Atropine improves insulin sensitivity in both lean and abdominally obese subjects.

BACKGROUND: Dysregulated autonomic nerve activity may contribute to the development of type 2 diabetes. The aim of this study was to assess the effects of an anticholinergic agent, atropine, and a cholinergic agent, physostigmine, on insulin sensitivity in lean and abdominally obese subjects. SUBJECTS AND METHODS: In a single-blinded three-way crossover study, six lean and six abdominally obese nondiabetic subjects [three males and three females in each group; age, 43.8 ± 14.8 vs. 46.8 ± 4.8 yr (mean ± sd); body mass index, 22.6 ± 1.7 vs. 28.8 ± 1.3 kg/m(2); and waist circumference, 85 ± 2 vs. 99 ± 6 cm, respectively] were given iv infusions with atropine (15 µg/kg bolus, 4 µg/kg · h infusion), physostigmine (0.12 µg/kg · min) or saline (0.9% NaCl) in a randomized treatment order. Infusions were started 30 min before and continued throughout a 120-min euglycemic (5.6 mm) hyperinsulinemic (40 mU/m(2) · min) clamp. RESULTS: Insulin sensitivity (M-value, i.e. glucose infusion rate divided by lean body mass) during the last 60 min of the clamp was higher during infusion with atropine than saline (9.2 ± 1.0 vs. 7.6 ± 1.0 mg/kg lean body mass · min, mean ± sem; P = 0.015) in all subjects. Physostigmine did not differ significantly from saline (8.2 ± 1.0). M-values were significantly higher in lean vs. obese [atropine, 11.6 ± 1.4 vs. 7.6 ± 1.3; physostigmine, 10.8 ± 1.3 vs. 6.3 ± 1.3; and saline, 9.1 ± 1.4 vs. 6.4 ± 1.3, respectively (all P < 0.05)], but the incremental effect of atropine vs. saline did not differ consistently between groups. CONCLUSION: Insulin sensitivity was higher during a short-term atropine infusion compared with saline in both lean and abdominally obese subjects. This insulin-sensitizing effect of cholinergic blockade is unexpected, and the underlying mechanisms should be further investigated.

Pressure pain thresholds and electromyographically defined muscular fatigue induced by a muscular endurance test in normal women.

OBJECTIVE: This study was undertaken to examine the relation between muscular tenderness measured as pressure pain thresholds (PPTs) and electromyographic (EMG) signs of fatigue before and after a local standardized static muscle contraction. DESIGN: Pressure pain thresholds were measured in the shoulder region before, immediately after, and 10 minutes after a standardized static endurance test while monitoring the EMG signs of local muscular fatigue and its recovery. The study did not address local biochemical issues. SETTING: The study was conducted at the Department of Rehabilitation, Lund University Hospital, Lund, Sweden. SUBJECTS: Twenty-five healthy female volunteers without musculoskeletal problems participated in this study. INTERVENTION: A static endurance test was performed, which consisted of a submaximal unilateral activation of the right trapezius and deltoid muscles for as long as possible. OUTCOME MEASURES: Bilateral PPTs over the trapezius and deltoid muscles were measured with an electronic pressure algometer. Established surface EMG parameters of local muscular fatigue were assessed. The Borg Rating of Perceived Exertion scale was used. RESULTS: The average endurance time was 330 seconds. Immediately after the test, significant bilateral increases in the normalized PPTs over both muscles were found, although the increase was more pronounced on the test side: over the right trapezius muscle by 13% (p <0.001), over the right deltoid muscle by 23% (p <0.001), and over the left trapezius and deltoid muscles by 6% (p = 0.04) and (p = 0.009), respectively. These increases persisted 10 minutes after the end of the test. The subjects developed significant signs of fatigue as defined by EMG criteria in both muscles on the right side during the test. The recovery from fatigue was approximately half complete 15 seconds after the end of the test and complete or almost complete 10 minutes thereafter. CONCLUSIONS: Pressure pain thresholds over shoulder muscles remained elevated up to 10 minutes after a unilateral static endurance test. This time course was completely different from that of EMG-defined muscle fatigue, which showed a fast recovery. These findings indicate that the mechanisms of recovery from fatigue and nociception are independent of each other. The bilateral PPT increases might be explained by central antinociceptive mechanisms activated by static muscle work.

