed weak correlation was based on limited data, we systematically investigated the relationship between genome size and SC length, using ten higher plant species. Two-dimensional spreads of SCs from primary microsporocytes at pachytene were prepared using a hypotonic bursting technique. The SC spreads were examined either by light or electron microscopy, and the lengths of at least ten complete sets of SCs were measured for each of the ten species. Additionally, the genome size of each species was determined from pollen tetrad protoplasts using flow cytometry. A strong correlation (r = 0.97) between total SC length and genome size was observed for higher plants, indicating a constant amount of DNA is associated with a given length of SC, at least when averaged over the whole genome.

Chromosomes↗
PubMed1985

The cell cycle dependence of thermotolerance. III. HeLa cells heated at 45.0 degrees C.

We have extended our studies on the cell cycle dependence of thermotolerance to include HeLa cells heated at 45.0 degrees C to compare the results to Chinese hamster ovary (CHO) cells. We found that asynchronous HeLa cells were more resistant to heat than CHO cells but showed a similar development and decay of thermotolerance. Flow cytometry (FCM) was used to study redistributions in the cell cycle after an initial heat dose. Cells heated for 35 min at 45.0 degrees C were delayed in G1 by about 7 h compared to controls, with delays in late S and G2/M phase also. The heat sensitivity varied through the cell cycle; G1 cells were the most resistant to heat, while S-phase cells were uniformly sensitive throughout S phase, and G2 cells were resistant. Thermotolerance could be induced and expressed in early or late S-phase cells, but to a lesser extent than for G1 cells. The results were similar in many respects to CHO cells, but there were significant differences.

Acclimatization↗
PubMed1984

The cell cycle dependence of thermotolerance. II. CHO cells heated at 45.0 degrees C.

This report extends our investigations of the cell cycle dependence of the expression of thermotolerance to include tolerance expressed by Chinese hamster ovary (CHO) cells exposed to 45.0 degrees C hyperthermia. We examined the response of asynchronous cells following exposure at 45.0 degrees C. A maximum in thermotolerance under these conditions was reached approximately 12 hr after a 15-min exposure to 45.0 degrees C hyperthermia and progressively decreased thereafter. Cells were delayed in S and G2 phase for 24 hr, after which time cell growth resumed. We then characterized the response of CHO cell populations synchronized in G1 or early or late S phase. We observed that the expression of tolerance depended on the position of cells in the cell cycle and was modulated by changes in the sensitivity of cells as they progressed through the cell cycle subsequent to the tolerance induction dose. We measured the variation in the sensitivity of these cells to 45.0 degrees C hyperthermia throughout the cell cycle and found substantial changes as cells progressed through S phase. Cells in early S phase were the most sensitive to heat at this temperature, and as these cells progressed through S phase, they became progressively more resistant. In addition, G1 cells were delayed for approximately 15 to 18 hr by a 15-min, 45.0 degrees C heat pulse, whereas S-phase cells were delayed to a lesser extent. The data presented in this report suggest that the induction of thermotolerance is relatively non-cell-cycle specific, but the magnitude of expression of tolerance depends on the position of cells in the cell cycle at the time of the subsequent challenge heat dose.

Animals↗
PubMed1983

The cell cycle dependence of thermotolerance. I. CHO cells heated at 42 degrees C.

We examined the dependence of heat killing and thermotolerance on the position and progression of Chinese hamster ovary (CHO) cells in the cell cycle. We measured cell cycle perturbations and survival of asynchronous and synchronized G1-, S-, and G2-phase cells resulting from continuous heating at 42.0 degrees C for up to 80 hr. Thermotolerance under these conditions was transient in nature, was dependent on the position of cells in the cell cycle, and occurred concurrently with a heat-induced delay of progression of G1- and G2-phase cells. When G1 cells were heated, survival decreased to 25% after 4 hr, at which time the thermotolerance was expressed. For G2 cells survival decreased initially at the same rate (T0 congruent to 3 hr) but thermotolerance was not expressed until approximately 12 hr, at which time the survival was 4%. The rate of decrease in survival was much more rapid for cells heated in mid-S phase (T0 congruent to 0.5 hr), and these cells did not express thermotolerance at a measurable level. Concurrent with the expression of thermotolerance, the progression of cells heated in G1 and G2 was delayed. Following the expression of tolerance, progression resumed at a rate approximately equal to the rate of decrease in survival of the G1 population. Cells heated in mid-S phase continued to progress through the cell cycle until they reached G2, where they were also delayed.

Animals↗
PubMed1981

Genetically controlled enzymatic variation in a southern, biracial, semi-rural community: the Bogalusa heart study.

Caucasian and Afro-American schoolchildren participating in a community-based cardiovascular screening program in Bogalusa, Louisiana are characterized with respect to phenotypic and allelic frequencies at 10 enzymatic and 3 nonenzymatic polymorphic loci. Biracial gene frequencies in Bogalusa are compared with those reported for other populations of similar ethnic composition. Intra- and interracial genetic distances within and between the racial subpopulations of Bogalusa and Seattle, Washington are compared. While the comparisons do not support the concept that northern, urban blacks have more Caucasian admixture than blacks residing in the rural South, they do suggest microdifferentiation of the two white populations.

Alleles↗