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Biomedical subjects

J E Morris

Publications and source records attributed to J E Morris.

At least 55 records · Page 3Linked to original sources

Effects of sulfur dioxide or ammonium sulfate exposure, alone or combined, for 4 or 8 months on normal and elastase-impaired rats.

Normal and lung-impaired rats were compared after exposure to SO2 and/or (NH4)2SO4 for 4 or 8 months, or for 8 months plus 3 months recovery. Young adult male Sprague-Dawley rats were pretreated intratracheally with either physiologic saline (normal lungs) or porcine pancreatic elastase (impaired lungs). Rats from each pretreatment group were exposed to filtered air (control), to SO2 (1 ppm) or (NH4)2SO4 (0.5 mg/m3), or to combined SO2 + (NH4)2SO4 for 5 hr/day, 5 days/week. Morphologic, physiologic, and immunologic criteria were evaluated. At 4 months cellular immunologic responsiveness was not impaired, but physiologic changes were detected. Morphologic changes were apparent in all time periods. Elastase-induced changes included greater lung volumes, emphysema, and alveolar interstitial fibrosis. Pollutant effects included bronchiolar epithelial hyperplasia and changes in alveolar mean chord length (MCL). Relative to controls, bronchiolar epithelial hyperplasia and MCL increased in saline/pollutant groups, but decreased in elastase/pollutant rats at 4 months. The pretreatment/pollutant interaction was not observed at 8 months. Elastase effects persisted throughout the recovery period. Pollutant effects were more transitory, although alveolar septal fibrosis was greater in saline/(NH4)2SO4 rats at 8 months. Pulmonary function changes associated with elastase included increases in residual volume, functional residual capacity, and the residual volume/total lung capacity ratios. The alveolar plateau of single-breath washout (N2 slope) was significantly steeper in elastase-treated rats but less steep in animals exposed to SO2 or to (NH4)2SO4 than in those exposed to air only.

Air Pollutants↗

Estradiol-stimulated turnover of heparan sulfate proteoglycan in mouse uterine epithelium.

Heparan sulfate proteoglycans (HSPGs) and dermatan sulfate/chondroitin sulfate proteoglycans may be extracted from the uterine epithelium of immature mice by a 1-min exposure of the luminal surface of excised uteri to 1% Nonidet P-40 detergent. In mice that are treated with estradiol there is a marked increase in free heparan sulfate glycosaminoglycan in the extract. (a) By Sepharose exclusion chromatography the [35S]sulfate-labeled major HSPG had a nominal Mr of 200-250 X 10(3), consisting of a core protein of about 80-90 X 10(3) Mr with about 8-10 heparan sulfate glycosaminoglycan chains (Mr = 13 X 10(3)). The HSPG had a lower bouyant density (less than 1.45 g/ml) than the dermatan sulfate/chondroitin sulfate proteoglycan and was heterogeneous, as was evident in the fact that HSPG attained equilibrium over a wide range of CsCl densities and also showed nonuniform interaction with octyl-Sepharose. (b) Virtually all of the major HSPG was removed when the epithelium was isolated by proteolysis, indicating a cell surface localization. A smaller, less prominent HSPG (nominal Mr = 80 X 10(3)) was synthesized during the first 2 h after isolation. (c) Label and chase experiments with and without chloroquine showed that virtually all of the free heparan sulfate glycosaminoglycan chains derived from endocytosis and lysosomal degradation of the plasma membrane-associated HSPG. We conclude that estradiol stimulates endocytosis of HSPG, predominantly from the basolateral epithelial surface and suggest that this HSPG turnover may reflect changes associated with blastocyst attachment and invasion of the endometrium.

Animals↗

Simplifying infusion chemotherapy: preliminary communication.

A low-intervention policy for Hickman catheter maintenance has been evaluated and found to be safe and cost effective. A simple, lightweight, disposable device has been used for 24-hour ambulatory home infusion. Implementation of this policy as part of a prospective randomized trial of single-agent chemotherapy in advanced breast cancer has demonstrated that slow intravenous infusion reduces the acute toxicity of epirubicin when compared with bolus injection.

Breast Neoplasms↗

Estradiol induces an accumulation of free heparan sulfate glycosaminoglycan chains in uterine epithelium.

