Non-CO2 greenhouse gases and climate change.

Earth's climate is warming as a result of anthropogenic emissions of greenhouse gases, particularly carbon dioxide (CO(2)) from fossil fuel combustion. Anthropogenic emissions of non-CO(2) greenhouse gases, such as methane, nitrous oxide and ozone-depleting substances (largely from sources other than fossil fuels), also contribute significantly to warming. Some non-CO(2) greenhouse gases have much shorter lifetimes than CO(2), so reducing their emissions offers an additional opportunity to lessen future climate change. Although it is clear that sustainably reducing the warming influence of greenhouse gases will be possible only with substantial cuts in emissions of CO(2), reducing non-CO(2) greenhouse gas emissions would be a relatively quick way of contributing to this goal.

In situ synthesis of oligonucleotide arrays by using surface tension.

This work describes the in situ synthesis of oligonucleotide arrays on glass surfaces. These arrays are composed of features defined and separated by differential surface tension (surface tension arrays). Specifically, photolithographic methods were used to create a series of spatially addressable, circular features containing an amino-terminated organosilane coupled to the glass through a siloxane linkage. Each feature is bounded by a perfluorosilanated surface. The differences in surface energies between the features and surrounding zones allow for chemical reactions to be readily localized within a defined site. The aminosilanation process was analyzed using contact angle, X-ray photoelectron spectroscopy (XPS), and time-of-flight/secondary ion mass spectroscopy (TOF-SIMS). The efficiency of phosphoramidite-based oligonucleotide synthesis on these surface tension arrays was measured by two methods. One method, termed step-yields-by-hybridization, indicates an average synthesis efficiency for all four (A,G,C,T) bases of 99.9 +/- 1.1%. Step yields measured for the individual amidite bases showed efficiencies of 98.8% (dT), 98.0% (dA), 97.0% (dC), and 97.6% (dG). The second method for determining the amidite coupling efficiencies was by capillary electrophoresis (CE) analysis. Homopolymers of dT (40- and 60mer), dA (40mer), and dC (40mer) were synthesized on an NH(4)OH labile linkage. After cleavage, the products were analyzed by CE. Synthesis efficiencies were calculated by comparison of the full-length product peak with the failure peaks. The calculated coupling efficiencies were 98.8% (dT), 96.8% (dA), and 96.7% (dC).

Is loss of attachment due to root planning and scaling in sites with minimal probing depths a statistical or real occurrence?

BACKGROUND: Following root planing and scaling many studies have implied an association between a loss of clinical attachment at sites with initially shallow pockets (1 to 3 mm) and gains in attachment level for deeper probing depths. However, these effects are also consistent with a statistical phenomenon referred to as regression towards the mean. This principle suggests that extreme values will moderate the next time they are recorded. The purpose of this report was to estimate the effect that regression towards the mean has on perceived changes in attachment level after root planing and scaling. METHODS: During the initial examination, 2 different investigators conducted 2 full-mouth probings. Two quadrants were randomly selected to be root planed and scaled until the root surfaces were smooth by tactile touch of an explorer. The 2 remaining quadrants were not treated. At 4 to 6 weeks after treatment, another full mouth probing was done. An examiner who was blind to the quadrants that had been scaled measured attachment level and probing depth after therapy. This study design provided periodontal measurements before and after root planing and scaling, measurements before and after a period of 4 to 6 weeks of no therapy, and duplicate measurements at the beginning of the study. RESULTS: Using the repeat examination when no true change could occur, shallow sites (< or =3 mm of probing depth) showed average negative differences between repeat attachment level measurements (-0.23 mm), which mimicked loss of periodontal attachment. Deep sites, (>6 mm) showed average positive values (0.40) mimicking gain in attachment level. These results suggest that regression towards the mean is a significant effect in this data set. Both shallow non-scaled and scaled sites had similar differences in repeat measures (-0.28 mm, -0.25 mm) which were also similar to and not statistically different from changes after therapy for both non-scaled (-0.21 mm) and scaled sites (-0.08 mm). Thus not only does this data set exhibit regression towards the mean, but it explains the majority of perceived loss of periodontal attachment after scaling at sites that have minimal probing depth. CONCLUSIONS: These results suggest that the majority of perceived loss of attachment due to scaling at sites of minimal probing depth that have been reported in many studies may be due a statistical phenomenon called regression towards the mean.

Opportunities for improving inpatient smoking cessation programs: a community hospital experience.

