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PubMed1997

Perinatal bile acid metabolism: analysis of urinary bile acids in pregnant women and newborns.

The metabolism of bile acids in 30 pregnant women was evaluated by analyzing the urinary composition of bile acids during late gestation (weeks 30-41) and again in these women and their newborn infants during the first week after delivery. The levels of individual bile acids were determined by gas chromatography-mass spectrometry after solvolysis and hydrolysis of bile acid conjugates. The mean total bile acid/creatinine ratio in pregnant women decreased from 1.22 micromol/mmol creatinine at 30-32 weeks of gestation to 0.15 micromol/mmol creatinine at 6-7 days after delivery. The mean percentage of 1beta-hydroxylated bile acids peaked at 27% at 3-4 days after delivery. In newborn infants, the mean total bile acid/creatinine ratio rapidly increased from 3.39 micromol/mmol creatinine at birth to 54.33 micromol/mmol creatinine at 7 days. During this period, large amounts (40-50%) of unsaturated ketonic bile acids, especially 7alpha,12alpha-dihydroxy-3-oxo-5beta-chol-1-en-24-oic acid and 7alpha,12alpha-dihydroxy-3-oxo-4-cholen-24-oic acid, were observed in the infants' urine. These data suggest that, during the perinatal period, the formation of polyhydroxylated and unsaturated ketonic bile acids probably represents a mechanism for the excretion of bile salts, and that the metabolism of bile acids in both the mother and the infant changes significantly after birth.

Adult↗
PubMed1992

[Noninvasive metabolic analysis of the diabetic cornea and lens: in vivo measurement].

In vivo measurement of metabolic changes in diabetic cornea and lens were performed using redox fluorometry in nonobese diabetic (NOD) mice. Autofluorescence from reduced pyridine nucleotides (PN) and oxidized flavoproteins (Fp) were measured, and the PN/Fp ratio was used as a tissue metabolism indicator. The PN/Fp ratios were significantly higher in the diabetic corneal endothelium. Morphometric analysis of the corneal endothelium using specular microscopy revealed no significant differences between the two groups. These results indicate that redox fluorometry is able to detect early metabolic changes in the corneal endothelium and lens epithelium, which are induced by diabetes mellitus. Activation of the polyol pathway may be responsible for the change. Corneal epithelia may be less susceptible to diabetic changes than the corneal endothelium and lens epithelium.

Animals↗
PubMed1988

A metabolic analysis of high myopia and senile macular degeneration.

Abnormal zinc and copper metabolism have been described in several retinal disorders affecting the retinal pigment epithelium. An infrequent presentation of senile macular degeneration in high myopia is well known. We examined the blood for zinc and copper and the urine for copper of three groups of patients, one consisting of high myopia, another with high myopia and primary retinal detachment and the other a group of patients with senile macular degeneration. When comparing the above groups of patients, we found statistically elevated values of serum zinc (p = less than 0.005) in the group of patients with senile macular degeneration. The significance of these findings, and a correlation with the pathogeneses of high myopia and senile macular degeneration is discussed.

Adult↗
PubMed2002

Disruption of maternal behavior by alcohol intoxication in the lactating rat: a behavioral and metabolic analysis.

BACKGROUND: Preweanling rats exhibit clear behavioral signs of distress after interacting with an alcohol-intoxicated dam. Interestingly, behavioral reactivity of infants to the experience of alcohol in the nursing context decreases as a function of repeated alcohol administrations to the mother. In this study, maternal activities were examined when dams were exposed to repeated administrations of a subnarcoleptic alcohol dose. Maternal changes in alcohol metabolism were also analyzed as a function of repeated exposures to the drug. METHODS: During postpartum days 3, 5, 7, 9, 11, and 13, nursing dams received an intragastric administration of either 2.5 g/kg of alcohol or water. Maternal behaviors were evaluated (experiment 1). Blood alcohol levels (BALs) of the dams were determined on postpartum day 16 after all mothers received either an intragastric (experiment 2) or an intraperitoneal (experiment 3) dose of alcohol. The doses used (2.5 g/kg intragastrically and 1.5 g/kg intraperitoneally) were chosen because they promote similar peak BALs in dams naive to alcohol. RESULTS: Maternal behaviors were strongly affected by the state of intoxication. Nevertheless, these disruptions clearly subsided with progression of alcohol-related experiences (experiment 1). Chromatographic analysis of alcohol metabolism indicated the development of tolerance in dams that had prior experience with alcohol (experiment 2). Changes in BALs as a function of prior experience with alcohol seemed related to first-pass alcohol metabolism rather than hepatic oxidative processes of the drug (experiments 2 and 3). CONCLUSIONS: When the dam first experiences a moderate state of alcohol intoxication, maternal behaviors are uniformly disrupted. Subsequent exposures to alcohol lead to maternal metabolic tolerance. In conjunction with previous studies, these data indicate that infantile reactivity to alcohol is dependent on how the members of the dam/pup dyad express or perceive ethanol's postabsorptive effects.

