Biomedical subjects

C A Edwards

Publications and source records attributed to C A Edwards.

At least 19 recordsLinked to original sources

PubMed2007

Parental perceptions of feeding practices in five European countries: an exploratory study.

OBJECTIVE: To gain an insight into parental perceptions of infant feeding practices in five European countries. DESIGN: An exploratory investigation using focus group discussions. Various aspects addressed included social and cultural setting for the consumption of food, infant feeding practice and behaviour, consumer health awareness and sources of information, and attitudes towards a healthy infant diet. SETTING: Focus group participants were recruited from centres in five countries, Germany, Italy, Scotland, Spain and Sweden, with three focus groups being conducted in each centre. SUBJECTS: A total of 108 parents with infants up to the age of 12 months participated in focus group discussions across these centres. METHODS: Focus groups were conducted with participants from centres in five countries. RESULTS: The majority of parents in this study chose to initiate breastfeeding and prepare infant food at home. Parents did not strictly adhere to infant feeding guidelines when introducing complementary foods into their infant's diets. There were cross-cultural differences in sources of information on infant feeding practice with the paediatrician in Germany, Italy and Spain. The health visitor in Scotland and the child welfare clinics in Sweden were the most popular sources. CONCLUSIONS: A number of cultural differences and similarities in attitudes towards infant feeding practice were revealed. This makes European wide approaches to promoting healthy infant feeding difficult as different infant feeding practices are influenced not only by parental perceptions but also by advice from health professionals and feeding guidelines. Further data need to be available on parents' attitudes and beliefs towards infant feeding practice to investigate further the rationale for differing beliefs and attitudes towards infant feeding practice. SPONSORSHIP: EU Fifth Framework QLRT 2002 02606.

Adult↗

PubMed2005

Influences of vermicomposts on field strawberries: part 2. Effects on soil microbiological and chemical properties.

The effects of applications of food waste and paper waste vermicomposts on some soil chemical and biological properties were evaluated in field plots planted with strawberries. Six-week old strawberries (Fragaria ananasa, var. Chandler) were transplanted into 4.5 m(2) raised beds under a plastic tunnel structure measuring 9.14 x 14.6 x 3.6 m. Vermicompost were applied at rates of 5 or 10 t ha(-1) supplemented with inorganic fertilizers to balance fertilizer recommendations for strawberries of 85-155-125 kg NPK ha(-1). Effects of vermicomposts on strawberry growth and yields have been reported previously [Arancon, N.Q., Edwards C.A., Bierman P., Welch, C., Metzger, J.D., 2004. The influence of vermicompost applications to strawberries: Part 1. Effects on growth and yield. Bioresource Technology 93:145-153]. Total extractable N, NH(4)-N, NO(3)-N and orthophosphates did not differ significantly between treatments, except on the last sampling date (harvest date) in which significantly greater amounts of NH(4)-N, NO(3)-N and orthophosphates (P <or= 0.05) were recorded in vermicompost-treated soils than in the controls. Two major results of vermicompost applications to soils were increases in dehydrogenase activity and microbial biomass-N which were not dose-dependent. Increased dehydrogenase activity and microbial biomass-N was correlated positively with the increased amounts of NH(4)-N, NO(3)-N and orthophosphates in the vermicompost-treated plots than in the controls. Increases in microbial populations and activities are key factors influencing rates of nutrient cycling, production of plant growth-regulating materials, and the build-up of plant resistance or tolerance to crop pathogen and nematode attacks.

Agriculture↗

PubMed2004

Influences of vermicomposts on field strawberries: 1. effects on growth and yields.

Vermicomposts processed commercially from food wastes and paper wastes were applied, to 4.5 m(2) field plots, under high plastic hoop tunnels, at rates of 5 or 10 tha(-1) to evaluate their effects on the growth and yields of strawberries (Fragaria ananasa) var. 'Chandler'. The vermicomposts were incorporated into the top 10 cm of soil and supplemented, based on chemical analyses, with amounts of inorganic NPK fertilizers calculated to equalize the initial fertilizer rates of 85-155-125 kgha(-1) NPK applied to the inorganic fertilizer plots. All treatments were replicated four times, in a completely randomized design, at two field sites on Doles silt loam or Hoytville silty clay loam at Piketon and Fremont, Ohio, respectively. Vermicompost applications increased strawberry growth and yields significantly; including increases of up to 37% in leaf areas, 37% in plant shoot biomass, 40% in numbers of flowers, 36% in numbers of plant runners and 35% in marketable fruit weights. These responses seemed not to be dose-dependent, since strawberries at one site grew fastest and yielded most in response to the 10 tha(-1) vermicompost application rate, whereas they responded positively and similarly to both the 5 and 10 tha(-1) rates of applications at the other site. These responses could not have been mediated by availability of macronutrients, since all plots were supplemented with inorganic fertilizers, to equalize macronutrient inputs for all treatments, but based on other research in our laboratory could have been due to production of plant growth regulators by microorganisms during vermicomposting.

