Metabolic Changes In Response Research Articles

Analysis of enzymopathies in the human red blood cells by constraint-based stoichiometric modeling approaches

The human red blood cell (RBC) metabolism is investigated by calculating steady state fluxes using constraint-based stoichiometric modeling approaches. For the normal RBC metabolism, flux balance analysis (FBA) is performed via optimization of various alternative objective functions, and the maximization of production of ATP and NADPH is found to be the primary objective of the RBC metabolism. FBA and two novel approaches, minimization of metabolic adjustment (MOMA) and regulatory on–off minimization (ROOM), which can describe the behavior of the metabolic networks in case of enzymopathies, are applied to observe the relative changes in the flux distribution of the deficient network. The deficiencies in several enzymes in RBC metabolism are investigated and the flux distributions are compared with the non-deficient FBA distribution to elucidate the metabolic changes in response to enzymopathies. It is found that the metabolism is mostly affected by the glucose-6-phosphate dehydrogenase (G6PDH) and phosphoglycerate kinase (PGK) enzymopathies, whereas the effects of the deficiency in DPGM on the metabolism are negligible. These stoichiometric modeling results are found to be in accordance with the experimental findings in the literature related to metabolic behavior of the human red blood cells, showing that human RBC metabolism can be modeled stoichiometrically.

Purinergic pathway suppresses the release of .NO and stimulates proteoglycan synthesis in chondrocyte/agarose constructs subjected to dynamic compression

Mechanical loading plays a fundamental role in the physiological and pathological processes of articular cartilage. The application of dynamic compression to chondrocytes cultured in agarose, downregulates the release of nitric oxide (NO) and enhances cell proliferation and proteoglycan synthesis. We hypothesize that the observed metabolic changes in response to dynamic compression involve a purinergic signaling pathway. Chondrocyte/agarose constructs were subjected to dynamic compression (15%, 1 Hz, 48 h) in the presence of antagonists for the purinergic pathway. Gadolinium was used as a putative inhibitor of stretch-activated calcium ion channels including adenosine 5'-triphosphate (ATP) release channels; suramin was employed as a P2 receptor antagonist and apyrase was used to catalyze the hydrolysis of extracellular ATP. The data presented demonstrate that in the absence of the inhibitor, dynamic compression suppressed .NO release. Treatment with gadolinium and suramin caused a compression-induced upregulation of .NO release, a response abolished with apyrase. Compression-induced stimulation of cell proliferation was reversed with gadolinium, suramin, or apyrase. By contrast, compression-induced stimulation of proteoglycan synthesis was abolished under all treatment conditions. Thus, the purinergic pathway is important in suppressing the release of .NO and stimulation of proteoglycan synthesis. Indeed, high levels of .NO could trigger a downstream catabolic response and mediate the compression-induced inhibition of cell proliferation. The current study demonstrates for the first time the importance of a purinergic pathway in mediating the metabolic response to dynamic compression and suppressing an inflammatory effect.

Carbohydrate Restriction, Dietary Energy Restriction and Weight Loss in Obese Mice: Metabolic Insights

Our objective was to compare weight loss, body composition, and key metabolic changes in response to carbohydrate restriction (modeling the Atkins diet) relative to dietary energy restriction (DER). Male C57BL/6 mice were randomized into five groups of 20. Group HC mice consumed a high carbohydrate diet and group HF consumed a high fat diet for 14 weeks. The remaining three groups consumed the HF diet for seven weeks to induce obesity and during weeks 8–14 were switched to either: a) an Atkins-like low carbohydrate diet similar to the induction phase of the Atkins diet; b) pair-feeding (PF) with the HC diet to match the energy intake of the Atkins group; or c) a high carbohydrate DER regimen providing 70% of the energy intake consumed by the HF group. At 14 weeks, body weights of the Atkins (40.3 ± 1.1 g) and HF (40.2 ± 1.1 g) groups were greater than the HC (33.0 ± 0.8 g), PF (31.5 ± 0.8 g) and DER (22.7 ± 0.6 g) groups. The Atkins (47.3 ± 1.2 %) and HF (46.8 ± 1.2 %) groups exhibited higher percent fat mass than the HC (35.4 ± 1.1 %), PF (33.9 ± 1.8 %), and DER (19.5 ± 0.8 %) groups. The Atkins and HF groups demonstrated lower bone mineral density than other groups. Relative to the DER group, the Atkins group displayed comparable serum ketone bodies but higher serum cholesterol, triglycerides, leptin, IGF-1, glucose and insulin. In contrast to DER, the Atkins-like diet did not cause weight loss or reduce serum markers associated with obesity-related diseases in mice. Funding provided in part by the Dept. Human Ecology and by N01-CO-12400.