Blastocyst implantation in the mouse uterus is triggered by an estrogen surge that induces a number of characteristic changes in biosynthetic activity of the endometrial stroma and epithelium, including a reduction in the thickness of the epithelial plasma membrane coat, immediately preceding attachment of the blastocyst. A brief exposure of the uterine luminal surface to 1% Nonidet-40 (NP-40) nonionic detergent extracted this surface coat along with membrane-associated and soluble [35S]sulfate-labeled proteoglycans specifically from the epithelium. Extraction from the underlying stroma appeared to have been prevented by the epithelial basal lamina, which was minimally affected by 1% NP-40, but was extracted by subsequent treatment with 1% NP-40 in 1 M KCl. About 70-80% of the [35S]sulfate in proteoglycans extracted from the uteri of ovariectomized mice with NP-40 was associated with heparan sulfate proteoglycans that eluted from Sepharose CL-4B in a peak at 0.55 Kd. All remaining radioactivity could be accounted for by dermatan sulfate/chondroitin sulfate proteoglycans. Specifically, in the NP-40 extract of uteri from ovariectomized mature mice and intact immature mice, but not in the culture medium or residual tissue, estradiol induced a pronounced increase in the relative amount of radioactivity in a 0.8 Kd peak, which appeared to contain only free heparan sulfate glycosaminoglycan chains. The same relative increase was seen in the NP-40 extract of uteri from intact 3.5-day pregnant mice. We conclude that estradiol induces cellular changes that result in an increased accumulation of epithelial heparan sulfate proteoglycan degradation product immediately preceding normal attachment of the blastocyst.

Animals↗

Proteoglycans synthesized by embryonic chicken retina in culture: composition and compartmentalization.

Characteristics of the chondroitin sulfate/dermatan sulfate proteoglycans (CS/DSPGs) and heparan sulfate proteoglycans (HSPGs) from retinas of 14-day chicken embryos were examined following specific lyase digestion of the HSPG and CS/DSPG glycosaminoglycans, respectively. On the basis of gel exclusion chromatography the prevalent CS/DSPGs in the tissue were above Mr 400 X 10(3) with two or three glycosaminoglycan chains of Mr 60-70 X 10(3). The HSPGs existed in two distinct populations in the tissue. Those in the dominant population appeared to be in the range of Mr 250-300 X 10(3) with 9 to 12 glycosaminoglycan chains of Mr 15-25 X 10(3). The other population consisted of free heparan sulfate chains of Mr 15-25 X 10(3). The HSPGs in the medium tended to be intermediate in size. To examine the distribution of proteoglycans, tissues were sequentially homogenized and extracted in saline and reextracted with 4 M guanidine HCl (GdnHCl) and Triton X-100 (TX), or they were washed in heparin solution and dissociated to single cells with trypsin before sequential extraction in saline and GdnHCl with TX. Through comparison of the results of these two extraction methods, CS/DSPGs were found to be almost entirely within the medium or matrix or loosely associated with the cell surface, and most HSPGs were associated with either the basal lamina or the plasma membrane. The single heparan sulfate glycosaminoglycan chains appeared to be intracellular degradation products. These results support reports that CS/DSPGs may be present in the retina interphotoreceptor matrix and that HSPGs may be present in regions of synaptogenesis, associated with cell membranes.

Animals↗

Development of mouse embryos in hanging drop culture.

Mouse blastocysts were cultured in hanging drops for up to 6 days in order to study development under conditions that avoid the distortion of embryos typically seen when they are allowed to attach to a glass or plastic surface. The survival rate of embryos in hanging drops was equal to that of embryos attached to culture dishes and superior to that of embryos suspended in gyrating flasks. Development of the embryonic portion was similar to that in vivo and on culture dishes but slower than in vivo; the egg cylinder stage was reached after 8-10 equivalent gestation days (4 to 6 days in culture), while that stage is reached at 5.5 to 6 days in vivo. The trophectoderm, however, developed in a unique manner. The cells migrated away from the inner cell mass (ICM), similar to embryos on a culture dish, but without a surface on which to spread they clustered distal to the ICM. In vivo, trophectoderm remained covering the ICM. By 5 days in hanging drop culture the embryos had developed a segmented appearance with trophoblast giant cells at the abembryonic pole, extraembryonic cells not covered by vacuolated endoderm in the central region, and embryonic endoderm surrounding a developing proamniotic cavity in embryonic ectoderm at the embryonic pole. These observations suggest that the trophectoderm is able to follow a developmental program independent of that in the embryonic portion and that its behavior is dominated by the different adhesive properties of the trophoblastic and embryonic cells.

Animals↗

Importance of low serum vitamin B12 and red cell folate concentrations in elderly hospital inpatients.

