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The effects of long-term smoking on mitochondrial DNA (mtDNA) deletions in hair follicles were investigated in subjects with different antioxidant capacity. Twenty-two male smokers with a smoking index of greater than 5 pack-years and without any known systemic diseases were recruited for this study. Forty healthy nonsmoking males were included as controls. We found that the concentrations of ascorbate and alpha-tocopherol and the activities of glutathione S-transferase (GST) and glutathione peroxidase in blood plasma were significantly decreased in smokers. The levels of glutathione and protein thiols in whole blood and the incidence of a 4,977 bp deletion of mtDNA (dmtDNA) in hair follicles were significantly increased in smokers. A significantly higher incidence of the 4,977 bp dmtDNA was found in smokers with plasma GST activity less than 5.66 U/l (OR = 7.2, P = 0.020). Using multiple covariate ANOVA and logistic regression, we found that age and low plasma GST activity were the only two risk factors for the 4,977 bp dmtDNA. These results suggest that smoking depletes antioxidants and causes mtDNA deletions and that plasma GST may play an important role in the preservation of the mitochondrial genome in tissue cells of smokers.
Environ Mol Mutagen 2002
PMID:Alterations of plasma antioxidants and mitochondrial DNA mutation in hair follicles of smokers. 1235 50

A number of morphological, physiological and phenological traits are known to improve the performance of rice challenged by drought. Root morphological traits and stress-induced response form important components of drought tolerance. Enhancing grain yield remains the principal objective of most breeding programs. Interaction between primary traits poses a formidable challenge while dealing with grain yield under stress. The evaluation of root morphology at three different growth stages and grain yield along with related characteristics under contrasting moisture regimes was made using nine backcrosses along with their parent and standard checks. The backcrosses invoved transgressant double haploid lines derived from IR64 and Azucena with IR64. Marked genotypic differences were observed for all root morphology as well as grain yield related characteristics across the sampling dates as revealed by individual and combined ANOVA. Among the nine backcrosses studied in this experiment, the BC1F2 population of P124 x IR64 were evaluated for forwarding based on their performance with respect to maximum root length and grain yield under both well-watered and low-moisture stress conditions. Sixty-nine plants - ten percent of the backcross population - were selectively genotyped using RAPD primers. Under well-watered conditions two RAPD markers showed strong linkage to QTLs for maximum root length evaluated under ww conditions. Two other markers could explain the considerable amount of variation in MRL under LMS. One of the markers identified under low-moisture stress conditions was also able to explain variability in maximum root length in the mean environment.
Cell Mol Biol Lett 2002
PMID:Tagging QTLs for maximum root length in rainfed lowland rice (Oryza sativa L.) using molecular markers. 1237 37

Combined morphological and molecular techniques were used to characterize variation in Elymus repens. We studied the morphological variability of E. repens in relation to the degree of its genetic differentiation, in order to unravel the causes of conspicuous intraspecific morphological variation. Four populations of E. repens from different habitats were analyzed for 35 morphological characters, and their genetic differentiation was assessed by Amplified Fragment Length Polymorphism (AFLP). Four pairs of selective primers were used to detect a total of 279 AFLP bands, of which 104 (37.28%) were polymorphic between populations. Cluster analysis based on AFLP fingerprint data showed that individuals were arranged in population-specific groups. The analyses of variance (ANOVA and AMOVA) indicated significant morphological and genetic differentiation among populations (P<0.01). This study revealed low levels of AFLP variation, which suggests that conspicuous morphological variation of E. repens is caused by plasticity. E. repens is an evolutionarily young species, of hybrid origin, in which microevolutionary processes continue. This study showed that common analysis of genetic diversity and morphology is a powerful tool in low-level taxonomy.
Cell Mol Biol Lett 2002
PMID:Morphological and AFLP variation of Elymus repens (L.) Gould (Poaceae). 1237 60

