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C McCaffrey

Publications and source records attributed to C McCaffrey.

7 recordsLinked to original sources
PubMed1992

In vitro preimplantation mouse embryo development with incubation temperatures of 37 and 39 degrees C.

Embryos from two strains of mice were used to assess the effect of incubation temperature on pronuclear and two-cell development to the morula/blastocyst (M/B) stage. Embryos from B6D2F2 and B6SJLF1 strains were cultured in medium M16 at either 37 or 39 degrees C until 120 hr post human chorionic gonadotropin (hCG) or 0, 24, or 48 hr at 37 degrees C and the remaining time at 39 degrees C. Overall M/B development for pronuclear embryos was 0.6, 0, 32.3, and 52.4% for 0-96, 24-72, 48-48, and 96-0 hr at 37 and 39 degrees C, respectively. Only 0-96 and 24-72 hr at 37 and 39 degrees C were not different (P greater than 0.10). Overall M/B development for two-cell embryos was 48.1, 78.1, and 98.0% for 0-72, 24-48, and 72-0 hr at 37 and 39 degrees C, respectively. Percentage development at each time was different (P less than .01) for each category. Additionally, the number of nuclei for morulae and blastocysts tended to be higher for embryos initiating culture at the two-cell stage compared to pronuclear embryos. The first cell cycle was most dramatically affected by a 2 degrees C increase in incubator temperature. More advanced embryos can tolerate slight increases in incubator temperature more readily than pronuclear embryos.

Animals↗
PubMed1992

Quinolone accumulation in Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.

The accumulation of quinolones by Escherichia coli JF568, Pseudomonas aeruginosa PAO1, and Staphylococcus aureus ATCC 29213 was measured by a modified fluorometric assay (J. S. Chapman and N. H. Georgopapadakou, Antimicrob. Agents Chemother. 33:27-29, 1989). The quinolones examined were fleroxacin, pefloxacin, norfloxacin, difloxacin, A56620, ciprofloxacin, ofloxacin, and Ro 09-1168. In all three organisms, uptake was complete in less than 5 min and was proportional to extracellular quinolone concentrations between 2 and 50 micrograms/ml, which is consistent with simple diffusion. Washing cells with quinolone-free buffer decreased accumulation by up to 70% in E. coli and P. aeruginosa but not in S. aureus. Similarly, incubation with the uncouplers 2,4-dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone increased accumulation up to fourfold in E. coli and P. aeruginosa, though not in S. aureus, suggesting endogenous, energy-dependent efflux. High quinolone hydrophobicity was generally associated with decreased accumulation in E. coli and P. aeruginosa (except in the case of pefloxacin) but was associated with increased accumulation in S. aureus (except in the case of difloxacin). Ciprofloxacin had the highest accumulation in E. coli and P. aeruginosa, while pefloxacin had the highest accumulation in S. aureus.

4-Quinolones↗
PubMed1991

Successful co-culture of 1-4-cell cattle ova to the morula or blastocyst stage.

Bovine ova (n = 326) collected at the 1-4-cell stage were cultured in TCM-199 + 10% foetal calf serum with or without oviducal cells. The bovine oviducal cells were collected and seeded either on the day of ovum recovery (BOC-0) or 3 days earlier (BOC-3). In Exp. 1, the effect of age of oviducal cells in co-culture on ovum development was examined. In the BOC-0 and BOC-3 treatments, respectively, 36/46 (78%) and 30/37 (81%) of ova developed to morulae or blastocysts, while no ova developed past the 8-16-cell stage in the absence of oviducal cells. In Exp. 2, the effect of age of oviducal cells and of physical contact between the oviducal cells and ova on ovum development was examined. In the BOC-0 and BOC-3 treatments, respectively, 29/42 (69%) and 23/43 (53%) of the ova developed to morulae or blastocysts, while 1/42 (2%) developed to the morula stage in the absence of oviducal cells. Physical separation of the ova using a microporous membrane inserted between the oviducal cells and the ova did not affect ovum development, with 26/42 (62%) and 22/42 (52%) of ova developing to morulae or blastocysts in the BOC-0 and BOC-3 treatments, respectively. A high proportion of the morulae and blastocysts in Exp. 1 (57/66, 86%) and Exp. 2 (67/100, 67%) were of quality grades 1 or 2, with mean nuclei counts of 85 for morulae and 111 for blastocysts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1979

Peritoneal clearance of creatinine and inulin during dialysis in dogs: effect of splanchnic vasodilators.

The peritoneal clearance of creatinine and inulin during isotonic peritoneal dialysis was studied in dogs before and after the administration of two vasodilators: isoproterenol and glucagon. One-liter exchanges with 15-min dwell times were used. Blood flow in the superior mesenteric artery was recorded with an electromagnetic flow probe and used as an index for total splanchnic flow. Intravenous isoproterenol (2.4 microgram/min) increased blood flow by 88%, but did not alter peritoneal clearance. When isoproterenol was given i.p. (0.5 microgram/ml dialysis fluid), blood flow increased by 81%, and inulin clearance rose by 26.8% (2.06 +/- 0.18 to 2.61 +/- 0.23 ml/min; P less than 0.05). Creatinine clearance increased by 17.5%, from 10.94 +/- 0.32 to 12.85 +/- 0.34 ml/min (P less than 0.05). When blood flow was returned to control levels with a clamp, clearances also returned to control levels. Glucagon given i.p. (1.0 microgram/ml) had no effect on any measured variable. Glucagon given i.v. at 10 microgram/min caused blood flow to rise by 81% and inulin clearance to rise by 25% from 1.88 +/- 0.16 to 2.35 +/- 0.19 ml/min (P less than 0.05). Cp, inulin clearance remained at control levels. Vasodilators seem to exercise two effects in augmenting clearance of small and middle-sized molecules: a direct permeability effect and a blood-flow related effect, possibly in increasing the surface area available for exchange and/or increasing permeability of the capillary endothelium.

Animals↗
PubMedDate not supplied

Introduction of unnatural amino acids into chalcone isomerase.

The active site cysteine residue of chalcone isomerase was rapidly and selectively modified under denaturing conditions with a variety of electrophilic reagents. These denatured and modified enzyme were renatured to produce enzyme derivatives containing a series of unnatural amino acids in the active site. Addition of methyl, ethyl, butyl, heptyl, and benzyl groups to the cysteine sulfur does not abolish catalytic activity, although the activity decreases as the steric bulk of the amino acid side-chain increases. Modification of the cysteine to introduce a charged homoglutamate or a neutral homoglutamine analogue results in retention of 22% of the catalytic activity. Addition of a methylthio group (SMe) to the cysteine residue of native chalcone isomerase preserves 85% of the catalytic activity measured with 2',4',4-trihydroxychalcone, 2',4',6',4-tetrahydroxychalcone, or 2'-hydroxy-4-methoxychalcone as substrates. The competitive inhibition constant for 4',4-dihydroxychalcone, the substrate inhibition constant for 2',4',4-trihydroxychalcone, and other steady-state kinetic parameters for the methanethiolated enzyme are very similar to those of the native enzyme. The strong binding of 4',4-dihydroxychalcone to the methanethiolated enzyme shows that there is no steric repulsion between this modified amino acid residue and the substrate analogue. This structure-activity study clearly demonstrates that the active site cysteine residue does not function as an acid-base or nucleophilic group in producing the catalysis or substrate inhibition observed with chalcone isomerase. The method presented in this paper allows for the rapid introduction of a series of unnatural amino acids into the active site as a means of probing the structure-function relationship.

Alkylation↗