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Biomedical subjects

B Norman

Publications and source records attributed to B Norman.

At least 37 records · Page 2Linked to original sources

AMP deaminase in skeletal muscle of healthy males quantitatively determined by new assay.

A new, sensitive assay for the quantitative determination of AMP deaminase activity in human skeletal muscle is presented. The method is based on the determination of the direct product of the AMP deaminase reaction, the formed IMP, by high performance liquid chromatography (HPLC). In order to evaluate the relationship between AMP deaminase activity on the one hand and the contractile and metabolic characteristics of the muscle and the physical performance on the other, muscle biopsies were taken from 20 male subjects. The subjects also performed a 30 s sprint test on a cycle ergometer. The inter-individual variation in AMP deaminase activity was large, ranging from 5.4 to 27.4 microkat g-1 dry muscle. AMP deaminase was positively correlated with phosphofructokinase (PFK), the marker of the glycolytic capacity of the muscle, but there was no correlation with enzymes of oxidative metabolism, such as 3-hydroxyacyl-CoA dehydrogenase and citrate synthase, or with the activity of myokinase and lactate dehydrogenase. There was no significant correlation between AMP deaminase activity and the proportion of the different muscle fibre types. A weak positive correlation was found between the sprint performance and the AMP deaminase activity. In conclusion, the HPLC assay was found to be a fast, sensitive and reliable method for the quantitative determination of AMP deaminase activity in muscle. A direct relationship between AMP deaminase activity on the one hand and glycolytic capacity and sprint performance on the other was found. However, no relationship to oxidative capacity or contractile properties was found.

AMP Deaminase↗

The effect of high-intensity training on purine metabolism in man.

The effect of intermittent high-intensity training on the activity of enzymes involved in purine metabolism and on the concentration of plasma purines following acute short-term intense exercise was investigated. Eleven subjects performed sprint training three times per week for 6 weeks. Muscle biopsies for determination of enzyme activities were obtained prior to and 24 h after the training period. After training, the activity of adenosine 5'-phosphate (AMP) deaminase was lower (P < 0.001) whereas the activities of hypoxanthine phosphoribosyl transferase (HPRT) and phosphofructokinase were significantly higher compared with pre-training levels. The higher activity of HPRT with training suggests an improved potential for rephosphorylation of intracellular hypoxanthine to inosine monophosphate (IMP) in the trained muscle. Before and after the training period the subjects performed four independent 2-min tests at intensities from a mean of 106 to 135% of VO2max. Venous blood was drawn prior to and after each test. The accumulation of plasma hypoxanthine following the four tests was lower following training compared with prior to training (P < 0.05). The accumulation of uric acid was significantly lower (46% of pre-training value) after the test performed at 135% of VO2max (P < 0.05). Based on the observed alterations in muscle enzyme activities and plasma purine accumulation, it is suggested that high intensity intermittent training leads to a lower release of purines from muscle to plasma following intense exercise and, thus, a reduced loss of muscle nucleotides.

AMP Deaminase↗

Decreased resting levels of adenine nucleotides in human skeletal muscle after high-intensity training.

The effect of high-intensity intermittent training on the adenine nucleotide content of skeletal muscle was studied. Eleven male subjects (group A) performed high-intensity intermittent training on a cycle ergometer three times per week for 6 wk, followed by 1 wk of the same kind of training with two sessions per day. Nine males (group B) exclusively performed 1 wk of training with two sessions per day. In group A, skeletal muscle total adenine nucleotide (TAN) levels decreased from 25.1 +/- 0.7 (SE) to 22.0 +/- 0.6 mmol/kg dry wt over the 6-wk period (P < 0.01). The subsequent intensive week did not further alter TAN levels. In group B, the intensive week of training reduced TAN levels from 25.1 +/- 0.5 to 19.4 +/- 0.6 mmol/kg dry wt (P < 0.001). The decrease was sustained 72 h after training (P < 0.001). During the intensive week, there was no change in plasma creatine kinase activity in either group A or group B. The plasma activity was, however, higher in group B than in group A on days 4 and 7 of the intensive week (P < 0.05). The results from this study indicate that high-intensity intermittent exercise causes a decrease in resting levels of skeletal muscle adenine nucleotide without a concomitant indication of muscle damage. A training-induced adaptation appears to occur with training by which a further loss of adenine nucleotides is prevented despite an increased training dose.

Adenine Nucleotides↗

Effects of selenium supplementation in cattle on aquatic ecosystems in northern California.

