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J E Morris

Publications and source records attributed to J E Morris.

At least 19 recordsLinked to original sources
PubMed1994

Estradiol-17 beta stimulates proliferation of uterine epithelial cells cultured with stromal cells but not cultured separately.

There is indirect evidence that the in vivo proliferative response of rodent uterine epithelium to estrogen requires interaction with the underlying stroma in pre- and post-pubescent animals. To examine this potential requirement directly, the proliferative response of epithelium to 17 beta-estradiol in the presence or absence of stroma was measured in vitro. Uterine epithelial and stromal cells were isolated separately from immature or adult mice, and were maintained as monocultures or cocultures in defined, serum-free medium with or without 8 x 10(-9) M 17 beta-estradiol. Incorporation of bromodeoxyuridine into the DNA was determined by immunolabeling to assay proliferation in individual cells. Cell morphology and immunolabeling of cytokeratin were used to distinguish epithelial from stromal cells. Treatment of cocultures with 17 beta-estradiol for 24 h increased the proliferation of epithelial cells relative to controls approximately threefold, whereas, in monocultures of epithelial or stromal cells 17 beta-estradiol decreased the number of bromodeoxyuridine-incorporating cells by approximately half. Furthermore, cell contact between epithelial and stromal cells was important for the effects of 17 beta-estradiol on cells in cocultures. Approximately three quarters of the 17 beta-estradiol-induced proliferation of epithelial cells in cocultures was produced by epithelial cells within colonies that were also contacting stromal cells. These results are consistent with the hypothesis that stromal cells mediate the estrogenic proliferative response, and provide evidence that this mediation involves cell contact or stroma-mediated changes in the microenvironment immediately around the epithelial cell.

Animals↗
PubMed1994

Specific stimulation of basal lamina heparan sulfate proteoglycan in mouse uterine epithelium by Matrigel and by transforming growth factor-beta 1.

The basal lamina of differentiated epithelium normally turns over only slowly unless stimulated by tissue repair and growth. We show here that one mechanism of this stimulation, as modeled by basal lamina proteoglycan synthesis, may be the release of basal lamina-bound transforming growth factor (TGF-beta). A large heparan sulfate proteoglycan (HSPG, 0.2 Kav on Sepharose CL-4B) that was extractable from mouse uterine epithelium with 4 M guanidine-HCl or 1 M KCl was recognized by a specific monoclonal antibody to the basal lamina HSPG, perlecan. This HSPG was metabolically inactive with respect to [35S]-sulfate labeling in pieces of whole uterus during 4 h of culture, but it was labeled in isolated cells under the same conditions, provided that the cells had been cultured at least 6 to 12 h before labeling. The rate of labeling was then constant during at least 4 days in culture in serum-containing medium. Cultures on Matrigel showed an enhanced [35S]-sulfate labeling specifically in the 0.2 Kav HSPG fraction. Partial stimulation was obtained with a serum-free medium extract of Matrigel, which fractionated on Sephadex G-50 in two components; a major one > 30 kDa and the other at about 15 to 25 kDa. The specific stimulation was mimicked by the addition of 10 ng/ml of TGF-beta 1, but there was no specific stimulation by basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulinlike growth factor-1 (IGF-1), or interleukin-1 (IL-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Distribution of heparan sulfate proteoglycans in embryonic chicken neural retina and isolated inner limiting membrane.

Quantitative distribution of proteoglycans was studied in retinal neural epithelium and its basement membrane (inner limiting membrane). Heparan sulfate proteoglycans (HSPGs) were primarily associated with both inner and outer plexiform (synaptic) layers, and inner limiting membrane (ILM), as determined by autoradiographs of lyase-digested cryosections. Based on distribution of 35S-sulfate-labeled proteoglycans, the isolated ILM contained on average approximately three fourths of its proteoglycans as HSPGs and one fourth as chondroitin sulfate/dermatan sulfate proteoglycans (CS/DSPGs), whereas the remaining retina contained approximately equal amounts of the two proteoglycans (PGs). Immunohistochemical staining indicates that the core proteins of the HSPGs in the ILM are distinct from those of the plexiform layers. The photoreceptor layer, which other studies have shown to contain much of the extracellular CS/DSPGs, was not examined. Enrichment of distinct HSPGs in the ILM and plexiform layers support the conclusion that the HSPGs may be intimately involved in the different developmental events characterizing the two regions: development and extension of ganglion cell axons in the former, synaptogenesis and neuronal function in the latter.

