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Biomedical subjects

H Green

Publications and source records attributed to H Green.

At least 181 records · Page 10Linked to original sources
PubMed1978

Adipose conversion of 3T3 cells depends on a serum factor.

The adipose conversion of 3T3-F442A cells depends on an adipogenic factor in serum. In the presence of this factor, cells grown to confluence become spherical, greatly increase the activity of their lipogenic enzymes, and accumulate triglyceride. In the absence of the adipogenic factor, the cells grow normally, but when they reach confluence they do not become spherical, do not accumulate triglyceride, and do not undergo any increase in activity of lipogenic enzymes. In cattle there is a great deal more of the adipogenic factor in the serum before birth than in the serum of grown animals. The nature of the adipogenic factor suggests that it may play an important role in the development of adipose tissue.

Adipose Tissue↗
PubMed1978

Relation of protein synthesis and transglutaminase activity to formation of the cross-linked envelope during terminal differentiation of the cultured human epidermal keratinocyte.

When serially cultivated human epidermal keratinocytes are placed in suspension culture they stop growing and form, beneath the plasma membrane, an insoluble envelope consisting of protein cross-linked by epsilon- (gamma-glutamyl)lysine. The formation of envelopes in suspended cells is preceded by a sharp decline in the rate of protein synthesis, and most envelopes appear only after the average rate of protein synthesis has fallen to a very low level. If protein synthesis is reduced over 98 percent with cycloheximide or emetine at the time that surface-grown cells are placed in suspension culture, cross-linked envelopes form in most of the cells. This shows that the precursor of the envelope and the cross-linking enzyme are already in the cytoplasm in most cells of growing surface cultures. The process of envelope formation by suspension cultures is actually accelerated by the inhibitors of protein synthesis; an increased number of cells with cross-linked envelopes is observable within 4-6 h after the addition of cycloheximide. The inhibitor also induces a large fraction of the cells of surface cultures to form enveloped within a few days. These findings suggest that arrest of protein synthesis leads to activation of the cross-linking process. Agents known to inhibit transglutaminase-mediated protein cross-linking-putrescine, iodoacetamide, and ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA)- also prevent envelope formation. Though the activity of the cross-linking transglutaminase depends on the presence of cellular Ca++, we have not been able to activate the cross-linking process by high external Ca++ concentration or ionophores.

Calcium↗
PubMed1978

Effects of continuous positive airway pressure after oleic acid-induced lung injury in dogs.

The physiologic effects of continuous positive airway pressure (CPAP) of 5,10,15, and 20 cm H2O during spontaneous ventilation were studied in six anesthetized dogs with simulated respiratory distress syndrome (RDS) induced by iv infusion of oleic acid and in three normal controls. After oleic acid, mean PaO2 dropped to 63.6 +/- 3.1 mm Hg while breathing 100% oxygen and mean shunt fraction was 48.3 +/- 3.0%. PaO2 and shunt fraction improved significantly at the two highest levels of CPAP (e.g.,PaO2 271.3 +/- 41.3 mm Hg and shunt fraction 17.8 +/-2.2% at 20 cm H2O CPAP). Mean mixed venous PO2 rose from 37.4 +/- 1.5 mm Hg with no CPAP TO 60.8 +/- 3.1 mm Hg at 20 cm H2O CPAP. Tissue oxygenation appeared to improve during CPAP, since cardiac output, oxygen delivery, and serum lactate were not significantly affected and mixed venous PO2 rose significantly. However, significant hypoventilation occurred at all but the lowest level of CPAP, mean PaCO2 rising from 44.1 +/- 1.8 mm Hg with no CPAP to 77.6 +/-6.8 mm Hg at 20 cm H2O CPAP. The hypoventilation during CPAP is consistent with increased work of breathing due to a combination of decreased lung compliance and increased dead space ventilation due to rapid, shallow breathing.

Animals↗
PubMed1978

Cell culture for the study of epithelial cells.

Keratinocytes depend on the support of fibroblasts or fibroblast products to grow from single cells into colonies. The essentials of a human stratified squamous epithelium can be constructed from single human epidermal keratinocytes and lethally irradiated fibroblasts. Established lines of mouse keratinocytes obtained from teratomas have many of the same properties. In this way it is possible to study many aspects of this epithelial tissue or organ under essentially cell culture conditions.

Animals↗
PubMed1977

Cultured epithelial cells of cornea, conjunctiva and skin: absence of marked intrinsic divergence of their differentiated states.

Keratinocytes of three different epithelia grown in cell culture express a large number of differentiation markers with either no differences or relatively small differences, depending on the species. Much of the distinctive phenotype of these epithelia in vivo must be due to external modulation and relatively little, at least in the case of the human, to permanent intrinsic divergence during development.

Animals↗
PubMed1977

Increasing activity of enzymes on pathway of triacylglycerol synthesis during adipose conversion of 3T3 cells.

