Effects of aerobic exercise training and yoga on the baroreflex in healthy elderly persons

It is unclear whether the age-associated reduction in baroreflex sensitivity is modifiable by exercise training. The effects of aerobic exercise training and yoga, a non-aerobic control intervention, on the baroreflex of elderly persons was determined. Baroreflex sensitivity was quantified by the alpha-index, at high frequency (HF; 0.15-0.35 Hz, reflecting parasympathetic activity) and mid-frequency (MF; 0.05-0.15 Hz, reflecting sympathetic activity as well), derived from spectral and cross-spectral analysis of spontaneous fluctuations in heart rate and blood pressure. Twenty-six (10 women) sedentary, healthy, normotensive elderly (mean 68 years, range 62-81 years) subjects were studied. Fourteen (4 women) of the sedentary elderly subjects completed 6 weeks of aerobic training, while the other 12 (6 women) subjects completed 6 weeks of yoga. Heart rate decreased following yoga (69 +/- 8 vs. 61 +/- 7 min-1, P < 0.05) but not aerobic training (66 +/- 8 vs. 63 +/- 9 min-1, P = 0.29). VO2 max increased by 11% following yoga (P < 0.01) and by 24% following aerobic training (P < 0.01). No significant change in alpha MF (6.5 +/- 3.5 vs. 6.2 +/- 3.0 ms mmHg-1, P = 0.69) or alpha HF (8.5 +/- 4.7 vs. 8.9 +/- 3.5 ms mmHg-1, P = 0.65) occurred after aerobic training. Following yoga, alpha HF (8.0 +/- 3.6 vs. 11.5 +/- 5.2 ms mmHg-1, P < 0.01) but not alpha MF (6.5 +/- 3.0 vs. 7.6 +/- 2.8 ms mmHg-1, P = 0.29) increased. Short-duration aerobic training does not modify the alpha-index at alpha MF or alpha HF in healthy normotensive elderly subjects. alpha HF but not alpha MF increased following yoga, suggesting that these parameters are measuring distinct aspects of the baroreflex that are separately modifiable.     

The inhibition of corrinoid-catalyzed oxidation of mercaptoethanol by methyl iodide: mechanistic implications

The cobalamin coenzymes (5'-deoxyadenosyl- and methylcobalamin) and their cobinamide counterparts (5'-deoxyadenosyl- and methylcobinamide) catalyze the oxidation of 2-mercaptoethanol to its disulfide with hydrogen peroxide formation under aerobic conditions. The reactions are blocked by methyl iodide. Inhibition by methyl iodide is apparently due to the formation of the trans dialkyl corrinoids: methyl(adenosyl)cobalamin, dimethylcobalamin, methyl(adenosyl)cobinamide, and dimethylcobinamide, respectively. When the reaction system is illuminated with visible light, inhibition is released and a dramatic enhancement in the rate of oxygen consumption occurs. For reactions catalyzed by adenosyl- and methylcobalamin and then inhibited by methyl iodide, the rates observed during photolysis approach those obtained with aquacobalamin. For reactions catalyzed by adenosyl- and methylcobinamide and then inhibited by methyl iodide, the rates observed during photlysis approach those obtained with diaquacobinamide. Thus, both trans axial carbon-cobalt bonds in the putative dialkyl corrinoid are homolyzed during photolysis. In contrast to these results, the catalysis of the aerobic oxidation of 2-mercaptoethanol by aquacobalamin is only weakly inhibited by methyl iodide. This observation suggests that aquacob(II)alamin is produced during the catalysis of this reaction. Superoxide, the anticipated product of the reaction between aquacob(II)alamin and dioxygen, is formed during aquacobalamin-catalyzed 2-mercaptoethanol oxidation since superoxide dismutase decreases the rate of oxygen consumption by 50%. However, the enzyme has no effect on oxygen uptake during reactions catalyzed by cobalamin coenzymes and their cobinamide counterparts. These corrinoid catalysts apparently transfer two electrons to dioxygen from cobalt(I) intermediates formed during the reactions. Nitrogenous bases inhibit corrinoid-catalyzed thiol oxidation by competing with 2-mercaptoethanol for axial-ligand coordination sites on the catalyst. In contrast to the inhibition observed with methyl iodide, visible light has no effect on the inhibition obtained with nitrogenous bases.     

