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Biomedical subjects

M E Palmer

Publications and source records attributed to M E Palmer.

At least 19 recordsLinked to original sources

Development of the chick chorioallantoic capillary plexus under normoxic and normobaric hypoxic and hyperoxic conditions: a morphometric study.

Fertile eggs from the domestic fowl were incubated under normobaric normoxic (21% O2), hypoxic (14% O2), and hyperoxic (40% O2) conditions in order to examine the influence of the prevailing oxygen level on the growth and maturation of the chorioallantoic membrane. Eggs were sampled at regular stages throughout incubation for morphometric analysis. Under normoxic conditions, maturation of the capillary plexus occurred in two distinct stages, both of which contributed to a reduction in the thickness of the air-blood barrier. Between days 6 and 10, the capillaries sprouted and fused to form a dense plexus. Subsequently, between days 10 and 14, this plexus invaginated into the chorionic epithelium. Differentiation of the chorioallantoic membrane appeared maximal by the end of this period. Hypoxia resulted in diminished growth of the embryo and chorioallantoic membrane, but in accelerated maturation of the capillary plexus. Hyperoxia had a less marked effect but appeared to retard the final invagination of the plexus, resulting in a thicker air-blood barrier.

Allantois

Effects of sulfate deprivation on the production of chondroitin/dermatan sulfate by cultures of skin fibroblasts from normal and diabetic individuals.

Human skin fibroblast monolayer cultures from two normal men, three Type I diabetic men, and one Type I diabetic woman were incubated with [3H]glucosamine in the presence of diminished concentrations of sulfate. Although total synthesis of [3H]chondroitin/dermatan glycosaminoglycans varied somewhat between cell lines, glycosaminoglycan production was not affected within any line when sulfate levels were decreased from 0.3 mM to 0.06 mM to 0.01 mM to 0 added sulfate. Lowering of sulfate concentrations resulted in diminished sulfation of chondroitin/dermatan in a progressive manner, so that overall sulfation dropped to as low as 19% for one of the lines. Sulfation of chondroitin to form chondroitin 4-sulfate and chondroitin 6-sulfate was progressively and equally affected by decreasing the sulfate concentration in the culture medium. However, sulfation to form dermatan sulfate was preserved to a greater degree, so that the relative proportion of dermatan sulfate to chondroitin sulfate increased. Essentially all the nonsulfated residues were susceptible to chondroitin AC lyase, indicating that little epimerization of glucuronic acid residues to iduronic acid had occurred in the absence of sulfation. These results confirm the previously described dependency of glucuronic/iduronic epimerization on sulfation, and indicate that sulfation of the iduronic acid-containing disaccharide residues of dermatan can take place with sulfate concentrations lower than those needed for 6-sulfation and 4-sulfation of the glucuronic acid-containing disaccharide residues of chondroitin. There were considerable differences among the six fibroblast lines in susceptibility to low sulfate medium and in the proportion of chondroitin 6-sulfate, chondroitin 4-sulfate, and dermatan sulfate. However, there was no pattern of differences between normals and diabetics.

Aggrecans

Production of [3H]hexosamine-labeled proteoglycans by cultures of normal and diabetic skin fibroblasts: dilution of exogenous [3H]glucosamine by endogenous hexosamine from glucose and other sources.

Human skin fibroblast monolayer cultures from two normal men, three Type I diabetic men, and one Type I diabetic woman were incubated with [3H]glucosamine and [35S]-sulfate for varying periods of time. Incorporation of 3H into macromolecules appearing in the medium was linear after approximately 45 min, and incorporation of 35S was linear after approximately 30 min. The amounts of 35S-proteoglycan formed by each of the cultures during 5-h incubations were compared and were found to be fairly similar for the six lines, varying from 0.08 to 0.14 nmol sulfate/microgram DNA. Isolated 3H,35S-glycosaminoglycans were then treated with chondroitin ABC lyase to characterize the location and degree of sulfation. Results indicated a considerable variation in completeness of chondroitin/dermatan sulfation and in proportions of 6-sulfation to 4-sulfation among the various lines. However these variations did not seem to be related to whether the cells were from normals or diabetics. 3H,35S-Labeled disaccharides were isolated and ratios of 3H to 35S determined in order to calculate the [3H]glucosamine dilution by endogenous glucosamine derived from glucose or other sources during the period of incubation. Dilutions varied widely from 160- to 635-fold among the different cell lines, but the variations did not seem to be related to whether the cells were from normals or diabetics.

Adult

Morphometric differences between the placental vasculature of non-smokers, smokers and ex-smokers.

The aim of this study was to determine whether cigarette smoking during pregnancy has an adverse effect upon the placenta's capacity for gaseous exchange. Using recently developed stereological techniques, in conjunction with perfusion fixation, computer-assisted measurements were made on the placentas of 15 non-smokers, 15 moderate smokers, 15 heavy smokers and 13 ex-smokers, 7 of whom stopped smoking during the course of the pregnancy. Compared with the placentas of non-smokers and of those who stopped before pregnancy, it was found that the placentas of smokers and of those who stopped after conception exhibited a reduced capillary volume fraction, and an increased thickness of the villous membrane. Although they must impair gaseous exchange across the placenta, these changes were less severe than suggested by previously published reports. Nonetheless it is clear that in order to prevent these changes women should stop smoking before conception rather than during the course of a pregnancy.

Adult

The chorioallantoic capillary plexus of the chicken egg: a microvascular corrosion casting study.