Agriculture↗

PubMed2004

Out of the Smokescreen: does an anti-smoking advertisement affect young women's perception of smoking in movies and their intention to smoke?

OBJECTIVE: To evaluate the effect of an anti-smoking advertisement on young women's perceptions of smoking in movies and their intention to smoke. SUBJECTS/ SETTING: 2038 females aged 12-17 years attending cinemas in New South Wales, Australia. DESIGN/ INTERVENTION: Quasi-experimental study of patrons, who were surveyed after having viewed a movie at their local cinema. The control group was surveyed during week 1 and the intervention group, during week 2. Before seeing the movie in week 2, a 30 second anti-smoking advertisement was shown, which featured a well known female actor drawing attention to the prevalence of smoking in movies. OUTCOMES: Attitude of current smokers and non-smokers to smoking in the movies; intention of current smokers and non-smokers to be smoking in 12 months time. RESULTS: Among non-smokers, 48.2% of the intervention subjects thought that the smoking in the movie they viewed was "not OK" compared with 28.3% of the control subjects (p < 0.0001). However, there was no difference among smokers in the intervention (26.4%) and control (16.9%) groups (p = 0.28). A higher percentage of current smokers in the intervention group indicated they were unlikely to smoke in 12 months time (47.8%) than smokers in the control condition (31.9%) (p = 0.03). For non-smokers, there was no difference in smoking intentions between conditions, with 95% saying they would be unlikely to be smoking in 12 months time. CONCLUSIONS: This "real world" study suggests that placing an anti-smoking advertisement before movies containing smoking scenes can help to "immunise" young women against the influences of film stars smoking.

Adolescent↗

PubMed2003

The DNA sequence and analysis of human chromosome 6.

Chromosome 6 is a metacentric chromosome that constitutes about 6% of the human genome. The finished sequence comprises 166,880,988 base pairs, representing the largest chromosome sequenced so far. The entire sequence has been subjected to high-quality manual annotation, resulting in the evidence-supported identification of 1,557 genes and 633 pseudogenes. Here we report that at least 96% of the protein-coding genes have been identified, as assessed by multi-species comparative sequence analysis, and provide evidence for the presence of further, otherwise unsupported exons/genes. Among these are genes directly implicated in cancer, schizophrenia, autoimmunity and many other diseases. Chromosome 6 harbours the largest transfer RNA gene cluster in the genome; we show that this cluster co-localizes with a region of high transcriptional activity. Within the essential immune loci of the major histocompatibility complex, we find HLA-B to be the most polymorphic gene on chromosome 6 and in the human genome.

Animals↗

PubMed2003

Starch fermentation by faecal bacteria of infants, toddlers and adults: importance for energy salvage.

OBJECTIVE: Little is known of the degree to which the colon salvages energy through starch fermentation in young children. Using a simulated colonic environment, we aimed to account for the fate of fermented raw and cooked starch in two groups of young children and in adults. DESIGN: A slurry was prepared from faecal samples from six infants (7-10 months), six toddlers (16-21 months) and seven adults (24-56 y). Each slurry was anaerobically incubated with raw or cooked maize starch in MacCartney bottles in a shaking water bath. Parallel incubations were stopped at 4 and 24 h. The headspace gas volume was analysed for CO(2) and methane. The culture supernatant was analysed for short-chain fatty acids (SCFA), lactate and residual starch. RESULTS: Different patterns of fermentation were seen at 4 and 24 h. For raw starch, the production of SCFA decreased with subject age at 4 h but not at 24 h. With both substrates at 4 h, toddler stools produced significantly more CO(2) than infants or adults, but there were no statistical differences at 24 h. Methane was detected in three adults only. Lactate was detected mainly at 4 h in children. CONCLUSIONS: The results suggest that fermentation, particularly of raw starch, is a more rapid process in young children than in adults. A highly efficient energy salvage process may occur in the colon of young children.