Potential photosensitizers for photodynamic therapy. IV. Photophysical and photochemical properties of azaporphyrin and azachlorin derivatives

The photophysical and photochemical properties of a 5-azaprotoporphyrin derivative ([5]AZPP), a zinc-15-azaporphyrin derivative (Zn-[15]AZIDP) and an E-Z isomeric mixture of a 5-azachlorin derivative ([5]AZCH) were studied in various solvents. The quantum yields of fluorescence phi F0, S1-T1 intersystem crossing phi T0 and singlet oxygen (1 delta g) formation phi delta were measured and the Stern-Volmer constants for the quenching of the S1 states by oxygen and the rate constants of quenching of O2(1 delta g) by the different azaporphyrinoid compounds were obtained. The fluorescence quantum yield (phi F0 = 0.23), the strong absorption in the red (lambda max = 674 nm, epsilon max = 66,000 M-1 cm-1) and the high value of the quantum yield for singlet oxygen (1 delta g) formation (phi delta = 0.65) observed for [5]AZCH recommend azachlorin derivatives as potential markers and photosensitizers for tumour therapy.     

Characterization of alpha 2-adrenoceptors mediating contraction of dog saphenous vein: identity with the human alpha 2A subtype

1. In the dog saphenous vein alpha 1- and alpha 2-adrenoceptors mediate noradrenaline-induced contractions in vitro. In order to study the alpha 2-adrenoceptor in isolation, alpha 1-adrenoceptors were inactivated by treatment of tissues with the alkylating agent phenoxybenzamine (3.0 microM for 30 min) in the presence of rauwolscine (1 microM) to protect alpha 2-adrenoceptors. 2. Noradrenaline-induced contractions of tissues treated with phenoxybenzamine were antagonized competitively by the selective alpha 2-adrenoceptor antagonist rauwolscine, pKB = 8.63 +/- 0.07 (means +/- s.e. mean; n = 3), consistent with an interaction at alpha 2-adrenoceptors. 3. Noradrenaline was a full agonist at alpha 2-adrenoceptors in dog saphenous vein. By use of the method of partial receptor alkylation and analysis of concentration-effect curve data by direct, operational model fitting methods, the affinity (pKA) and efficacy (tau) were 5.74 +/- 0.07 and 7.50 +/- 1.05, respectively (n = 6). Nine other agonists which were examined each had affinities higher than noradrenaline, but with the exception of the imidazoline, A-54741 (5,6-dihydroxy-1,2,3,4-tetrahydro-1-naphthyl-imidazoline) had relatively lower efficacies. 4. To compare the alpha 2-adrenoceptor in dog saphenous vein to the human recombinant subtypes, the affinities of twenty-one compounds were estimated in functional studies in the dog saphenous vein and in radioligand binding studies for the human alpha 2A, alpha 2B and alpha 2C receptor subtypes expressed in Chinese hamster lung (CHL) cells. 5. Of twenty-one compounds examined in ligand binding studies, only nine had greater than ten fold selectivity for one human receptor subtype over either of the other two. These compounds were A-54741, oxymetazoline, guanfacine, guanabenz, prazosin, spiroxatrine, tolazoline, WB 4101 and idazoxan. In dog saphenous vein, their affinities (pKA and pKB for agonists and antagonists respectively) were: A-54741 (pKA = 8.03 +/- 0.05), oxymetazoline (pKA = 7.67 +/- 0.09), guanfacine (pKA = 6.79 +/- 0.03); guanabenz (pKA = 7.02 +/- 0.13); prazosin (pKB = 5.19 +/- 0.08), spiroxatrine (pKB = 6.59 +/- 0.04), tolazoline (pKB = 6.21 +/- 0.07), WB 4101 (pKB = 7.42 +/- 0.09) and idazoxan (pKB = 7.11 +/- 0.08). 6. Comparisons of affinity estimates for these nine compounds at the receptor in dog saphenous vein and at the human recombinant subtypes suggest that the vascular receptor is most similar to the h alpha 2A subtype; correlation coefficients (r) were 0.82 (h alpha 2A), 0.24 (h alpha 2B) and 0.04 (h alpha 2C).     

