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PubMed2002

Effect of prostaglandins on parasympathetic neurons in the rat lumbosacral spinal cord.

Prostaglandin E(2)(PGE(2)) elicits a variety of effects by activating four subtypes of receptors, EP1, EP2, EP3 and EP4. We examined receptor subtypes mediating the effects of PGE(2) on parasympathetic preganglionic neurons that regulate the activity of pelvic visceral organs. In tonic parasympathetic preganglionic neurons in neonatal rat spinal slices, PGE(2) increased the firing frequency to depolarizing current pulses, induced after-discharges and inhibited spike after-hyperpolarization. PGE(2) did not affect phasic preganglionic neurons. An EP1 agonist inhibited after-hyperpolarizations and induced after-discharges, whereas EP4 agonist reduced after-hyperpolarization and increased evoked firing but did not induce after-discharges. EP2 and EP3 agonists were inactive. These results indicate that PGE(2) acting via EP1 and/or EP4 receptors modulates the excitability and/or excitatory synaptic input to tonic parasympathetic preganglionic neurons.

Action Potentials↗
PubMed1999

Enteral infusion of sodium 2-ketoisocaproate in endotoxic rats.

OBJECTIVE: In view of our previous finding that the intravenous infusion of 2-ketoisocaproate (KIC) improved survival in septic rats, we endeavored to determine whether the enteral infusion of KIC improves survival in endotoxic rats, and, if so, the mechanism of this effect. SUBJECTS: Eighty-five rats were given 15 mg/kg of Escherichia coli lipopolysaccharide (026:B6). INTERVENTIONS: KIC, sodium pyruvate (PYR), or sodium bicarbonate (HCO3) was infused continuously intragastrically at 18.75 mmol/kg/day. MEASUREMENTS AND MAIN RESULTS: KIC administration increased circulating concentrations of KIC and ketone bodies. Survival rates were: KIC 17/32; PYR 2/22; and HCO3 8/31. The significant improvement in survival with KIC, in contrast with HCO3 (p<.04) or PYR (p<.002), points to an effect specific to KIC rather than to ketoacids generally, and argues against an antioxidant mechanism to explain improved survival with enteral administration. To determine whether altered nitric oxide production was responsible, plasma nitrite plus nitrate concentrations were measured sequentially in rats given a lower dose of lipopolysaccharide plus continuous intragastric KIC, PYR, or HCO3. All rats exhibited pronounced increases in plasma nitrite plus nitrate concentrations, peaking at 8 hrs, but both KIC and PYR caused greater increases than HCO3. Thus, differences in nitric oxide production cannot account for the different effects of PYR and KIC on survival. However, KIC infusion for 8 hrs substantially increased ketone bodies in blood and liver, in comparison with the infusion of HCO3 or PYR. CONCLUSION: Continuous enteral infusion of KIC improves survival in endotoxemia, probably by its conversion to ketone bodies, which serve as an alternative energy substrate.

Animals↗
PubMed2000

High-intensity dynamic human muscle performance enhanced by a metabolic intervention.