BACKGROUND: This cohort study examined the role of smoking during hospitalization, duration of cessation counseling, patient awareness of the hospital's smoke-free policy, belief that smoking is associated with a current symptom or disease, and the presence of withdrawal symptoms with 12-month smoking cessation among inpatients enrolled in a smoking cessation program. METHODS: Inpatients in four community hospitals (N = 1,317) participated in a smoking cessation intervention consisting of face-to-face counseling at baseline and four follow-up counseling phone calls. Patients were classified as nonsmokers only if they reported not smoking at both the 6- and the 12-month interviews. All patients lost to follow-up were considered smokers. RESULTS: At 1 year the smoking cessation rate was 22.5%. Cessation was independently associated with reporting no smoking during hospitalization, noting no withdrawal symptoms at baseline, and believing that a current illness or symptom is related to smoking. Length of counseling interview and awareness of the hospital's smoke-free policy were not independently associated with cessation. CONCLUSIONS. Smoking cessation programs and hospital policies that decrease smoking during hospitalization, address withdrawal symptoms during hospitalization, and make clear the connection between a patient's health and cigarette smoking may increase the effectiveness of their smoking cessation efforts.

In vitro and in vivo evidence for actin association of the naphthylphthalamic acid-binding protein from zucchini hypocotyls.

The N-1-naphthylphthalamic acid (NPA)-binding protein is part of the auxin efflux carrier, the protein complex that controls polar auxin transport in plant tissues. This study tested the hypothesis that the NPA-binding protein (NBP) is associated with the actin cytoskeleton in vitro and that an intact actin cytoskeleton is required for polar auxin transport in vivo. Cytoskeletal polymerization was altered in extracts of zucchini hypocotyls with reagents that stabilized either the polymeric or monomeric forms of actin or tubulin. Phalloidin treatment altered actin polymerization, as demonstrated by immunoblot analyses following native and denaturing electrophoresis. Phalloidin increased both filamentous actin (F-actin) and NPA-binding activity, while cytochalasin D and Tris decreased both F-actin and NPA-binding activity in cytoskeletal pellets. The microtubule stabilizing drug taxol increased pelletable tubulin, but did not alter either the amount of pelletable actin or NPA-binding activity. Treatment of etiolated zucchini hypocotyls with cytochalasin D decreased the amount of auxin transport and its regulation by NPA. These experimental results are consistent with an in vitro actin cytoskeletal association of the NPA-binding protein and with the requirement of an intact actin cytoskeleton for maximal polar auxin transport in vivo.

A high-density screening format for encoded combinatorial libraries: assay miniaturization and its application to enzymatic reactions.

A novel, miniaturized high-throughput screening format is described for assay of combinatorial libraries generated on beads. This approach, which is ideally suited to encoded libraries synthesized on beads, utilizes the photolytic cleavage of individual compounds into a high-density well array (>6500 wells within a standard 96-well microtiter plate footprint) with well volumes as low as 0.37 microl. As a model study, an encoded dipeptide library (324 members) acylated with isobutyl succinate was assayed using this format to search for potential inhibitors of matrilysin, a member of the matrix metalloproteinase superfamily. In situ release of compounds from solid support was accomplished by photochemical cleavage after beads and enzyme were distributed to the wells. After the addition of a fluorogenic substrate to the array, the extent of enzyme inhibition and identification of active compounds was quantitated by imaging of the fluorescence emission upon uv irradiation. The structure-activity relationship data generated from the identified inhibitors in this study corroborate previous findings, thus validating the utility of this approach as a means of high-throughput screening of bead-based libraries.

High affinity type I interleukin 1 receptor antagonists discovered by screening recombinant peptide libraries.

Two families of peptides that specifically bind the extracellular domain of the human type I interleukin I (IL-1) receptor were identified from recombinant peptide display libraries. Peptides from one of these families blocked binding of IL-lalpha to the type I IL-1 receptor with IC50 values of 45-140 microM. Affinity-selective screening of variants of these peptides produced ligands of much higher affinity (IC50 approximately 2 nM). These peptides block IL-1-driven responses in human and monkey cells; they do not bind the human type II IL-1 receptor or the murine type I IL-1 receptor. This is the first example (that we know of) of a high affinity peptide that binds to a cytokine receptor and acts as a cytokine antagonist.

A Net Sink for Atmospheric CH3Br in the East Pacific Ocean.