When preadipose 3T3 cells pass from growing to resting state, they increase their rate of synthesis of triacylglycerol from precursors and convert to adipose cells. The behavior of two enzymes involved in the synthesis of triacylglycerol, glycerophosphate acyltransferase, and malic enzyme have been examined during the adipose conversion of 3T3-F442A in surface culture and in suspension culture stabilized with methylcellulose. Glycerophosphate acyltransferase activity rises sharply during the conversion and reaches a level of 80 times higher than that of another 3T3 subline in which practically no adipose conversion takes place (3T3-C2). The activity of malic enzyme also rises during the adipose conversion of 3T3-F442A and reaches a level of 15-fold higher than that of 3T3-C2. The activity of glycerophosphate acyltransferase responds more sharply than that of malic enzyme but the rise in activity is not sustained as long, so that the relative levels of the two enzymes change during the conversion. The adipose conversion appears to be the result of increases in the activity of the synthesizing enzymes, brought about by either of the two most physiological methods available for arresting the growth of cultured cells.

Adipose Tissue↗
PubMed1977

Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes.

The culture lifetime of epidermal cells of newborn humans is increased from 50 to 150 generations by adding to the medium epidermal growth factor, a polypeptide mitogen. EGF seems to delay senescence of the cells by maintaining them in a state further removed from terminal differentiation. This effect is revealed by a greater ability of the cells to survive subculture and initiate new colonies, but not necessarily by an increased growth rate.

Cell Differentiation↗
PubMed1977

The relative amounts of the cytoplasmic RNA species in normal, transformed and senescent cultured cell lines.

We have examined the relative quantities of 18S and 28S rRNA, 4S RNA and poly (A) + mRNA in the following cultured cells: the mouse fibroblast lines 3T3 and 3T6 in the resting (contact inhibited) and growing (sparse) states, 3T3 clones transformed with SV40 (SV3T3) and with both SV40 and polyoma SV-Py 3T3), hamster lung fibrobalsts (v79), human cervical carcinoma cells (HeLa), and human diploid fibroblasts at early and late passage. The relative quantities of the RNA species were determined by labeling the cells to equilibrium with 32PO4 and measuring the amount of label in each RNA species. The ratio of mRNA to rRNA varied form 1.1% to 2.7% in the different cell lines, the more rapidly growing cell lines usually giving a higher ratio. In cells experiencing growth limitation either by contact inhibition or due to senescence, the ratio of mRNA to rRNA was about 30% lower than in the corresponding cells in the growing state. In most cell lines the ratio of 4S RNA to 18S rRNA was between 0.8 and 1.2, but in seescent fibroblasts, this ratio increased to greater than 1.7. Senescent fibroblasts also contained much more total RNA per unit of DNA than the same cells at early passage or than 3T6 or 3T3 cells.

Animals↗
PubMed1977

The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein.

A small proportion of the protein of stratum corneum of human epidermal callus is insoluble even when boiled in solutions containing sodium dodecylsulfate and a reducing agent. This protein is present in the cornified envelope, a structure located beneath the plasma membrane. When cornified envelopes were dissolved by exhaustive proteolytic digestion and the products analyzed by chromatography, approximately 18% of the total lysine residues were found as the cross-linking dipeptide epsilon-(gamma-glutamyl) lysine. Labeled cornified envelope protein was synthesized by human epidermal keratinocytes allowed to differentiate terminally in culture. The extent of cross-linking, determined from the proportion of radioactive lysine in epsilon-(gamma-glutamyl) lysine after exhaustive proteolysis, was similar to that in stratum corneum. The properties of the cornified envelopes (insolubility in detergent and reducing agents, and solubility following proteolytic digestion) are readily explained by a structure consisting of a cross-linked protein lattice.

Cell Differentiation↗
PubMed1977

Properties of an epithelial cell type in culture: the epidermal keratinocyte and its dependence on products of the fibroblast.

Keratinocytes of stratified squamous epithelium can be grown serially in culture and retain the various markers typical of their form of differentiation. In order to form colonies at each transfer, the keratinocytes must be suitably supported by fibroblasts. Established keratinocyte lines of teratomal origin show this dependence, as do diploid strains of finite culture life derived from human skin. For at least some keratinocyte lines, this requirement can be satisfied by soluble products elaborated by the fibroblasts. It is suggested that epithelial cells in general may not be independent cell types and that their poor cultivability may be due to failure to provide suitable fibroblast support. The existence of a number of established lines of epithelial origin that can grow without such support and of lines of fibroblastic origin which cannot support keratinocytes suggests that both epithelial dependence and the fibroblast supporting function can sometimes be lost in established cell lines.

Cell Adhesion↗
PubMed1976

Differentiation of the epidermal keratinocyte in cell culture: formation of the cornified envelope.

