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Methanoaricia dendrobranchiata Blake (Polychaeta; Orbiniidae) occurs in large numbers in association with communities of the mussel Bathymodiolus childressi at hydrocarbon seeps on the Louisiana Slope of the Gulf of Mexico. Its microhabitat can be strongly hypoxic (oxygen is often undetectable) and sulfidic (sulfide concentrations can reach millimolar levels), which may seriously challenge aerobic metabolism. We describe a suite of adaptations to its low-oxygen environment. The worms are capable of regulating their rate of oxygen consumption down to partial pressures of approximately 870 Pa oxygen. This capability correlates with a large gill surface area, a small diffusion distance from sea water to blood, a very high hemoglobin oxygen-affinity (P50=27.8 Pa at 10°C and pH 7.6) and a Bohr effect that is pronounced at high oxygen saturations. When fully saturated, the hemoglobin binds sufficient oxygen for only 31 min of aerobic metabolism. However, these polychaetes can withstand extended periods of anoxia both in the absence and presence of 1 mmoll-1 sulfide (TL50=approx. 5.5 and 4 days, respectively).
- hypoxia
- anoxia-tolerance
- sulphide-tolerance
- functional properties
- cold seep
- oxygen consumption
- physiology
- haemoglobin
- orbiniid
- polychaete
- Methanoaricia dendrobranchiata
Metazoan communities thriving on local chemolithoautotrophic and methanotrophic primary production were discovered in 1984 by the Geochemical and Environmental Research Group (GERG) on the Louisiana Slope of the Gulf of Mexico (Kennicutt et al., 1985; Brooks et al., 1987). Extensive beds of the methanotrophic symbiont-containing mussel Bathymodiolus childressi occur at many sites in the area (Childress et al., 1986; MacDonald et al., 1990a, b; Nix et al., 1995). Methanoaricia dendrobranchiata Blake (Polychaeta; Orbiniidae) occurs with the mussels at most of the sites and is particularity abundant in the mussel bed around Brine Pool NR1 (Macdonald et al., 1990c), where it occurs either individually or in tight aggregations of as many as 150 worms (Smith et al., 2000; Hourdez et al., 2001). The worms often extend the anterior part of their body above the mussels into the overlying water (Fisher, 1996; Hourdez et al., 2001).
At Brine Pool NR1, the mussel bed rings the edge of the hypersaline pool on unconsolidated sediments saturated with brine. The oxygen concentration in water sampled between the mussels averaged 39 μmol l-1 and was often undetectable by gas chromatography (<5 μmol l-1; Smith et al., 2000). Although no sulfide could be detected in the brine pool itself, millimolar levels are present in the brine-saturated sediments under the outer edge of the mussel bed (Smith et al., 2000). The combination of sulfidic and hypoxic conditions presents a major respiratory challenge for active metazoans (Somero et al., 1989; Grieshaber and Völkel, 1998). Similar conditions characterize the microhabitats of many hydrothermal vent communities (Johnson et al., 1988; Fisher et al., 1988a, b; Sarrazin and Juniper, 1999).
Several studies on hydrothermal vent polychaetes have shown larger mass-specific gill surface areas and shorter diffusion distances between the environment and the body fluids than in related genera from other environments (Jouin and Gaill, 1990; Hourdez and Jouin-Toulmond, 1998). The gills of M. dendrobranchiata are also hypertrophied compared with those of other orbiniids and resemble those of the vent polychaetes (Hourdez et al., 2001). This characteristic and the short diffusion distance across the gill epithelia have been interpreted as adaptations for life in a hypoxic environment (Jouin and Gaill, 1990; Jouin-Toulmond et al., 1996; Hourdez and Jouin-Toulmond, 1998; Hourdez et al., 2001).
In addition to these anatomical adaptations, the respiratory pigments of the vent polychaetes and vestimentiferan species studied to date have very high oxygen affinities and strong Bohr effects that may facilitate oxygen uploading in the gills at low oxygen tensions as well as its release in the tissues (Arp et al., 1990; Toulmond et al., 1990; Hourdez et al., 1999). M. dendrobranchiata contains an extracellular hemoglobin in its blood vessels (Hourdez et al., 2001). In annelids, such extracellular hemoglobins are usually giant hexagonal bilayer (HBL) molecules, made up of 12 large subunits (dodecamers) composed of globin chains. These dodecamers are held together by polypeptide chains known as linkers (for a review, see Lamy et al., 1996).
We studied aspects of the respiratory physiology of M. dendrobranchiata from the cold seeps of the Gulf of Mexico. The relationship between oxygen consumption rate and environmental oxygen concentration was determined together with tolerance of anoxia in both the presence and absence of sulfide. We also determined the concentration of hemoglobin in the worms and describe the molecular and functional properties of the purified molecule. Finally, these findings are discussed in the context of the animal's habitat and behavior.
Materials and methods
Animal collection
Specimens of Methanoaricia dendrobranchiata Blake were collected together with mussels at the Brine Pool NR-1 site on the Louisiana Slope in the Gulf of Mexico (23°43′24″N, 91°16′30″W at 650 m depth; MacDonald et al., 1990c; Smith et al., 2000) in July 1998 using the scoop collector on the arm of the submersible Johnson Sea-Link I. The animals were placed in a temperature-insulated box on the submarine for recovery by the RV Edwin Link, where they were transferred to aquaria of chilled (8 °C) aerated sea water for between 1 and 7 days before use in oxygen consumption and anoxia experiments. There were no apparent adverse effects of maintenance at ambient pressure or normoxia on M. dendrobranchiata, which have been maintained alive in the laboratory, with mussels, for over a year.
Oxygen consumption rates
The oxygen consumption rates of individual worms were determined in water-jacketed, gas-tight chambers (Fisher et al., 1985) at 8.5±0.5°C using Clark-type oxygen probes (Orion 835 dissolved oxygen meter) (Fig. 1). Oxygen concentrations in the chamber were recorded every 5 min for 16-17 h. After the experiment, the worms were rapidly dried with paper tissues and frozen in cryovials for transport to Pennsylvania State University. To determine the wet mass of the worm, the cryovial was weighed before and after removing the worm, Mass-specific oxygen consumption rates were calculated for each 20-min interval (using five data points) and plotted against the mean oxygen concentrations for that time interval.
Anoxia and sulfide-tolerance
Worms were maintained anaerobically in water containing 0, 0.06 or 1 mmol l-1 sulfide to determine their tolerance to anoxia under these conditions. Oxygen-free sea water was obtained by bubbling with nitrogen for 30 min and then transferred to a nitrogen atmosphere in a glove bag in a cold room at 8 °C, where all subsequent manipulations were conducted. The absence of detectable oxygen was confirmed with a dissolved oxygen meter (Orion 835). Preweighed washed crystals of sodium sulfide were then added, and the sulfide concentrations were measured by gas chromatography (Childress et al., 1984). Individual worms were incubated separately in sealed 30 ml glass scintillation vials secondarily contained in a 500 ml gas-tight container filled with nitrogen-equilibrated water (to dampen minor temperature fluctuations and maintain anaerobic conditions). Eleven vials containing one worm each were incubated anaerobically under each experimental condition, and an additional 22 worms from the same collection were kept under normoxia in individual vials that were open to the atmosphere (controls). Survival was checked every 12 h until the ship reached port (7 days).
To evaluate the time corresponding to a 50 % survival rate (TL50), the following survival curve was fitted to the data: 1 where s is the percentage of survivors, t is time and b is a sigmoidicity coefficient. Iterations were repeated using the program DeltaGraph 4.5 until the parameters TL50 and b did not vary by more than 1 %.
Extraction and purification of hemoglobin
Because of the very small blood vessels, it was not possible to sample pure blood intravascularly. Approximately 75 worms from a single collection were batch-frozen 1 h after recovery for shipment to the laboratory and hemoglobin extraction. In the laboratory, the worms were homogenized with 100ml of extraction buffer (50 mmoll-1 Tris, pH8, 1 mmoll-1 EDTA and 1 μmoll-1 phenylmethanesulfonic fluoride, PMSF). The homogenate was centrifuged for 20 min at 20 000 g to remove the coarse debris. The bright red supernatant was then centrifuged at 100 000 g for 5 h to pellet the hemoglobin. The pellet containing the hemoglobin was resuspended in 1 ml of the extraction buffer, and the hemoglobin was purified by fast protein liquid chromatography (FPLC) on a Superose 6 column (10 mm×300 mm; Pharmacia Inc., 5-5000 kDa separation range) using 400 mmoll-1 NaCl, 2.95 mmoll-1, KCl, 32 mmoll-1 MgSO4, 11 mmoll-1 CaCl2 and 50 mmoll-1 Hepes at pH 7.0 as the elution buffer. The elution rate was typically 0.5 ml min-1, and the elutant was monitored at 280 nm. The molecular mass of the collected fractions was estimated using a sample of Riftia pachyptila blood, which contains four peaks of known molecular mass (Zal et al., 1996). The presence of hemoglobin in colored fractions was verified on the basis of peak absorptions near 414 nm.