Hemodynamic and metabolic responses to activation, deactivation and epileptic discharges

To investigate the coupling between the hemodynamic and metabolic changes following functional brain activation as well as interictal epileptiform discharges (IEDs), blood oxygenation level dependent (BOLD), perfusion and oxygen consumption responses to a unilateral distal motor task and interictal epileptiform discharges (IEDs) were examined via continuous EEG-fMRI. Seven epilepsy patients performed a periodic (1 Hz) right-hand pinch grip using ∼8% of their maximum voluntary contraction, a paradigm previously shown to produce contralateral MI neuronal excitation and ipsilateral MI neuronal inhibition. A multi-slice interleaved pulsed arterial spin labeling and T2*-weighted gradient echo sequence was employed to quantify cerebral blood flow (CBF) and BOLD changes. EEG was recorded throughout the imaging session and reviewed to identify the IEDs. During the motor task, BOLD, CBF and cerebral metabolic rate of oxygen consumption (CMRO2) signals increased in the contra- and decreased in the ipsilateral primary motor cortex. The relative changes in CMRO2 and CBF were linearly related, with a slope of 0.46 ± 0.05. The ratio of contra- to ipsilateral CBF changes was smaller in the present group of epilepsy patients than in the healthy subjects examined previously. IEDs produced both increases and decreases in BOLD and CBF signals. In the two case studies for which the estimation criteria were met, the coupling ratio between IED-induced CMRO2 and CBF changes was estimated at 0.48 ± 0.17. These findings provide evidence for a preserved coupling between hemodynamic and metabolic changes in response to both functional activation and, for the two case studies available, in response to interictal epileptiform activity.

Endogenous Fluorescence Spectroscopy of Cell Suspensions for Chemopreventive Drug Monitoring¶

ABSTRACTCancer chemopreventive agents such as N‐4‐(hydroxyphenyl)‐retinamide (4HPR) are thought to prevent cancers by suppressing growth or inducing apoptosis in precancerous cells. Mechanisms by which these drugs affect cells are often not known, and the means to monitor their effects is not available. In this study endogenous fluorescence spectroscopy was used to measure metabolic changes in response to treatment with 4HPR in ovarian and bladder cancer cell lines. Fluorescence signals consistent with nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD) and tryptophan were measured to monitor cellular activity through redox status and protein content. Cells were treated with varying concentrations of 4HPR and measured in a stable environment with a sensitive fluorescence spectrometer. Results suggest that redox signal of all cells changed in a similar dose‐dependant manner but started at different baseline levels. Redox signal changes depended primarily on changes consistent with NADH fluorescence, whereas the FAD fluorescence remained relatively constant. Similarly, tryptophan fluorescence decreased with increased drug treatment, suggesting a decrease in protein production. Given that each cell line has been shown to have a different apoptotic response to 4HPR, fluorescence redox values along with changes in tryptophan fluorescence may be a response as well as an endpoint marker for chemopreventive drugs.