PURPOSE: The purpose of this study was to quantify the effects of a metabolic treatment on human muscle dynamic performance (strength, work, and fatigue) measured under conditions of acute, exhaustive high-intensity anaerobic isokinetic exercise. METHODS: Unilateral prefatigue and postfatigue peak torque and work values were measured in the quadriceps femoris of 13 subjects using a computer-controlled isokinetic dynamometer, over a 23-d interval. The two experimental treatments were: 1) a glycine and L-arginine salt of alpha-ketoisocaproic acid calcium ("GAKIC"); and 2) isocaloric sucrose (control). Based on a randomized double-blind cross-over repeated measures design, measurements were made before and during an exhaustive anaerobic fatigue protocol to calculate a Fatigue Resistance Index (FRI = [peri-exhaustion torque]\[baseline peak torque]), as well as total work. RESULTS: The FRI and total work for each of the exhaustion sets measured at 0, 5, and 15 min after oral GAKIC treatment were greater than values obtained for isocaloric control treatment (P < 0.02). GAKIC treatment increased the mean resistance to fatigue (FRI) up to 28% over isocaloric control. Overall gain in total muscle work attributable to GAKIC was 10.5 +/- 0.8% greater than control, sustained for at least 15 min. After 24 h, both GAKIC and control concentric forces returned to the same absolute values (P > 0.05): mean FRI = 0.42 +/- 0.05 and mean total work = 4600 +/- 280 J. There were no significant differences attributable to random order of testing. CONCLUSIONS: Compared with isocaloric carbohydrate, oral GAKIC treatment increased muscle torque and work sustained during intense acute anaerobic dynamic exercise; additionally, it increased overall muscle performance by delaying muscle fatigue during the early phases of anaerobic dynamic exercise.

Adult↗
PubMed2005

Osteoblastic phenotype expression of MC3T3-E1 cells cultured on polymer surfaces.

BACKGROUND: Current efforts in bone tissue engineering have as one focus the search for a scaffold material that will support osteoblast proliferation, matrix mineralization, and, ultimately, bone formation. The goal is to develop a bone substitute that is functionally equivalent to autograft bone. Previously published reports have shown that osteoblasts exhibit varying rates and degrees of proliferation and mineralization when grown on different surfaces. METHODS: This study presents a histologic and biomolecular analysis of MC3T3-E1 murine preosteoblast cells grown on poly(lactide-co-glycolide) (PLGA) versus poly(-caprolactone) (PCL), two commonly studied scaffold polymers. MC3T3-E1 cells were cultured on slides coated with either PLGA or PCL, and on uncoated glass slides as control, with six slides in each group. After 6 weeks in culture, the cells were stained for osteocalcin, alkaline phosphatase activity, and matrix mineralization. In addition, to assess the effects of the surface material on phenotypic expression at the molecular level, MC3T3-E1 cells were cultured on polymer-coated 24-well plates for 4 days, and analyzed by reverse transcription polymerase chain reaction for the expression of osteocalcin and alkaline phosphatase. RESULTS: The results showed that three groups of slides stained positively for osteocalcin at 6 weeks. However, markedly less alkaline phosphatase activity and mineralization were observed on the cells grown on PCL. Real-time polymerase chain reaction assays subsequently revealed decreased expression of both markers by cells cultured on PCL compared with PLGA. CONCLUSIONS: These results suggest that PCL does not support the full expression of an osteoblastic phenotype by MC3T3-E1 cells. PCL, therefore, is less desirable as a scaffold polymer in bone tissue engineering in so far as supporting bone formation is concerned. However, because PCL has favorable handling characteristics and strength, modifications of PCL may prompt further investigation.

3T3 Cells↗
PubMed1986

Characterization of Flavobacterium species by analysis of volatile fatty acid production.

Seventy-four Flavobacterium strains were characterized by gas-liquid chromatographic analysis of volatile fatty acids produced in the culture medium. Principal components analysis permitted the graphic representation of the relative positions of the different strains, and aggregation according to the variance enabled a hierarchical classification to be established. The study revealed three subgroups each for F. meningosepticum and F. odoratum. Our F. breve, Flavobacterium sp. group IIb and F. multivorum strains appeared to be homogeneous. These results tallied with those of previous studies on DNA base composition and reassociation, electrophoretic protein profiles and cellular fatty acid composition.

Butyrates↗
PubMed1989

Site-directed inhibition of Haemophilus influenzae malate dehydrogenase.