Hemoglobin content
To assess the significance of the hemoglobin for oxygen storage, 61 worms ranging widely in size (0.08-1.95 g) were individually frozen on board ship. In the laboratory, they were weighed and homogenized individually, using a ground-glass tissue homogenizer, in 1 mmoll-1 Tris buffer, pH 8, containing 1 mmoll-1 EDTA and 1 μmoll-1 PMSF to prevent hydrolysis by proteases. The homogenate was centrifuged at 10 000 g for 15 min to remove cellular debris, and the supernatant was used for heme concentration measurements after appropriate dilution of the samples in the extraction buffer. Potassium ferricyanide solution (10 mmoll-1; 10 ml) was added to 2 ml of this diluted solution, which had been incubated for 5 min at room temperature to oxidize the hemoglobin completely to methemoglobin. Potassium cyanide (50 mmoll-1; 10 ml) was then added to obtain the cyan-methemoglobin derivative, which was quantified on the basis of the absorbance of the solution at 540 nm and the most appropriate published extinction coefficient of 11.0l mmol-1 cm-1 (Van Assendelft, 1970).
Negative-stain transmission electron microscopy of purified, whole hemoglobin molecules
Images of negatively stained M. dendrobranchiata hemoglobin molecules were recorded by transmission electron microscopy to assess their molecular organization following a method modified from Valentine et al. (1968). Diluted purified hemoglobin (3 μl; 0.037 mmoll-1 heme in 100 mmoll-1 Tris buffer at pH 6.85) was applied to a very thin (10-30 nm) carbon foil supported on a standard 3.05 mm copper electron microscopy grid. After allowing 1 min for the hemoglobin molecules to adhere to the carbon foil, 6μ l of 2 % (m/v) uranyl acetate solution was applied to stain the molecules. After 10 s, the grid with the stained sample was blotted with Whatman filter paper, leaving only a very thin wet layer on the carbon foil, which was allowed to dry in air. The negatively stained specimens were examined using a Philips CM120 electron microscope at 100 keV, and images were recorded under minimum-dose conditions (Kenney et al., 1995) at 40 000× magnification.
The transmission electron microscopy images of the negatively stained hemoglobin molecules were analyzed to reveal the symmetry of their macromolecular structure. The images were digitized, and individual hemoglobin molecules were selected and centered by reference free alignment using the program SPIDER (Frank et al., 1996). The rotational frequency analysis of SPIDER-processed images was performed using the Medical Research Council image-processing programs (Crowther et al., 1996), and the rotational power spectra of the images were calculated.
Electro-spray ionization mass spectrometry
Electro-spray ionization mass spectrometry (ESI-MS) was used under denaturing conditions to determine the molecular masses of the chains and covalently bound subunits present in the native hemoglobin and in the 210 kDa fraction (see Results). Three different treatments were applied to this hemoglobin sample to analyze its structure, as described previously (Green et al., 1996, 1999): (i) carboxyamidomethylation (Cam) to determine the number of free cysteines, (ii) reduction with dithiothreitol (DTT) to determine the composition of dimeric species and the reduced masses of the chains, and (iii) reduction plus Cam (reduced/Cam), to determine the total number of cysteine residues in each chain. The DTT reduction was carried out at a concentration of 5 mmoll-1 for 2 min at 20 °C. Briefly, samples were introduced at a rate of 5 μl min-1 into the electrospray source of a Quattro LC mass spectrometer (Micromass UK Ltd). They were analyzed at a concentration of 0.25-0.5 mg ml-1 in 1:1 (by volume) acetonitrile:water containing 0.2 % formic acid. The scan range was m/z 600-2500, and the cone voltage (counter electrode to skimmer voltage) ramp was from 30 V at m/z 600 to 100 V at m/z 2500 (m is mass; z is charge). Mass scale calibration employed the multiply charged series of ions from horse heart myoglobin (Mr 16 951.5 Da; Sigma, M-1882). Data were processed using the Maximum Entropy (MaxEnt)-based software supplied with the instruments. Molecular masses are based on the atomic masses of the elements: C=12.011, H=1.00794, N=14.00674, O=15.9994 and S=32.066 (IUPAC).
Oxygen equilibrium curves
Equilibrium curves were obtained with a diffusion chamber apparatus (Sick and Gersonde, 1969) modified as described previously (Weber, 1981). Briefly, small (4 μl) samples of purified hemoglobin were equilibrated with pure (>99.998 %) N2 and O2 and mixtures of these gases and air prepared using Wösthoff pumps. The pH of the hemoglobin solutions was varied by adding 6 μl of 1 moll-1 Bis-Tris Propane buffer of a range of pH values to 100 μl aliquots of the sample, and water was added to bring the total volume to 120 μl. The pH was measured in duplicate with a blood gas analyzer (BMS2, Radiometer) on 50 μl subsamples. Oxygen equilibrium and pH measurements were carried out at 10, 20 and 30 °C (±0.1 °C).
Values of P50 (the PO2 at which the hemoglobin is half-saturated with oxygen) and n50 (cooperativity at P50) were derived from linear regression Hill plots of log[Y/(1-Y)]=f(logP02) for S (saturation values) of 30-70 %. The Bohr factor (Φ) was calculated as Φ=ΔlogP50/ΔpH, and the apparent heat of oxygenation was calculated asΔ Hobs=2.303RlogP50/[1/T1)-(1/T2)], where R is the gas constant and T1 and T2 are the absolute temperatures. The value thus obtained (ΔHobs) comprises the intrinsic heat of oxygenation (ΔHintr), the heat of solution of O2 (ΔHsol, approximately 13 kJ mol-1) and contributions from oxygenation-linked processes such as the heats of reaction with protons and other effectors. In the absence of oxygenation-linked binding of protons (when Φ=0) and other allosteric effectors,Δ Hintr=ΔHobs-ΔHsol (see Wyman et al., 1977).
Values reported as means ± standard deviation (S.D.) throughout this manuscript.
Rates of oxygen consumption
Fig. 2A shows the reduction in oxygen concentration from 4 to 0.1 mgl-1 (partial pressure decreasing from 10 000 to 260 Pa) recorded in the incubation chamber for a worm 1.24 g in wet mass. The calculated oxygen consumption rates for this experiment are shown in Fig. 2B. The oxygen consumption rate is relatively constant (0.66±0.08 μl O2 min-1 g-1 wetmass, N=75 points) for oxygen levels between normoxia and 0.5 mg l-1 (PO2=1300 Pa). Below the critical value of 0.5 mg l-1 (PO2=1300 Pa), oxygen consumption rate decreases gradually and ceases at a concentration of 0.1 mg l-1 (PO2=260 Pa). The small variations in the mean oxygen consumption rate at the beginning of the incubation are interpreted as periods of activity and rest.
A logistic curve was found to describe the data best on the basis of theχ 2-value: 2 where [O2] is in mg l-1 and c is the value of [O2] at which the oxygen uptake rate (V̇2) is zero, and a and b are constants. When PO2 tends to infinity, V̇O2 tends to a/b, which is the maximum oxygen consumption rate (V̇O2max). The constants a and b describe the initial slope and, thereby, how fast the plateau is reached. For the worm depicted in Fig. 2B, a=6.146, b=8.705 ml μl-1 O2 and c=0.106μ l O2 ml-1. The maximum oxygen consumption rate, V̇O2max, is then 0.706 μl O2 min-1 g-1 wet mass.
Separate linear regressions for points below 0.4 mg O2 l-1 and points above 1.5 mg O2 l-1 allowed us to estimate the critical pressure (the intercept of the two lines). For the worm in Fig. 2, the intercept gives a critical pressure Pc of 0.27 mg O2 l-1 (700 Pa).
These measurements were made for seven worms ranging from 0.64 to 1.87 g in wet mass. For two of these experiments, the oxygen concentration did not fall below 1 mg l-1. In these cases, we used the linear portion of the oxygen concentration as a function of time curve (Fig. 2A) to calculate the oxygen consumption rate. For the five other experiments, the shape of the curve was always the same, and the χ2-value supported the use of the above equation for each data set (P<0.01). The Pc values ranged from 0.25 to 0.38 mg O2l-1 in the five experiments (PO2=650-990 Pa) and oxygen consumption ceased at 0.09-0.11 mg O2l-1 (PO2=230-280 Pa).
Fig. 3 shows the relationship between respiratory rate and animal wet mass. The oxygen consumption rate follows a power function trend described by the equation: 3 where V̇O2 is the oxygen consumption rate (in μl O2 min-1 g-1) and M is the wet mass of the worm (in g). This curve fits the data with a correlation coefficient r2 of 0.457 (P<0.1). This lack of significance is not surprising because the wet mass does not span an order of magnitude. The mean oxygen consumption rate for all the experiments is 0.479±0.160 μl O2 min-1 g-1.