Adult↗

PubMed2003

Dietary fibre in infancy and childhood.

There is very little evidence for the effects of dietary fibre in young children and current dietary guidelines are based on assumptions and data extrapolated from studies in adults. The first years of life may be critical for the establishment of a healthy colonic microflora, as well as good eating habits. The lack of clear and well-founded guidelines for the intake of dietary fibre in childhood may hinder both factors. The fears that a high-fibre diet in children < 5 years of age will lead to growth faltering and mineral imbalance are not well supported in the literature, especially for children in the developed world. Indeed, with the rising levels of obesity, fibre intake may be of benefit in reducing energy intake. A low-fibre diet may also be implicated in the aetiology of childhood constipation and appendicitis. The latest proposals for the definition of dietary fibre include oligosaccharides, which may act as prebiotics. There are potential health benefits of including oligosaccharides in the diets of infants and children, but more research is needed to consider the long-term effects. The immature intestine of the infant may also result in a greater amount of starch entering the colon during weaning, and this starch would now be considered dietary fibre under the new definitions. Much new research is needed to allow adequate recommendations for the intake of dietary fibre in childhood based on data collected in appropriate age-groups.

Bacteria↗

PubMed2003

Relationship between birth weight and adult lung function: controlling for maternal factors.

BACKGROUND: There is conflicting evidence on the "fetal origins hypothesis" of association between birth weight and adult lung function. This may be due to failure to control for confounding maternal factors influencing birth weight. In the present study access to birth details for adults aged 45-50 years who were documented as children to have asthma, wheezy bronchitis, or no respiratory symptoms provided an opportunity to investigate this association, controlling for maternal factors. METHODS: In 2001 the cohort was assessed for current lung function, smoking status, and respiratory symptoms. Birth details obtained from the Aberdeen Maternity and Neonatal Databank recorded birth weight, gestation, parity, and mother's age and height. RESULTS: 381 subjects aged 45-50 years were traced and tested for lung function; 323 (85%) had birth details available. A significant linear trend (p<0.01) was observed between birth weight and current forced expiratory volume in 1 second (FEV(1)) and forced vital capacity (FVC) values (adjusted for height, age, sex, weight, deprivation category (Depcat), childhood group, and smoking status). This trend remained significant after adjusting birth weight for gestation, parity, sex, mother's height and weight (p = 0.01). The relationship between birth weight and FEV(1) and FVC remained significant when adjusted for smoking history. There was no association between birth weight and current wheezing symptoms. CONCLUSION: There is a positive linear trend between birth weight, adjusted for maternal factors, and lung function in adulthood. The strength of this association supports the "fetal origins hypothesis" that impairment of fetal growth is a significant influence on adult lung function.

Adult↗

PubMed2002

The influence of earthworm-processed pig manure on the growth and productivity of marigolds.

The effects of additions of earthworm-processed pig manure (vermicompost) on the growth and productivity of French marigold (Tagetes patula) plants were evaluated under glasshouse conditions. Marigolds were germinated and grown in a standard commercial greenhouse container medium (Metro-Mix 360), substituted with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% (by volume) pig manure vermicompost. The control consisted of Metro-Mix 360 alone without vermicompost. Plants were supplied regularly with a complete mineral nutrient solution for 121 days. The greatest vegetative growth resulted from substitution of Metro-Mix 360 with 30% and 40% pig manure vermicompost, and the lowest growth was in the potting mixtures containing 90% and 100% vermicompost. Most flower buds occurred in the potting mixtures containing 40% pig manure vermicompost (19.4 buds), and fewest in the potting mixtures containing 100% vermicompost. Marigolds grown in Metro-Mix 360 substituted with 90% and 100% pig manure vermicompost had the fewest and smallest flowers. After substitution of Metro-Mix 360 with 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90% vermicompost, the marigold roots were larger than those of plants grown in the Metro-Mix 360 controls. Substitution of Metro-Mix 360 with any concentration of pig manure vermicompost, with all needed nutrients provided, increased the overall nitrate-nitrogen concentrations of the marigold leaf tissues at flowering stage. Some of the marigold growth and productivity enhancement, resulting from substitution of Metro-Mix 360 with pig manure vermicompost, may be explained by nutritional factors; However, other, factors, such as plant-growth regulators and humates, might have also been involved since all plants were supplied regularly with all required nutrients.