Methimazole, thyroid hormone replacement, and lipogenic enzyme gene expression in broilers

The purpose of this experiment was to determine the possible relationship between certain indices of lipid metabolism and specific gene expression in chickens fed methimazole to simulate hypothyroidism. Male broiler chickens ( Gallus gallus) growing from 7 to 28 days of age were fed diets containing 18% crude protein and either 0 or 1 g methimazole per kilogram of diet. At 28 days, these two groups were further subdivided into groups receiving 18% crude protein diets containing either 0 or 1 mg triiodothyronine (T 3) per kilogram. Birds were sampled at 28, 30, and 33 days. Measurements taken included in vitro lipogenesis (IVL), malic enzyme (ME) activity, isocitrate dehydrogenase, aspartate amino transferase, and the expression of the genes for ME, fatty acid synthase (FAS), and acetyl coenzyme carboxylase (ACC). Hypothyroidism decreased IVL and ME at 28 days of age; however, T 3 supplementation for 2 days restored both IVL and ME. Paradoxically, continuing T 3 replenishment for an additional 3 days decreased IVL but did not decrease ME activity. In contrast, supplemental T 3 decreased IVL in euthyroid birds, regardless of the dosing interval, but had no effect on ME activity. Although methimazole decreased ME gene expression, there was only a transitory relationship between enzyme activity and gene expression when plasma T 3 was restored with exogenous T 3. These data may help to explain some of the apparent reported dichotomies in lipid metabolism elicited by changes in the thyroid state of animals. In addition, most metabolic changes in response to feeding T 3 occurred within 2 to 5 days, suggesting that changes in intermediary metabolism preceded morphological changes. In conclusion, the thyroid state of the animal will determine responses to exogenous T 3.

Exposure to appetitive food stimuli markedly activates the human brain

Objective: The increased incidence of obesity most likely reflects changes in the environment that had made food more available and palatable. Here we assess the response of the human brain to the presentation of appetitive food stimuli during food presentation using PET and FDG. Method: Metabolic changes in response to food presentation were done in 12 healthy normal body weight subjects who were food deprived before the study. Results: Food presentation significantly increased metabolism in the whole brain (24%, P < 0.01) and these changes were largest in superior temporal, anterior insula, and orbitofrontal cortices. The increases in the right orbitofrontal cortex were the ones that correlated significantly with the increases in self-reports of hunger and desire for food. Discussion: The marked increase in brain metabolism by the presentation of food provides evidence of the high sensitivity of the human brain to food stimuli. This high sensitivity coupled with the ubiquitousness of food stimuli in the environment is likely to contribute to the epidemic of obesity. In particular, the activation of the right orbitofrontal cortex, a brain region involved with drive, may underlie the motivation to procure food, which may be subjectively experienced as “desire for food” and “hunger” when exposed to food stimuli.

Endogenous Fluorescence Spectroscopy of Cell Suspensions for Chemopreventive Drug Monitoring

Cancer chemopreventive agents such as N-4-(hydroxyphenyl)retinamide (4HPR) are thought to prevent cancers by suppressing growth or inducing apoptosis in precancerous cells. Mechanisms by which these drugs affect cells are often not known, and the means to monitor their effects is not available. In this study endogenous fluorescence spectroscopy was used to measure metabolic changes in response to treatment with 4HPR in ovarian and bladder cancer cell lines. Fluorescence signals consistent with nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD) and tryptophan were measured to monitor cellular activity through redox status and protein content. Cells were treated with varying concentrations of 4HPR and measured in a stable environment with a sensitive fluorescence spectrometer. Results suggest that redox signal of all cells changed in a similar dose-dependant manner but started at different baseline levels. Redox signal changes depended primarily on changes consistent with NADH fluorescence, whereas the FAD fluorescence remained relatively constant. Similarly, tryptophan fluorescence decreased with increased drug treatment, suggesting a decrease in protein production. Given that each cell line has been shown to have a different apoptotic response to 4HPR, fluorescence redox values along with changes in tryptophan fluorescence may be a response as well as an endpoint marker for chemopreventive drugs.