Animals↗
PubMed1994

Micronuclei induced by radon and its progeny in deep-lung fibroblasts of rats in vivo and in vitro.

Genotoxic damage induced by radon and its progeny was investigated using the micronucleus assay in deep-lung fibroblasts to compare the response induced in vitro with that induced from inhalation of radon and its progeny in vivo. Male Wistar rats were exposed to 0, 115, 213 and 323 working-level months (WLM) of radon and its progeny by inhalation. After sacrifice, the cells were isolated and grown in culture, and the frequency of micronuclei was determined. A linear increase in the frequency of micronuclei was measured as a function of exposure [micronuclei/1000 binucleated cells = (29 +/- 9) + (0.47 +/- 0.04) WLM]. To compare exposure in WLM to dose in mGy, and to study how cell proliferation influences the way inhalation of radon and its progeny induces micronuclei, lung fibroblasts were isolated and exposed in vitro to graded doses from radon and its progeny after either 16 or 96 h in tissue culture. Cell cycle stage at the time of exposure was determined using flow cytometry. Primary lung fibroblasts exposed as either nondividing or dividing cells showed dose-dependent increases in micronuclei [micronuclei/1000 binucleated cells = (33 +/- 40) + (593 +/- 68)D and micronuclei/1000 binucleated cells = (27 +/- 69) + (757 +/- 88)D, respectively, where D is dose in Gy]. Results showed no significant influence (P = 0.20) of cell proliferation at the time of exposure on the frequency of micronuclei induced by radon and its progeny. Comparing dose-response relationships for nondividing cells to the exposure response for cells exposed by inhalation of radon and its progeny, it was estimated that a 1-WLM exposure in vivo caused the same amount of cytogenetic damage as produced by 0.79 mGy in vitro. In vivo/in vitro research using the micronucleus assay in lung fibroblasts serves as a powerful tool to estimate effective dose to cells in the respiratory tract after inhalation of radon and its progeny. Such studies form the basis for understanding the relationship between exposure, dose and biological damage.

Administration, Inhalation↗
PubMed1993

Proteoglycans and the modulation of cell adhesion by steric exclusion.

The hypothesis that cell aggregation may be driven by linear polymers in the matrix, particularly glycosaminoglycans, is revisited in light of more recent evidence. A model is proposed that extends the concept of steric exclusion to include a role in determining the directionality of cell migration and neurite extension. Recent literature is reviewed to support the conclusion that in living tissues the theoretical conditions for driving aggregation and migration by steric exclusion are met. The ability of a linear polymer to exclude cells is a function of its viscosity, which is optimum with glycosaminoglycans similar to chondroitin sulfate. It is ineffective with low viscosity glycosaminoglycans such as most heparin or heparan sulfate. Hyaluronic acid, a massive polymer, excludes cells poorly when present as an open matrix gel but forms an effective exclusion barrier when attached to the cell surface. According to a model for steric exclusion in organogenesis, when cells have a glycocalyx of linear polymer, they should disperse and migrate down a viscosity gradient of excluding matrix polymer; when they shed or internalize their surface coat in the continued presence of matrix, they should be excluded into a smaller volume and thus stimulated to aggregate.

Animals↗
PubMed1993

Molecular confirmation of alpha 1-antitrypsin genotypes in newborn dried blood specimens.