To determine the functional importance of the low B12 and red cell folate concentrations repeatedly observed in the elderly 200 consecutive patients admitted to a geriatric unit were studied. Forty six of the patients had low serum concentrations of B12 (15), red cell folate (26), or both (five). Serum B12 and red cell folate concentrations correlated with mean cell volume, and serum B12 correlated with the neutrophil lobe count. Bone marrow deoxyuridine suppression was abnormal in 35% of the patients with low vitamin concentrations, but 55% of those with abnormal deoxyuridine suppression had morphologically normal bone marrow, and 73% had a normal mean cell volume. In patients with low vitamin values the deoxyuridine suppressed value correlated with the haemoglobin concentration and neutrophil lobe count. Thus synthesis of thymidylate was impaired by vitamin B12 or folate deficiency in at least 8% of newly admitted elderly patients, many of whom had normal blood counts despite the biochemical disturbance affecting haemopoiesis. A nutritionally depleted diet may have been responsible for many of the low vitamin values.

Aged↗

Isolation of the major chondroitin sulfate/dermatan sulfate and heparan sulfate proteoglycans from embryonic chicken retina.

A technique is presented for the preparation of three major proteoglycans from 14-day embryonic chicken retinas following their culture overnight with [35S]sulfate and either [3H]glucosamine or [3H]serine. Homogenization of the tissue in saline permitted extraction of heterogeneous soluble proteoglycans separately from most of the heparan sulfate proteoglycans. The latter were extracted from the 140,000g pellet with 0.5% Triton X-100 in 8 M urea. The medium plus the saline and urea-detergent extracts were separated from low-molecular-weight contaminants, and fractionated into two peaks of radioactivity on Sephacryl S-300 in saline with 3 M urea and 0.5% Triton X-100. The proteoglycans were isolated directly from these fractions on DEAE-Sephacel, and subjected to ultrafiltration concentration and then further purification on cesium chloride density gradient centrifugation in 4 M guanidine hydrochloride. A further step involving cetylpyridinium chloride precipitation was examined, but it resulted in essentially no further purification. The fractionations separated a large chondroitin sulfate/dermatan sulfate proteoglycan from the culture medium that was excluded from S-300 and of low buoyant density; a large heparan sulfate proteoglycan from the urea-detergent extract that was also excluded from S-300 and of low buoyant density; and two smaller and possibly related heparan sulfate proteoglycans. One was found in the medium and showed low to intermediate buoyant density; the other was isolated from the urea-detergent extract and showed a significantly higher buoyant density, associated with a lower protein content. The saline extract contained both of the two larger proteoglycans and only minor amounts of the smaller molecules.

Animals↗

A comparison of developmental changes in surface charge in mouse blastocysts and uterine epithelium using DEAE beads and dextran sulfate in vitro.

The ability of 3.5-day mouse blastocysts and vesicles prepared from maternal uterine epithelium to adhere to surfaces by charge interactions was compared by observing their adhesion to DEAE-Sephadex beads in the presence of increasing concentrations of dextran sulfate. The adhesion frequency for both the blastocysts and epithelium declined in a manner suggesting that predominantly ionic sites were being titrated, but differences between the two tissues in characteristics of the titration curve and susceptibility to neuraminidase digestion indicated that nonionic interactions were relatively more important for blastocysts. Because the threshold concentration of dextran sulfate required to initiate displacement of uterine epithelium from the DEAE beads was at least 4X that required to initiate the displacement of blastocysts, we argue that the uterine epithelium had at least 4X more interactive charged groups on its surface than the blastocysts. These differences were even more pronounced 4.5 days after mating, a time when attachment to the uterine epithelium is normally first seen in vivo. Blastocysts isolated at this time showed a marked increase in resistance to polyanion competition, but the epithelium showed a nearly 50% decline in surface negative charge that was not compensated by nonionic mechanisms. These observations support the conclusion that the initial adhesion of blastocysts in vivo is accompanied by a reduction in negativity of the uterine epithelial surface and by the formation of new trophoblast cell surfaces that adhere by nonelectrostatic interactions.

Agglutination↗

Low buoyant density proteoglycans from saline and dissociative extracts of embryonic chicken retinas.