Methylcobalamin treatment of Bell's palsy

Sixty patients with Bell's palsy were included in an open randomized trial. Patients were assigned into three treatment groups: steroid (group 1), methylcobalamin (group 2) and methylcobalamin + steroid (group 3). Comparison between the three groups was based on the number of days needed to attain full recovery, facial nerve scores, and improvement of concomitant symptoms. The time required for complete recovery of facial nerve function was significantly shorter ( p < 0.001) in the methylcobalamin (mean of 1.95 +/- 0.51 weeks) and methylcobalamin plus steroid groups (mean of 2.05 +/- 1.23 weeks) than in the steroid group (mean of 9.60 +/- 7.79 weeks). The facial nerve score after 1-3 weeks of treatment was significantly more severe (p < 0.001) in the steroid group compared to the methylcobalamin and methylcobalamin plus steroid groups. The improvement of concomitant symptoms was better in the methylcobalamin treated groups than the group treated with steroid alone.     

Developmental variations in factors related to initial and increased levels of adolescent drug involvement

Effects of two 9-month sports programmes (four or two sessions per week) on level of daily physical activity (PA), fat mass (FM), and physical fitness were assessed in children with spastic cerebral palsy (CP; n = 20, 9.2 +/- 1.4 yr), randomly assigned to an experimental and control group after matching. Four sessions per week tended to increase PA ratio (24-h energy expenditure/sleeping (resting) energy expenditure) after 9 months from 1.34 +/- 0.25 to 1.55 +/- 0.18 (P = 0.07; not different versus controls). FM increased continuously in the control group (after 9 months + 1.1 +/- 1.6 kg, P < 0.05), whereas the experimental groups showed no changes. Training (respectively four and two sessions) increased peak aerobic power 35% (P < 0.01; P < 0.05 versus controls) and 21% (P < 0.01; P = 0.17 versus controls). Results also suggest that training has a favourable effect on isokinetic muscle strength. No training-related effects were found on anaerobic power. It was concluded that although aerobic training has a limited effect on PA in children with CP, it may prevent deterioration in body composition and muscle strength. Furthermore, training has a favourable effect on peak aerobic power.     

Effect of short- and long-term treatment with omeprazole on the absorption and serum levels of cobalamin

AIMS: To evaluate absorption of protein-bound and unbound cyanocobalamin before and during treatment with omeprazole, and cobalamin levels in patients on long-term treatment with omeprazole. METHODS: In eight former duodenal ulcer patients absorption of unbound and protein-bound cobalamin was determined by measuring 24-h urinary excretion of unbound 58Co-cyancobalamin or protein-bound 57Co-cyanocobalamin during a modified Schilling test. Tests were performed before and during treatment with 20 mg and 40 mg omeprazole daily for 9 days. Serum cobalamin levels were assessed in 25 patients with gastro-oesophageal reflux disease (GERD) before and during long-term maintenance therapy with omeprazole. Mean treatment duration was 56 months (range 36-81 months). RESULTS: Urinary excretion of unbound cobalamin was unchanged with both dosages of omeprazole. Excretion of 57Co-cyanocobalamin, however, decreased significantly during treatment with both 20 mg omeprazole (mean +/- S.E.M.: 1.31 +/- 0.20 vs. 0.54 +/- 0.17%; P < 0.02) and 40 mg omeprazole (1.25 +/- 0.26 vs. 0.29 +/- 0.06%; P < 0.02). Mean serum cobalamin levels (+/- S.E.M.) before and during therapy with omeprazole in GERD patients were 298 +/- 27 and 261 +/- 16 pg/mL (normal range 180-900 pg/mL), respectively (P = N.S.). CONCLUSIONS: Absorption of protein-bound, but not unbound, cyanocobalamin is decreased when measured by a modified Schilling test during treatment with omeprazole. However, no change in serum cobalamin levels was observed in patients with GERD after treatment with omeprazole for up to 7 years.     