Deficiency of alpha 1-antitrypsin (alpha 1AT), a common hereditary disorder of Caucasians, is associated with an increased risk for early-onset chronic obstructive pulmonary disease and childhood liver dysfunction. The two most common deficiency variants, PiS and PiS, are both single base-pair substitutions causing amino acid modifications, although neither mutation creates or destroys a naturally occurring restriction site. Dried blood specimens (DBS) submitted to the New York State Department of Health for mandated newborn screening tests were tested for alpha 1AT activity using a fluorometric elastase inhibition assay. A second DBS from specimens determined to be alpha 1AT deficient was phenotyped on an agarose isoelectric focusing gel. Genotypic confirmation was performed by amplifying, directly from a DBS, the regions of the DNA containing the S and Z mutation. The Z mutation was analyzed with a modified primer designed to create an artificial restriction site in the normal allele. TaqI digestion produces two bands, a 157- and a 22-bp fragment. The single base substitution in PiS individuals eliminates this TaqI restriction site, thus showing the same 179-bp fragment before and after digestion. A primer mismatch placed close to the S mutation creates a restriction site in the normal allele, producing a 100-bp product after TaqI digestion. The restriction site is abolished in individuals that carry the S mutation, with a 121-bp product observed before and after digestion. Of 11,081 specimens screened, 3 PiS neonates, all Caucasian, were detected by these methodologies for an estimated incidence of 1:2019 in the Caucasian or 1:3694 in the general population in New York State.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1992

Immunophenotypic characterization of owl monkey peripheral blood mononuclear cells.

A two-phase study was initiated to delineate the peripheral blood lymphocyte populations present in owl monkeys and to correlate those populations with immune response and parasitism during malaria infection. The goal of phase I of the study was to elucidate a monoclonal antibody panel that could be used to characterize peripheral blood mononuclear cell (PBMC) populations with flow cytometric techniques. Forty-two monoclonal antibodies (reported to be reactive with human and macaque lymphocyte antigens) were screened for activity to owl monkey PBMC. Eleven monoclonals were found to react: anti-H42A (MHC Class II DP-like); anti-TH14B (MHC Class II DR-like); and anti-TH81A5 (MHC Class II DQ-like); anti-H58A (MHC Class I); anti-DH59B (granulocyte and monocyte); anti-B1 (B cell); anti-T4 (CD4); anti-Leu3a (CD4); anti-Leu11a (CD16); anti-60.3 (CD18); and anti-OKM1 (NK and monocyte). In a preliminary retrospective study correlating antibody titers, parasitemia values, and MHC Class I and Class II marker profiles on PBMC to test antigens used in malaria vaccine trials, a significant negative association was observed between cells bearing MHC Class II molecules and the other elements of the comparison. In summary, an appropriate panel of monoclonal antibodies has been identified for characterizing PBMC in owl monkeys, and preliminary studies indicate a possible association between clinical outcome and expressed phenotypic PBMC markers.

Animals↗
PubMed1992

Changes in histochemical distribution of cell surface heparan sulfate proteoglycan in mouse uterus during the estrous cycle and early pregnancy.

We have investigated the changes in immunolocalization of a cell surface heparan sulfate proteoglycan (HSPG) in the mouse uterus during the estrous cycle and at the time of implantation in early pregnancy. A monoclonal antibody prepared against syndecan, a cell surface HSPG from mouse mammary epithelium (gift of Dr. M. Bernfield), was reacted with unfixed and fixed frozen sections of uteri from normally cycling, 3.5 and 4.5 days pregnant, and estradiol-treated immature and ovariectomized mature mice. A polyclonal antibody prepared against basal lamina HSPG from Engelbreth-Holm-Swarm (EHS) tumor cells (gift of Dr. John Hassell) was used as a positive control. The latter showed no variation during the estrous cycle or early pregnancy. Localization of syndecan in uterine epithelium changed from basolateral to predominantly basal as the cycle progressed from metestrus toward estrus. A similar pattern was seen in immature and ovariectomized mature females that had received estradiol injections. With the onset of pregnancy, the basolateral localization became progressively less intense from 3.5 days through 4.5 days of pregnancy. Thus, cell surface HSPG distribution is modulated by hormonally dependent changes in cycling and pregnant mice, supporting previous suggestions that early pregnancy in mice is accompanied by a turnover and rearrangement of uterine epithelial cell surface.

Animals↗
PubMed1991

Evaluation of changes in diatom mobility after exposure to 16-Hz electromagnetic fields.