Alcoholic Intoxication↗
PubMed1990

Metabolic analysis of reepithelializing rabbit cornea using phosphorus-31 nuclear magnetic resonance spectroscopy.

To investigate metabolic differences between the central and peripheral cornea the latter including the limbal area, corneas were dissected and examined using phosphorus-31 (31P) nuclear magnetic resonance spectroscopy. Since most 31P signals originate from the epithelium, 31P spectra of the cornea primarily represent the metabolic state of the epithelium. The spectra of the peripheral cornea showed all phosphorus resonances detected in the whole cornea; in contrast, the central cornea showed no phosphocreatine and glycerophosrylethanolamine, and only low levels of ATP. These results indicate that there is a higher metabolic activity in the peripheral epithelium, especially in the limbal area, than in the central epithelium. To evaluate the metabolic state of corneal epithelium during regeneration, we also examined corneas reepithelializing after 7 mm of central epithelial tissue had been removed by mechanical scraping. Rabbits were killed 24 and 48 h after scraping. The reepithelializing corneas clearly showed an increase in ATP, phosphocreatine, and sugar phosphates with time, although phosphorylcholine remained depressed. These findings suggest that the reepithelializing cornea has an elevated level of energy production and that it may have reached a higher steady state, thereby indicating accelerated metabolism of the epithelium during regeneration.

Animals↗
PubMed1993

Gestational and non-gestational trophoblastic neoplasms: new developments in DNA analysis, metabolic function, diagnosis and treatment.

Recent technological advances have made it possible to further define important molecular biological characteristics of gestational and non-gestational trophoblast neoplasm, as they relate to genetic origin and metabolic, endocrine, and diagnostic functions. Exciting new work with DNA analysis has defined origins of placental site trophoblastic tumors, a variant of choriocarcinoma not previously well understood. Coexistent hydatidiform mole and normal gestation have been diagnostically defined and carried out to viability. Finally, refinement of classification systems and expanded tumor markers offer promise of improved salvage in gestational neoplasms and an enlarging number of non-gestational neoplasms.

DNA, Neoplasm↗
PubMed2001

Novel amidohydrolytic reactions in oxidative pyrimidine metabolism: analysis of the barbiturase reaction and discovery of a novel enzyme, ureidomalonase.

Amidohydrolytic reactions in oxidative pyrimidine metabolism were investigated in detail. Barbiturase has been reported to catalyze the amidohydrolysis of barbituric acid to urea and malonic acid. However, purification of the enzyme revealed that it catalyzes the ring-opening of barbituric acid to ureidomalonic acid. The existence of a consecutive enzyme named ureidomalonase, which hydrolyzes ureidomalonic acid to urea and malonic acid, was also discovered during the purification of barbiturase.

Amides↗
PubMed1990

The evolution of focal ischemic damage: a metabolic analysis.

Focal cerebral ischemia in the rat was induced by left middle cerebral artery occlusion. The area of ischemia was determined by infusion of a qualitative perfusion indicator, neutral red. The temporal evolution of alterations in regional energy metabolism was assessed by direct microquantitative histochemical analysis of high-energy phosphates, glucose, glycogen, and lactate content of the tissue. Perfusion analyses demonstrated a perifocal region of diminished, but not absent perfusion up to 6 hr after occlusion. By 24 hr, there was an abrupt demarcation between perfused and nonperfused regions. Profound metabolic alterations were seen as early as 20 min after occlusion. Although there was an area of intermediate metabolic derangement in the more medial portions of the lateral ipsilateral cortex up to 6 hr, by 24 hr there was an abrupt transition from normal to abnormal cortex. No evidence of metabolic recovery was seen in this model of permanent occlusion.

Adenosine Triphosphate↗
PubMed2000

Bone metabolic analysis in patients with primary hyperparathyroidism.