A polymorphic trinucleotide repeat (CGG/GCC) within the human Reelin gene (RELN) was examined as a candidate gene for autism spectrum disorders (ASDs). This gene encodes a large extracellular matrix protein that orchestrates neuronal positioning during corticogenesis. The CGG-repeat within the 5' untranslated region of RELN exon 1 was examined in 126 multiple-incidence families. The number of CGG repeats varied from three to 16 in affected individuals and controls, with no expansion or contraction observed during maternal (n = 291) or paternal (n = 287) transmissions in families with autistic probands. Although the frequencies of the RELN alleles and genotypes in affected children were not different from those in the comparison group, a family-based association test (FBAT) showed that the larger RELN alleles (> or = 11 repeats) were transmitted more often than expected to affected children (S = 43, E(S) = 34.5, P = 0.035); this was particularly the case for the 13-repeat RELN allele (S = 22, E(S) = 16, P = 0.034). Affected sib-pair (ASP) analysis found no evidence of excess sharing of RELN alleles in affected siblings. The impact of genotypes with large alleles (> or = 11 repeats) on the phenotypes in individuals with ASD was analyzed by ANOVA in a subset of the families for which results of the Autism Diagnostic Interview-Revised were available. Children with large RELN alleles did not show any difference in scores for questions related to the core symptoms of autistic disorder, but there was a tendency for children with at least one large RELN allele to have an earlier age at first phrase (chi(2) = 3.538, P = 0.06). Thus, although the case-control and affected sib-pair findings did not support a role for RELN in susceptibility to ASD, the more powerful family-based association study demonstrated that RELN alleles with larger numbers of CGG repeats may play a role in the etiology of some cases of ASD, especially in children without delayed phrase speech.
Mol Psychiatry 2002
PMID:Reelin gene alleles and susceptibility to autism spectrum disorders. 1239 38

The amount of radical scavenging activity in muscle is unknown. The present study examines whether electron spin resonance (ESR) could measure and distinguish antioxidant capacity in muscle with different contractile and metabolic characteristics. Specimens of the soleus, plantaris, gastrocnemius (deep/surface portions), heart and diaphragm were obtained from female Wistar rats (n=7; 12 weeks old). Scavenging activity against superoxide anions in these specimens were determined by ESR using a spin-trapping chemical (5,5-dimethyl-1-pyrroline-N-oxide). The ESR signal intensity of reaction mixtures containing muscle tissues was significantly lower in the heart, soleus, diaphragm and deep portion of the gastrocnemius than in the plataris and surface portion of the gastrocnemius. Thus, the amount of scavenging activity converted into superoxide dismutase activity was the highest in the heart, and higher in the soleus, diaphragm and deep portion of the gastrocnemius than in other muscles (ANOVA, P<0.01). In addition, scavenging activity significantly correlated with citrate synthase activity (r=0.72, P<0.01, n=42) and myoglobin content (r=0.63, P<0.01, n=42). These findings suggested that ESR and spin-trapping can be detect differences in free radical scavenging activity among muscle tissues with different metabolic characteristics.
Comp Biochem Physiol B Biochem Mol Biol 2003 Feb
PMID:Antioxidant capacity in rat skeletal muscle tissues determined by electron spin resonance. 1256 99

We tested the hypothesis that transcranial oximetry by fast scanning near infrared spectroscopy can detect local desaturation of hemoglobin in arterial vessels of cerebral circulation with impaired blood supply. A total of 74 near infrared spectroscopy recordings were taken from the intact skull of humans. Perfusion of the hemisphere under the detector was assessed in one of four groups: (1) healthy volunteer; (2) patient, unaffected side; (3) patient, affected side with intact collateral blood supply; (4) patient, affected side, impaired collateral blood supply. Transcranial saturation was 0.90+/-0.01 (all values reported as mean+/-S.E.) in healthy volunteers (n=24), 0.92+/-0.008 in the unaffected hemisphere of patients (n=23), 0.92+/-0.001 in the affected side if collateral supply with blood was intact (n=16). There was no statistical significance between these groups. Saturation in affected hemispheres with impaired collateral blood supply (n=9) was 0.81+/-0.028, which was significantly different from all other groups (P<0.05, one way-ANOVA). We conclude, that transcranial pulse oximetry can detect local hypoxia if collateral blood supply fails.
Comp Biochem Physiol A Mol Integr Physiol 2003 Mar
PMID:Transcranial oximetry using fast near infrared spectroscopy can detect failure of collateral blood supply in humans. 1260 Jun 62