The potential impact on aquatic ecosystems of supplementing the diets of beef cattle with selenium (Se) was studied on 4 northern California ranches. All study sites included an area of concentrated use by cattle that had diets supplemented with Se. In each case, a stream flowed through the site and provided a control sampling area upstream and a treated sampling area downstream. Specimens of water, sediment, algae, aquatic plants, aquatic invertebrates, and fish were analyzed fluorometrically for total Se content. Significant differences in Se concentration were not found between specimens from upstream control areas and those from downstream areas subjected to use by Se-treated cattle. Evidence was not found that Se supplementation in cattle at maximal permitted concentrations caused Se accumulation in associated aquatic ecosystems.

Analysis of Variance↗

Assignment of structures to oligosaccharides produced by enzymic degradation of a beta-D-glucan from barley by 1H- and 13C-n.m.r. spectroscopy.

The structures of one tri-(1), two tetra-(2 and 3), and one hexa-saccharide (4) produced by treatment of barley flour, after removal of the starch components, with a fungal beta-D-glucanase (Finizyme) have been assigned on the basis of 1H- and 13C-n.m.r. data as follows: beta-D-Glcp-(1----3)-beta-D-Glcp-(1----4)-D-Glcp (1), beta-D-Glcp-(1----4)-beta-D-Glcp-(1----3)-beta-D-Glcp-(1----4)-D-Glcp (2), beta-D-Glcp-(1----3)-beta-D-Glcp-(1----4)-beta-D-Glcp-(1----4)-D-Glcp (3), and beta-D-Xylp-(1----4)-[alpha-L-Araf-(1----3)]-[alpha-L-Ara f-(1----2)-beta-D-Xylp-(1----4)-beta-D-Xylp- (1----4)-D-Xylp (4).

Carbohydrate Sequence↗

Small accumulation of inosine monophosphate (IMP) despite high lactate levels in latissimus dorsi during transplantation.

The effects of prolonged ischaemia and subsequent reperfusion during and after reconstructive microsurgery on energy metabolism were studied. Repeated skeletal muscle biopsies were taken and analysed for high energy phosphates and their degradation products by high performance liquid chromatography and for lactate by a fluorometric procedure. Moderate changes in adenine nucleotides occurred during the first 4 h of ischaemia. After 6 h of ischaemia, when the creatine phosphate store was almost depleted and the lactate level had increased to 111 mmol kg-1 dry muscle, ATP content decreased and inosine monophosphate started to accumulate. The inosine monophosphate accumulation was however small, in spite of a high lactate level, which suggests that the increase in H+ associated with lactate formation is not important for the activation of AMP-deaminase during the present conditions. In spite of the accelerating metabolic deterioration during the later period of ischaemia, the reperfusion of the muscle resulted in a rapid normalization of all the studied metabolites, thereby indicating a rapid restoration of the muscle energy stores.

AMP Deaminase↗

The effect of lidocaine on myocardial ischemia with asanguinous reperfusion. An in vitro study.

The effect of lidocaine on the ischemic nor-mothermic rat heart was studied in a Langendorff preparation. Ventricular fibrillation, total retrograde coronary flow and effluent lactate concentration were monitored in preischemia (control), ischemia (20 min) and reperfusion (20 min). Myocardial metabolites were determined in specimens excised at termination of reperfusion. Six hearts were infused with lidocaine in Ringer solution at onset of ischemia (group A) and six with only Ringer solution (group B). Sinus rhythm proceeded directly to diastolic arrest after 17 sec in group A, while all group B hearts showed ventricular fibrillation before arrest at 174 sec. Effluent lactate concentration was reduced in group A during the first 10 min of ischemia, but not subsequently. After 10 min of reperfusion, coronary flow was reduced by 12% in group A and 20% in group B. ATP was higher and ADP, AMP and IMP were lower in group A than in group B after 20 min of reperfusion. Creatine phosphate showed no intergroup difference, but creatine was higher in group B. Cardiac arrest with lidocaine thus reduced lactate formation during ischemia and lessened high-energy phosphate depletion after reperfusion.

Adenine Nucleotides↗

Ectopic hyperprolactinemia resulting from an ovarian teratoma.

Detection of an elevated prolactin level in the nonpregnant patient usually directs attention to the investigation of the pituitary gland. We describe a patient with secondary amenorrhea and galactorrhea in whom such an elevated prolactin level did initially lead to the discovery of a pituitary mass that was treated first unsuccessfully with surgery, and then successfully with medical therapy. After medical therapy normal menstruation continued, galactorrhea recurred intermittently, and the prolactin level rose again and remained in the approximately 36-micrograms/L range. However, 2 years later an ovarian teratoma was discovered incidentally and was removed surgically. On pathological evaluation the teratoma was noted to include prolactin-containing tissue, and immediately after surgery the patient's prolactin level and responsiveness to stimulatory testing returned to normal. To our knowledge this is the first case report of the association of clinically evident hyperprolactinemia with an ovarian cystic teratoma that contained prolactin-producing tissue.

