Search PubMed⌕ Search

Biomedical subjects

F Green

Publications and source records attributed to F Green.

At least 73 records · Page 4Linked to original sources

Adaptation of the Nelson-Somogyi reducing-sugar assay to a microassay using microtiter plates.

The Nelson-Somogyi assay for reducing sugars was adapted to microtiter plates. The primary advantages of this modified assay are (i) smaller sample and reagent volumes, (ii) elimination of boiling and filtration steps, (iii) automated measurement with a dual-wavelength scanning TLC densitometer, (iv) increased range and reproducibility, and (v) automated colorimetric readings by reflectance rather than absorbance.

Animals↗

Reversible forskolin-induced impairment of sucrase-isomaltase mRNA levels, biosynthesis, and transport to the brush border membrane in Caco-2 cells.

Hybridization analysis of mRNA with a cDNA probe for human sucrase-isomaltase, pulse-chase experiments with L-[35S]-methionine followed by SDS-PAGE, and immunofluorescence detection of sucrase-isomaltase were used to analyze the level(s) at which forskolin interferes with the expression of the enzyme in Caco-2 cells in culture. Three effects are observed in Caco-2 cells treated with forskolin: 1) a marked decrease in the level of sucrase-isomaltase mRNA, 2) a marked decrease in the biosynthesis of the enzyme without any alteration of its stability, and 3) an almost total inhibition of its transport to the brush border membrane. All three effects are reversible when the drug is removed from the culture medium, though this reversibility is asynchronous: transport to the brush border membrane resumes after 24 h, sucrase-isomaltase mRNA levels are back to the normal after 5 days, whereas the biosynthesis of the enzyme, although increasing progressively, remains lower than in control cells, even 10 days after removal of the drug. The possibility that some effects are directly dependent on cAMP and others a consequence of changes in glucose metabolism is discussed.

Biological Transport↗

Apolipoprotein (apo) E genotypes by polymerase chain reaction and allele-specific oligonucleotide probes: no detectable linkage disequilibrium between apo E and apo CII.

Allelic sequence variation in the apolipoprotein (apo) E gene has been analysed by means of synthetic oligonucleotide probes that detect single base pair substitutions in the codons for amino acid positions 112 and 158, substitutions that are responsible for the common isoforms. Use of the polymerase chain reaction procedure to amplify a sequence of 330 base pairs of the human apo E gene has permitted the development of a robust method for apo E genotyping. This technique has been used to determine the apo E genotype in 95 individuals in whom the genotype for an apo CII TaqI restriction fragment length polymorphism has also been determined. No strong linkage disequilibrium between the two gene loci was detected. This suggests that the metabolic effects of variation in the apo E and apo CII genes, as detected by the polymorphisms used here, would operate in a statistically independent manner.

Adult↗

Studies on the cellular mechanisms of action of positive and negative inotropic agents.

We have reviewed the mechanism by which drugs that elevate cyclic AMP level modify myocardial contractility. We have presented preliminary evidence about the mechanism by which muscarinic agonists antagonize the effects of these drugs. Finally, we suggest that the protein phosphorylation experiments, particularly if done in dispersed myocytes, could be an efficient and cost-effective method of screening drugs which may act by elevating intracellular levels of cyclic AMP.

Acetylcholine↗

Control of plasma fibrinogen levels.

It is clear that the control of plasma fibrinogen levels is complex, involving not only many environmental factors such as alcohol intake, smoking habit, age, obesity and the acute phase response, but also genetic factors as shown by the association of the Bcl I RFLP of the beta-fibrinogen gene with plasma fibrinogen levels. The advent of recombinant DNA technology has made the dissection of the different factors controlling plasma fibrinogen levels a valid proposition, and great progress is already being made. The goals of this research are twofold. First, it may be possible to develop DNA tests to identify individuals who, on the basis of their genotype, are at high risk of ischaemic heart disease. Once identified, the subsequent risk of these individuals can be reduced by modifying life-style or by drug therapy to reduce other known risk factors such as cholesterol levels. Second, once the mechanisms controlling fibrinogen concentration are better understood at the molecular level, it may be possible to develop directed therapeutic strategies that will reduce fibrinogen synthesis in a specific manner, an approach that is not possible at present. In the future, such pharmacological agents may have as wide an impact on reducing ischaemic heart disease as cholesterol-lowering drugs do today.

Base Sequence↗

Resectable bronchogenic carcinoma presenting with hypercalcemia: tumor-associated granulomatous reaction and probable production of 1,25-dihydroxyvitamin D.

A patient with hypercalcemia, increased levels of 1,25-dihydroxyvitamin D, and a resectable squamous cell bronchogenic carcinoma surrounded by numerous non-caseating epithelioid granulomas, achieved normocalcemia, decreased levels of 1,25-dihydroxyvitamin D, and no evidence of tumor recurrence at 30 months following complete resection of his tumor. We suggest that an excess production of 1,25-dihydroxyvitamin D, through the granulomatous reaction around the tumor, was the mechanism of hypercalcemia. To our knowledge, no such mechanism of hypercalcemia has been previously reported with bronchogenic carcinoma. Furthermore, a review of the literature reveals that in cases of bronchogenic carcinoma, hypercalcemia is almost always associated with large and unresectable tumors, with a median survival after the discovery of hypercalcemia of only one month. This case, then, is unique because it describes an unusual mechanism of hypercalcemia with bronchogenic carcinoma and it emphasizes the rare occurrence of the potentially curable patient presenting with bronchogenic carcinoma complicated by hypercalcemia.

