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Biomedical subjects

P Feng

Publications and source records attributed to P Feng.

134 records · Page 8Linked to original sources

Activation of protein kinase C potentiates cyclic AMP production and stimulates steroidogenesis in differentiated ovarian granulosa cells.

Gonadotropin-stimulated steroidogenesis in the differentiating ovarian granulosa cell is mediated through the activation of cAMP-dependent protein kinase, and is also modulated by calcium-dependent mechanisms. Granulosa cells contain calcium-activated, phospholipid-dependent protein kinase (C kinase), and show an increase in phosphatidylinositol turnover in response to GnRH agonist analogs. To evaluate the role of C kinase in ovarian steroidogenesis, the potent phorbol ester, TPA, and the permeant diacylglycerol, OAG, were used to activate C kinase in granulosa cells from PMSG-treated immature rats. Both TPA and OAG caused dose-dependent stimulation of progesterone production without affecting intra- or extracellular cAMP levels. However, the maximum steroid responses to these compounds were less than those stimulated by cAMP. The ED50 for TPA-stimulated progesterone production was 3 nM, which is close to the known Km for activation of C kinase. Stimulation of steroidogenesis was only observed with biologically-active phorbol esters and permeant diacylglycerols such as OAG and DOG. Exposure of granulosa cells to phospholipase C also increased progesterone production in a dose-dependent manner without changing the cAMP content. Although TPA and OAG did not increase basal cAMP production, both agents enhanced the cAMP responses stimulated by hCG and forskolin; likewise, phospholipase C alone did not change cAMP production but caused a dose-dependent increase in the cAMP responses to hCG and forskolin. These results demonstrate that activation of C kinase promotes steroidogenesis in ovarian granulosa cells, and potentiates the activation of adenylate cyclase by hCG and forskolin. Such findings support the possibility that the calcium, phospholipid-dependent enzyme could be involved in the regulation of progesterone production by hormonal ligands such as gonadotropins and GnRH.

Animals↗

Gonadotropin-releasing hormone as a modulator of ovarian function.

GnRH and its agonist analogs exert direct inhibitory and stimulatory effects on the ovaries of animals from several species. In the immature follicle, GnRH inhibits the actions of FSH on an integrated array of biochemical responses that lead to follicular development and corpus luteum formation. GnRH also suppresses gonadotropin action in mature follicles, and stimulates certain ovarian processes such as steroidogenesis and oocyte maturation. The inhibitory and stimulatory effects of GnRH are mediated through the binding of the peptide to high-affinity receptors in granulosa and thecal cells. Recent studies have shown that GnRH action in the ovary is dependent upon calcium mobilization and probably operates through stimulation of phospholipid turnover and activation of protein kinase C.

Animals↗

Formation of 12-[18-O]oxo-cis-10, cis-15-phytodienoic acid from 13[18-O]hydroperoxylinolenic acid by hydroperoxide cyclase.

13-[18-O]Hydroperoxylinolenic acid was permitted to react with an extract of flaxseed acetone powder containing hydroperoxide cyclase activity. The resulting product, 12-oxo-cis-10,cis-15-phytodienoic acid (12-oxo-PDA), contained 18O in the carbonyl oxygen at carbon 12, suggesting that an epoxide was an intermediate in the hyderoperoxide cyclase reaction. A substrate specificity study showed that a cis double bond beta, gamma to the conjugated hydroperoxide group was essential for the substrate to be converted to a cyclic product by hydroperoxide cyclase.

Carbon Isotopes↗

Direct identification of Yersinia enterocolitica in blood by polymerase chain reaction amplification.

Primers based on the nucleotide sequence of the virF gene in the pYV plasmid and the chromosomal ail gene were used in polymerase chain reaction (PCR) amplifications to directly identify Yersinia enterocolitica in blood. Approximately 500 bacteria seeded into 100 microL of blood can be extracted and amplified by PCR to yield positive results. PCR analyses of seven Y. enterocolitica isolates previously implicated in blood contaminations showed that only one isolate harbored the plasmid-borne virF gene; however, all seven isolates were identified effectively by the PCR product amplified from the chromosomal gene. The PCR assay has the potential for use in the identification of Y. enterocolitica contamination in stored units of blood or in the rapid diagnosis of transfusion-related bacteremia caused by Y.

Base Sequence↗

Escherichia coli serotype O157:H7: novel vehicles of infection and emergence of phenotypic variants.

Escherichia coli serotype O157:H7 was only recognized as a human pathogen a little more than a decade ago, yet it has become a major foodborne pathogen. In the United States, the severity of serotype O157:H7 infections in the young and the elderly has had a tremendous impact on human health, the food industry, and federal regulations regarding food safety. The implication of acidic foods as vehicles of infection has dispelled the concept that low-pH foods are safe. Further, the association of nonbovine products with outbreaks suggests that other vehicles of transmission may exist for this pathogen. In laboratory diagnosis, most microbiologic assays rely on a single phenotype to selectively isolate this pathogen. However, the increasing evidence that phenotypic variations exist among isolates in this serogroup may eventually necessitate modifications in assay procedures to detect them.

Animals↗

Emergence of rapid methods for identifying microbial pathogens in foods.

Because of the complexities of food analysis, conventional microbiological methods must use time-consuming enrichment steps for culturing viable bacterial cells in foods. With rapid advancements in technology, however, numerous so-called rapid methods were introduced into the field of food microbiology in a relatively short time. Culture methods that were once used to obtain profiles for bacterial identification were simplified or automated. Many microbiological procedures were also streamlined or automated to reduce assay time, labor, and materials. Nucleic acid-based assays are used to identify gene sequences in foodborne bacteria, and antibody-based assays are used in numerous formats to detect bacterial pathogens and toxins in foods. The difficulties of analyzing food, however, remain challenging, and rapid methods need to be evaluated thoroughly before they are used for routine food analysis.

Food Microbiology↗

Comparison of sorbitol MacConkey and hemorrhagic coli agars for recovery of Escherichia coli O157:H7 from brie, ice cream, and whole milk.

The relative efficacies of hemorrhagic coli (HC) agar and several formulations of sorbitol MacConkey (SorMac) agar, with and without 0.1% (w/v) 4-methyllumbelliferyl-beta-D-glucuronide (MUG), in recovering unstressed and heat-stressed Escherichia coli O157:H7 from Brie cheese, ice cream, and whole milk were determined. Recovery of unstressed E. coli O157:H7 was determined quantitatively by spread-plating diluted samples onto different agars and performing plate counts. Recovery of stressed E. coli O157:H7 was determined qualitatively by enriching samples in modified trypticase soy broth, streaking the incubated enrichments, and isolating E. coli O157:H7 colonies from the agars. HC agar and the SorMac agar formulations did not differ significantly in their ability to recover unstressed E. coli O157:H7 from ice cream and whole milk; however, HC agar recovered significantly more unstressed E. coli O157:H7 from Brie cheese than did the SorMac agar formulations. Bacteriological Analytical Manual and Oxoid SorMac agar formulations made from individual ingredients, did not differ significantly in recovering unstressed E. coli O157:H7 from Brie cheese. The efficiency of the commercially available Oxoid SorMac agar could not be determined because of overgrowth by indigenous microflora. HC and SorMac agars did not differ significantly in recovering stressed E. coli O157:H7 from Brie cheese, ice cream, and whole milk. MUG had no apparent effect on recovery of either stressed or unstressed E. coli O157:H7 from the dairy foods examined.

Agar↗