It is estimated that about 2.5 million people only in the United States are affected by epilepsy. Labelled red blood cells (RBC) and plasma proteins (PP) are used for several evaluations in nuclear medicine and drugs affecting those labelings have previously been described. The aim of this study was to evaluate whether the most popular antiseizure drugs interfere with the 99mTc labeling process of RBC and PP. Heparinized blood withdrawn from Wistar rats was incubated with phenobarbital (0.2, 2, 20, 200, 2,000 microg/ml), phenytoin (0.15, 1.5, 15, 150, 1,500 microg/ml), carbamazepine (0.7, 7, 70 microg/ml), clonazepam (0.5, 5, 50, 500 microg/ml) or valproic acid (0.5, 5, 50, 500 microg/ml) for I hr. Stannous chloride (SnCl2), in two different concentrations (0.012 or 1.2 microg/ml) and 99mTc were added. Plasma and cellular fractions were isolated by centrifugation, soluble and insoluble fractions were separated by trichloroacetic acid precipitation. The percentage of radioactivity was calculated for each fraction. Statistical analysis was performed with ANOVA and Dunnet tests. The analysis of the results has shown that phenobarbital (2,000 microg/ml) and clonazepam (50 microg/ml) significantly have reduced the RBC labeling efficiency when it was used the optimal SnCl2 concentration (1.2 microg/ml) and clonazepam (5, 50 microg/ml) has significantly decreased the PP labeling efficiency with 99mTc. Phenytoin (1,500 microg/ml) has decreased the RBC labeling efficiency when the experiments were carried out with a small SnCl2 concentration (0.012 microg/ml). We can suggest that with this in vitro assay, at the therapeutic level of phenytoin, phenobarbital, carbamazepine and valproic acid will not interfere on the 99mTc labeling process of RBC. Interference is displayed at higher phenobarbital concentrations (2,000 microg/ml). However, humans do not tolerate this concentration. On the other hand, a decreased RBC and PP labeling efficiency with 99mTc may be expected for clonazepam at therapeutic levels.
Cell Mol Biol (Noisy-le-grand) 2002 Nov
PMID:Assessment of the effect of antiseizure drugs on the labeling process of red blood cells and plasma proteins with technetium-99m. 1261 78

Creatine has been reported to be an effective ergogenic aid for athletes. However, concerns have been raised regarding the long-term safety of creatine supplementation. This study examined the effects of long-term creatine supplementation on a 69-item panel of serum, whole blood, and urinary markers of clinical health status in athletes. Over a 21-month period, 98 Division IA college football players were administered in an open label manner creatine or non-creatine containing supplements following training sessions. Subjects who ingested creatine were administered 15.75 g/day of creatine monohydrate for 5 days and an average of 5 g/day thereafter in 5-10 g/day doses. Fasting blood and 24-h urine samples were collected at 0, 1, 1.5, 4, 6, 10, 12, 17, and 21 months of training. A comprehensive quantitative clinical chemistry panel was determined on serum and whole blood samples (metabolic markers, muscle and liver enzymes, electrolytes, lipid profiles, hematological markers, and lymphocytes). In addition, urine samples were quantitatively and qualitative analyzed to assess clinical status and renal function. At the end of the study, subjects were categorized into groups that did not take creatine (n = 44) and subjects who took creatine for 0-6 months (mean 4.4 +/- 1.8 months, n = 12), 7-12 months (mean 9.3 +/- 2.0 months, n = 25), and 12-21 months (mean 19.3 +/- 2.4 months, n = 17). Baseline and the subjects' final blood and urine samples were analyzed by MANOVA and 2 x 2 repeated measures ANOVA univariate tests. MANOVA revealed no significant differences (p = 0.51) among groups in the 54-item panel of quantitative blood and urine markers assessed. Univariate analysis revealed no clinically significant interactions among groups in markers of clinical status. In addition, no apparent differences were observed among groups in the 15-item panel of qualitative urine markers. Results indicate that long-term creatine supplementation (up to 21-months) does not appear to adversely effect markers of health status in athletes undergoing intense training in comparison to athletes who do not take creatine.
Mol Cell Biochem 2003 Feb
PMID:Long-term creatine supplementation does not significantly affect clinical markers of health in athletes. 1270 16