Aged↗

Isolation of a cDNA probe for a human jejunal brush-border hydrolase, sucrase-isomaltase, and assignment of the gene locus to chromosome 3.

We report the nucleotide sequence and derived amino acid sequence of a cDNA clone encoding most of the N-terminal, isomaltase region of human sucrase-isomaltase (SI). A plasmid containing this cDNA, pS12, identifies a 6-kb mRNA found in human jejunum and the human colon carcinoma cell line Caco-2. This human SI cDNA shows extensive overall homology with recently published rabbit SI cDNA. Using pS12 to probe DNA from a panel of somatic cell hybrids, we have assigned the gene encoding human SI to chromosome 3.

Amino Acid Sequence↗

Acquired immunity to Trichophyton mentagrophytes in thymus-grafted or peritoneal exudate cell-injected nude rats.

Congenitally athymic "nude" (RNU/RNU) rats were grafted with syngeneic neonatal thymus glands and 30 days later cutaneously inoculated with Trichophyton mentagrophytes. Nude rats and thymus-grafted nude rats were susceptible to infection with T. mentagrophytes but only thymus-grafted nude rats cleared the dermatophytosis. Clearance of the fungal infection took twice as long (approximately 60 days) in thymus-grafted nude rats when compared with heterozygous euthymic (+/RNU) controls (approximately 35 days). Following clearance of the primary dermatophytosis, peritoneal exudate cells (PEC) were adoptively transferred from either thymus-grafted or heterozygous rats to nude rats. Eight of 12 PEC-recipient nude rats acquired the capacity to resolve T. mentagrophytes-induced dermatophytosis (mean clearance time was approximately 40 days). All heterozygous (+/RNU) rats, thymus-grafted nude rats (4/4), and 8 of 12 PEC-recipient nude rats, which had cleared a primary dermatophytosis also expressed delayed-type hypersensitivity and elevated serum antibody titers to trichophytin antigen. These results demonstrate that immunity to T. mentagrophytes can be acquired in congenitally athymic nude rats following thymus grafting or injection of PEC from syngeneic +/RNU rats; however, injection of PEC from trichophytin-sensitized +/RNU donor rats to nude recipient rats did not result in transfer of specific adoptive immunity to T. mentagrophytes. Interestingly, PEC transfer from non-sensitized +/RNU rats was comparable to thymus grafting in effecting clearance of T. mentagrophytes dermatophytosis. These results suggest that acquired immunity to T. mentagrophytes in the rat is T cell-dependent, and that the absence of functional T lymphocytes and not an epithelial defect results in chronic dermatophytosis in the nude rat.

Animals↗

A procedure for isolation and partial purification of guinea pig lung mast cells.

Pulmonary mast cells were obtained from guinea pig lung using a combination of enzymatic digestion of tissue, centrifugal elutriation, and density gradient centrifugation on Percoll. In the initial procedure, lung tissue was enzymatically digested with collagenase and elastase in four 30 min incubations. Typically, monodispersed cell suspensions contained 4% mast cells. Further purification of these lung mast cells using elutriation and Percoll gradients consistently yielded mast cells of 40-78% (mean 51%) purity. These cells were morphologically intact, viable and found to be functional as determined by histamine release evoked by antigen and anti-guinea pig IgG1 antibody.

Animals↗

Influenza virus infection in mice after exposure to coal dust and diesel engine emissions.

Influenza virus infection initiated after aerosol exposure of CD-1, white Swiss mice for durations of 1, 3, and 6 months to respirable particulates maintained at 2 mg/m3 of either coal dust (CD), diesel engine emissions (DEE), a combination of both (CD/DEE), or to filtered air (control) was studied. The course of infection in mice previously exposed for 1 month to various particulates did not differ appreciably among the four animal groups with respect to mortality, virus growth in lungs, interferon levels, or hemagglutinin antibody response. In mice exposed for 3 and 6 months to different particulates, the mortality response was similar among all animal groups. However, the percentage of animals showing lung consolidation was significantly higher in the 3-month groups exposed to DEE (96.5%) and CD/DEE (97%) than in the control (61.2%); in the 6-month groups, the percentages were twice that of the control for both DEE- and CD/DEE-exposed animals. Complementing these observations of both 3- and 6-month-exposed animals was the higher virus growth levels attained in the DEE and CD/DEE animals with concomitant depressed interferon levels which were the inverse of findings noted in the control group. Hemagglutinin-antibody levels in particulate-exposed animals, especially at the 6-month interval, were fourfold less than the control. Histopathologic examination of lungs revealed no qualitative differences in the inflammatory response at any one specified time interval of exposure to influenza virus among the control and particulate-exposed animal groups. However, there were differences in severity of reaction in relation to the particulate component of the exposures. Focal macular collections of pigment-laden macrophages were seen only in DEE and CD/DEE but not in CD animals after 3- and 6-month exposures. The findings of this study indicated that the severity of influenza virus infection is more pronounced in mice exposed to diesel engine emissions than in control animals and it is not appreciably accentuated by coal dust.

Animals↗