Compartmental analysis of colonic transit reveals abnormalities in constipated patients with normal transit

Previous studies had concluded that the treadmill velocity-endurance time hyperbolic relationship for runs could be accuratly approached with a regression at condition that bouts of exercise duration were included between 2 and 12 min. This regression allows to calculate the critical speed (CS) defined as the slope of the regression of work (distance) on time to exhaustion, the anaerobic running capacity (ARC) being the intercept of this line (Monod & Scherrer, 1965). The purpose of this investigation was to give practical indication concerning the choice of the velocities in reference to the maximal aerobic speed (MAS i.e. the minimum speed which elicits VO2max). Subjects were fourteen elite male long-distance runners (27 +/- 3 years old; VO2max = 74.9 +/- 2.9 ml.kg-1.min-1, MAS = 22.4 +/- 0.8 km.h-1, CS = 19.3 +/- 0.7 km.h-1 and 86.2 +/- 1.5% MAS). tlim 100 values (321 +/- 83 s) were negatively correlated with MAS (r = -0.538, p < 0.05) and with CS (km.h-1) (r = -0.644, p < 0.01). tlim 90 (1015 +/- 266 s) was positively correlated with CS when expressed in % MAS (r = 0.645, p < 0.01) and not when expressed in km.h-1 (r = -0.095, P > 0.05). tlim 105 (176 +/- 40 s) only was correlated with ARC (r = 0.526, p < 0.05). These data demonstrate that running time to exhaustion at 100 and 105% of MAS in a homogeneous elite male long-distance runners group is inversely related to MAS. Moreover, tlim 90 is positively correlated with CS (%MAS) but neither with tlim 100 and 105 nor with maximal aerobic speed. So from a practical point of view, the velocities chosen to determine the critical speed, would be closed to the maximal aerobic speed (time to exhaustion around 6 min), taking into account that the tlim 105 is correlated with the anaerobic capacity, whereas tlim 90 is correlated with the critical speed.     

Predictive value of antepartum ultrasound examination for anomalies in twin gestations

RATIONALE AND OBJECTIVES: We investigated the potential of manganese (III) mesoporphyrin (Mn-mesoporphyrin) as a hepatobiliary contrast agent for magnetic resonance (MR) imaging in rabbits given VX-2 carcinoma liver implants. METHODS: Rabbits given VX-2 carcinoma liver implants (n = 8) were imaged before and after the intravenous (i.v.) administration of 0.04 mmol/kg Mn-mesoporphyrin. MR images were correlated with gross-specimen cross-sections. The distribution of Mn in various tissues following i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin was determined using atomic absorption analysis. A standard panel of serum chemistries was followed over 7 days in six rabbits following this same dose of Mn-mesoporphyrin and compared with chemistries from two control rabbits. RESULTS: I.v. administration of 0.04 mmol/kg (25 mg/kg) Mn-mesoporphyrin resulted in improvement of tumor-to-liver contrast, with enhancement of normal liver (99.7 +/- 14.7%) and the gallbladder (442 +/- 116%), but not VX-2 tumor tissue (14.8 +/- 13.9%), (n = 8, p = .05). Analysis of tissue Mn levels 100 min after i.v. Mn-mesoporphyrin injection demonstrated preferential distribution of Mn to normal liver tissue (57.8 +/- 15.3 micrograms Mn/g) compared with VX-2 tumor (4.28 +/- 1.48 micrograms Mn/g). No significant change was found in the serum chemistries of six normal rabbits over a 7-day period after the i.v. administration of 0.04 mmol/kg Mn-mesoporphyrin. CONCLUSION: I.v. Mn-mesoporphyrin improved lesion-to-liver contrast because of preferential distribution of Mn-mesoporphyrin to normal liver parenchyma and bile.     

The coenzyme B12 derivative N1-methyl-5'-deoxyadenosylcobalamin: synthesis, characterization, stability towards dimroth rearrangement, and Co-C thermolysis product and kinetic studies

The mechanism of Co-C heterolysis of adenosylcobalamin (AdoCbl) is studied, specifically the effect of a positive charge produced by methylation (CH3+, a "sticky proton" analog of H+ addition) to the most basic adenine N1 nitrogen site in AdoCbl. This is accomplished by the synthesis, characterization and Co-C thermolysis in the presence of the radical trap TEMPO of the N1 methylated AdoCbl derivative, [N1-methyl-5'-deoxyadenosycobalamin]+, 5. A variety of needed additional syntheses and other control experiments are also reported, including effective purification methods using ultrafiltration membranes, procedures that may prove to be the more generally useful part of the synthetic work reported. The thermolysis results with 5 are unequivocal in showing that a positive charge at the N1 nitrogen in the adenosyl group of AdoCbl has no effect within +/- 8% on the products or rates of Co-C thermal cleavage. The data are most consistent with a recently suggested, new mechanism for AdoCbl Co-C heterolysis at less acidic pH values, one not involving the protonation of the beta-ribose oxygen that is known to occur under strongly acidic conditions. In addition, the present study illustrates some of the positive features, and also some of the pitfalls, in the approach where CH3+ is used as a "sticky proton" analog in mechanistic studies.     