Hostility differentiates the brain metabolic effects of nicotine

The brain mechanisms underlying the cause of nicotine dependence are unknown, however, hostility traits are associated with increased susceptibility to nicotine dependence. We used FDG PET to measure brain metabolic response to nicotine administered by patch while the subject performed the Bushman aggression task in 86 high- and low-hostility subjects. Low-hostility trait subjects demonstrated no significant change in brain metabolic response to nicotine. In marked contrast, high-hostility non-smokers subjects demonstrated dramatic metabolic changes to low dose (3.5 mg patch) as did high-hostility smokers to high dose nicotine (21 mg patch) throughout the brain bilaterally ( p<0.025). Correlational analyses demonstrated greater metabolic changes in response to nicotine in subjects with greatest hostility trait measures. The observed differences were not a consequence of plasma nicotine or cotinine levels. These metabolic changes were not observed when subjects performed a sustained attentional task (continuous performance task; CPT). Behaviorally, high-hostility subjects had higher ratings of anger, impatience, irritability and nervousness, and lower ratings of happiness, relaxation and curiosity than low-hostility subjects. Smokers had significantly greater scores on impatience and restlessness than non-smokers. This PET study demonstrates a conspicuous lack of the brain metabolic response to nicotine in low-hostility non-smokers in contrast to a dramatic brain response to nicotine in high hostility subjects. This biological difference in brain metabolic response to nicotine between high and low hostility trait subjects may explain differences in susceptibility to nicotine dependence.

Age-related differences in metabolic adaptations following resistance training in women

The purpose of this study was to determine whether aging alters changes in resting metabolic rate, body composition and insulin sensitivity in response to resistance training (RT) in women. The impact of a 6-month controlled RT program on fat-free mass (FFM), resting energy expenditure (REE), and glucose disposal was examined in 19 younger (27.8±3.5 yr; range 18–35) and 12 older (66.6±4.9 yr, range 55–70) non-obese caucasian women. For younger women, the RT program increased body weight (59.1±6.2 to 60.8±6.1 kg, p<0.05) due to an increase in FFM (39.2±3.7–40.4±3.2 kg, p<0.05). REE increased from 1379±114 to 1451±140 kcal day −1, p<0.0001), and glucose disposal (from 364.1±91.1 to 402.1±87.8 mg min −1, p<0.05). Neither fat mass nor V ̇ O 2 max changed significantly. On the other hand, older women lost fat mass (24.0±4.6–23.2±5.4 kg, p<0.05) and tended to gain FFM (from 37.3±4.0–38.0±4.3 kg, +1.9%; p=0.057), whereas no change occurred in body weight, REE, V ̇ O 2 max or glucose disposal. Thus, younger women showed greater metabolic changes in body composition, REE and insulin sensitivity in response to RT than older women. These results suggest an age-related alteration in metabolic changes in response to a 6-month RT program in previously untrained women.

Methimzole and thyroid hormone replacement in broilers

Seven-day-old chickens were fed diets containing 18% crude protein+0 or 1 g methimazole/kg to produce either euthyroid or hypothyroid groups of birds at 28 days of age. These two groups were then offered diets containing either 0 or 1 mg triiodothyronine (T 3)/kg diet. Birds were sampled at 0, 2, 5, and 8 days following the onset of the T 3 treatment. Measurements taken at these intervals included in vitro hepatic lipogenesis (IVL), growth and feed consumption, hepatic enzyme activities (malic enzyme, ME; isocitrate dehydrogenase, ICD; and aspartate amino transferase, AAT), plasma hormones (T 3; thyroxine, T 4; insulin like growth factors I, IGF-I; and insulin like growth factors II, IGF-II) and metabolites (glucose; fatty acids, NEFA; triglyerides; uric acid). Hypothyroidism decreased IVL and ME at 28 days of age; however, T 3 supplementation for 2 days restored both IVL and ME. Paradoxically, continuing T 3 replenishment for an additional 3–6 days decreased IVL without affecting ME activity. In contrast, supplemental T 3 decreased IVL in euthyroid birds, regardless of the dosing interval, but had no effect on ME activity. Methimazole decreased plasma T 3, T 4, uric acid, and IGF-I, but did not affect IGF-II at 28 days. Giving T 3 to birds previously on methimazole increased plasma IGF-I as did feeding a control diet. Supplemental T 3 increased NEFA in both euthyroid and hypothyroid birds, but only for a short period following the initiation of supplementation (2 days post-supplementation). These data may help to explain some of the apparent reported dichotomies in lipid metabolism elicited by changes in the thyroid state of animals. In addition, most metabolic changes in response to feeding T 3 occurred within 2–5 days, suggesting that changes in intermediary metabolism preceded morphological changes. In conclusion, the thyroid state of the animal will determine responses to exogenous T 3.