Human epidermal keratinocytes grow from single cells into stratified colonies. Cells in the upper layers of the colonies lose their ability to divide and begin terminal differentiation. In this process, there develops a cornified cell envelope that remains insoluble after heating in solutions of sodium dodecylsulfate and beta-mercaptoethanol. The insolubility of the cornified envelope depends upon proteins, since after treatment with proteolytic enzymes, the envelope becomes soluble in the detergent. Cells with cornified envelopes can be identified under the light microscope either in living colonies or following fixation and silver nitrate staining. Keratinocytes of the basal layer move in a characteristic way, but cornified cells do not move at all and form an immobile upper layer in the colonies. Keratinocytes disaggregated from growing colonies are of differing size and density, and can be separated on isopycnic gradients of Ficoll. The DNA-synthesizing cells are small (mean diameter 14 mum). The nonmultiplying cells are large and have a protein content proportionate to their size. Their final diameter may exceed 30 microns (volume increase greater than 10 fold). Cornified envelopes are found in some of the large cells but in none of the small. In growing colonies, usually 5-10% of the cells have cornified envelopes. The fraction is reduced in colonies growing in the presence of epidermal growth factor. Strain XB, an established keratinocyte line of mouse teratomal origin, also forms cornified envelopes, but the kinetics of the process are different, indicating that the program of terminal differentiation is not initiated at corresponding times in the two cell types.

Cell Adhesion↗
PubMed1976

Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells.

When their growth is arrested in culture, susceptible 3T3 fibroblasts differentiate into adipose cells. Different clones form adipose cells with different frequency, depending upon the proportion of susceptible cells they contain. In cultures grown from small inocula, the fat cells appear in clusters formed by colonies of susceptible cells. Study of these clusters indicates the infrequent occurrence of cellular transitions from insusceptible to susceptible state. Beginning with a clone converting to adipose cells with a vary low frequency, it has been possible, by serial selection, to generate subclones which convert with a high frequency. This evolution is due to spontaneous heritable changes affecting susceptibility to the adipose conversion. Presumably, they involve the control of triglyceride synthesis. Early stages of the adipose conversion may be recognized in stained cultures. When triglyceride first begins to accumulate, the highly extended and flattened processes of the cells are probably similar to those of nonfatty cells in the same cultures. As the adipose conversion proceeds, the processes thicken and retract; the cells eventually acquire the rounded shape of the more mature adipose cells.

Adipose Tissue↗
PubMed1976

Changes in RNA in relation to growth of the fibroblast. IV. Alterations in theproduction and processing of mRNA and rRNA in resting and growing cells.

In previous reports, it was shown that both the concentration and rate of production of rRNA and mRNA were greater in growing than in resting 3T6 fibroblasts. Studies on isolated nuclei indicated that ribosomal RNA production is apparently controlled at the level of transcription. In contrast, hnRNA, the putative precursor of mRNA, appeared to be synthesized at the same rate in resting and growing cells. This finding was unexpected and has been tested in several ways. In this report, we show by an independent method that the relative rate of production of mRNA compared to hnRNA is several-fold higher in growing than in resting cells. However, the kinetics of processing of mRNA appear unchanged. This result suggests either that mRNA arises from a small subfraction of hnRNA or that the efficiency of processing of the hnRNA precursor is an important control mechanism which determines mRNA production ingrowing and resting states. Comparison of the initial rates of labeling of hnRNA and cytoplasmic message gives the efficiency with which the cytoplasmic mRNA is produced from nucleoplasmic RNA. The very low efficiency (3-4% in growing and 1-2% in resting cells) suggests that not every hnRNA molecule gives rise to a cytoplasmic message. In contrast to the similar kinetics of mRNA production in resting and growing states, processing of ribosomal RNA is much slower in the resting state and the emergence time for 28S RNA from nucleolus is greatly lengthened.

Cell Division↗
PubMed1976

Time-motion and physiological assessments of ice hockey performance.

Ice hockey performance was studied during 10 contests to assess time-motion characteristics and associated physiological changes that occur for each position. Depending on the position played, the actual playing time per game for forwards and defensemen ranged between 20.7 and 28.0 min. The number of shifts ranged from 14 to 21 with an average playing time of 85.4 s/shift. Further analysis of each shift indicated that there was an average of 2.3 play stoppages which averaged 27.1 s, producing a continuous playing time of 39.7 s. Telemetered recordings of heart rate during each shift revealed sustained rates of between 170 and 174 beats/min over the three periods of the game. For both the forwards and defensemen, values for blood lactates were highest during the first and second periods (x = 78.0 and 66.1 mg/100 ml), then declined considerably during the third period (x = 44.5 mg/100 ml). Blood glucose showed a similar tendency (x = 139.3, 133.7, 114.1 mg/100 ml), while hematocrit and total protein showed little tendency to change.

Adult↗