Metabolic profile of high intensity intermittent exercises

To evaluate the magnitude of the stress on the aerobic and the anaerobic energy release systems during high intensity bicycle training, two commonly used protocols (IE1 and IE2) were examined during bicycling. IE1 consisted of one set of 6-7 bouts of 20-s exercise at an intensity of approximately 170% of the subject's maximal oxygen uptake (VO2max) with a 10-s rest between each bout. IE2 involved one set of 4-5 bouts of 30-s exercise at an intensity of approximately 200% of the subject's VO2max and a 2-min rest between each bout. The accumulated oxygen deficit of IE1 (69 +/- 8 ml.kg-1, mean +/- SD) was significantly higher than that of IE2 (46 +/- 12 ml.kg-1, N = 9, p < 0.01). The accumulated oxygen deficit of IE1 was not significantly different from the maximal accumulated oxygen deficit (the anaerobic capacity) of the subjects (69 +/- 10 ml.kg-1), whereas the corresponding value for IE2 was less than the subjects' maximal accumulated oxygen deficit (P < 0.01). The peak oxygen uptake during the last 10 s of the IE1 (55 +/- 6 ml.kg-1.min-1) was not significantly less than the VO2max of the subjects (57 +/- 6 ml.kg-1.min-1). The peak oxygen uptake during the last 10 s of IE2 (47 +/- 8 ml.kg-1.min-1) was lower than the VO2max (P < 0.01). In conclusion, this study showed that intermittent exercise defined by the IE1 protocol may tax both the anaerobic and aerobic energy releasing systems almost maximally.     

Physiological correlates of 3-kilometer running performance in male children

The purpose of this study was to examine the influence of body fatness, aerobic and anaerobic ability on 3-km running performance in 19 physically active boys (mean +/- SD, age = 10.4 +/- 0.9 yrs). The sum of six skinfolds, VO2 at 8.04 and 9.65 km.hr-1, and VO2max were measured in the laboratory. Run time for 3 km was assessed twice on separate days on a 200-meter indoor track. Prior to each run, every child performed two 55-meter sprints and two vertical jumps. Mean +/- SD values for the sum of skinfolds, %VO2max at each running speed, VO2max and 3-km run time were: 33.9 +/- 14.9 mm; 70.6 +/- 6.6% and 81.0 +/- 7.9%; 54.6 +/- 5.0 ml.kg-1.min-1; 16.41 +/- 2.58 min, respectively. Significant (p < 0.05) correlations were observed between the following variables and run time: sum of skinfolds (r = 0.72); vertical jump (r = 0.67); sprint time (r = 0.59); VO2max (r = 0.61); and, %VO2max at each treadmill speed (r = 0.79 and r = 0.75, respectively). Stepwise multiple regression analysis indicated that the combination of the %VO2max at 8.04 km.hr-1 and vertical jump accounted for 83% (adjusted R2) of the variance in running time (SEE = 1.06 min, p < 0.05). This study suggests that 3-km run time in physically active boys is influenced by aerobic and anaerobic indices as well as body fatness, supporting the notion that children, compared to adults, are not metabolic specialists.     