Previous studies of Haemophilus organisms documented the importance of an NAD+-dependent malate dehydrogenase in the incomplete tricarboxylic acid cycle present in these organisms. Selective interactions occurring at the coenzyme and substrate binding sites of a purified Haemophilus influenzae malate dehydrogenase were investigated. Coenzyme-competitive inhibition by adenosine derivatives demonstrated the presence of regions in the coenzyme binding site that interacted with the adenosine and pyrophosphate moieties of the coenzyme. Positive chainlength effects in the coenzyme-competitive inhibition by aliphatic carboxylic acids indicated the presence of a hydrophobic region at this site that was close to the pyrophosphate region. Seven analogues of NAD+ that were structurally altered in either the pyridine or purine ring were evaluated as selective inhibitors of the enzyme. The three most effective inhibitors of the purified malate dehydrogenase inhibited the growth of H. influenzae when the organism was grown on a limiting concentration of NAD+.

Adenosine Diphosphate↗
PubMed1970

Structure-activity correlations with compounds related to abscisic acid.

Inhibition of cell expansion of excised embryonic axes of Phaseolus vulgaris was used to evaluate the growth-inhibiting activity of abscisic acid and related compounds. None of the 13 compounds tested was as active as abscisic acid. 4-Hydroxyisophorone, a substance representative of the abscisic acid ring system was essentially inactive; cis, trans-3-methylsorbic acid, a compound resembling the side chain of abscisic acid, had low activity; and cis, trans-beta-ionylideneacetic acid was one-sixth as active. Loss of the ring double bond results in a drastic decrease in biological activity. Comparison of our results with those reported previously leads to the suggestion that the double bond of the cyclohexyl moiety may have an important function in determining the degree of activity of cis, trans-ionylideneacetic acids. Two modes of action are discussed. It seems possible that the ring double bond is involved in covalent bonding in binding of the abscisic acid analogue to macromolecules. This may require formation of an intermediate epoxide. It can also be argued that stereochemical differences between cyclohexane derivatives are important factors in determining the degree of biological activity.

Acetates↗
PubMed1996

Effects of H/D substitution on thermal vibrations in piperazinium hexanoate-h11, d11.

The crystal structures of piperazinium hexanoate-h11, 1/2C4H12N2(2+).C6H11O2-, and piperazinium hexanoate-d11, 1/2C4H12N2(2+).C6D11O2-, have been determined from neutron diffraction data collected at 15 K. Nuclear anisotropic displacement parameters have been analyzed to obtain the internal molecular displacements of the H and D nuclei, given by (u(obs)2)-(u(ext)2) where (u(ext)2) is the contribution assuming all H/D to be carried rigidly on the vibrating molecular framework consisting of the heavier nuclei. In both crystal structures the cation ring is well fitted by the rigid-body model and the anion chain by a model with two rigid segments. In the piperazinium cations the corresponding protons in the two structures have about the same internal vibrational directions and magnitudes except for the two N--H protons, perhaps owing to differences in N--H...O hydrogen bonding. The internal vibrations of corresponding H/D in the h11 and d11 anions have approximately the same vibrational directions. The internal mean-square displacements of the H nuclei are systematically greater than the values of the corresponding D nuclei by an average factor 1.7(3). For both anions, normal-mode analyses have been carried out using the force fields derived from ab initio quantum-mechanical calculations with HF/3-21G and HF/6-31G** basis sets. The values of the resultant H/D internal displacements for C--H(D) bond stretching and methylene out-of-plane vibrations are in good agreement with experiment. However, with either basis set, theory predicts methylene in-plane mean-square displacements significantly greater than the experimental values.

Caproates↗
PubMed1975

Influence of prostaglandins E1, E2, and arachidonate on melanosomes in melanocytes and keratinocytes of anagen hair bulbs in vitro.

Anagen hair bulbs were collected and maintained in organ culture for periods up to 24 hr. Prostaglandin E1, E2 and arachidonate were introduced to the cultures and stimulated a series of cellular events characterized by peripheral orientation of microfilaments in the dendritic processes of melanocytes, complexing of melanosomes; and degradation of melanosomes within keratinocytes. These events were dependent on the concentration of prostaglandins. In addition, the most potent agent for these events was arachidonate. The action of arachidonate was hypothesized to occur through the production of endogenous prostaglandins since arachidonate was ineffective in the presence of inhibitors of prostaglandin synthetase.