The effect of a 16-Hz electromagnetic field on the mobility of the diatom Amphora coffeaformis was examined on agar plates that contained no added calcium and also on agar plates containing 0.25 or 2.5 mM exogenous Ca2+. Exposure conditions consisted of an ac field of 16 Hz with an amplitude of 20.9 microT parallel to the horizontal component of the dc field (BH = 20.9 microT, where BV = 0). To assess results, the percentage of diatoms that moved a distance greater than their body length was determined. We observed the field-associated increase in diatom motion at 0.25 mM Ca++, which was previously reported in the literature. Although the magnitude of the effect at 16 Hz was significant, the percentage of cells that moved was not sufficiently reproducible to allow examination for frequency dependence.

Calcium↗
PubMed1990

Exposure of mammalian cells to 60-Hz magnetic or electric fields: analysis of DNA repair of induced, single-strand breaks.

DNA damage was induced in isolated human peripheral lymphocytes by exposure at 5 Gy to 60Co radiation. Cells were permitted to repair the DNA damage while exposed to 60-Hz fields or while sham-exposed. Exposed cells were subjected to magnetic (B) or electric (E) fields, alone or in combination, throughout their allotted repair time. Repair was stopped at specific times, and the cells were immediately lysed and then analyzed for the presence of DNA single-strand breaks (SSB) by the alkaline-elution technique. Fifty to 75 percent of the induced SSB were repaired 20 min after exposure, and most of the remaining damage was repaired after 180 min. Cells were exposed to a 60-Hz ac B field of 1 mT; an E field of 1 or 20 V/m; or combined E and B fields of 0.2 V/m and 0.05 mT, 6 V/m and 0.6 mT, or 20 V/m and 1 mT. None of the exposures was observed to affect significantly the repair of DNA SSB.

Cobalt Radioisotopes↗
PubMed1990

An outbreak of foodborne botulism associated with contaminated hazelnut yoghurt.

The largest recorded outbreak of foodborne botulism in the United Kingdom occurred in June 1989. A total of 27 patients was affected; one patient died. Twenty-five of the patients had eaten one brand of hazelnut yoghurt in the week before the onset of symptoms. This yoghurt contained hazelnut conserve sweetened with aspartame rather than sugar. Clostridium botulinum type B toxin was detected in a blown can of hazelnut conserve, opened and unopened cartons of hazelnut yoghurt, and one faecal specimen. Cl. botulinum type B was subsequently cultured from both opened and unopened cartons of the hazelnut yoghurt and from one faecal specimen. Investigations indicated that the processing of the conserve was inadequate to destroy Cl. botulinum spores. Control measures included the cessation of all yoghurt production by the implicated producer, the withdrawal of the firm's yoghurts from sale, the recall of cans of the hazelnut conserve, and advice to the general public to avoid the consumption of all hazelnut yoghurts.

Adolescent↗
PubMed1990

Evidence for an effect of ELF electromagnetic fields on human pineal gland function.

A study was carried out to determine possible effects of 60-Hz electromagnetic-field exposure on pineal gland function in humans. Overnight excretion of urinary 6-hydroxymelatonin sulfate (6-OHMS), a stable urinary metabolite of the pineal hormone melatonin, was used to assess pineal gland function in 42 volunteers who used standard (conventional) or modified continuous polymer wire (CPW) electric blankets for approximately 8 weeks. Volunteers using conventional electric blankets showed no variations in 6-OHMS excretion as either a group or individuals during the study period. Serving as their own controls, 7 of 28 volunteers using the CPW blankets showed statistically significant changes in their mean nighttime 6-OHMS excretion. The CPW blankets switched on and off approximately twice as often when in service and produced magnetic fields that were 50% stronger than those from the conventional electric blankets. On the basis of these findings, we hypothesize that periodic exposure to pulsed DC or extremely low frequency electric or magnetic fields of sufficient intensity and duration can affect pineal gland function in certain individuals.

Adult↗
PubMed1989

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↗
PubMed1988

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↗
PubMed1988

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↗
PubMed1988

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↗
PubMed1987

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↗