Contribution of phosphocreatine and aerobic metabolism to energy supply during repeated sprint exercise

This study examined the contribution of phosphocreatine (PCr) and aerobic metabolism during repeated bouts of sprint exercise. Eight male subjects performed two cycle ergometer sprints separated by 4 min of recovery during two separate main trials. Sprint 1 lasted 30 s during both main trials, whereas sprint 2 lasted either 10 or 30 s. Muscle biopsies were obtained at rest, immediately after the first 30-s sprint, after 3.8 min of recovery, and after the second 10- and 30-s sprints. At the end of sprint 1, PCr was 16.9 +/- 1.4% of the resting value, and muscle pH dropped to 6.69 +/- 0.02. After 3.8 min of recovery, muscle pH remained unchanged (6.80 +/- 0.03), but PCr was resynthesized to 78.7 +/- 3.3% of the resting value. PCr during sprint 2 was almost completely utilized in the first 10 s and remained unchanged thereafter. High correlations were found between the percentage of PCr resynthesis and the percentage recovery of power output and pedaling speed during the initial 10 s of sprint 2 (r = 0.84, P < 0.05 and r = 0.91, P < 0.01). The anaerobic ATP turnover, as calculated from changes in ATP, PCr, and lactate, was 235 +/- 9 mmol/kg dry muscle during the first sprint but was decreased to 139 +/- 7 mmol/kg dry muscle during the second 30-s sprint, mainly as a result of a approximately 45% decrease in glycolysis. Despite this approximately 41% reduction in anaerobic energy, the total work done during the second 30-s sprint was reduced by only approximately 18%. This mismatch between anaerobic energy release and power output during sprint 2 was partly compensated for by an increased contribution of aerobic metabolism, as calculated from the increase in oxygen uptake during sprint 2 (2.68 +/- 0.10 vs. 3.17 +/- 0.13 l/min; sprint 1 vs. sprint 2; P < 0.01). These data suggest that aerobic metabolism provides a significant part (approximately 49%) of the energy during the second sprint, whereas PCr availability is important for high power output during the initial 10 s.     

Experiences with the application of QSAR in the routine of the notification procedure

IL-8 is a recently described chemokine that increases polymorphonuclear neutrophil infiltration and has been implicated in inflammatory pathology. This study assesses monocyte (M phi) interleukin-8 (IL-8) levels in severe trauma patients (injury severity score > 16) who have elevated levels of M phi cell-associated tumor necrosis factor alpha (TNF alpha), a major marker for systemic inflammatory response syndrome after injury. We demonstrate elevated (p = .0007) levels of M phi IL-8 only in those trauma patients who also have increased (p = .0001) M phi-secreted TNF alpha whereas the patients having normal M phi-secreted TNF alpha levels have normal or even decreased M phi IL-8 production. There is no association between M phi IL-8 production and cell-associated TNF alpha levels. M phi induction by Fc gamma RI cross-linking, a common induction pathway in trauma patients' M phi that increases the production of both cell-associated and secreted TNF alpha, can also increase (p = .0022) M phi IL-8 levels. Therefore, post-trauma elevation of M phi IL-8 levels may be associated with increased secreted TNF alpha resulting from, at least in part, Fc gamma RI cross-linking stimulation in vivo.     

Conformation of manganese(II)-nucleotide complexes bound to rabbit muscle creatine kinase: 13C NMR measurements using [2-13C]ATP and [2-13C]ADP

Conformations of cation-nucleotide complexes bound to rabbit muscle creatine kinase were investigated by measuring paramagnetic effects on 13C spin relaxation in E.Mn[2-13C]ATP and E.Mn[2-13C]ADP at three different frequencies, viz., 50, 75, and 125 MHz, and as a function of temperature in the range of 7-35 degrees C (at 75 MHz). Arrhenius plots of the temperature dependencies of relaxation rates show a positive slope with low activation energies of 1.3 +/- 0.2 kcal/mol and 2.0 +/- 0.2 kcal/mol for E.Mn ATP and E.MnADP, respectively. The relaxation rates of both complexes show strong frequency dependence, indicating that these rates are not exchange limited. Analysis of the data yields Mn(II)-2C distances of 10.0 +/- 0.5 A for E.MnATP and 8.6 +/- 0.5 A for E.MnADP. These data were interpreted, along with previously published information, on the location of the cation with respect to the phosphate chain [Jarori, G. K., Ray, B.D., & Nageswara Rao, B. D. (1985) Biochemistry 24, 3487-3494], and on the adenosine conformation [Murali, N., Jarori, G. K., & Nageswara Rao, B. D. (1993) Biochemistry 32, 12941-12948] in these complexes. The Mn(II)-2C distances depend on the orientation of the phosphate chain relative to the adenosine moiety. Conformational searches were performed by varying the two torsion angles, phi 1 (C4'-C5'-O5'-P alpha), and phi 2 (C5'-O5'-P alpha-O alpha beta), along with CHARMm energy computations, in order to determine acceptable conformations compatible with the distances determined. The significant difference in the Mn(II)-2C distances in E.MnATP and E.MnADP is indicative of the structural alterations occurring at the active site as the enzyme turns over.     