Surgical treatment for primary hyperparathyroidism (PHPT) improves not only the calcium and phosphate metabolism but also the bone metabolism. This study was conducted to analyze the bone metabolism after PHPT operations. Bone mineral density (BMD) was measured by dual-photon absorptiometry in 50 patients before and after operation. Osteocalcin (OC) and alkaline-phosphatase activity (Alp) in serum were measured before and after surgery as markers of bone formation, and urinary deoxypiridinorine (DPD) as an index of osteoclast activity. The 50 patients under study were 40 women (80%) and ten men (20%). Increases in BMD at the lumbar spine were remarkable at three months following operation. Slow but steady progress was made until six months, reaching a plateau thereafter. The increase in BMD of lumbar spine was approximately 10%. Urinary DPD was the most sensitive among the three bone metabolic markers. Although serum Alp and OC remained high after operation, urinary DPD was normalized earlier. The discrepancy of bone formation and resorption was shown after operation and this contributed to the increases in BMD in the first six months.

Absorptiometry, Photon↗
PubMed1993

Metabolic analysis of latissimus dorsi coupled to a mock circulation.

A mock circulation system has been used to examine the metabolic and hemodynamic responses of untrained and trained latissimus dorsi muscle in a normal animal model. The metabolic response of untrained latissimus dorsi to differing stimulation regimes runs parallel to its mechanical performance. The ratio of power generated to oxidative capacity (a measure of metabolic efficiency) was maintained to a greater extent in muscle trained for 5 months subjected to specific fatigue tests, falling by only 20% (as opposed to 80% observed in untrained control muscle). This approach to studying metabolic and hemodynamic performance may have relevance when skeletal muscle is used for cardiac assistance.

Adenosine Triphosphate↗
PubMed1984

Nondestructive metabolic analysis of a cornea with the use of phosphorus nuclear magnetic resonance.

The viability of corneal donor material for penetrating keratoplasty depends on the metabolic status of the tissue; application of nondestructive nuclear magnetic resonance (NMR) techniques to the evaluation of donor tissue metabolism may provide information that would allow improved selection of donor material. The NMR spectrum generated from a single intact cornea permits qualitative and quantitative analysis of the following phosphatic metabolites: the sugar phosphates, inorganic orthophosphate, the alpha- and beta-phosphates of adenosine diphosphate and adenosine triphosphate, and the gamma-phosphate of adenosine triphosphate. Furthermore, the intracorneal pH (6.8) can be monitored from the resonance shift position of inorganic orthophosphate and can serve as an additional indicator of metabolic viability. To our knowledge, this study is the first to demonstrate the feasibility of the use of phosphorus NMR to monitor the metabolic status of a single intact cornea preserved in culture medium.

Adenosine Diphosphate↗
PubMed1997

Functional and metabolic analysis of cerebral ischemia using magnetoencephalography and proton magnetic resonance spectroscopy.

The details of the relationship between brain function and metabolism in brain infarcts have not been studied. Using magnetoencephalography (MEG) and proton magnetic resonance spectroscopic imaging (1H MRSI), we localized sources of abnormal magnetic activities in ischemic brain regions and biochemical changes in suspected lesions showing pathological characteristics. Twelve patients with ischemic stroke were examined and the results of MEG and 1H MRSI were superimposed onto the corresponding MR images. The signal intensities of N-acetyl (NA) and lactate (Lac) were measured in the lesions with highly concentrated dipoles of slow wave activity. Eleven of 12 cases had increased slow wave activity in the cortical areas adjacent to the infarcts; 1 case was excluded because the infarct was too small (<1 cm in diameter). The signal intensity of NA in the regions with the highest slow wave activity was significantly reduced and was well correlated with the dipole density of slow waves. Though Lac was mildly accumulated in the lesions, the Lac level had no correlation with slow wave magnetic activity. The remaining and metabolically active cortical tissue showing NA signal produced the abnormal slow wave activity under lactic acidosis (mild accumulation of Lac).

Adult↗
PubMed1988

[Effect of buflomedil on cerebral acetylcholine, neuroactive amino acids contents and energy metabolism: analysis using spontaneously hypertensive rat (SHR)].

Effects of continuous oral administration of Buflomedil on acetylcholine (ACh) and neuroactive amino acids contents and energy metabolism in the brain were investigated using Wistar Kyoto rat (WKY) and spontaneously hypertensive rat (SHR). In both WKY and SHR, continuous oral administration of Buflomedil (30 mg/kg x 7 days) had no effect on ACh content in all cerebral areas examined. On the other hand, continuous oral administration of Buflomedil to SHR induced a significant increase in the contents of glutamic acid and taurine in the striatum and hypothalamus, while it resulted in a significant decrease in the contents of glycine and taurine in the midbrain and hippocampus. Glucose and ATP contents in the striatum of SHR showed a significant increase following the continuous oral administration of Buflomedil. These results suggest that Buflomedil may be a drug capable of improving various neuropsychiatric symptoms associated with cerebrovascular insufficiency by modifying the content of neuroactive amino acids and accelerating energy metabolism in the brain.

