The influence of smoking and of oral and transdermal nicotine on blood pressure, and haematology and coagulation indices

Nicotine is helpful in stopping smoking but its influence on cardiovascular risk factors is incomplete. Our aim was to determine its effect on blood pressure, routine haematology indices, and coagulation indices relevant to thrombosis. Eighteen subjects were seen whilst smoking (cotinine levels 1119 +/- 414 ng/ml), again after stopping smoking but while using nicotine chewing gum and/or skin patches (392 +/- 198 ng/ml), and again when not using nicotine (cotinine undetectable). There were no significant changes in blood pressures, platelet count, mean platelet volume, viscosity or anti-thrombin III. However, white blood cell count (p = 0.003), lymphocyte count (p = 0.016), red blood cell count (p = 0.02), haemoglobin (p <0.001), fibrinogen (p <0.001) and von Willebrand factor (p = 0.001) all fell between the first and second samples (when still using nicotine) but not between the second and third samples (when off nicotine). Oral and/or transdermal nicotine does not influence blood pressure or the haematology and coagulation indices we have measured.     

Reconstitution of an active magnesium chelatase enzyme complex from the bchI, -D, and -H gene products of the green sulfur bacterium Chlorobium vibrioforme expressed in Escherichia coli

Magnesium-protoporphyrin chelatase, the first enzyme unique to the (bacterio)chlorophyll-specific branch of the porphyrin biosynthetic pathway, catalyzes the insertion of Mg2+ into protoporphyrin IX. Three genes, designated bchI, -D, and -H, from the strictly anaerobic and obligately phototrophic green sulfur bacterium Chlorobium vibrioforme show a significant level of homology to the magnesium chelatase-encoding genes bchI, -D, and -H and chlI, -D, and -H of Rhodobacter sphaeroides and Synechocystis strain PCC6803, respectively. These three genes were expressed in Escherichia coli; the subsequent purification of overproduced BchI and -H proteins on an Ni2+-agarose affinity column and denaturation of insoluble BchD protein in 6 M urea were required for reconstitution of Mg-chelatase activity in vitro. This work therefore establishes that the magnesium chelatase of C. vibrioforme is similar to the magnesium chelatases of the distantly related bacteria R. sphaeroides and Synechocystis strain PCC6803 with respect to number of subunits and ATP requirement. In addition, reconstitution of an active heterologous magnesium chelatase enzyme complex was obtained by combining the C. vibrioforme BchI and -D proteins and the Synechocystis strain PCC6803 ChlH protein. Furthermore, two versions, with respect to the N-terminal start of the bchI gene product, were expressed in E. coli, yielding ca. 38- and ca. 42-kDa versions of the BchI protein, both of which proved to be active. Western blot analysis of these proteins indicated that two forms of BchI, corresponding to the 38- and the 42-kDa expressed proteins, are also present in C. vibrioforme.     

Bone density and cyclic ovarian function in trained runners and active controls

This study was conducted to determine whether rigorous exercise training adversely affects ovarian hormone levels and bone health in cyclically menstruating trained runners. Ovarian hormones, bone mineral density (BMD), body composition, 3-d diet records, 3-d estimated energy expenditure, and menstrual histories were evaluated in 10 trained collegiate runners and 10 moderately active controls. The trained runners had lower total body calcium per kg of soft lean tissue measured by DEXA (P = 0.045). Half of the trained runners had experienced stress fractures compared with only one of the moderately active controls. The trained runners' lumbar (L2-L4) BMD (1.178 g.cm-2) was not significantly different from that of the active controls (1.283 g.cm-2) (P = 0.074) but, for all subjects combined, there wasa significant inverse relation between L2-L4 BMD and distance run per week (P = 0.036). Further, adding age, body weight, percent body fat, daily energy intake, and daily calcium intake to a stepwise multiple regression analysis did not significantly improve predictive precision. The trained runners consumed nearly twice the amount of calcium (1089 mg.d-1 vs 641 mg.d-1, respectively; P = 0.036), while intake of other nutrients did not differ significantly between groups. Urinary estrone conjugates (E1C) were lower in the trained runners during the early follicular phase (P = 0.028), while pregnanediol-3-glucuronide (PdG) was not significantly different between groups during the luteal phase (P = 0.213). Thus, it appears that lower estrogen production, especially during the early follicular phase, and not progesterone, is associated with lower whole body calcium per kg of soft lean tissue and, probably, L2-L4 BMD. Results of this study also suggest that regular menstrual cycles do not imply normal ovarian hormone function in young women who are engaged in either recreational or competitive running.     