Anoxia and sulfide-tolerance
Fig. 4 shows the percentage survival as a function of time under normoxia, anoxia, anoxia + 60μ moll-1 sulfide and anoxia + 1 mmoll-1 sulfide. None of the normoxic (control) animals died during the experiment, while only a single worm survived for 7 days under anoxia. The mortality curves are sigmoidal. Curve-fitting yield TL50 values (time to reach 50 % survival) of 99.1, 119.6 and 133.5h, respectively, for anoxia + 1 mmoll-1 sulfide, anoxia + 60 μmoll-1 sulfide and anoxia without sulfide. Analysis of covariance (ANCOVA) of the linearized data indicated that the effect of the treatment was highly significant, and pairwise comparisons confirmed that all TL50 values were significantly different (P<0.01).
Hemoglobin content
Fig. 5 shows the hemoglobin content as a function of wet mass for the 61 worms. The masses ranged from 0.008 to 1.95 g and the hemoglobin (Hb) contents from 143 to 373 nmol Hb g-1 wet mass. The individual mass-specific hemoglobin content (Q) increases with the size, reaching a plateau at approximately 250 nmol g-1 wet mass. The mean mass-specific hemoglobin content is 237±52 nmol Hb g-1 wet mass (N=61). The following equation fits the data, with a correlation coefficient r=0.388 (N=61, P<0.01): 4 Assuming complete saturation of hemoglobin and a constant oxygen consumption rate, the temporal significance of the oxygen store (A; in minutes) can be derived as Q/V̇O2 (equations 4 and 3), which yields: 5 For a 1 g worm, the fully saturated hemoglobin would then represent an oxygen store allowing 31 min of aerobic metabolism. A corresponding calculation using the mean oxygen consumption rate instead of equation 3 gives a very similar result (32 min).
Purification of the hemoglobin
The extracellular hemoglobin was purified by FPLC. Absorbances at 280 nm (which characterize proteins) and 414 nm (the Soret band, which characterizes hemoglobins) show two hemoglobin fractions with masses of approximately 3.5×106 Da and 210 kDa. The second fraction, which was very small (elution profile not shown), is referred to as `dodecamers' (D in the figures) and is considered to be a dissociation product of the first fraction of whole hemoglobin molecules (W-Hb) (for an example, see Zal et al., 1996). Support for this designation is presented below.
Image analysis of transmission electron microscopy pictures of whole hemoglobin molecules
The transmission electron microscopy pictures of the whole hemoglobin molecules (Fig. 6A,B) showed a strong peak at the sixth rotational harmonic, indicating a sixfold rotation symmetry (Fig. 6C). This information was used to produce a rotationally Fourier-filtered image. The result is a typical top (sixfold axis) view of a hexagonal bilayer (HBL) (Fig. 6D).
Structure of the hemoglobin
Under denaturing conditions, ESI-MS of the native hemoglobin revealed three groups of peaks with molecular masses of approximately 16, 32 and 49 kDa. In the first group, four peaks can be distinguished: a1, a2, a3 and a4, with masses of 15 824.8, 15 835.1, 15 864.5 and 16 255.4±1.5 Da and relative intensities of 0.40:0.34:0.19:0.17, respectively (N=5) (Fig. 7A; Table 1). A single peak (Dm) occurred at 31 972.6±3.0 Da (N=5), while the third group consisted of four peaks (LD1-LD4) between 48 715.5 and 48 804.9 Da (Fig. 7, inset in A). Careful examination of the ESI spectra from several preparations of W-Hb analyzed under various conditions failed to reveal the presence of components that could correspond to trimers or tetramers of globin chains. ESI-MS on the 210 kDa fraction revealed peaks corresponding to a1-a4 and Dm only (no peaks of higher mass were seen).
Under mild reducing conditions (1 and 2 min in the presence of 5 mmol l-1 DTT at 22 °C) (Fig. 7B), the masses of a1-a3 remained unchanged, while the mass of a4 decreased by 119.5 Da, suggesting that this chain is cysteinylated. Concomitantly, the intensity of the Dm peak decreased and the group of peaks around 49 kDa disappeared. In addition, several new peaks appeared: b, c and d at 15 373.4, 16 600.6 and 17 172.9 Da, respectively (±1.5 Da, N=6), and L1, L2, L3, L4 and L5 at 24 358.7, 24 379.0, 24 404.2, 26 279.3 and 27 687.1 Da, respectively (±3.0 Da, N=4). These results imply that Dm is a disulphide-bonded dimer composed of monomers b and c, since b+c-2H=31 972.0, which is the mass determined for Dm in the unreduced hemoglobin (Green et al., 1996). Furthermore, twice the masses of L1, L2 and L3 minus 2H are within experimental error of the masses of LD1, LD2 and LD4 (Fig. 7B), implying that L1-L3 exist as disulfide-linked dimers in the native hemoglobin. L1-L5 have masses that are typical of linker chains. However, no peaks were observed in the native hemoglobin that could correspond to dimers of L4 and L5. Moreover, while L1-L3 remained present upon reduction, L4 and L5 disappeared, with the concomitant appearance of several relatively minor peaks in the range 18-23 kDa. Peak d cannot be assigned to a higher-mass component in the W-Hb. It is possible that it is one of the components in a dimer or higher-order multimer that was not detected in the denatured W-Hb. It could also correspond to a fragment of a linker, made more fragile after reduction.
After carboxyamidomethylation (Cam) with iodoacetamide, the masses of a1-a3 remained unchanged, indicating that they contain no free cysteine. However, the mass of Dm increased by 2 Cam units, implying that it contains two free cysteine residues. The total number of cysteine residues in each chain and the reduced masses of the chains were determined from the reduced/Cam hemoglobin, as shown in Table 1 and Fig. 7C. Strong support for LD1-LD4 being dimeric forms of L1-L3 is provided by the reduced/Cam results as follows. L1-L3 have masses typical of linker chains and appear to exist as dimers in the native hemoglobin. The peak at 48 715.5 Da is probably a dimer made of two L1 chains (2L1-22H=48 710.2 Da) and the peak at 48 755.1 Da could be a dimer made of two L2 chains, or one L1 chain and one L3 chain (2L2-22H=48 757.8 Da and L1+L3-22H=48 757.1 Da, respectively). The peak at 48 777.4 Da is probably a dimer of L2 and L3 (L2+L3-22H=48 780.9 Da) and, finally, the peak at 48 804.9 Da is probably a dimer of two L3 chains (2L3-22H=48 804.0 Da). All these calculated masses are well within the experimental error for the measured masses of the peaks.
Functional properties of the hemoglobin
In Fig. 8, the effects of pH on the affinity (P50) and cooperativity (n50) of the intact hemoglobin (W-Hb) and dodecamers (D) at 10, 20 and 30 °C are shown. W-Hb and D exhibit the same oxygen affinities at pH 7.6 for 10 °C (P50=27.8 Pa), at pH 7.4 for 20 °C (P50=82 Pa) and at pH 7.1 for 30 °C (P50=271 Pa). At lower pH values, D shows progressively higher affinities than W-Hb, whereas the reverse applies at higher pH.
Both hemoglobins express Bohr effects in the range pH 6.7-7.5. The Bohr factors in W-Hb vary inversely with temperature Φ=-0.48, -0.44 and -0.35 at 10, 20 and 30 °C, respectively, and exceed those in D (Φ=-0.11, -0.13 and -0.11, respectively).
W-Hb and D also differ in cooperativity coefficients, n50. In W-Hb, the coefficient is higher, with a maximum at pH 7.5, and appears to vary inversely with temperature (n50=2.5 at 10 °C, n50=2.0 at 20° C and n50=1.8 at 30 °C). In D the value of n50 (1.2) is almost independent of pH.
Extended Hill plots of oxygen equilibria of the intact hemoglobin molecules (W-Hb) at pH 6.77 and 7.60 are shown in Fig. 9. The allosteric and other parameters derived from analysis of the equilibria in terms of the two-state Monod—Wyman—Changeux (MWC) allosteric model (as described by Weber et al., 1995) are given in Table 2. The correspondence between the cooperativity value at half-saturation (n50) and the maximum cooperativity (nmax) and between P50 and the median oxygen affinity Pm (Table 2) reflects symmetrical oxygen-binding curves and permits the use of P50 values to analyse allosteric interactions. The control mechanism of the Bohr effect in this hemoglobin is primarily an increase in the oxygen-binding affinity of the hemoglobin in the oxygenated R (`relaxed') state (KR) as pH rises, while the affinity (KT) of the deoxygenated T (`tense') state remains relatively constant (Table 2). This raises the free energy of cooperativity (▵G) (Table 2) and the KT/KR ratio with increasing pH. The analyses indicate the presence of 3-5 interacting oxygen-binding sites (Q) in the functional subunits of the intact hemoglobin molecules.