The chorioallantoic membrane of the avian egg serves as the principal organ of respiratory gaseous exchange for the embryo until close to hatching. It lies closely apposed to the inner shell membrane and contains an extremely dense capillary plexus supplied by the allantoic blood vessels. This study applied the microvascular corrosion casting technique to investigate the three-dimensional arrangement of the plexus at various stages of incubation. Casts were produced between days 6 and 14 of incubation, and their appearances were compared with those obtained from traditionally sectioned material and from freeze-cleaved specimens. By day 6 the capillary network was remarkably profuse but showed considerable regional variation in vessel density. In some areas there were only short capillary buds whereas in other areas fusion had taken place so that a true plexus was formed. By day 10 the capillaries had become confluent to such a degree that the cast consisted of a thin sheet of resin perforated only by an array of small irregularly shaped orifices. These corresponded closely in size to the intervening columns of chorionic epithelial cells seen in the sectioned material. It is clear from the appearances of the casts that the capillary surface density becomes maximal at approximately day 10 of incubation. From then on in incubation any increase in the diffusing capacity of the chorioallantoic membrane must be the result of either an increase in its overall surface area, or a decrease in the thickness of the air-blood barrier.

Allantois

Activation of N-methyl-D-aspartate-sensitive glutamate receptors stimulates arachidonic acid release in primary cultures of cerebellar granule cells.

In cultured granule cells prelabeled with [3H]arachidonate the activation of excitatory amino acid receptors by various agonists results in a dose-dependent stimulation of [3H]arachidonic acid release. Glutamate and aspartate were the most potent agonists, whereas N-methyl-D-aspartate, kainate and quisqualate were less potent. Other neurotransmitter receptor agonists--GABA, baclofen and norepinephrine--were inactive, while carbachol induced only a slight effect. Since the transmitter-mediated release of [3H]arachidonate was blocked by phencyclidine, a selective inhibitor of NMDA-sensitive glutamate receptors, it can be inferred that the effects of all other receptor agonists were indirectly mediated via the release of glutamate from granule cells. Aspartate-evoked release was Ca2+-dependent and was abolished by the glutamate receptor inhibitors: Mg2+ ions and 2-amino-5-phosphonovalerate. The inhibitors of phospholipase A2, quinacrine and p-bromophenacyl bromide, decreased the release of [3H]arachidonate in a dose-related manner.

Animals

Formation of dermatan sulfate by cultured human skin fibroblasts. Effects of sulfate concentration on proportions of dermatan/chondroitin.

[3H,35S]Dermatan/chondroitin sulfate glycosaminoglycans produced during culture of fibroblasts in medium containing varying concentrations of sulfate were tested for their susceptibility to chondroitin ABC lyase and chondroitin AC lyase. Chondroitin ABC lyase completely degraded [3H]hexosamine-labeled and [35S] sulfate-labeled dermatan/chondroitin sulfate to disaccharides. Chondroitin AC lyase treatment of the labeled glycosaminoglycans produced different results. With this enzyme, dermatan/chondroitin sulfate formed at high concentrations of sulfate yielded small glycosaminoglycans and larger oligosaccharides but almost no disaccharide. This indicated that the dermatan/chondroitin sulfate co-polymer contained mostly iduronic acid with only an occasional glucuronic acid. As the medium sulfate concentration was progressively lowered, there was a concomitant increase in the susceptibility to degradation by chondroitin AC lyase. Thus, the labeled glycosaminoglycans formed at the lowest concentration of sulfate yielded small oligosaccharides including substantial amounts of disaccharide. The smaller chondroitin AC lyase-resistant [3H,35S]dermatan/chondroitin sulfate oligosaccharides were analyzed by gel filtration. Results indicated that, in general, the iduronic acid-containing disaccharide residues present in the undersulfated [3H,35S]glycosaminoglycan were sulfated, whereas the glucuronic acid-containing disaccharide residues were non-sulfated. This work confirms earlier reports that there is a relationship between epimerization and sulfation. Moreover, it demonstrates that medium sulfate concentration is critical in determining the proportions of dermatan to chondroitin (iduronic/glucuronic acid) produced by cultured cells.

Adult

The use of lead methacrylate 2-ethylhexanoate to create a radio-opaque resin suitable for microvascular corrosion casting.

A radio-opaque methacrylate-based resin suitable for microvascular corrosion casting can be prepared by substituting lead methacrylate 2-ethylhexanoate for a proportion of the methyl methacrylate monomer normally used. Best results are achieved when conventional methacrylate resin is injected first to fill the vascular tree, and this is chased with the radio-opaque resin. The pattern of arterial and arteriolar distribution can then subsequently be followed by means of a radiographic survey of the cast, and this is of value prior to sampling from vascular systems which show functional subdivisions that cannot be correlated with any surface features. The radio-opacity of the resin and the quantity injected can be varied easily to suit individual requirements.

Acrylates

The influence of mode of fixation on morphometrical data derived from terminal villi in the human placenta at term: a comparison of immersion and perfusion fixation.

The placenta is a highly vascular organ which is subjected to a variable degree of compression during delivery. Partial collapse of the fetal capillary network inevitably results, and this has significant effect upon the maternofetal diffusion distance. Fixation via a dual perfusion system operating at physiological pressures can redistend the vascular tree to its antenatal state. Morphometrical analysis provides a volume fraction for the fetal capillaries within the terminal villi of 38.4 per cent, a figure which approximates closely to values obtained from in situ biopsy of the placenta. The harmonic mean diffusion distance was found to be 3.6 micron. Perfusion fixation resulted in a lower coefficient of variation for many parameters compared with traditional immersion fixation. By negating the unpredictable influence of the placental transfusion, it is considered that perfusion fixation will make true interplacental variations easier to detect.

Female