Age-dependent change in metabolic response to photic stimulation of the primary visual cortex in infants: functional magnetic resonance imaging study.

The blood oxygen level-dependent (BOLD) response to photic stimulation in the primary visual cortex (V1) reverses from positive to negative around 8 weeks of age. This phenomenon may be caused by increased oxygen consumption during stimulation as the result of a rapid increase of synaptic density at this age. To test this hypothesis, we applied existing mathematic models of BOLD signals to the experimental data from infants. When the stimulus-related increments of cerebral blood flow and cerebral blood volume were fixed at 60% and 20%, respectively, the mean estimated increment of the cerebral metabolic rate of oxygen of the V1 in the elder infant group (57.1% +/- 8.8%) was twice as large as that in the younger infant group (32.2% +/- 4.7%), which corresponds to the reported difference in synaptic density. The present data confirmed that a change in oxygen consumption could explain a transition from a positive to a negative BOLD response.

Effects of moderate hyperoxia on oxygen consumption during submaximal and maximal exercise.

The present study examined the effect of hyperoxia on oxygen uptake (VO(2)) and on maximal oxygen uptake (VO(2max)) during incremental exercise (IE) and constant work rate exercise (CWRE). Ten subjects performed IE on a bicycle ergometer under normoxic and hyperoxic conditions (30% oxygen). They also performed four 12-min bouts of CWRE at 40, 55, 70 and 85% of normoxic VO(2max) (ex1, ex2, ex3 and ex4, respectively) in normoxia and in hyperoxia. VO(2max) was significantly improved by 15.0 (15.2)% under hyperoxia, while performance (maximum workload, W(max)) was improved by only +4.5 (3.0)%. During IE, the slope of the linear regression relating VO(2) to work rate was significantly steeper in hyperoxia than in normoxia [10.80 (0.88) vs 10.06 (0.66) ml x min(-1) x W(-1)]. During CWRE, we found a higher VO(2) at ex1, ex2, ex3 and ex4, and a higher VO(2) slow component at ex4 under hyperoxia. We have shown that breathing hyperoxic gas increases VO(2max), but to an extent that is difficult to explain by an increase in oxygen supply alone. Changes in metabolic response, fibre type recruitment and VO(2) of non-exercising tissue could explain the additional VO(2) for a given submaximal work rate under hyperoxia.

Genetic variation at the uncoupling protein 1, 2 and 3 loci and the response to long-term overfeeding.