Involvement of Fas/Fas ligand system-mediated apoptosis in the development of concanavalin A-induced hepatitis

A series of 2H- and 13C-labeled glutamates were used as substrates for coenzyme B12-dependent glutamate mutase, which equilibrates (S)-glutamate with (2S,3S)-3-methylaspartate. These compounds contained the isotopes at C-2, C-3, or C-4 of the carbon chain: [2-2H], [3,3-2H2], [4,4-2H2], [2,3,3,4,4-2H5], [2-13C], [3-13C], and [4-13C]glutamate. Each reaction was monitored by electron paramagnetic resonance (EPR) spectroscopy and revealed a similar signal characterized by g'xy = 2.1, g'z = 1.985, and A' = 5.0 mT. The interpretation of the spectral data was aided by simulations which gave close agreement with experiment. This approach underpinned the idea of the formation of a radical pair, consisting of cob(II)alamin interacting with an organic radical at a distance of 6.6 +/- 0.9 A. Comparison of the hyperfine couplings observed with unlabeled glutamate with those from the labeled glutamates enabled a principal contributor to the radical pair to be identified as the 4-glutamyl radical. These findings support the currently accepted mechanism for the glutamate mutase reaction, i.e., the process is initiated through hydrogen atom abstraction from C-4 of glutamate by the 5'-deoxyadenosyl radical, which is derived by homolysis of the Co-C sigma-bond of coenzyme B12.     

Ventilatory and cardiovascular responses to inspired He-O2 during exercise in chronic obstructive pulmonary disease

Blunted maximum cardiac output and systemic O2 extraction could constitute primary limits to exercise in severe chronic obstructive pulmonary disease (COPD) or they could simply reflect cessation of exercise because of abnormal pulmonary mechanics. To determine which is the case, eight consecutive patients with severe COPD (FEV1 = 0. 56 +/- 0.04 L, mean +/- SEM), five of whom had alpha1-antiprotease deficiency, performed two incremental cycling tests while breathing N2-O2 or He-O2. Expired gases and V E were measured, and radial and pulmonary arterial blood was simultaneously sampled each minute. Peak exercise V E was higher with He-O2 than with N2-O2 (25.5 +/- 2. 2 versus 19.3 +/- 1.5 L/min, p = 0.002) and PaCO2 was lower (42 +/- 2 versus 46 +/- 2 mm Hg, p = 0.0003). V O2max improved only modestly (594 +/- 75 versus 514 +/- 54 ml/min, p = 0.04), and was accompanied by an increase in peak exercise CaO2 (18.7 +/- 0.9 versus 17.6 +/- 0. 9 ml/dl, p = 0.02). Peak Fick cardiac output was decreased (39 +/- 3% pred) and CvO2 was elevated (130 +/- 10% pred), and neither improved with He-O2 (p > 0.05 for each). Abnormal peak exercise cardiac output and systemic O2 extraction in severe COPD cannot be fully accounted for by limiting pulmonary mechanics and may contribute to exercise intolerance.     

Antisense oligonucleotides against alpha1E reduce R-type calcium currents in cerebellar granule cells