The oxygen affinities of both hemoglobins show strong temperature-dependence (Figs 8, 10) at pH 6.8 and 7.6. The temperature effect is smaller in W-Hb than in D (▵Hobs=-58 and -66 kJ mol-1 for W-Hb at pH 6.8 and 7.6, respectively, and -77 kJ mol-1 for D at both pH values).
In contrast to the marked effect of 100 mmol l-1 Ca2+, which increases the oxygen affinity of W-Hb (▵logP50=0.28) (Fig. 11), D lacks sensitivity to Ca2+.
Methanoaricia dendrobranchiata is found in high densities at cold-seep sites often in association with mussels on the Upper Louisiana Slope (Blake, 2000; Smith et al., 2000). While this environment is characterized by a high local production (a strong trophic attractor), it provides harsh chemical conditions for the survival of metazoans (low oxygen and high sulfide concentrations). To survive and thrive under these conditions, the polychaetes must be adapted to take up oxygen at low environmental partial pressures and to tolerate anoxia and sulfide exposure.
Hemoglobin structure
Intact Methanoaricia dendrobranchiata hemoglobin has a native mass and appearance (as seen in transmission electron microscopy) typical of giant HBL annelid hemoglobins (for a review, see Lamy et al., 1996). On the basis of the masses of its dissociation products, it contains both globin and linker chains, as expected for HBL hemoglobins (for a review, see Lamy et al., 1996). There appear to be five linkers, although three of them (L1-L3) behave quite differently from the other two (L4 and L5) when reduced. Linkers L1-L3 possess 11 cysteine residues, 10 involved in intra-chain disulfide bonds and one involved in a disulfide bond between two linker chains (Table 1). All the linkers are dimeric, as in Arenicola marina (Zal et al., 1997), and are stable as monomers under reducing conditions. However, the other two linkers (L4 and L5), if indeed they are linkers, only appeared as monomers under mild reducing conditions and were never observed as either monomers or dimers in the native hemoglobin. Upon further reduction, they disappeared, and a number of components in the mass range 18-23 kDa were observed; these then disappeared after a longer reduction time. At this time, monomeric L1-L3 were still present, suggesting that L4 and L5 are significantly less stable than L1-L3. Both monomeric (a1-a4) and dimeric (b and c) globins are present in M. dendrobranchiata hemoglobin. The globin dimers were unexpected because dimers have previously been considered to be typical of hirudinean (leech) and vestimentiferan hemoglobins in contrast to all other studied polychaetes, which possess globin trimers (for a review, see Lamy et al., 1996). This is also the first record of an annelid globin found to be cysteinylated. The origin of d remains unknown: it is probably involved in higher aggregation states, not detected by ESI-MS, or corresponds to a fragment of a linker, probably L4 or L5. A similar situation, where some subunits do not seem to ionize properly under non-reducing conditions, was encountered in Eudistylia vancouverii chlorocruorin (Green et al., 1998).
Functional properties
Both intact molecules and dodecamers exhibit similar and very high oxygen affinities but differ in other oxygen-binding characteristics (cooperativity, Bohr effect, sensitivity to Ca2+ and sensitivity to temperature). Their affinities are among the highest reported in annelids and probably represent adaptations to the animal's low-oxygen environment (Toulmond et al., 1990; Weber, 1978a, 1980; Toulmond, 1992; Hourdez et al., 1999, 2000; Weber and Vinogradov, 2001). The increase in KR with rising pH (Fig. 9) is opposite to the situation for human hemoglobin (in which the Bohr effect is due to variation in KT; Tyuma et al., 1973), but similar to that in the polychaete Arenicola marina, where it increases the Bohr factor at high oxygen saturation and favors oxygen saturation of blood entering the relatively alkaline gills (Weber, 1980; Toulmond, 1992).
The Bohr effect in the intact molecules (W-Hb) is similar to that of most extracellular hemoglobins (Weber, 1978a) but is greatly reduced in the dodecamer, as previously reported for other annelids (for a review, see Lamy et al., 1996).
In contrast to similar blood Ca2+ concentrations among vestimentiferans and other annelids studied (7-13 mmol l-1), blood Mg2+ levels vary widely among species (from 2 to 44 mmol l-1) (Oglesby, 1978; Sanders and Childress, 1991) and thus may affect the functional properties of annelid hemoglobins in vivo. Ca2+ increases the oxygen affinity of intact M. dendrobranchiata hemoglobin but has no effect on the dodecamers, suggesting that the effect of group IIA cations such as Ca2+ and Mg2+ involves the linkers. This agrees with other studies on HBL hemoglobin and most studies of dodecamers, with the exception of Lumbricus terrestris hemoglobin dodecamers, which are sensitive to group IIA cations (for a review, see Lamy et al., 1996). Lumbricus terrestris hemoglobin has been shown to have Ca2+-binding sites with a variety of affinities that mainly involve linkers (Kuchumov et al., 2000). The low-affinity sites are likely to be involved in the effects of group IIA cations on the structure and function of hemoglobin (Kuchumov et al., 2000).
The cooperativity coefficients of M. dendrobranchiata hemoglobin are within the range previously reported for HBL hemoglobins (Weber and Vinogradov, 2001). The value for the intact molecule (2.5) is approximately twice that of the dodecamer (1.2), as seen in Lumbricus terrestris hemoglobin (n≈5 and n≈2, respectively, at pH 7.2; Krebs et al., 1996). The difference suggests a participation of the linker chains in heterotropic interactions in the molecule, as in other HBL hemoglobins (for a review, see Lamy et al., 1996).
The temperature-sensitivity of the intact hemoglobin (▵H=-66 kJ mol-1 at pH 7.6) is similar to that of the polychete Abarenicola clarapedii, another annelid that lives in a fairly stenothermal environment, but greater than that of Arenicola marina (▵H=-22 kJ mol-1; Weber, 1972), which experiences larger temperature variations in its natural habitat. Given that dissociation of Bohr protons is endothermic and reduces the net heat liberated upon oxygenation of the hemes, the lower temperature effect in the whole molecules than in the dodecamers (Fig. 10) accords with the larger Bohr effect in the former. Analogously, ▵Hobs of the intact molecules is lower at pH 6.8 than at pH 7.6, where the Bohr effect is smaller (see Fig. 8).
Adaptations to the environment
The high affinity and strong Bohr effect observed in the vascular hemoglobin probably have adaptive significance in a low-oxygen environment, favoring both the binding of oxygen at the respiratory surfaces and its release in the tissues, where pH decreases as a result of the production of acid excretory products.
The oxygen consumption rates of M. dendrobranchiata are within the lower end of the normal range of respiratory rates reported for polychaetes (Childress and Mickel, 1985; Weber, 1978b). The worms are very good oxyregulators and are able to maintain a constant metabolic rate as environmental concentrations fall to 0.3 mg O2 1-1 (where Po2=870 Pa) (Fig. 2). At lower partial pressures, the oxygen consumption rate drops sharply, and oxygen consumption ceases below 0.1 mg O2 1-1 (Po2=260 Pa).
Strong oxyregulation extending to very low oxygen concentrations is normally correlated with one or more of the following adaptations: (i) a large gas-exchange surface, (ii) a short diffusion distance between the environment and the body fluids to facilitate gas diffusion, (iii) the presence of a respiratory pigment to increase the oxygen-carrying capacity of the circulating body fluid(s), (iv) a high oxygen affinity of the respiratory pigments to increase oxygen loading at low oxygen tension, (v) a high ventilatory rate to renew the medium as it becomes depleted of oxygen, and (vi) a high circulatory rate. M. dendrobranchiata have a large gill surface area and reduced diffusion distances across their respiratory surface as a result of intra-epidermal blood vessels in the gills (Hourdez et al., 2001). Furthermore, the worms are capable of greatly extending their gills when exposed to hypoxic and anoxic conditions (S. H., personal observation). The presence of cilia on the surface of the gills (Hourdez et al., 2001) facilitates renewal of the diffusion-limited water layer. These worms do not ventilate their gills, and no information is available about ciliary movement under hypoxia. However, mussels from their natural habitat can create strong, small-scale currents.
M. dendrobranchiata hemoglobin has a very high oxygen affinity under physiological conditions; an oxygen partial pressure of 100 Pa, which is far below Pc, saturates the hemoglobin to 96 % at 10 °C in vitro. However, the oxygen consumption rate of M. dendrobranchiata decreases when the oxygen partial pressure falls below 870 Pa, probably as a result of diffusion-limitation driven by the decreased oxygen concentration gradient across their gills. However, this value (870 Pa) is low, indicating that the high oxygen affinity partially offsets the lack of ventilatory capability. The low Pc value reflects a high degree of oxyregulation, which is similar to that in Riftia pachyptila but slightly greater than those in the crab Bythograea thermydron and the clam Calyptogena magnifica, all from deep-sea hydrothermal-vent habitats (Childress and Fisher, 1992). Many deep-sea hydrothermal-vent and cold-seep microhabitats are characterized by low levels of dissolved oxygen, and endemic animals often show similar capacities to oxyregulate, delaying the onset of anaerobic metabolism.