Collagen fiber orientation is one aspect of the microstructure of bone that influences its mechanical properties. While the spatial distribution of preferentially oriented collagen is hypothesized to reflect the effects of loading during the process of aging, its variability in a modern human sample is essentially unknown. In a large sample (n = 67) of autopsied adults, the variability of collagen fiber orientation in the mid-shaft femur was examined in relation to age and sex. Montaged images of entire 100 microm thick cross-sections were obtained using circularly polarized light microscopy (CPLM) under standardized illuminating conditions. An automated image-analyzing routine divided images into 48 segments according to anatomical position. Average gray values (varying with orientation) were quantified for each segment, and one-way ANOVA with Tukey HSD post hoc tests were applied to assess differences between segments. Collagen fiber orientation appeared to be nonrandomly distributed across the mid-shaft femur sample; however, no single "human" pattern was identified. Individual variation, unexplainable by age, sex, or body size, exceeded population-level trends. Differences between age and sex groups suggest there is a strong correspondence between collagen fiber orientation and tissue-type distributions. The minimal consistencies demonstrated here may reflect mechanical forces induced at the femoral mid-shaft. However, the myriad of other factors that may influence collagen fiber orientation patterning, including growth trajectories, metabolic and nutritional status, and disease states, must be explored further. Only then, in conjunction with studies of other structural and material properties of bone, will we be able to elucidate the linkages between microstructure and functional adaptation in the human mid-shaft femur.
Anat Rec A Discov Mol Cell Evol Biol 2003 May
PMID:Preferred collagen fiber orientation in the human mid-shaft femur. 1270 1

This study was designed to evaluate the neuroprotective effect of the cannabinoid agonist WIN-55212 after inducing acute severe asphyxia in newborn rats. The left common carotid artery was ligated in anaesthetised 7-day-old Wistar rats, which were then asphyxiated by inhaling 100% nitrogen for 10 min. Pups recovering from asphyxia were s.c. administered vehicle (n=23), WIN-55212 (0.1 mg/kg, n=18), or WIN-55212 plus the CB1 receptor antagonist SR141716 (3 mg/kg, n=10). Pups undergoing a sham operation served as controls (n=12). Coronal sections of the brain were obtained on the 14th day after surgery and observed under light microscope after Nissl or Fluoro-Jade B (FJB) staining, to respectively quantify surviving or degenerating neurones in the CA1 area of the hippocampus and parietal cortex. Acute asphyxia led to early neurone loss amounting to 19% in the hippocampus and 29% in the cortex (both ANOVA P<0.05 vs. control). Delayed neurone loss occurred in the proportions 13% in the hippocampus and 20% in the cortex (both ANOVA P<0.05 vs. control). Neuronal loss was fully prevented by WIN-55212 administration. Co-administration of SR141716 failed to modify the protective effect of WIN-55212 on early neuronal death, but abolished the WIN-55212-induced prevention of delayed neuronal death. We conclude that when administered after acute severe asphyxia in newborn rats, WIN-55212 shows a neuroprotective effect, reducing both early and delayed neurone loss. This effect is achieved through two parallel CB1-dependent and -independent mechanisms.
Brain Res Mol Brain Res 2003 Jun 10
PMID:Neuroprotection by the cannabinoid agonist WIN-55212 in an in vivo newborn rat model of acute severe asphyxia. 1282 23


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