Cysteinyl and substrate radical formation in active site mutant E441Q of Escherichia coli class I ribonucleotide reductase.

All classes of ribonucleotide reductase are proposed to have a common reaction mechanism involving a transient cysteine thiyl radical that initiates catalysis by abstracting the 3'-hydrogen atom of the substrate nucleotide. In the class Ia ribonucleotide reductase system of Escherichia coli, we recently trapped two kinetically coupled transient radicals in a reaction involving the engineered E441Q R1 protein, wild-type R2 protein, and substrate (Persson, A. L., Eriksson, M., Katterle, B., Pötsch, S., Sahlin, M., and Sjöberg, B.-M. (1997) J. Biol. Chem. 272, 31533-31541). Using isotopically labeled R1 protein or substrate, we now demonstrate that the early radical intermediate is a cysteinyl radical, possibly in weak magnetic interaction with the diiron site of protein R2, and that the second radical intermediate is a carbon-centered substrate radical with hyperfine coupling to two almost identical protons. This is the first report of a cysteinyl free radical in ribonucleotide reductase that is a kinetically coupled precursor of an identified substrate radical. We suggest that the cysteinyl radical is localized to the active site residue, Cys439, which is conserved in all classes of ribonucleotide reductase, and which, in the three-dimensional structure of protein R1, is positioned to abstract the 3'-hydrogen atom of the substrate. We also suggest that the substrate radical is localized to the 3'-position of the ribose moiety, the first substrate radical intermediate in the postulated reaction mechanism.

A new mechanism-based radical intermediate in a mutant R1 protein affecting the catalytically essential Glu441 in Escherichia coli ribonucleotide reductase.

The invariant active site residue Glu441 in protein R1 of ribonucleotide reductase from Escherichia coli has been engineered to alanine, aspartic acid, and glutamic acid. Each mutant protein was structurally and enzymatically characterized. Glu441 contributes to substrate binding, and a carboxylate side chain at position 441 is essential for catalysis. The most intriguing results are the suicidal mechanism-based reaction intermediates observed when R1 E441Q is incubated with protein R2 and natural substrates (CDP and GDP). In a consecutive reaction sequence, we observe at least three clearly discernible steps: (i) a rapid decay (k1 >/= 1.2 s-1) of the catalytically essential tyrosyl radical of protein R2 concomitant with formation of an early transient radical intermediate species, (ii) a slower decay (k2 = 0.03 s-1) of the early intermediate concomitant with formation of another intermediate with a triplet EPR signal, and (iii) decay (k3 = 0.004 s-1) of the latter concomitant with formation of a characteristic substrate degradation product. The characteristics of the triplet EPR signal are compatible with a substrate radical intermediate (most likely localized at the 3'-position of the ribose moiety of the substrate nucleotide) postulated to occur in the wild type reaction mechanism as well.

Lack of bacterial invasion in experimental periodontitis.

The present study in the beagle dog was performed to analyze whether micro-organisms from a subgingival microbiota could be translocated into or had the potential to invade the pocket epithelium and the gingival connective tissue during a phase of rapid breakdown of the attachment apparatus. An attempt was also made to assess whether tetracycline therapy suppressed the subgingival microbiota and changed the size and quality of the lesions in the gingival tissue. 5 inbred beagle dogs were used. Throughout the period of experimentation, the animals were fed a soft diet permitting gross accumulation of plaque and calculus. No mechanical plaque control measures were performed during the course of the study. On day 0, a 120-day period of periodontal tissue breakdown was initiated at the right mandibular 3rd and 4th premolars by tying cotton floss ligatures around the neck of these teeth. The process of tissue breakdown at the mandibular left 3rd and 4th premolars was started 30 days later. The ligatures were replaced once every 2 weeks during the subsequent 4-month period. On experimental day 120, the first biopsy was performed and gingival tissue sections prepared for light and electron microscopic assessment of a series of histometric characteristics. On day 120, a 30-day period of tetracycline (per os) administration was initiated. Each dog was given a dose of 500 mg tetracycline twice daily. On day 150, the biopsy procedure was repeated in the mandibular left premolar regions.(ABSTRACT TRUNCATED AT 250 WORDS)