Sulfide and anoxia-tolerance
Although the high mass-specific gill surface area favors oxygen uptake, it cannot selectively favor the entry of oxygen without permitting the inward diffusion of potentially harmful sulfide (Somero et al., 1989), which often occurs at substantial concentrations in the mussel beds (Smith et al., 2000; Nix et al., 1995). Sulfide further compromises respiration in most animals by inhibiting respiration at the level of cytochrome c oxidase and hinders binding of oxygen to most hemoglobins (Somero et al., 1989). The sulfide-tolerance under anoxia of these worms compares with that of Marenzelleria cf. wireni and is the highest found among the species studied to date (Fig. 12; Groenendaal, 1980; Schiedeck et al., 1997; Gamenick et al., 1998). The hemoglobin-bound oxygen store is not enough to sustain aerobic metabolism for more than 31 min. However, worms faced with prolonged anoxia switch to anaerobiosis and can survive for days even in the presence of sulfide.
Cold seeps, like hydrothermal vents, are habitats characterized by very high productivity and biomass compared with the generally nutrient-poor deep sea. The high level of endemism and low species-richness of these communities suggest that these environments are also physiologically challenging and that they require specific adaptations. We suggest that the ability of Methanoaricia dendrobranchiata to colonize their extreme environment is due at least in part to the adaptations reported here, such as their capacity to oxyregulate, which is due to the presence of a high-affinity hemoglobin, and their tolerance to anoxia and sulfide. These adaptations allow the worm to forage in the sulfidic, organic-rich sediments between and below the mussels and to exploit a habitat with little or no competition and limited predation (MacAvoy et al., 2002).
This work was supported primarily by subcontract L100094 to the Mineral Management Service project RF-6899 and the Minerals Management Service, Gulf of Mexico Regional OCS Office, through contract number 1435-01-96-CT30813, and the Danish Natural Science Research Council. We thank Anny Bang, Århus, Denmark, for technical assistance and Catherine Venien-Bryan, Laboratory of Molecular Biophysics, Oxford, UK, for the use of the electron microscope facilities. This work would not have been possible without the expert assistance of the captain and crew of R/V Edwin Link and the pilot and crew of the DSRV Johnson Sea-Link.
- © The Company of Biologists Limited 2002


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"What is a Spiritual Hospital? by Emma Bragdon"
Painting in Main Assembly Hall; Photo by Karen Leffler www.spiritfotos.com
I was sitting in the Blessing Room at John of God's sanctuary, the Casa de Dom Inacio, preparing for the day's "work" of meditation and prayer. The Brazilian man sharing my bench was intensely concentrated, reading a 'Prayer of Compassion' in Portuguese. He kindly offered me a copy of the folder he was reading. It was titled, "Hospital Espiritual do Mundo" (Spiritual Hospital of the World) with the subtitle, "Casa de Dom Inacio de Loyola" (The House of Saint Ignatius of Loyola) under which was written, "Medium: Joao de Deus" (The Medium: John of God).
I realized, we don't have "Spiritual Hospitals" in the USA and Europe. At least, I have never heard of any. In fact, I don't know of any other such specialized hospital in any part of the world.
We have Ear, Nose and Throat Hospitals for people with problems in these areas. We have Cancer Hospitals for people who want to get rid of cancer, and Mental Hospitals for people who want to resolve psychological problems.
Is a Spiritual Hospital designed to help a person heal a problem in his or her spirit? Do we come here because our spirits are sick and our spirits need medication, bandaging, therapy or surgery?
No. Our spirits are eternal, never become ill, and never die. It is the small self, the ego, the Soul (after its excursions in and out of the body through lifetimes) and our emotional reaction to life that become burdened and out of balance, leading to confusion, fragmentation and unhappiness. Thus, the small self loses sight of and disconnects from the essence of spirit…infinite love and wisdom.
At a Spiritual Hospital we come to heal from such disconnection, to connect fully to our own spirit, aka the Higher Self. We come also to connect with the Transcendent, that which is much larger than ourselves, whom some call God—or the great Choreographer. We come to refresh ourselves in the river of Infinite Love that is the source of all religions, and thus experience the Oneness that goes beyond religious and cultural differences.
In a Medical Conference in 2008 Amrit Goswami, PhD (Physics) revealed that when a person reconnects with the Transcendent, and becomes absorbed in that spiritual experience, then he or she, as a result, can have an immediate extraordinary healing. (Not "WILL" have, but "CAN" have.) Spontaneous remission of cancer is just one example of what may happen. To repeat, the main ingredient is to surrender fully to the experience of the Transcendent.
Many visitors to the Casa de Dom Inacio do have spontaneous healing of the physical body—probably because they connect with the Transcendent so fully in the "Current". In the Current rooms dedicated to prayer and meditation we are asked to close our eyes, and not cross arms or legs so that the stream of good feelings stimulated by prayer can flow through us, joining each of us in an unbroken chain of positive intention and energy. We sit for 2,3,4 even 5 hours at a stretch. Those who sit in the current rooms often report having contact with Jesus, Mary, Angels, Saints and other supra-conscious beings with pure Souls who live in the realm of spirit and do not have physical bodies.
"Surrendering to the Transcendent is an essential step. It opens up a portal for Grace and more resources to come through. It definitely can shift one's attention from more egoic identification to a vaster sense of God as who one is. However, it seems that the karma and the momentum of habits in one's life continue to demand attention and healing, which may involve the need to change one's behavior and/or way of thinking. That can often take a long time, so complete healing is not instantaneous.
The Current has been an incredible way not only to experience this Great Vast Love but also for that Love to penetrate any areas where there is still that sense of separation and the emotional pain and trauma within that sense of separation. That to me is where healing happens. I also think healing happens as a result of direct intervention from the non-physical assistance that is very specific and works on different levels."—a member of the Casa community
The chain of connection in the current certainly explains why most visitors report feeling happy here, and often say "The Casa is my spiritual home." It is set up to bring us back to our innermost selves, our true home where we sense our connection to our extended spiritual family, which includes ascended masters.
On a practical level, guided meditations given in the first current room in English and Portuguese facilitate the special work of inner transformation. Current room leaders remind us to forgive, to make amends to those we have hurt, to remember to appreciate life, to treat others in a charitable way, and not only recall our mission for this life, but re-new our commitment to it. This guidance helps us to let go and let the energy we call "God" move through us, unobstructed.
John of God as the Casa medium, medical intuitive, and psychic surgeon facilitates contact with Spirit in other ways. When we stand in front of him for a consultation John is incorporating a being with a very evolved level of consciousness and compassion. As this "Entity" he has the eyes to see though us-- body, mind, and soul-- and suggest treatment. He consults, free of charge, with all visitors who want to see him three days a week and he stays those days until the last person is seen. He sometimes does physical interventions on those that request it; more often his treatments involve "psychic surgery" in which benevolent spirits perform interventions that positively effect healing on the physical as well as subtle bodies. This often includes helping disincarnate entities who come to the Casa for help as well. John is a master of "Energy Medicine", and the Casa is uniquely prepared to assist both disincarnates and incarnate beings who are ready to evolve spiritually.
One day, unexpectedly, I had a stroke. The left side of my body became paralyzed and I couldn't speak. My husband and I prayed to John of God and the Entities of the Casa to help. That night, I sensed an invisible being doing acupuncture up and down my left arm. I felt needles going in and out of my body—but no physical person was there. As this happened, the sensations of the left side of my body began to return. By the morning, I was a bit weak, and needed rest, but I was no longer paralyzed, and my ability to think and talk had returned.—a hotel manager in the Casa community
No matter how phenomenal the healing, John of God always says, "I do not heal, God is the one who heals." His lifelong dedication to be of service to others healing through mediating with Higher Forces is a model of humility, compassion and generosity.
Photo by Karen Leffler www.spiritfotos.com
Thus, the Casa, as a spiritual hospital, restores us to our eternal Spirit, our Higher Self and our guardian spirits. We are given the opportunity for deeper connection with spirituality and the discarnate beings that can help humankind. We are freed of negative thought forms and entities that have interfered with our ability to connect with the purest aspects of our spiritual nature. We are allowed to identify more intensely with the Spiritual self that we are, that may have been dormant throughout lifetimes. We remember that at our core, we are Love and have access to profound Truth and wisdom through the guidance of benevolent Spirits who are with us to help us.