To evaluate the effects of uncoupling protein (UCP) 1, UCP2 and UCP3 gene variants on body composition and metabolic changes in response to chronic overfeeding and the recovery after the period of overfeeding. Twenty-four normal weight men (21+/-2 y), who constituted 12 pairs of identical twins, ate a 4.2 MJ/day energy surplus, 6 days a week, during a period of 100 days. The subjects were asked to return to the laboratory for testing at 4 months and for a final examination 5 y after completion of the overfeeding protocol. Resting metabolic rate (RMR) measurements were performed before and after overfeeding. A 4.2 MJ test meal was consumed, after which calorimetric measurements were continued for 240 min. Total body fat was assessed by hydrodensitometry and total subcutaneous fat by the sum of eight skinfolds. Polymorphisms were typed by PCR and PCR-RFLP-techniques. Thyroid stimulating hormone (TSH) concentrations after a thyrotropin releasing hormone (TRH) injection were measured by radioimmunoassay (RIA). The changes in body weight and adiposity were not different between UCP1 Bcl I, UCP2 alanine to valine (A55V), UCP2 insertion/deletion (I/D) or UCP3 Rsa I genotypes. However, the recovery from overfeeding was worse among G-allele carriers of the UCP1 Bcl I, I allele non-carriers of the UCP2 I/D, AV heterozygote subjects of the UCP2 A55V and CC subjects of the UCP3 Rsa I polymorphisms. RMR was lower both before (P=0.01) and after (P=0.001) overfeeding in subjects with the CC genotype of the UCP3 Rsa I polymorphism. Moreover, after overfeeding, the UCP2 A55V heterozygote and UCP3 Rsa I CC homozygote subjects had significantly higher respiratory quotient (RQ) values at rest (P<0.01) and during the meal test (P from<0.01 to<0.05). Also mean plasma TSH concentrations 20, 30 and 45 min after the TRH injection increased more with overfeeding among UCP2 A55V (P<0.005) and UCP3 Rsa I CC (P=0.017) subjects. These data suggest that UCP polymorphisms may play a role in the recovery from the overfeeding by regulating substrate oxidation in response to long-term caloric surplus. The association of the UCP2 A55V and UCP3 Rsa I CC genotypes with a greater increase in the TSH response to TRH load could reflect a compensatory mechanism counteracting the effects of overfeeding. A longer period of exposure to chronic positive energy balance conditions may be necessary before sequence variation in UCP2 and UCP3 makes an impact on thyroid metabolism to influence body mass and composition changes.

Glucocorticoid receptor Bcl I variant is associated with an increased atherogenic profile in response to long-term overfeeding

The effect of the glucocorticoid receptor (GRL) gene Bcl I polymorphism on body composition and metabolic changes in response to overfeeding was studied. Twenty-four men (mean age 21±2 years) who constituted 12 pairs of identical twins ate a 4.2 MJ/day energy surplus, 6 days a week, during a period of 100 days. The GRL Bcl I marker was identified by Southern Blot technique. Total body fat was assessed by hydrodensitometry and abdominal fat areas were measured by computed tomography. Fasting plasma glucose and insulin during an oral glucose tolerance test (OGTT) were assayed. The insulin and glucose areas were computed using the trapezoidal method. Triglyceride and cholesterol concentrations in plasma and lipoprotein fractions were determined enzymatically. The results showed that overfeeding induced a greater increase in body weight ( p=0.002) in the 2.3/2.3 kb ( n=12) than in the 4.5/2.3 kb ( n=12) subjects. In addition, plasma levels of total ( p=0.007) and low-density lipoprotein (LDL) cholesterol ( p=0.003), as well as systolic blood pressure ( p=0.036) increased more in the 2.3/2.3 kb than in the 4.5/2.3 kb subjects. The 2.3/2.3 kb genotype was also associated with a greater increase in abdominal visceral fat ( p=0.040) compared to the 4.5/2.3 kb genotype. In conclusion, 2.3/2.3 kb subjects of the GRL Bcl I polymorphism experience greater increases in body weight, blood pressure and cholesterol levels as well as visceral fat than 4.5/2.3 kb subjects in response to overfeeding. These data suggest that overfeeding induces an atherogenic profile in subjects who are homozygotes for the 2.3 kb allele.

Somatotropin and adipose tissue metabolism: substrate and temporal effects.