Surface waters along a cruise track in the East Pacific Ocean were undersaturated in methyl bromide (CH(3)Br) in most areas except for coastal and upwelling regions, with saturation anomalies ranging from + 100 percent in coastal waters to -50 percent in open ocean areas, representing a regionally weighted mean of -16 (-13 to -20) percent. The partial lifetime of atmospheric CH(3)Br with respect to calculated oceanic degradation along this cruise track is 3.0 (2.9 to 3.6) years. The global, mean dry mole fraction of CH3Br in the atmosphere was 9.8 +/- 0.6 parts per trillion, with an interhemispheric ratio of 1.31 +/- 0.08. These data indicate that approximately 8 percent (0.2 parts per trillion) of the observed interhemispheric difference in atmospheric CH3Br could be attributed to an uneven global distribution of oceanic sources and sinks.

Properties of mutant acetolactate synthases resistant to triazolopyrimidine sulfonanilide.

Triazolopyrimidine sulfanilides are a class of highly active herbicides whose primary target is acetolactate synthase. Spontaneous mutants of tobacco (Nicotiana tabacum) (KS-43) and cotton (Gossypium hirsutum) (PS-3 and DO-2) resistant to triazolopyrimidine sulfonanilide were selected in tissue culture. Acetolactate synthase partially purified from the three mutants were 80- to 1000-fold less sensitive to inhibition by the compound compared with the corresponding wild-type enzyme. The mutants also varied in the cross-resistance pattern to other acetolactate synthase inhibiting herbicides in the sulfonylurea, imidazolinone, and pyrimidyl-oxy-benzoate chemical families. Thus, acetolactate synthase from KS-43, PS-3, and DO-2 cultures have different mutations. The affinities for pyruvate, thiamine pyrophosphate, as well as the activity of the mutant enzymes were found to be comparable to the corresponding wild-type enzymes. However, the enzyme from PS-3 was highly resistant to feedback inhibition by valine and leucine. In contrast, acetolactate synthase from KS-43 and DO-2 were inhibited by valine and leucine to nearly the same extent as the wild-type enzymes. Also, PS-3 cultures accumulated much higher levels of the branched chain amino acids compared to the wild-type cotton culture. The mutation in the PS-3 enzyme has therefore rendered it insensitive to feedback regulation by valine and leucine.

Pulsatile growth hormone, insulin-like growth factors and antler development in red deer (Cervus elaphus scoticus) stags.

Plasma samples taken every 30 min over a 26-h period each month from six 4- to 15-month-old red deer stags were analysed for GH. In addition, two samples taken at 10.00 and 22.00 h were analysed for insulin-like growth factor-I (IGF-I) and insulin-like growth factor-II (IGF-II). A concentrate diet was available ad libitum. Food intake, body weight and antler status were recorded. Concentrations of GH were analysed using the PULSAR peak detection routine. Secretion of GH was pulsatile in every month of sampling, but the pattern of pulsatility differed seasonally. During the autumn and early winter (April-June in the Southern hemisphere) GH pulses were frequent and of low amplitude. In contrast, GH pulses in spring (August-September) were of high amplitude and high frequency resulting in a high mean level of GH circulating in the plasma. In early summer (November) the GH pulse amplitude was much lower and pulse frequency fell. There was a rise in GH pulse frequency not accompanied by an increase in GH pulse amplitude in summer (December-January). GH pulse amplitude seemed to be the main determinant of mean GH plasma level. Secretion of IGF-I was raised 1 month after peak monthly mean GH secretion. There was little consistent relationship between concentrations of IGF-II and mean daily GH. Concentrations of GH correlated positively and significantly with liveweight gain and antler growth rate with a delay of 1 month. Significantly positive correlations between concentrations of IGF-I, liveweight gain and antler growth rate were observed. It is considered that the spring and summer (September-December) seasonal acceleration of liveweight gain and antler development in stags could be a consequence of high winter/early spring (August-September) GH pulse frequency and amplitude resulting in increased concentrations of IGF-I, particularly in October.

The effects of pancreatectomy on the plasma concentrations of insulin-like growth factors 1 and 2 in the sheep fetus.