Adult↗

Relationship of recovery from intensive exercise to the oxidative potential of skeletal muscle.

This study examined if there was a relationship between the aerobic-oxidative potential of skeletal muscle and the metabolic and force recovery after intense exercise. Eleven male subjects performed three bouts of unilateral knee extensions using an isokinetic device. Sixty seconds of rest separated bouts. Muscle biopsies were taken from the vastus lateralis prior to exercise, immediately after bout 2 and before bout 3. Samples were analysed for adenosine triphosphate (ATP), adenosine diphosphate (ADP), inosine monophosphate (IMP), creatine phosphate (CP) and lactate contents and citrate synthase (CS) activity. Peak torque at the end of bout 2 was 45% of initial peak torque of bout 1 (IPT1). With recovery, initial peak torque of bout 3 (IPT3) was 81% of IPT1. Peak torque after recovery (IPT3/IPT1) was related to CS activity (r = 0.69). ATP, CP and ATP/ADP decreased with exercise. ADP, IMP and lactate increased. With recovery, ATP and CP remained depressed. IMP and lactate remained elevated. ATP/ADP and ADP returned towards 'normal', but only the latter attained resting levels. When analysing the individual responses the following correlations were found. After recovery, ATP/ADP (r = 0.57), ATP/ADP relative to rest (r = 0.71), lactate (r = -0.62), CP (r = 0.75) and CP relative to rest (r = 0.83) were related to CS activity. The changes in lactate (r = -0.76) and CP (r = 0.79) during recovery (bout 3-bout 2) were also related to CS activity. The results suggest that the recovery of force and the 'normalization' of metabolite contents after short-term, intense exercise are dependent on the aerobic-oxidative potential of skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗

Regulation of the mouse alpha A-crystallin gene: isolation of a cDNA encoding a protein that binds to a cis sequence motif shared with the major histocompatibility complex class I gene and other genes.

We have shown by site-directed mutagenesis that the sequence between positions -69 and -40 of the mouse alpha A-crystallin gene is crucial for tissue-specific gene expression in a transfected mouse lens epithelial cell line transformed with the early region of simian virus 40. Gel retardation experiments with synthetic oligodeoxynucleotides revealed a mouse lens nuclear protein which bound specifically to the palindromic sequence 5'-GGGAAATCCC-3' at positions -66 to -57 in the alpha A-crystallin promoter. By screening a bacteriophage lambda gt11 expression library of the transformed lens cells, we isolated a 2.5-kilobase-pair cDNA encoding a fusion protein which bound to this sequence and to the regulatory element of the major histocompatibility complex (MHC) class I gene. This cDNA hybridized to a 10-kilobase-pair polyadenylated RNA present in many different tissues, including lens. It encoded a protein, tentatively called alpha A-CRYBP1, containing at least two zinc fingers. alpha A-CRYBP1 is either homologous or very similar to the human nuclear proteins MBP-1 (Baldwin et al., Mol. Cell. Biol. 10:1406-1414, 1990), PRDII-BFI (Fan and Maniatis, Genes Dev. 4:29-42, 1990), and HIV-EP1 (Maekawa et al., J. Biol. Chem. 264:14591-14593, 1989), which bind to regulatory elements of the MHC class I, beta interferon, and human immunodeficiency virus genes, respectively. Our results suggest that the lens-specific alpha A-crystallin, MHC class I, beta interferon and other genes have a similar cis-acting DNA regulatory motif that shares alpha A-CRYBPI, MBP-1, PRDII-BF1, HIV-EP1, or other closely related proteins as trans-acting factors.

Amino Acid Sequence↗

Pregnancy, abruptio placentae and cocaine.

Widespread cocaine use has caused concern about possible harmful effects during pregnancy. Several studies have suggested an increased incidence of abruptio placentae and stillbirths with cocaine use during pregnancy. This study examined what percentage of pregnant women in a lower socioeconomic population use cocaine, as well as positive urine cocaine screens in those with abruptio placentae and intrauterine fetal death (IUFD). Thirty patients were enrolled in the control group, eight in the abruptio placentae group and five in the IUFD group. One of 30 controls, one of eight in the abruptio placentae group and none of the IUFD group urines were positive for cocaine. A higher percentage of those in the abruptio placentae group had a history of abruptio placentae or fetal death, suggesting possible prior drug use. This group also had little, if any, prenatal care. Cocaine abusers may show signs of abuse in their obstetrical histories.

Abruptio Placentae↗

Increased IMP content in glycogen-depleted muscle fibres during submaximal exercise in man.