"My intuition said I must come here for two months, so I came. Every day I sit in the current I feel new sensations in my heart. Actually it pulsates and many times it feels like it is on fire. Always, there is tremendous movement of energy. I often feel overwhelmed. Friday tears were streaming down my face all morning. But, I know this is important because my heart is opening. I am feeling my real feelings, and letting go of what other people told me I should feel and think. I am also sensing what my true direction in life is—what will give me real joy to do. I am so glad I came."—a 30 year old man, visiting the Casa
Along with the Current, the presence of Higher Beings, and John of God, the community of the Casa welcomes visitors with good food, blessed water and soup, a sacred waterfall for purification, sunshine, and laughter with the town comedienne, Grainne 'McEntity' (McEntee). Visitors relax in simple, clean Inns that function as recovery wards for those resting after surgery. Amble through town and you enjoy exotic birds' singing, extraordinary flowers, gentle horses that walk freely in the street, and more than one ice cream shop. A way to say good-bye after a chat is, "Fica com Deus", i.e., "Be with God". These elements of life, although not absolutely necessary, certainly support the work of the spiritual hospital.
Grainne 'McEntity' (McEntee) the town comedienne, practicing "Ascension". Photo: Emma Bragdon
Given the success of the Casa de Dom Inacio in Brazil—that participants are healing from cancer and all manner of physical and mental diseases—we could consider the Casa as a model to use elsewhere. After all, it is called, "Spiritual Hospital of the World". That doesn't mean it has to be the only one. We can bring components of the Casa back home and see what fits the needs of our own communities. We can't find a John of God anywhere else, but we can find good medical intuitives and energy workers who are capable of assisting others in recovering health.
God knows, our fractured world needs the healing and positive transformation that occurs in "spiritual hospitals". Although there are dedicated health professionals who care about their patients, most of the hospitals in the USA are managing disease for profit. That system is prohibitively expensive and excludes those who can't afford it. The Casa de Dom Inacio is teaching us how to have wellness for free for everyone, and the healthcare comes with liberal doses of spiritually enlightening experiences.
Certainly, one aspect of John of God's mission is to demonstrate how a good spiritual hospital functions in Brazil. It is up to us to seek contact with the inspired beings of Light who can help us seed such hospitals where they will be welcomed in other parts of the world. Both the Light-beings of the Spiritual realm and the Light-workers of the physical realm will be working together to create and maintain this new kind of hospital.
Emma Bragdon, PhD. (EmmaBragdon.com) leads groups to the Casa de Dom Inacio several times a year. She is the Director of the Foundation for Energy Therapies, a charitable educational and research organization. She has published a book about the Casa, "Spiritual Alliances", as well as produced a highly acclaimed professional documentary film, "I do not Heal, God is the One who Heals" Contact Emma Bragdon firstname.lastname@example.com


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The President's Advisory Group advises the Superintendent/President (S/P) about issues of college-wide importance. It ensures that college members and the S/P are informed of institutional perspectives on major initiatives that impact the campus community and/or will be submitted to the Board of Trustees. For example, the President's Advisory Group will:
- Represent institutional perspective on issues of college-wide importance
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The President's Advisory Group regular recurring meetings are held on the 2nd and 4th Tuesday of each month with exceptions. The first meeting of Fall 2019 is scheduled for August 13, from 2 - 4 p.m. Meeting schedules are subject to change.
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Present Environmental Science Events
We mix modern ideas with deep business data and world technology teams to deliver business options. Veri-park, the ticketless parking fee systems specialist, has launched a complicated vehicle counting technology that can be integrated with SWARCO and third-occasion Variable Message Signs (VMS) to raised direct visitors by means of busy town centres.
Unlike traditional commercial real property the place monthly utility bills equivalent to energy, water, gas, internet and different things, Atlanta Tech Village includes that at no additional price. India's analytics business is predicted to the touch US$ 16 billion by 2025 from the present US$ 2 billion, as per the Nationwide Association of Software and Companies Companies (Nasscom).
Boston Dynamics' robots can lurch like a human, sprint on all fours like a canine and navigate round small spaces like a spider. In her present position as head of gross sales for Ireland, she leads a workforce of digital consultants to top Irish firms within the travel, retail and CPG area and is also responsible for driving the YouTube strategy within the Irish market.
That is why we're so excited to host among the finest Chinese companies and buyers at Disrupt SF… Learn More. With as we speak's know-how, the aftermath is being despatched all over the world in seconds. The objective of the conference is to figure out the position of environmental analysis in current-day society and politics, by finding out how environments, sustainability, and technology are linked in contemporary culture and politics.
The Office of Naval Research's (ONR) biennial 2017 Naval Future Power Science and Technology (S&T) EXPO will take place July 20-21, 2017. By conducting analysis, PSI works on lengthy-time period options for major challenges going through society, business and science.


text: man.endnotes.are.numbered boolean man.endnotes.are.numbered Number endnotes? 1 Description If the value of man.endnotes.are.numbered is non-zero (the default), then for each non-empty A "non-empty" notesource is one that looks like this: <ulink url="http://docbook.sf.net/snapshot/xsl/doc/manpages/">manpages</ulink> an "empty" notesource is on that looks like this: <ulink url="http://docbook.sf.net/snapshot/xsl/doc/manpages/"/> "notesource": a number (in square brackets) is displayed inline after the rendered inline contents (if any) of the notesource the contents of the notesource are included in a numbered list of endnotes that is generated at the end of each man page; the number for each endnote corresponds to the inline number for the notesource with which it is associated The default heading for the list of endnotes is NOTES. To output a different heading, set a value for the man.endnotes.section.heading parameter. The endnotes list is also displayed (but without numbers) if the value of man.endnotes.list.enabled is non-zero. If the value of man.endnotes.are.numbered is zero, numbering of endnotess is suppressed; only inline contents (if any) of the notesource are displayed inline. If you are thinking about disabling endnote numbering by setting the value of man.endnotes.are.numbered to zero, before you do so, first take some time to carefully consider the information needs and experiences of your users. The square-bracketed numbers displayed inline after notesources may seem obstrusive and aesthetically unpleasingAs far as notesources that are links, ytou might think it would be better to just display URLs for non-empty links inline, after their content, rather than displaying square-bracketed numbers all over the place. But it's not better. In fact, it's not even practical, because many (most) URLs for links are too long to be displayed inline. They end up overflowing the right margin. You can set a non-zero value for man.break.after.slash parameter to deal with that, but it could be argued that what you end up with is at least as ugly, and definitely more obstrusive, then having short square-bracketed numbers displayed inline., but in a text-only output format, the numbered-notesources/endnotes-listing mechanism is the only practical way to handle this kind of content. Also, users of "text based" browsers such as lynx will already be accustomed to seeing inline numbers for links. And various "man to html" applications, such as the widely used man2html (VH-Man2html) application, can automatically turn URLs into "real" HTML hyperlinks in output. So leaving man.endnotes.are.numbered at its default (non-zero) value ensures that no information is lost in your man-page output. It just gets "rearranged". The handling of empty links is not affected by this parameter. Empty links are handled simply by displaying their URLs inline. Empty links are never auto-numbered. If you disable endnotes numbering, you should probably also set man.font.links to an empty value (to disable font formatting for links. Related Parameters man.endnotes.list.enabled, man.font.links


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What if you could turn into your driveway and a sensor recognized your vehicle, lit your home's exterior, opened the garage door, and cued a welcome scene of lights, climate and even music. Or, as the heat soared on a southwestern summer day, your shades dropped and your climate system went into an eco-mode, and all you did was receive a text alert that this was complete. If every action caused a reaction, your home would be smart; if you could manage this at home or away, it would be even smarter still. Welcome to connected living Sierra Integrated Systems style.
Project Information
A Centralized Platform allows for total home or office control, from any room. Control the exterior lights from your master bedroom turn off the TV in the kids room from the kitchen.


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Not so long ago, there were two major competing theories in the United States regarding the origins of life: evolution and creationism. Evolution represented science's opinion of how the universe began, and creationism offered the religious explanation. It was pretty cut and dry. Then came "intelligent design."
The intelligent design (ID) movement claims that life as we know it could not have developed through random natural processes -- that only the guidance of an intelligent power can explain the complexity and diversity that we see today.
How does intelligent design explain the origins of life? Does it fit the criteria of a scientific theory, or is it religious dogma in a scientific wrapper? In this article, we'll examine intelligent design and the controversy it has generated.
What is Intelligent Design?
Intelligent design (ID) states that the universe and its inhabitants could not have evolved by the "blind chance" set forth in Darwinism. Its arguments are mostly concerned with what it considers to be holes in the theory of evolution, and it claims that these holes scientifically prove the presence of an "intelligent designer" in nature.
Unlike creationism, ID does not state that God is the intelligent designer. It only says that there is clear evidence in nature of intelligent design. The designer in ID could be God, but it could also be an extraterrestrial race or some other supernatural force. Also, ID does not draw its arguments directly from the Christian Bible.