Arachidonic Acids↗
PubMed2003

Combined effects of prostaglandin E receptor subtype EP1 and subtype EP4 antagonists on intestinal tumorigenesis in adenomatous polyposis coli gene knockout mice.

Previous studies have shown that prostaglandin E(2) (PGE(2)) is involved in intestinal carcinogenesis through its binding to the PGE(2) receptor subtypes EP(1) and EP(4) and activation of downstream pathways. ONO-8711 and ONO-AE2-227, prostaglandin E receptor subtype EP(1)- and EP(4)-selective antagonists, respectively, are known to suppress formation of intestinal polyps in adenomatous polyposis coli gene-deficient mice. The present study was designed to investigate the combined effects of EP(1) and EP(4) antagonists on spontaneous polyp formation in APC1309 mice in order to determine the contribution of each receptor to intestinal tumorigenesis. APC1309 mice were treated with 400 ppm of ONO-8711 alone, 400 ppm of ONO-AE2-227 alone or both in combination in the diet for 6 weeks. The mean area of polyps found in the intestine, calculated as the longer diameter x the shorter diameter x pi, was reduced by 12%, 43% (P < 0.01) and 56% (P < 0.01) of the mean control value (8.8 mm(2)) in the ONO-8711 alone, ONO-AE2-227 alone and combination treatment groups, respectively, suggesting clear additive effects of the combination. The same additive tendency for suppression was also observed with respect to the numbers of polyps in the intestine. Polyp size reduction was more remarkable with the EP(4) antagonist, while the number reduction was more pronounced with the EP(1) antagonist. Our results indicate that EP(1) and EP(4) may have separate intrinsic roles and, to some extent, contribute to polyp formation independently. Thus, combination treatment has potential for the chemoprevention of colon carcinogenesis.

Animals↗
PubMed2005

Suppression of azoxymethane-induced colon cancer development in rats by a prostaglandin E receptor EP1-selective antagonist.

Prostaglandin E(2) is involved in colon carcinogenesis through its binding to the PGE(2) receptor subtypes EP(1), EP(2), EP(3) and EP(4). We have demonstrated that administration of ONO-8711, an EP(1)-selective antagonist, suppresses development of AOM-induced ACF in C57BL/6 mice and F344 rats. ONO-8711 also reduced the numbers of intestinal polyps in Min mice. In the present study, we investigated the long-term effects of ONO-8711 on colon cancer development in rats treated with AOM. Male F344 rats were injected subcutaneously with AOM (15 mg/kg body weight) once a week for the first 2 weeks to develop colon cancer. Administration of 400 or 800 p.p.m. ONO-8711 in their diets for 32 weeks reduced the incidence, multiplicity and volume of colon carcinomas. The incidence of colon adenocarcinomas in AOM-treated rats was 97, 83 and 76% (P < 0.05) in the 0, 400 and 800 p.p.m. of ONO-8711 groups, respectively. The multiplicity of adenocarcinomas was also decreased significantly, being 3.31 +/- 0.33, 2.34 +/- 0.27 (P < 0.05) and 2.06 +/- 0.34 (P < 0.01) with 0, 400 and 800 p.p.m. of ONO-8711, respectively. Moreover, treatment with 800 p.p.m. ONO-8711 reduced the mean volume of adenocarcinomas to 49% (P < 0.05) of the value for the AOM treatment alone. Furthermore, the BrdU labeling index was decreased significantly in colon cancer cells by 800 p.p.m. ONO-8711. These results confirm that EP(1) is involved in colon carcinogenesis and that EP(1)-selective antagonists might be promising candidates for colon cancer chemopreventive agents.

Animals↗