The purpose of these studies was to determine the time course for changes in feed intake, blood metabolites, and lipogenic activity in adipose tissue in response to the initiation of porcine somatotropin (pST) treatment and following withdrawal from treatment in barrows. An initial study was conducted to determine the impact of chronic pST treatment (4 wk of daily injection; 0 vs 4 mg/d) on adipose tissue lipid metabolism in barrows (initial weight 67 kg). Feed efficiency was improved 27%, backfat thickness was decreased 43%, and glucose and lactate oxidation and incorporation into lipid in adipose tissue was reduced 70 to 86% in pST-treated pigs. Palmitate esterification was decreased 44%, whereas palmitate oxidation was unaffected. In vitro metabolism of lactate, glucose, and palmitate in liver slices was not affected by pST treatment. The time-course for changes in intake and adipose tissue metabolism in response to 7 d of pST (0 vs 4 mg/d) treatment and 7 d of withdrawal was examined in subsequent studies in barrows (initial weight 75 kg). Feed intake during pST treatment was significantly (P < .05) less than in control pigs within 24 h of the initiation of treatment and remained low through 3 d after withdrawal. Adipose tissue biopsies were obtained on d 0, 1, 2, 4, and 7 of the treatment phase and on d 2, 4, and 7 after withdrawal from 7 d of treatment. Maximal inhibition of lipogenesis by pST treatment in adipose tissue in vitro was observed on d 4 (-68%) and d 7 (-69%). Similarly, fatty acid synthase activity declined during the treatment period, with the greatest change noted on d 7 (-26%). After withdrawal from treatment, lipogenesis gradually increased, returning to control values 7 d after withdrawal. Levels of IGF-I began to increase from d 1 to d 7 of treatment, continually decreased during withdrawal, and were normalized by the end of the withdrawal period. Plasma urea nitrogen concentrations decreased during treatment, increased during the withdrawal phase, and were normalized 4 d after the last pST treatment. Overall results indicate that most of the metabolic changes in response to pST occur within 1 wk of treatment and return to pretreatment values after 7 d of withdrawal from treatment.

Sequential changes in the metabolic response in severely septic patients during the first 23 days after the onset of peritonitis.

To quantify the sequential changes in metabolic response occurring in patients with severe sepsis after the onset of peritonitis. Understanding the changes in energy expenditure and body composition is essential for the optimal management of severely septic patients; however, they have not been quantified in the context of modern surgical care. Twelve patients with severe sepsis secondary to peritonitis (median APACHE II score = 21.5) had measurements of energy expenditure and body composition as soon as they were hemodynamically stable and 5, 10, and 21 days later. Sequential measurements of acute-phase proteins and cytokine responses were also made. Resting energy expenditure rose to 49% above predicted and remained elevated throughout the study period. Total energy expenditure was 1.25 x resting energy expenditure. Body fat was oxidized when energy intake was insufficient to achieve energy balance. There was a positive fluid balance of 12.5 1 over the first 2 days after onset of sepsis; thereafter, body water changes closely paralleled body weight changes and were largely accounted for by changes in extracellular water. During the 21 -day study period, there was a loss of 1.21 kg (13%) of total body protein. During the first 10 days, 67% of the protein lost came from skeletal muscle, but after this time it was predominantly from viscera. Intracellular potassium levels were low but did not deteriorate further after hemodynamic stability had been reached. There was a reprioritization of hepatic protein synthesis that was obligatory and independent of changes in total body protein. The cytokine responses demonstrated the complexity, redundancy, and overlap of mediators. The period of hypermetabolism in severely septic patients is similar to that previously described, but the fluid changes are larger and the protein loss is greater. Protein loss early on is predominantly from muscle, thereafter from viscera. Fat loss can be prevented and cell function preserved once hemodynamic stability is achieved.

The gastrointestinal tract: An underestimated organ as demonstrated in an experimental LVAD pig model