Animals↗

PubMed2002

The influence of humic acids derived from earthworm-processed organic wastes on plant growth.

Some effects of humic acids, formed during the breakdown of organic wastes by earthworms (vermicomposting), on plant growth were evaluated. In the first experiment, humic acids were extracted from pig manure vermicompost using the classic alkali/acid fractionation procedure and mixed with a soilless container medium (Metro-Mix 360), to provide a range of 0, 50, 100, 150, 200, 250, 500, 1,000, 2,000, and 4,000 mg of humate per kg of dry weight of container medium, and tomato seedlings were grown in the mixtures. In the second experiment, humates extracted from pig manure and food wastes vermicomposts were mixed with vermiculite to provide a range of 0, 50, 125, 250, 500, 1,000, and 4,000 mg of humate per kg of dry weight of the container medium, and cucumber seedlings were grown in the mixtures. Both tomato and cucumber seedlings were watered daily with a solution containing all nutrients required to ensure that any differences in growth responses were not nutrient-mediated. The incorporation of both types of vermicompost-derived humic acids, into either type of soilless plant growth media, increased the growth of tomato and cucumber plants significantly, in terms of plant heights, leaf areas, shoot and root dry weights. Plant growth increased with increasing concentrations of humic acids incorporated into the medium up to a certain proportion, but this differed according to the plant species, the source of the vermicompost, and the nature of the container medium. Plant growth tended to be increased by treatments of the plants with 50-500 mg/kg humic acids, but often decreased significantly when the concentrations of humic acids derived in the container medium exceeded 500-1,000 mg/kg. These growth responses were most probably due to hormone-like activity of humic acids from the vermicomposts or could have been due to plant growth hormones adsorbed onto the humates.

Animals↗

PubMed2002

Initial characterization of the Streptococcus gordonii htpX gene.

Examination of the Streptococcus gordonii chromosomal region, which lies immediately upstream of the glucosyltransferase positive regulatory determinant rgg, revealed two open reading frames. Based on nucleotide sequences, these genes were similar to the Listeria monocytogenes lemA gene, which is involved in antigen presentation, and the Escherichia coli htpX heat shock gene, which has an unknown function. Northern hybridization analysis indicated that S. gordonii lemA and htpX genes were associated with a ca. 1.7-kb polycistronic transcript. Although levels of the lemA/htpX transcript did not increase in response to heat to levels seen with dnaK controls, insertional inactivation of htpX resulted in changes in adhesiveness, cellular morphology and detergent-extractable surface antigens in cells grown at 41 degrees C, implying that htpX may be involved in surface protein expression. Insertional inactivation of lemA and htpX indicated that, despite their proximity to rgg and the structural gene, gtfG, these upstream genes do not affect S. gordonii glucosyltransferase activity.

Amino Acid Sequence↗

PubMed2002

Intestinal flora during the first months of life: new perspectives.

Increasing awareness that the human intestinal flora is a major factor in health and disease has led to different strategies to manipulate the flora to promote health. The complex microflora of the adult is difficult to change in the long term. There is greater impact of diet on the infant microflora. Manipulation of the flora particularly with probiotics has shown promising results in the prevention and treatment of diarrhoea and allergy. Before attempting to change the flora of the infant population in general, a greater understanding of the gut bacterial colonisation process is required. The critical stages of gut colonisation are after birth and during weaning. Lactic acid bacteria dominate the flora of the breast-fed infant. The formula-fed infant has a more diverse flora. The faeces of the breast-fed infant contain mainly acetic and lactic acid whereas the formula fed-infant has mainly acetic and propionic acid. Butyric acid is not a significant component in either group. The formula-fed infant also has higher faecal ammonia and other potentially harmful bacterial products. The composition of the microflora diversifies shortly before and particularly after weaning. The flora of the formula-fed infant develops more quickly than that of the breast-fed infant. Before embarking on any strategy to change the flora, the following questions should be considered: Should we retain a breast-fed style flora with limited ability to ferment complex carbohydrates? Can pro- and prebiotics achieve a flora with adult characteristics but with more lactic acid bacteria in weaned infants? Are there any health risks associated with such manipulations of the flora?