To study the relationship between glycogen depletion and IMP accumulation in different fibre types, single fibres were dissected from biopsies taken at rest and after one hour of exercise at 70% of maximal oxygen uptake. These fibres were analysed histochemically for glycogen and fibre types and pooled into classes of type I or type II fibres with low, medium or high glycogen content, in a total of six classes. These pools were analysed for ATP, ADP, AMP and IMP contents by high performance liquid chromatography. The contents of ATP, ADP and AMP at rest, and immediately after exercise, were not significantly different between the six fibre classes. The IMP content in glycogen-depleted fibres obtained after exercise was, however, higher than in pools of glycogen-filled fibres obtained both at rest and after exercise. In conclusion, the elevated IMP content in glycogen-depleted but not in glycogen-filled type I and type II muscle fibres during prolonged submaximal exercise indicates a decreased ATP regeneration rate in glycogen-depleted fibres, which may be a factor limiting exercise duration during prolonged submaximal exercise.

Adenosine Diphosphate↗

Conservation of delta-crystallin gene structure between ducks and chickens.

A cloned chicken delta-crystallin cDNA was used to identify two putative delta-crystallin genes in the duck by Southern blot hybridization. A DNA fragment containing most of one of these genes was isolated from a library made in bacteriophage lambda Charon 28A containing genomic DNA from 14-day-old embryonic ducks. Electron microscopy, partial gene sequencing, primer extension analysis using duck mRNA, and comparison with the well-characterized chicken delta-crystallin genes suggest that our cloned duck delta-crystallin gene, like the chicken delta-crystallin genes, is 8-10 kb long and contains 17 exons. Hybridization and sequencing data show great similarity between the homologous 5' untranslated and coding exons of the duck and chicken delta-crystallin genes. Overall, the homologous introns also appear to have approximately 30% sequence similarity, and have been subject to deletion/insertion events. Our partial characterization of duck delta-crystallin gene sequences suggests that this avian and reptilian crystallin family has been conserved during evolution, as have the other crystallin gene families that are expressed in the eye lens.

Animals↗

ATP and IMP in single human muscle fibres after high intensity exercise.

Within a muscle there is large variation in the activity of various enzymes among single fibres. It is reasonable, therefore, to expect a corresponding variation in their metabolic response to exercise. This was examined by obtaining muscle biopsies from five men at rest and after intense short-term exercise consisting of three bouts of 30 knee extensions each and intervened by 60 s rest. Freeze-dried dissected single fibres identified as type 1 or type 2 were analysed for ATP and IMP contents by liquid chromatography. Rest. Average ATP tended to be higher in type 2 than in type 1 fibres. The ATP range was 14-30 and 14-32 mmol X kg-1 dry muscle (dm) in type 1 and 2 fibres, respectively. IMP was less than 1 mmol X kg-1 dm in most fibres and similar in types 1 and 2. Exercise. Muscle force decreased 70% during exercise. The average decrease in ATP content was significant for both fibre types with somewhat larger response for type 2 (20%) than type 1 (10%) fibres. The ATP range was 10-28 and 10-30 mmol X kg-1 dm in type 1 and 2 fibres, respectively. Average IMP content increased substantially in both fibre types, more so for type 2, and the range for individual fibres was 0-13 (type 1) and 0-21 (type 2) mmol X kg-1 dm. Conclusion. After exhaustive short-term exercise, a large variation in ATP and IMP contents was evident among single type 1 and type 2 fibres. None of the fibres, however, showed an ATP content lower than 10 mmol X kg-1 dm.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Monophosphate↗

ATP breakdown products in human skeletal muscle during prolonged exercise to exhaustion.

To study changes in muscle energy state during prolonged exercise, especially in relation to fatigue, muscle biopsies were obtained from seven healthy males working until exhaustion on a cycle ergometer at 68% (63-74%) of their maximal oxygen uptake. Biopsies were taken at rest, after 15 and 45 min of exercise and at exhaustion, and analysed for ATP, ADP, AMP, inosine monophosphate (IMP) and hypoxanthine content by high performance liquid chromatography (HPLC), and for creatine phosphate (CP), lactate and glycogen by enzymatic fluorometric techniques. Glycogen content at exhaustion was approximately 30% of the pre-exercise level. The CP content decreased steeply during the first 15 min of exercise (P less than 0.01) and continued to decrease during the rest of the exercise period (P less than 0.05). Pronounced increases in contents of IMP (64% P less than 0.001) and hypoxanthine (69%, P less than 0.05) were found when exhaustion was approaching. Furthermore, energy charge [EC; (ATP + 0.5 ADP)/(ATP + ADP + AMP)] was decreased at exhaustion (P less than 0.05). The increases in IMP and hypoxanthine which occurred when exhaustion was approaching during prolonged submaximal exercise together with the decrease in EC during this phase of exercise suggest a failure of the exercising skeletal muscle to regenerate ATP at exhaustion.

Adenosine Diphosphate↗