Genotypic effects on norepinephrine-induced changes in thermogenesis, metabolic hormones, and metabolites in newborn calves

Heat production was measured in newborn Angus-, Brahman-, and Tuli-sired calves born to Angus (n = 20) and Brahman (n = 26) dams, before (thermoneutral metabolic rate, TMR) and after norepinephrine (NE) infusion (peak metabolic rate, PMR), to assess genotypic effects on nonshivering thermogenesis in brown adipose tissue. Calves were fed pooled colostrum, fitted with jugular catheters, and placed in a temperature-controlled (37 degrees C) water immersion system. Heat production, determined by indirect calorimetry, and tympanic temperature were measured continuously in calves from approximately 3 to 6 h of age. Blood samples were collected at birth and at 0, 5, 20, 40, 60, 80, 100, and 120 min relative to NE infusion (35 micrograms.min-1.kg BW-1 for 4 min), and plasma was analyzed for metabolites (glucose, NEFA, and urea nitrogen [PUN]) and hormones (cortisol, triiodothyronine [T3] and thyroxine [T4]). Weight-specific TMR (cal.min-1.kg-1) was not affected by breed of sire or dam, although weight-specific PMR (cal.min-1.kg-1) was lower (P < .01) in Brahman-sired calves than in Angus- or Tuli-sired calves and was lower (P < .001) in calves born to Brahman rather than Angus dams. The reduction in weight-specific PMR due to the maternal Brahman influence was sire-breed dependent, and the reduction was largest (P < .01) for Tuli-sired (34.3%), intermediate (P < .05) for Brahman-sired (15.1%), and lowest (P > .25) for Angus-sired calves (4.1%). The PMR:TMR ratio was 1.80 and 2.21 +/- .06 in calves born to Brahman and Angus dams, respectively. Peak tympanic temperature was .6 degree C lower (P < .01) in calves born to Brahman rather than Angus dams. At birth, plasma NEFA concentrations were higher (P < .001) and glucose tended (P = .13) to be higher in calves born to Brahman rather than Angus dams. Cortisol, T3, and T4 concentrations at birth were higher in calves born to Brahman dams than in those born to Angus dams. These results suggest that calves born to Brahman dams may have less thermogenically active brown adipose tissue than calves born to Angus dams, which may contribute to the relative cold intolerance of calves with Bos indicus inheritance.     

Human NELL-1 expressed in unilateral coronal synostosis

Surgical correction of unilateral coronal synostosis offers a unique opportunity to examine the molecular differences between an abnormal and a normal cranial suture. We isolated and identified a cDNA fragment whose expression was up-regulated in the premature fusing and fused coronal sutures, as compared with normal coronal sutures. The nucleotide sequence of the full-length cDNA of this gene, human NELL-1, has approximately 61% homology with the chicken Nel gene. Both chicken Nel and human NELL-1 are comprised of six epidermal growth factor-like repeats. The human NELL-1 messages were localized primarily in the mesenchymal cells and osteoblasts at the osteogenic front, along the parasutural bone margins, and within the condensing mesenchymal cells of newly formed bone in sites of premature sutural fusion. Human multiorgan tissue mRNA blot showed that NELL-1 was specifically expressed in fetal brain but not in fetal kidney, liver, or lung. We also showed that Nell-1 was expressed in rat calvarial osteoprogenitor cells and was largely absent in rat tibiae and fibroblast cell cultures. In conclusion, our data suggest that the NELL-1 gene is preferentially expressed in cranial intramembranous bone and neural tissue (both of neural crest cell origin) and is up-regulated during unilateral premature closure of the coronal suture. The precise role of this gene is unknown.     

Narrow band fiber-optic phase-shifted Fabry-Perot Bragg grating filters for atmospheric water vapor lidar measurements

A unique ultranarrowband fiber-optic phase-shifted Fabry-Perot Bragg grating filter for atmospheric water vapor lidar measurements was designed, fabricated, and successfully tested. Customized optical fiber Bragg gratings were fabricated so that two transmission filter peaks occurred: one (89% transmission, 8 pm FWHM) near the 946-nm water vapor absorption line and the other peak (80% transmission, 4 pm FWHM) at a region of no absorption. Both transmission peaks were within a 2.66-nm stop band. Demonstration of tension tuning to the 946.0003-nm water vapor line was achieved, and the performance characterization of custom-made optical fiber Bragg grating filters are presented. These measurements are successfully compared to theoretical calculations using a piecewise-matrix form of the coupled-mode equations.