Bacteria↗

PubMed2002

Production of 13C labelled pea flour for use in human digestion and fermentation studies.

Stable isotope breath tests offer a new approach to the study of digestion and fermentation of carbohydrates in man. In this study, 13C labelled peas were grown by pulsing 250 ml 13CO2 into a sealed growth chamber. A second pulse was added to a portion of the peas to increase the 13C enrichment. This generated pea flour with an enrichment of 2.36 at.% excess (range 2.09-2.71 n = 3) and 8.64 atom % excess (range 7.37-9.78 n = 3) respectively. This represented incorporation of an absolute yield of 3.8% of the 13CO2 into peas in the 'once-labelled' treatment and 7.5% in the 'twice-labelled' treatment. Ingestion of a mixture of the labelled pea flour (300 mg) by two volunteers generated measurable 13CO2 excretion for breath test analysis. The profile of breath 13CO2 enrichment increased to a maximum within three hours after consuming the pea flour followed by a decrease almost back to baseline by 13 hours. Breath 13CO2 appeared to rise again after this apparent nadir at 13 hours until the end of the sampling period. Mathematical analysis of the data suggested that two peaks best described the profile of breath 13CO2 up to 13 hours. A third peak was necessary to describe the late rise in breath 13CO2 enrichment. This use of 13C enriched pea flour may provide a useful non invasive method for measurement of digestion and fermentation in vivo.

Adult↗

PubMed2001

The physical maps for sequencing human chromosomes 1, 6, 9, 10, 13, 20 and X.

We constructed maps for eight chromosomes (1, 6, 9, 10, 13, 20, X and (previously) 22), representing one-third of the genome, by building landmark maps, isolating bacterial clones and assembling contigs. By this approach, we could establish the long-range organization of the maps early in the project, and all contig extension, gap closure and problem-solving was simplified by containment within local regions. The maps currently represent more than 94% of the euchromatic (gene-containing) regions of these chromosomes in 176 contigs, and contain 96% of the chromosome-specific markers in the human gene map. By measuring the remaining gaps, we can assess chromosome length and coverage in sequenced clones.

Chromosomes, Human, Pair 1↗

PubMed2001

Specific down-regulation of an engineered human cyclin D1 promoter by a novel DNA-binding ligand in intact cells.

Cyclin D1 is expressed at abnormally high levels in many cancers and has been specifically implicated in the development of breast cancer. In this report we have extensively analyzed the cyclin D1 promoter in a variety of cancer cell lines that overexpress the protein and identified two critical regulatory elements (CREs), a previously identified CRE at -52 and a novel site at -30. In vivo footprinting experiments demonstrated factors binding at both sites. We have used a novel DNA-binding ligand, GL020924, to target the site at -30 (-30-21) of the cyclin D1 promoter in MCF7 breast cancer cells. A binding site for this novel molecule was constructed by mutating 2 bp of the wild-type cyclin D1 promoter at the -30-21 site. Treatment with GL020924 specifically inhibited expression of the targeted cyclin D1 promoter construct in MCF7 cells in a concentration-dependent manner, thus validating the -30-21 site as a target for minor groove-binding ligands. In addition, this result validates our approach to regulating the expression of genes implicated in disease by targeting small DNA-binding ligands to key regulatory elements in the promoters of those genes.

Binding Sites↗

PubMed2001

Pig manure vermicompost as a component of a horticultural bedding plant medium: effects on physicochemical properties and plant growth.