But while it does acknowledge the possibility of slight evolutionary changes within a species, it does not recognize the possibility of one species evolving from another nor the chance of highly complex biological systems resulting from natural selection. ID proponents have stated that they aim to debunk Darwinism as the dominant origin theory and to remove naturalism -- the belief that everything around us can be explained by natural causes -- from both science and culture.
The Science of Intelligent Design
While the intelligent design movement has, at present, made more of a mark on politics than on science, it nonetheless defines itself as a scientific movement and sets forth various arguments to support that claim. The science of intelligent design is very controversial -- the scientific community does not recognize its methods as scientific -- and its arguments do not always form a cohesive vision of the scientific evidence for design. Instead, this evidence consists of the work of several scientists all setting forth their own theories in support of an intelligent, supernatural designer at work in nature. The common thread in the science of ID lies mostly in the structure of the work, all of which adheres to a double goal: To disprove Darwinism and to prove design in nature.
Some intelligent design arguments apply to both goals, and most of them are interconnected; but in the interest of structure, we will group them here according to which aim they most directly address. In the following sections, we'll examine the more prominent intelligent design claims, including:
- To disprove Darwinism: Irreducible complexity; Specified complexity; Law of Conservation of Information
- To prove design: Three-stage "Explanatory Filter"
ID Goal: To Disprove Darwinism
Michael Behe: Irreducible complexity
Irreducible complexity essentially states that there are biological structures that could not have evolved from a simpler state. A cell, for example, is composed of hundreds of complex molecular machines. Without any one of those machines, the cell would not work. So the cell is irreducibly complex: It couldn't have evolved from a simpler state because it couldn't have worked in a simpler state, and natural selection can only choose among traits that are already functioning.
Behe offers the example of a mousetrap, which typically has five parts: a wooden base to support the contraption, a metal hammer to pound the mouse, a spring to power the hammer, a catch to release the spring and a metal bar that holds back the hammer. Without any one of these parts, the device is useless. Therefore, a mouse trap is irreducibly complex.
In biology, Behe sees the bacterial flagellum as an irreducibly complex system.
A flagellum works as a propeller to help some bacteria get around. This "propeller" contains about 30 different proteins that make it work -- some act as a motor, some act as a stator and others act as a bushing to guide the driveshaft through the bacterial membrane. Without almost any one of these 30 proteins, the entire system breaks down [ref].
The scientific community responds to irreducible complexity by stating that while it is true that natural selection can only choose among traits that are already functioning, the traits don't have to be functioning in their current form. They could have been serving other purposes when they were chosen as advantageous for their current function.
In the example of the mouse trap, scientists point out that if you remove the catch and metal bar, you've got a tie clip. If you remove the spring, you've got a nice keychain. They also claim that science has already discovered that a group of the proteins that make up bacterial flagella is used by certain bacteria for an entirely different function: It acts as a sort of "molecular pump" in the bacterial membrane.
Biologist Kenneth Miller states, "The point, which science has long understood, is that bits and pieces of supposedly irreducibly complex machines may have different -- but still useful -- functions ... Evolution produces complex biochemical machines by copying, modifying, and combining proteins previously used for other functions" [ref].
William Dembski: Specified complexity
Specified complexity in a system means it could not have occurred by chance and it is not the result of any natural law that says it must be the way it is. A biological system exhibits specified complexity if it meets three criteria:
- Its makeup is not merely the result of a natural law.
- Its makeup is complex.
- Its makeup reflects an "independently given pattern or specification."
For clarification, Dembski refers us to the example of detecting an extraterrestrial radio signal in the movie "Contact."
The radio astronomers in "Contact" detected design in a radio signal when they discovered its pulses reflected all and only the prime numbers from 2 to 101. It was not the result of necessity -- there is no law that requires radio signals to transmit in that pattern; it was complex -- the series of signals was long and so was unlikely to take that particular form by chance; and the series of signals reflected an objective pattern -- one that was specified in mathematics long before the astronomers received the radio signal. This signal had specified complexity, and Ellie Arroway and crew took this as evidence of intelligent design.
According to Dembski, a biological system is clearly designed if it exhibits specified complexity.
The scientific community sees this argument as inherently flawed. It points out that Dembski sets forth a negative hypothesis: Anything not created by chance or law must be designed. But scientists claim that chance, law and design are not mutually exclusive, and they are not the only possibilities. So the process of elimination cannot be applied. And in any event, they say, science does not accept the process of elimination as proof of anything. The scientific method requires a positive hypotheses -- you cannot prove one thing simply by disproving another.
Another objection to Dembski's method of detecting design is that it appears to necessitate prior knowledge of the specified pattern. If the radio astronomers in "Contact" had no knowledge of the natural laws governing radio signals and did not recognize consecutive prime numbers as a mathematical sequence, they would never have detected a pattern. When dealing with something like DNA, scientists claim, there are no externally recognizable patterns and therefore no way to detect if a pattern occurs by chance or was "independently given." Dembski's process of detecting design presupposes design [ref].
William Dembski: Law of Conservation of Information
The Law of Conservation of Information was created by William Dembski and involves some very detailed and complex mathematical equations. At its most basic, Dembski's law states that nature cannot create new information (as in information contained in DNA); it can only work with the information it already has. Therefore, a more complex species -- one that contains more information -- could not have evolved from a less complex species [ref].
The scientific community believes that Dembski is repackaging the creationist argument that the theory of evolution violates the second law of thermodynamics, which states that there is a tendency in nature for complexity to decrease. It claims that science has understood for a long time that this theory applies "only to closed systems, and biological systems are not closed" [ref].
ID Goal: To Prove Design
William Dembski has come up with what he considers a fool-proof method of detecting design. This method is a process of elimination that asks three questions of anything found in nature:
- Does a law explain it?
- Does chance explain it?
- Does design explain it?
This is similar to the process of detecting specified complexity. To explain the filtering process in common language, Dembski uses the example of a 1985 civil case tried in the New Jersey Supreme Court. The case involved county clerk Nicholas Caputo, who was accused by the Republican party of rigging elections by always putting the name of the Democratic candidate at the top of the ballot. According to Dembski, this is known to increase a candidate's chances of winning. Dembski asserts that the court must have considered the three options in his Explanatory Filter in order to determine whether Caputo had intentionally placed Democrats in the first position on the ballots.
First, the court had to determine whether the placement occurred by law. That is, did Caputo unknowingly subject the process to a law of odds that explains the coincidence? Did he believe he was using a truly random method to determine placement when in fact the method was flawed and was sure to result with the Democratic name in position one? If the answer was no, the court would move to question two and ask whether Caputo's method was in fact random. Was it by pure chance that the democratic candidate always ended up on position one? If the court discovered a pattern -- i.e. the first position on the ballot was always occupied by the candidate from a single political party -- then it could not be the result of chance. So, if Caputo's method was not truly random, and it was already determined that it was not the result of a law that kicked in because of a mistakenly flawed method, then it must be the result of design. In other words, the only option left is that Caputo knew he was cheating: The Democratic name always ended up at the top of the ballot by design.
Dembski explains that we actually use this method all the time, probably without even knowing it. It is only a matter of quantifying the process in order to make it scientific instead of merely instinctive. In its quantitative form -- the Explanatory Filter -- it can be applied to scientific questions as successfully as it is applied to questions that arise in everyday life.
Using this method, Dembski argues, will never result in a false positive for design. However, he notes that there could be a problem of false negatives:
One difficulty is that intelligent causes can mimic law and chance, thereby rendering their actions indistinguishable from these unintelligent causes. It takes an intelligent cause to know an intelligent cause, but if we don't know enough, we'll miss it [ref].
The response by the scientific community to Dembski's three-pronged approach to identifying design is essentially the same as its response to his argument for specified complexity. Most scientists note that it is not, in fact, a positive test for design, but in fact a negative test for eliminating chance and necessity. The process of elimination can not lead to any definitive conclusion in the world of science.
Overall, the most significant objection by the scientific community to intelligent design as a scientific theory is that it not empirical. Scientists cannot test for the presence of design, nor can they disprove the presence of design. By its very nature, scientists claim, intelligent design is not a scientific argument but a philosophical one.
Controversy: Creationism in Disguise?
The intelligent design movement has created quite a stir in the United States. ID proponents claim that their theory is scientifically sound and is not based on any religious beliefs, and it should be taught alongside evolution in public-school science classes. The scientific community claims that intelligent design is not scientific at all and is in fact a metaphysical theory that should be taught in philosophy class, not science class.
People who object to the inclusion of intelligent design in science instruction are concerned not only about what the scientific community considers to be bad science, but also about what the leaders of the ID movement have said to their followers. A mission statement of the Discovery Institute's Center for Science and Culture (then the Center for the Renewal of Science and Culture) states:
Discovery Institute's Center for the Renewal of Science and Culture seeks nothing less than the overthrow of materialism and its cultural legacies. Bringing together leading scholars from the natural sciences and those from the humanities and social sciences, the Center explores how new developments in biology, physics and cognitive science raise serious doubts about scientific materialism and have re-opened the case for a broadly theistic understanding of nature [ref].