The effects of hypoinsulinaemia and altered metabolite concentrations on the fetal plasma concentrations of insulin-like growth factors (IGF) have been investigated in chronically catheterized fetal sheep made insulin deficient by pancreatic ablation. Fetal pancreatectomy reduced significantly the plasma IGF-1 concentration and increased plasma IGF-2 activity in comparison with the values observed in sham operated fetuses. Mean plasma IGF-1 concentrations in the sham operated and pancreatectomized fetuses were 18.6 +/- 3.1 ng/ml (n = 7) and 13.4 +/- 1.4 ng/ml (n = 13) respectively. When all the data were combined, there was a significant positive correlation between the plasma concentrations of IGF-1 and insulin in utero. The mean IGF-2 activity was 2349 +/- 83 ng/ml (n = 7) in the sham operated fetuses and 3800 +/- 532 ng/ml in the pancreatectomized animals (n = 13). Plasma IGF-2 activity was correlated positively with plasma glucose, fructose and alpha-amino nitrogen levels and inversely related to the plasma insulin concentration in utero. These observations demonstrate that the fetal pancreas is essential for normal IGF production in the fetus and suggest that insulin, substrate availability and the IGFs may interact in the regulation of fetal growth.

The somatotrophic axis in young steers: influence of nutritional status on pulsatile release of growth hormone and circulating concentrations of insulin-like growth factor 1.

The relationship between plasma GH profiles and circulating concentrations of insulin-like growth factor 1 (IGF-1) at three different planes of nutrition, chosen to represent a high, medium and low level of nutrition (3%, 1.8% and 1% dry matter of liveweight per day) was studied in 15 young Angus steers. All steers were maintained on 3% dry matter for 5 weeks, then on one of the three nutritional planes for 4 weeks and then all were returned to 3% dry matter for 3 weeks. Blood was sampled through jugular catheters at 15-min intervals for 25 h at the end of each phase of the study and additional samples were taken on 2 days each week. Pulsatile release of GH occurred episodically with a diurnal increase during night and morning hours only in steers on high nutritional intakes. Reduced feeding at both the medium and the low plane abolished the diurnal rhythm and significantly increased mean plasma GH concentrations, the amplitude of GH pulses and the area under the GH profiles. Baseline concentrations of GH and pulse frequency did not change through nutritional manipulation. Upon realimentation, plasma GH concentrations decreased in both previously undernourished groups, with those fed 1% dry matter still having increased levels 10 days after refeeding. Plasma IGF-1 concentrations showed no periodicity. With nutritional deprivation, a decrease in IGF-1 concentration was observed only at negative energy balance (1% group). In this group plasma IGF-1 concentrations were progressively restored within 1 week of realimentation.(ABSTRACT TRUNCATED AT 250 WORDS)

Circulating insulin-like growth factor-binding proteins in fetal, neonatal and adult sheep.

A specific radioimmunoassay for insulin-like growth factor I (IGF-I) has been used to study the distribution of circulating IGF-I on its carrier proteins in sheep. Serum or plasma samples obtained from fetal, neonatal and adult sheep were chromatographed at neutral pH on Sephadex G-200 columns. IGF-I eluted primarily in the 20 000-100 000 dalton region and to a lesser extent in the 20 000-100 000 dalton region in the adult. In the fetus, IGF-I eluted primarily in the 20 000-50 000 dalton region. Studies in the neonatal lamb showed IGF-I activity to fluctuate between different size binding proteins but by 7 days after birth the pattern was similar to that of the adult. When labelled IGF-I and IGF-II were added to serum which was chromatographed over Sephadex G-200 at a neutral pH, binding was displayed in the 50 000-100 000 dalton region for both adult and fetus. However in the fetus, but not in the adult, binding of 125I-labelled IGF-II was also found at greater than 200 000 daltons. These studies demonstrate that the distribution of IGF-binding proteins varies between the adult and the fetal lamb. The appearance of the 150 000 dalton binding protein is coincidental with the appearance of the somatogenic receptor in the infant liver and is consistent with the GH-dependent nature of this form of binding protein in other species. The marked developmental changes in the circulating form of IGF-I in the perinatal lamb may be of functional significance and lead to altered biological availability of circulating IGF-I.

Circulating insulin-like growth factor-I and -II concentrations are not dependent on pituitary influences in the midgestation fetal sheep.

Studies were performed to investigate the possible role of pituitary factors on the regulation of circulating concentrations of insulin-like growth factor-I and -II in the midgestation sheep fetus. Four fetuses were decapitated at 59-64 days of gestation and fetal serum obtained at sacrifice at 90-102 days of gestation. Insulin-like growth factor-I and -II concentrations were similar in these samples to those from 6 control fetuses (83-102 days). A further 4 fetuses were studied following electrolytic destruction of the median eminence of the hypothalamus at 108-110 days of gestation. Four sham operated controls were also studied. Circulating growth hormone concentrations were markedly reduced (P less than 0.01) by destruction of the median eminence. However neither insulin-like growth factor-I nor -II levels differed from those of sham operated fetuses. We conclude that, in the midgestation fetal sheep, growth hormone is not essential for the maintenance of circulating concentrations of insulin-like growth factor-I or -II.