This experiment was designed to characterize the physical, chemical and microbial properties of a standard commercial horticultural, greenhouse container, bedding plant medium (Metro-Mix 360), that had been substituted with a range of increasing concentrations (0%, 5%, 10%, 25%, 50% and 100% by volume) of pig manure vermicompost and to relate these properties to plant growth responses. The growth trials used tomatoes (Lycopersicon esculentum Mill.), grown in the substituted media for 31 days under glasshouse conditions, with seedling growth recorded in 20 pots for each treatment. Half of the tomato seedlings (10 pots per treatment) were watered daily with liquid inorganic fertilizer while the other half received water only. The percentage total porosity, percentage air space, pH and ammonium concentrations of the container medium all decreased significantly, after substitution of Metro-Mix 360 with equivalent amounts of pig manure vermicompost; whereas bulk density, container capacity, electrical conductivity, overall microbial activity and nitrate concentrations, all increased with increasing substitutions of vermicompost. The growth of tomato seedlings in the potting mixtures containing 100% pig manure vermicompost was reduced, possibly as a result of high soluble salt concentrations in the vermicompost and poorer porosity and aeration. The growth of tomato seedlings was greatest after substitution of Metro-Mix 360 with between 25% and 50% pig manure vermicompost, with more growth occurring in combinations of pig manure vermicompost treated regularly with a liquid fertilizer solution than in those with no fertilizer applied. Some of the growth enhancement in these mixtures seemed to be related to the combined effects of improved porosity, aeration and water retention in the medium and the high nitrate content of the substrate, which produced an increased uptake of nitrogen by the plant tissues, resulting in increased plant growth. When the tomato seedlings were watered daily with liquid inorganic fertilizer, substitution of Metro-Mix 360 with a very small amount (5%) of pig manure vermicompost resulted in a significant increase in the growth of tomato seedlings. Such effects could not be attributed solely to the nutritional or physical properties of the pig manure vermicompost. Therefore, it seems likely that the pig manure vermicompost provided other biological inputs, such as plant growth regulators into the container medium, that still need to be identified fully.

Animals↗

PubMed2001

Cytotoxic effects of cleansing solutions recommended for chemical lavage of pulp exposures.

PURPOSE: To evaluate the in vitro cytotoxic effects of three cleansing solutions used for chemical lavage of pulp exposures. MATERIALS AND METHODS: The immortalized odontoblast cell line (MDPC-23) was plated (30,000 cells/cm2) and incubated for 72 hrs in 24-well dishes. After counting the cell number under inverted light microscopy, 20 microl of the experimental and control solutions were added to 980 microl of fresh culture medium. Then, hydrogen peroxide (3%, H2O2), sodium hypochlorite (6%, NaOCl) or calcium hydroxide-saline solution (5g of Ca(OH)2 in 10 ml of sterile distilled water) were added to wells for experimental Groups 1, 2 and 3, respectively. The positive and negative control groups received Syntac Sprint bonding agent (SS) and phosphate buffered saline (PBS), respectively. Following incubation for 120 min the cell number was counted again, the cell morphology was evaluated by scanning electron microscopy (SEM) and the cell metabolism was determined by the methyltetrazolium (MTT) assay. The scores obtained from cell counting and MTT assay were analyzed with an ANOVA followed by Fisher's PLSD tests. RESULTS: H2O2, NaOCl solutions, and SS bonding agent were more cytotoxic than Ca(OH)2 or PBS. In the groups with H2O2 or SS, only a few cells remained attached to the bottom of wells. The difference between these two groups was not statistically significant. H2O2, NaOCl and SS depressed the mitochondrial enzyme response by 97.7%, 97.3%, and 95.0%, respectively. On the other hand, Ca(OH)2 depressed the metabolic activity of cells by only 5%. While H2O2, NaOCl and SS caused extreme changes on the cell morphology, neither Ca(OH)2 nor PBS promoted dramatic changes in the cell morphology.

Analysis of Variance↗

PubMed2000

Caveolar structure and protein sorting are maintained in NIH 3T3 cells independent of glycosphingolipid depletion.

Glycosphingolipids have been proposed to be critical components of clustered lipids within cell membranes that serve as rafts for the attachment and sorting of proteins to the cell membrane. Density gradient centrifugation was used to isolate and to ascertain the lipid composition of caveolin-enriched membranes. These membranes demonstrated a significant enrichment of sphingolipids and cholesterol containing up to 20 and 30%, respectively, of the cellular glucosylceramide and lactosylceramide. A specific inhibitor of glucosylceramide synthase, d-threo-1-phenyl-2-palmitoyl-3-pyrrolidino-propanol, was used to test the hypothesis that glycosphingolipids are required for the sorting of proteins to caveolae. When NIH 3T3 cells were depleted of their glucosylceramide based glycosphingolipid mass, the caveolar structure remained intact as determined by electron microscopy and confocal microscopy. The caveolar proteins caveolin and annexin II sorted normally to caveolae, as determined by immunoblotting and confocal microscopy. When the GPI-linked protein B61 was inducibly expressed in these cells, sorting to caveolar membranes occurred normally, even in the presence of glucosylceramide depletion. These observations suggest that protein sorting to caveolae in fibroblasts occurs independently of glycosphingolipid synthesis.

3T3 Cells↗