Acetylcholine↗
PubMed1984

Ex vivo metabolic analysis of eye bank corneas using phosphorus nuclear magnetic resonance.

Application of nondestructive and noninvasive nuclear magnetic resonance (NMR) techniques to the evaluation of corneal donor tissue metabolism may provide information that would allow improved selection of donor material for keratoplasty. A phosphorus 31 NMR spectrum was generated from a single intact human cornea, evaluating phosphatic metabolites qualitatively and quantitatively. The NMR analyses were conducted on four corneas (from patients aged from 73 to 75 years). Intracorneal pH (7.2) was monitored from the resonance shift position of inorganic orthophosphate (Pi). A numerical index (1.15) of tissue energy status was determined by comparing the relative tissue content of high-field, high-energy phosphate signals (adenosine triphosphate, adenosine diphosphate, nicotinamide-adenine dinucleotide, and the uridine diphospho-hexoses) to that of the low-field, low-energy phosphate signals (Pi, sugar phosphates, and phosphodiesters). This is the first demonstration of the feasibility of using nondestructive 31P NMR to monitor the metabolic status of a single intact eye bank cornea as would be required in the evaluation of eye bank tissue before keratoplastic procedures.

Adenosine Diphosphate↗
PubMed2005

Microfluidic chip toward cellular ATP and ATP-conjugated metabolic analysis with bioluminescence detection.

In this article, a microfluidic platform integrating capillary electrophoresis and bioluminescence (BL) detection that was fabricated in poly(dimethylsiloxane) (PDMS) with lab-on-a-chip technology was demonstrated for cellular metabolic analyses. A microchannels network, "cross combining with Y", was designed to perform on-chip sample preparation, separation, and BL detection of ATP and ATP-conjugated metabolites, using firefly luciferin-luciferase BL system. A dynamic modification of the channel wall of PDMS proved to be crucial to reverse the direction of electroosmotic flow (EOF), which was uniquely achieved by a prewash cycle with a cationic surfactant didodecyldimethylammonium bromide. The influences of surfactant on the EOF and BL reaction were also investigated. Quantitative analyses revealed a dynamic linear range over 2 orders of magnitude for ATP, with a detection limit down to submicromolar (midattomole). The method was validated by measuring cellular ATP of E. coli. with direct on-chip cell lysis. Further work was emphasized on ATP-conjugated metabolite analysis, using galactose as an example. Assays of galactose in human urine samples confirmed the reliability of the protocol, which revealed good prospect of this platform for ATP-conjugated submetabolomic profiling.

Adenosine Triphosphate↗
PubMed2006

Metabolic analysis of adaptive evolution for in silico-designed lactate-producing strains.

Experimental evolution is now frequently applied to many biological systems to achieve desired objectives. To obtain optimized performance for metabolite production, a successful strategy has been recently developed that couples metabolic engineering techniques with laboratory evolution of microorganisms. Previously, we reported the growth characteristics of three lactate-producing, adaptively evolved Escherichia coli mutant strains designed by the OptKnock computational algorithm. Here, we describe the use of (13)C-labeled experiments and mass distribution measurements to study the evolutionary effects on the fluxome of these differently designed strains. Metabolic flux ratios and intracellular flux distributions as well as physiological data were used to elucidate metabolic responses over the course of adaptive evolution and metabolic differences among strains. The study of 3 unevolved and 12 evolved engineered strains as well as a wild-type strain suggests that evolution resulted in remarkable improvements in both substrate utilization rate and the proportion of glycolytic flux to total glucose utilization flux. Among three strain designs, the most significant increases in the fraction of glucose catabolized through glycolysis (>50%) and the glycolytic fluxes (>twofold) were observed in phosphotransacetylase and phosphofructokinase 1 (PFK1) double deletion (pta- pfkA) strains, which were likely attributed to the dramatic evolutionary increase in gene expression and catalytic activity of the minor PFK encoded by pfkB. These fluxomic studies also revealed the important role of acetate synthetic pathway in anaerobic lactate production. Moreover, flux analysis suggested that independent of genetic background, optimal relative flux distributions in cells could be achieved faster than physiological parameters such as nutrient utilization rate.

Acetate-CoA Ligase↗