Insulin-like growth factor 1 (IGF-1) antler-stimulating hormone?

We have investigated the possibility that IGF-1 may play a role in the regulation of antler development. Plasma IGF-1 concentrations were measured throughout the first period of development of the pedicle and first antler of red deer (Cervus elaphus) to determine whether a relationship existed between growth of antler cartilage (velvet antler) and IGF-1. We report that plasma levels of IGF-1 are significantly elevated during the velvet antler growing phase relative to the other phases of pedicle and first antler development and a strong positive correlation exists between antler growth rate and circulating concentrations of IGF-1. As IGF-1 has been demonstrated to influence cartilage growth, we suggest that IGF-1 is a candidate as an antler stimulating hormone.

Circulating thyrotropin in the ovine fetus: evidence for pulsatile release and the effect of hypothermia in utero.

The secretion of thyrotropin (TSH) has been investigated in the chronically catheterized ovine fetus (term 145-150 days). Forty-two random plasma samples from 25 fetuses (86-149 days of gestation) were measured for TSH concentrations by radioimmunoassay. Plasma TSH concentrations were highest in the youngest fetuses [86-110 days, 3.9 +/- (SD) 5.5 microU/ml, n = 13]. Thereafter TSH concentrations declined to 0.4 +/- 0.6 microU/ml (n = 13, p less than 0.05) at 130-150 days of gestation. However, serial sampling at 15-20 min intervals for 180 min from 14 individual fetuses (91-139 days) showed that TSH was secreted in a markedly exaggerated pulsatile manner compared to that observed after birth. The mean amplitude of TSH pulses fell (p less than 0.005) from 5.9 +/- 8.1 microU/ml in the fetuses to 2.1 +/- 1.1 microU/ml in five neonatal lambs (6-22 days) and to 1.5 +/- 0.4 microU/ml in three adult nonpregnant ewes. The mean pulse frequency for the 14 fetuses was 0.7 +/- 0.3 pulses/h and was reduced (p less than 0.001) to 0.3 +/- 0.1 pulses/h in lambs and to 0.3 +/- 0.1 pulses/h in the ewes. In the neonate, hypothermia is a potent stimulus to TSH release. To examine the ontogeny of this response, the temperature of the fetus in utero (106-127 days of gestation) was lowered by circulating water (14-18 degrees C) at either a fast or slow rate through a coil placed either externally around the fetus or internally in the fetal esophagus and stomach.(ABSTRACT TRUNCATED AT 250 WORDS)

Parturition-related changes in insulin-like growth factors-I and -II in the perinatal lamb.

Specific radioligand assays for the two somatomedins, insulin-like growth factor (IGF)-I and IGF-II, have been used to study the ontogeny of somatomedin secretion in the perinatal lamb. Plasma samples were obtained from sheep fetuses from 54 days of gestation to term (147 days) and from neonatal lambs. All samples were first extracted in acid-ethanol to remove the somatomedin-binding proteins. Concentrations of IGF-I measured by radioimmunoassay were lower (P less than 0.01) in the fetus than in the adult sheep (0.96 +/- 0.17 (S.D.) units/ml, n = 11). Fetal IGF-I values rose (P less than 0.01) from 0.29 +/- 0.15 units/ml (n = 6) at 50-80 days to 0.79 +/- 0.18 units/ml (n = 13) at 140-150 days. While values were similar 0-2 days after birth, they rose (P less than 0.01) to 2.4 +/- 1.3 units/ml (n = 15) 3-7 days after birth. By 60 days they had fallen to adult values. In contrast, IGF-II levels measured by rat placental membrane radioreceptor assay were higher (P less than 0.001) in the fetus (2.71 +/- 1.06 units/ml, n = 18) than in the adult (1.0 +/- 0.17 units/ml) and showed no gestational trend between 50 and 140 days of gestation. Longitudinal studies showed a fall (P less than 0.01) in IGF-II values starting several days before birth. By 12 h after birth, IGF-II concentrations were similar to the adult and showed no subsequent postnatal change. These results demonstrate that IGF-I and IGF-II are not secreted in parallel in the perinatal lamb. There are major changes in the regulation of both IGF-I and IGF-II in relationship to birth. It is suggested that the high fetal IGF-II concentrations appear to be maintained by a stimulus withdrawn before birth. The postnatal rise in IGF-I may be related to the increase in hepatic somatogenic receptors at this age.