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R Sager

Publications and source records attributed to R Sager.

159 records · Page 9Linked to original sources

A genetic map of non-Mandelian genes in Chlamydomonas.

A group of eight non-Mendelian genes have been shown by recombination analysis to be linked into a linear structure or chromosome. Similar genetic maps of gene order and relative distances between genes have been constructed by two methods, one based on additivity of recombination frequencies, the other on frequency of reciprocal recombination with a postulated attachment point. The data indicate that the progeny are diploid for this linkage group, and that the strands are distributed in a precisely oriented manner at mitosis. Evidence is discussed in support of the view that this linkage group is located in chloroplast DNA.

Chloroplasts↗

Cytoplasmic and chloroplast ribosomes of Chlamydomonas: ultracentrifugal characterization.

Ribosomes isolated from the cytoplasmic and chloroplast fractions of Chlamydomonas were characterized in the ultracentrifuge. The cytoplasmic ribosomes belong to the 80S class of ribosomes, and, like animal ribosomes, dissociate to 60, 50, and 40S subunits. However, like the ribosomes of microorganisms, they contain smaller RNA's, 24 and 16S, and require 0.01 mole of magnesium ions per liter for stability. Chloroplast ribosomes are 70S like those of higher plants but are very unstable. A stable 50S subunit has been observed.

Chloroplasts↗

Trophoblast viability in perfused term placental tissue and explant cultures limited to 7-24 hours.

Human term-placental culture techniques such as villous explant or dual perfusion are commonly used to study trophoblast function under control and experimentally manipulated conditions. We have compared trophoblast viability during perfusion and in explants cultured under various conditions by monitoring glucose consumption, protein synthesis and secretion, expression of differentiation-specific genes, induction of stress proteins and apoptotic cell death. The tissue was obtained from term-placentae of uncomplicated pregnancies after elective Caesarean delivery. We observed a severe loss of trophoblast viability in explants irrespective of the culture conditions used. Over 7 h of culture the amount of the differentiation specific placental hormones hCG, hPL and leptin accumulated in the medium dropped significantly. Analysis of their expression by semi-quantitative and real-time RT-PCR revealed that the down-regulation of expression occurred at the transcriptional level. This transcriptional repression was accompanied by induction of the stress-proteins RTP and BiP/GRP78. Analysis of apoptotic cell death by TUNEL assay and immunohistochemical detection of the caspase-3-specific degradation product of cytokeratin 18 revealed prominent cell death after 7 h of culture. These results are in contrast to the findings obtained in perfused placental tissue where, after 7 h of culture, hormone secretion, expression of stress proteins and cell death were similar as in native tissue. This difference between villous explant incubation and dual perfusion is also reflected by a significantly higher consumption of glucose in perfused tissue.

Apoptosis↗

Glucose consumption and lactate production of human placental tissue under different conditions of in vitro incubation.

OBJECTIVE: To assess the glycolytic activity of human placental tissue in the third trimester as measured by glucose consumption and lactate production under different conditions of in vitro incubation. METHOD: An incubation technique was used to study the metabolic activity of the human placenta by comparing large blocks (0.3 cm3, T1) and small fragments (explants, 0.03 cm3, T2). Placentas were obtained from premature (28-33 weeks) and term (39-41 weeks) deliveries. In addition, different experimental conditions were used to investigate the influence of incubation medium (Earle's buffer and cell culture medium NCTC = 135), medium oxygen pressure (PO2) (400 and 30 mmHg), and regional sampling of placental tissue (central, intermediate, and peripheral). All media contained glucose (1 g/L). The tissue (2-3 g/25 mL medium, pH 7.2-7.4) was incubated for 3 hours at 37C. 3H-inulin was used for the determination of the extracellular space. RESULTS: Incubation of both tissue forms yielded a higher metabolic activity as measured by glucose consumption and lactate production when incubated with Earle's buffer compared with incubations with NCTC medium. In general, the metabolic activity was consistently higher for small fragments compared with the large blocks. Extracellular space values found for large fragments (25-31%) were significantly lower than for small pieces (39-46%), indicating that an equilibration of the medium with the extracellular space is inadequate with large fragments. Incubation with small fragments showed that 1) there is a tendency for higher metabolic activity during incubation with lower PO2, 2) the metabolically most active part of the placenta is the intermediate tissue region, and 3) placental metabolic activity was significantly higher at 28-33 weeks (n = 5) than at term (n = 6). These differences were not seen with large fragment incubations. CONCLUSION: The smaller tissue fragments are preferable for the in vitro incubation study of placental glucose metabolism. Apparently, there are differences in the metabolic activity with regard to the placental tissue region and gestational age.

Culture Media↗

Effect of physiologic perfusion-fixation on the morphometrically evaluated dimensions of the term placental cotyledon.

OBJECTIVE: To estimate the in vivo dimensions of the fetal villous tree of the normal term placenta. METHODS: Dual-circuit perfusion-fixation of a cotyledon from eight normal term placentas was performed with random intra-cotyledon tissue sampling. Stereologic methods were used to derive estimates of villous (intermediate and terminal) surface area and volume, and star volume (a measure of villous volume). RESULTS: Villous surface area (mean 20.9 m2 [standard deviation 1.8]), capillary surface area (12.8 m2 [1.5]), villous volume (469 mL [40]), and capillary volume (80 mL [10]) values were all approximately 50% higher than reported previously. Star volume estimates ranged from 480 to 1350 microns 3. CONCLUSION: Tissue perfusion-fixation more accurately reconstructs the in vivo state, resulting in higher reference values than previously thought for the fetal villous tree dimensions. Up to one-quarter of fetoplacental blood volume is likely to be accommodated within the placenta at term.

Birth Weight↗