Although hemodynamic stability and renal function are important and are monitored closely in patients with implanted left ventricular assist devices (LVAD), the gastrointestinal tract may be underestimated in the early postoperative period with regard to adequate perfusion. We investigated renal, intestinal, and whole body metabolic changes in response to variations in LVAD flow and inspired oxygen concentration (FiO2). Left ventricular assist devices were implanted in 10 adult pigs (weight, 55 +/- 1.76 kg). Renal vein (RV), superior mesenteric vein (SMV), and pulmonary artery (PA) blood oxygen saturation and lactate concentration were measured and used as tissue perfusion markers. These measurements were made at baseline and after changes in LVAD flow or FiO2. Oxygen saturation in the PA, SMV, and RV decreased significantly after a reduction in LVAD flow (P < 0.05), with a greater reduction in the SMV than in the PA and RV (p < 0.05 at LVAD flow 3.5L/min; p < 0.01 at LVAD flow 2.0 and 1.0 L/min). The lactate concentration in the PA and SMV increased significantly (p < 0.01) with decreased flow, with a greater increase in the SMV than in the PA (p< 0.05), whereas it remained unchanged in the RV. Oxygen saturation in the PA, SMV, and RV decreased significantly after a reduction in FiO2 (p < 0.05). Lactate concentration in the PA, SMV, and RV increased significantly at FiO2 of 0.10 (p < 0.05). Lactate concentration in the PA and SMV was significantly higher than that in the RV at Fi)2 of 0.10 (p < 0.01). The results show that the gastrointestinal tract is at high risk during low perfusion or low FiO2, whereas the kidneys' metabolic function appears to be less disturbed. In clinical practice, this emphasizes the need to ensure adequate blood flow and respiratory function, especially after extubation, in patients with implanted LVAD. This might avoid intestinal ischemia and subsequent endotoxemia. Gastrointestinal tonometry may help in the assessment of intestinal perfusion.

Hormone dependence of breast cancer cells and the effects of tamoxifen and estrogen:31P NMR studies

Many breast tumors appear to progress from estrogen-dependent growth to a more malignant phenotype characterized by estrogen-independent growth, antiestrogen resistance, and a high metastatic potential. Utilizing 31P NMR spectroscopy on human breast cancer cells growing in vitro, we have investigated the effects of 17 beta-estradiol and tamoxifen on the metabolic/bioenergetic spectra of a series of human breast cancer cells that vary in their estrogen and antiestrogen responsiveness. A comparison of baseline spectra associates higher levels of phosphodiesters and UDP-glucosides (e.g. UDP-glucose, UDP-N-acetylglucosamine), and lower phosphocholine/glycerylphosphocholine and phosphocholine/phosphoethanolamine ratios, with the acquisition of estrogen-independent growth in estrogen receptor expressing cells. No metabolic changes are clearly associated with the metastatic phenotype. Whilst estrogen treatment produces no consistently significant spectral changes in any of the cell lines, the estrogen-independent and estrogen-responsive MCF7/MIII cell line responds to tamoxifen treatment by significantly increasing all spectral resonances 30%-40% above baseline values. This may reflect a tamoxifen-induced change to a more differentiated or apoptotic phenotype, or an attempt by the cells to reverse the inhibitory effects of the drug. The ability to detect metabolic changes in response to tamoxifen by NMR spectroscopy may provide a novel means to identify those tumors that are responsive to antiestrogen therapy.

Serum and Saliva Cortisol Responses and Blood Lactate Accumulation during Incremental Exercise Testing

In six male physically active subjects the adrenocortical and metabolic changes in response to incremental exercise testing were investigated. Blood and saliva samples were taken at rest, at the end of every workload (duration 4 min with 50 W increment), immediately and 10 min after 1 min all out spurt on the electrically braked cycle ergometer. Both saliva and serum cortisol were measured as well as blood lactate. The cortisol response in serum and in saliva showed similar dynamics (r = 0.86, p less than 0.001, n = 50) at submaximal work. At maximal work the serum cortisol concentration showed a transitory decrease, which was not manifested in saliva. It is hypothesized that a factor related to the metabolic acidosis masks the actual adrenocortical response in the serum but not in saliva. Correlation analysis revealed a positive relation between lactate and cortisol in serum (r = 0.56, p less than 0.01, n = 50) and saliva (r = 0.70, p less than 0.01, n = 50). Apparently, salivary cortisol closely reflects plasma free cortisol level, presenting advantage over total cortisol measurements. Moreover salivary measurement will permit studies in their authentic settings and should assist attempts to understand the nature of the adrenocortical function in exercise.