Many neurons of the central nervous system display multiple high voltage-activated Ca2+ currents, pharmacologically classified as L-, N-, P-, Q-, and R-type. Of these current types, the R-type is the least understood. The leading candidate for the molecular correlate of R-type currents in cerebellar granule cells is the alpha1E subunit, which yields Ca2+ currents very similar to the R-type when expressed in heterologous systems. As a complementary approach, we tested whether antisense oligonucleotides against alpha1E could decrease the expression of R-type current in rat cerebellar granule neurons in culture. Cells were supplemented with either antisense or sense oligonucleotides and whole-cell patch clamp recordings were obtained after 6-8 days in vitro. Incubation with alpha1E antisense oligonucleotide caused a 52.5% decrease in the peak R-type current density, from -10 +/- 0.6 picoamperes/picofarad (pA/pF) (n = 6) in the untreated controls to -4.8 +/- 0.8 pA/pF (n = 11) (P < 0.01). In contrast, no significant changes in the current expression were seen in sense oligonucleotide-treated cells (-11.3 +/- 3.2 pA/pF). The specificity of the alpha1E antisense oligonucleotides was supported by the lack of change in estimates of the P/Q current amplitude. Furthermore, antisense and sense oligonucleotides against alpha1A did not affect R-type current expression (-11.5 +/- 1.7 and -11.7 +/- 1.7 pA/pF, respectively), whereas the alpha1A antisense oligonucleotide significantly reduced whole cell currents under conditions in which P/Q current is dominant. Our results support the hypothesis that members of the E class of alpha1 subunits support the high voltage-activated R-type current in cerebellar granule cells.     

Small bowel tonometry is more accurate than gastric tonometry in detecting gut ischemia

Gastric tonometer PCO2 measurement may help identify gut ischemia in critically ill patients but is frequently associated with large measurement errors. We tested the hypothesis that small bowel tonometer PCO2 measurement yields more accurate information. In 10 anesthetized, mechanically ventilated pigs subject to progressive hemorrhage, we measured gut oxygen delivery and consumption. We also measured tonometer PCO2 minus arterial PCO2 (DeltaPCO2) and calculated the corresponding intracellular pH from tonometers placed in the stomach and jejunum. We found that the correlation coefficient (r2) for biphasic gut oxygen delivery-DeltaPCO2 relationships was 0.29 +/- 0.52 for the gastric tonometer vs. 0.76 +/- 0.25 for the small bowel tonometer (P < 0.05). In addition, the critical gastric tonometer DeltaPCO2 was excessively high and variable (62.9 +/- 39.6) compared with the critical small bowel tonometer DeltaPCO2 (17.0 +/- 15.0, P < 0.01). Small bowel tonometer PCO2 was closely correlated with superior mesenteric vein PCO2 (r2 = 0.81, P < 0.001), whereas gastric tonometer PCO2 was not (r2 = -0.13, P = not significant). We conclude that measurement of gastric tonometer PCO2 yields excessively noisy and inaccurate data on the onset of gut anaerobic metabolism in hemorrhagic shock. Small bowel tonometer PCO2 is less noisy and, as a result, is superior in detecting gut hypoperfusion and the onset of anaerobic metabolism.     

Expression of inhibin alpha and inhibin/activin beta A subunits in the granulosa layer of the large preovulatory follicles of the hen

The expression of the mRNA for the inhibin/activin subunits (alpha and beta A) in the granulosa layer of the five largest preovulatory follicles of the hen was investigated. Total RNA from the granulosa layer of the F5 (the fifth largest) to F1 (the largest) follicles was extracted and analyzed by Northern blot analysis using homologous chicken inhibin alpha and beta A subunit cDNA probes. RNA loading was quantified by a cDNA probe of bovine 18S rRNA. Results showed that for the chicken inhibin alpha subunit mRNA signals (n = 3), the mean relative intensity for the F1, F2, F3, and F4 follicles was 0.50 +/- 0.10 ( +/- SEM,), 0.52 +/- 0.08, 0.59 +/- 0.06, and 0.81 +/- 0.04, respectively, compared to a mean relative intensity of 1.00 (p < 0.05) for the F5 follicle. For the beta A subunit mRNA signals (n = 3), the mean relative intensity for the F5, F4, F3, and F2 follicles was 0.25 +/- 0.06, 0.28 +/- 0.15, 0.40 +/- 0.17, and 0.48 +/- 0.10 (p < 0.05) for the F1 follicle. The inhibin alpha subunit was also estimated to be more abundantly expressed among follicles in the granulosa layer than was the beta A subunit. Our data indicate that the expression of inhibin alpha and beta A subunits is differentially regulated in the hen granulosa layer during follicular development. Expression of the alpha subunit is reduced with follicular development whereas inhibin beta A subunit expression is dramatically enhanced. In addition, the granulosa layer of the large preovulatory follicles may produce more inhibin alpha subunit than beta A subunit, and the F1 follicle may be the primary source of the beta A subunit for dimeric inhibin and/or activin in the hen.