Retinas were labeled in culture with [3H]glucosamine or [3H]leucine and [35S]sulfate and extracted sequentially with physiologically balanced saline and 4 M guanidine HCl. They were dialyzed into associative conditions (0.5 M NaCl) and chromatographed on agarose columns. Under these conditions, some of the proteoglycans were associated in massive complexes that showed low buoyant densities when centrifuged in CsCl density gradients under dissociative conditions (4 M guanidine HCl). Much of the label in these complexes was in molecules other than proteoglycans. Most of the proteoglycans, however, were included on the agarose columns, where they appeared to be constitutionally of low buoyant density. They resisted attempts to separate potential low buoyant density contaminants from the major proteoglycans by direct CsCl density gradient centrifugation or by the fractionation of saline or 8 M urea extracts on diethylaminoethyl-Sephacel. The diethylaminoethyl-Sephacel fractions were either subjected to CsCl density gradient centrifugation or were chromatographed on Sephacryl S-300, in both cases before and after alkaline cleavage, to confirm the presence of typical O-linked glycosaminoglycans. The medium and balanced salt extracts were enriched in chondroitin sulfate and other sulfated macromolecules, possibly highly sulfated oligosaccharides, that resisted digestion by chondroitinase ABC but were electrophoretically less mobile than heparan sulfate. Guanidine HCl or urea extracts of the residues were mixtures of high and low density proteoglycans that were enriched in heparan sulfate.

Animals↗

Hematologic and immunologic effects of pulsed microwaves in mice.

Mice were exposed in the far field in an anechoic chamber to 2,880-MHz pulsed microwaves 3 to 7.5 h daily, 5 days/week for 60 to 360 h. Three experiments were performed at average power densities of 5 mW/cm2 and six at 10 mW/cm2, corresponding to averaged specific absorption rates (SARs) of 2.25 and 4.50 mW/g, respectively. Each experiment consisted of eight mice, with a concurrently sham-exposed group of eight. In two of three studies at 5 mW/cm2, there was a significant increase in bone marrow cellularity in the microwave-exposed groups compared to the sham-exposed groups. Significant differences were occasionally seen in erythrocyte, leukocyte, and platelet values from microwave-exposed groups, but were not consistently observed. In one of six groups exposed at 10 mW/cm2, mean bone marrow cellularity was reduced significantly in the microwave-exposed mice; in another group, the lymphocyte count was increased. In only one exposure (10 mW/cm2 for 360 h) was any significant effect noted on serum proteins: a reduction to 5.1 +/- 0.3 g/dl in the exposed versus 5.6 +/- 0.4 g/dl in the sham-exposed mice. This was due to a decrease in alpha and beta globulins, with no effect on albumin or gamma globulin concentrations. No effect on bone marrow granulocyte/macrophage colony-forming units (CFU) was revealed following exposure of mice to pulsed microwaves at 5 mW/cm2. In one of four exposures at 10 mW/cm2, there was a significant increase in CFU-agar colonies. No significant effects of exposures at 10 mW/cm2 were observed on in vivo and in vitro assays of cell-mediated immune functions. No exposure-related histopathologic lesions were found from examination of several tissues and organs. Results of these series of exposures of mice at SARs of 2.25 and 4.50 mW/g indicated no consistent effects on the hematologic, immunologic, or histopathologic variables examined.

Animals↗

Adhesion of mouse blastocysts to uterine epithelium in culture: a requirement for mutual surface interactions.

Blastocysts readily adhered to inert materials in culture, but they resisted adhesion to living cells even after several days under conditions which encouraged cell aggregation. As far as could be determined by observing their spreading behavior on polylysine- and polyglutamate-coated dishes, the mechanism of adhesion of blastocysts to inert surfaces was similar to that of freshly dissociated cells and cell lines. However, their adhesion to vesicles of isolated uterine epithelium, which was encouraged by hanging drop culture, was by a different mechanism that involved microvilli on both the embryonic and maternal surfaces. This interactive step, which was similar to that seen during attachment in vivo, was followed by a brief period of close trophoblast-epithelial contact which led ultimately to phagocytosis of sloughed epithelium. Blastocysts showed a clear preference for adhesion to cultured epithelium in vesicles that had begun to collapse. In this case the cells showed a columnar profile with sharply defined microvillous apexes, unlike the flattened cells in fully expanded vesicles or on culture dishes. We conclude that the preimplantation adhesion of mouse blastocysts requires specific changes on both the embryonic and maternal surfaces to overcome the mutual nonadhesiveness typical of epithelia. The relatively rapid adhesion of blastocysts to a culture dish, on the other hand, is more typical of the well-known spreading behavior of cells on a highly attractive surface.

Animals↗

Pathologic effects in rodents exposed to sodium combustion products.

Three species of rodents exhibited respiratory distress when exposed to aerosols of sodium combustion products, the major constituent of which was shown to be sodium carbonate. At the higher doses, animals died during or shortly after 2 hr exposures. Lesions, primarily involving the pharynx and larynx, included accumulation of mucus, vesiculation, and mucosal edema. Other lesions included edema and vesiculation of the anterior trachea, hemorrhage in the lungs, and severe gastric tympany. Basal epithelial cells of the posterior pharynx and anterior trachea had enlarged mitochondria following exposures of 1 hr or more.

Animals↗