At the same time, statements from ID movement leaders call greatly on religion. Phillip E. Johnson of the Discovery Institute, speaking of the purpose of the ID movement, has said:
The objective ... is to convince people that Darwinism is inherently atheistic, thus shifting the debate from creationism vs. evolution to the existence of God vs. the non-existence of God [ref].
In the July/August, 1999, issue of Touchstone Magazine, William Dembski says, "... intelligent design is just the Logos theology of John's Gospel restated in the idiom of information theory." And later, in 2005, he makes the following remarks:
... intelligent design should be understood as the evidence that God has placed in nature to show that the physical world is the product of intelligence and not simply the result of mindless material forces. This evidence is available to all apart from the special revelation of God in salvation history as recounted in Scripture.
... To be sure, creationists who support intelligent design think it does not go far enough in elucidating the Christian understanding of creation. And they are right!
... Even so, there is an immediate payoff to intelligent design: it destroys the atheistic legacy of Darwinian evolution. Intelligent design makes it impossible to be an intellectually fulfilled atheist. This gives intelligent design incredible traction as a tool for apologetics, opening up the God-question to individuals who think that science has buried God [ref].
There are also widespread claims that the majority of the Discovery Institute's funding comes from Christian fundamentalist organizations and individuals, noting especially the millions of dollars donated by philanthropist Howard Ahmanson, an evangelical Christian, and hundreds of thousands of dollars provided by the Maclellan Foundation, which seeks to "serve strategic international and national organizations committed to furthering the Kingdom of Christ ... by providing financial and leadership resources to extend the Kingdom of God to every tribe, nation, people, and tongue" [ref]. The Discovery Institute responds by saying that the avenues of its research are determined by its board of directors, not its financial supporters.
All of this, along with questions as to the scientific validity of intelligent design arguments, has led many to believe that the intelligent design movement is a calculated approach to get a creationist view of the origins of life into public-school science classes -- that it is in fact religion disguised as science. Proponents of ID insist that this is not the case -- that intelligent design is separate from creationism and is based entirely on scientific evidence.
Origins and Progress of the ID Movement
In the 18th and 19th centuries and until the introduction of Darwin's theory of evolution, the "argument from design" was the prevailing view of the origins of the natural world.
In 1802, this view was crystallized in William Paley's watchmaker analogy. It goes something like this: If you find a watch in the middle of a field, you note that it is a complex object that serves a particular purpose. It has many different parts that all work together to tell time. When you see the watch, you automatically understand it is the product of design, not chance. It follows that we should assume the same of natural world when it displays complex processes that meet a particular need.
The argument from design reigned until Darwin published "The Origin of Species" in 1859. Biological science responded overwhelmingly to Darwin's evidence and quickly adopted evolution as the prevailing explanation of the development of the universe and life. And by 1940, almost all the biologists in the world believed that natural selection was driving force behind evolution.
Then, in 1991, law professor Phillip E. Johnson effectively launched the intelligent-design movement with his best-selling book, "Darwin on Trial." The movement quickly gained momentum in the United States. In 1996, the Discovery Institute, a think tank based in Seattle, launched the Center for the Renewal of Science and Culture (CRSC). The CRSC's original mission statement included studying and promoting intelligent design as a scientific theory.
Politically, the ID movement has made incredible strides in a short amount of time. In 1999, just eight years after the movement really took off, the Kansas Board of Education voted to remove evolution from Kansas schools' science curriculum, and the decision was widely attributed to campaigning by proponents of intelligent design. In 2004, the Dover Area School Board in Pennsylvania decided to require that all public schools in the district teach ID alongside evolution in science classes. A year later, a U.S. District Court Judge ruled that the requirement was unconstitutional.
For more information on intelligent design, creation, evolution and related topics, check out the links on the next page.
Related HowStuffWorks Articles
More Great Links
- Dembski, William A., Ph.D. "The Explanatory Filter: A three-part filter for understanding how to separate and identify cause from intelligent design." http://www.origins.org/articles/dembski_explanfilter.html
- Dembski, William A., Ph.D. "The Intelligent Design Movement." http://www.origins.org/articles/dembski_idesignmovement.html
- "Intelligent Design?" Natural History, April 2002. http://actionbioscience.org/evolution/nhmag.html
- The Maclellan Foundation. http://www.maclellanfdn.org/about/home.asp
- Miller, Kenneth R. "Darwin's Pope?" Harvard Divinity Bulletin. http://www.hds.harvard.edu/news/bulletin_mag/articles/33-2_miller.html
- Orr, Allen H. "Master Planned." The New Yorker, May 30, 2005.
- "Politicized Scholars Put Evolution on the Defensive." The New York Times, August 21, 2005. http://www.nytimes.com/2005/08/21/national/21evolve.html
- The Skeptic's Dictionary: Intelligent Design. http://skepdic.com/intelligentdesign.html


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I was chastised recently (and rightly so) for failure to respond to an e-mail sent by a colleague. It did set me thinking, however, about a possible analogy between e-mail and teaching. This may be old hat, so apologies if I've reinvented the round thing with tyres.
People tend to make the assumption when an e-mail has been sent that it has been received, but there may be several ways in which this doesn't hold true. Firstly, it may not have been delivered at all – wrongly addressed perhaps (there's a new student same surname, same initial just started at Leicester and they've kindly forwarded mail misdirected to them, though they need not have done so). Secondly, there may be some technical glitch – machine failure, necessary disconnection of PC whilst moving house/office, etc. Thirdly, the message may have made it successfully to the mailbox, it may even have been opened, but not to the grey matter of the recipient.
It's this third example of the lack of – meaningful – receipt that struck me as most analagous to teaching. We can stand at the front of a lecture theatre "sending off e-mails" of the finest quality in the form of our lecture content, but there is no guarantee that this content is being received. Even if the student is present, and has signed the register to prove it, the message we are sharing may not elicit the response that was intended/expected. There may just be too much going on in their mental mailbox for our nuggets to be deemed to have the worth we attached to them.
I'm still working on how the analogy stretches to the other versions – failure to turn up to a session would be in there, as an example of the second sort, I guess. Any suggestions?


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Deadline for applications: 1 April 2014.
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text:
Lately, my eldest will utter these words when I put him to bed. It used to be hubbs who would put him to bed, where after the usual night routine (brush teeth, pee, had scott, emulsion & vitamins, wear his socks, say his prayers), he will sing a song of JH's choice. Among JH's favourite songs are:
- Summer of 69
- Dream
- Only You
- Can't Help Falling In Love (He called this 'Wise Man' song. LOL)
- Eternal Flame
- Wonderful World
- Eelderwisse
- Moon River
- Sunshine
But hubbs had been working late for awhile now, so most of the days I gotta put that cheeky monkey to bed. I don't know how it happen but I must have slept on his bed with him a couple of times, and somehow it just because something that he's attached to, even if its for 5 minutes. It just so happened that around this time, we've established Junna's sleeping habit, so we can just put her in her crib (after her last feed around 9pm), and she'll go to sleep on her own, and bless me some time to spend with his gorgor before the day ends.
It may seem like a very little gesture but I know it means the world to him whenever I spend a few minutes with him on bed before sleep. He will choose the sides he'd like to sleep on, ask me to hug him tighter, share the blacket with me, tell me what he wants for his bento breakkie etc…I don't know when I start to appreciate this precious little time with him, but it just overwhelms me whenever I think about how fast he's growing up and one day, he wouldn't want me to sleep on his bed anymore.
So for now, my dear baby Ju, I will sleep with you for as long as you want me to.


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For some reason, I thought How to Lose Friends & Alienate People was a Run Fatboy Run-level Simon Pegg vehicle, so I didn't rush to see it, and came in with very low expectations. But it's actually pretty good -- oh, it's no Hot Fuzz; I'll admit, but it's funny a lot of the time, and the "only a non-elite life is worth living" message doesn't become dull and blunt until near the obvious end -- so I was happily surprised.
It's based on a memoir by Toby Young -- apparently, quite loosely based, since Young crawled back to England at the end of his adventure, which the Pegg character does not -- and is of the time-honored "young man from the sticks comes to the Big City, is tempted by its decadent charms (and its even more decadent women) but eventually chooses the path of righteousness and a Good Woman" formula, with Megan Fox as the bad girl (a dumb-as-rocks up-and-coming movie star) and Kirsten Dunst as the good girl (Pegg's colleague and eventual boss).
Pegg is the publisher and editor of a scurrilous, low-rent London satire magazine when Jeff Bridges (playing not-Graydon Carter and running not-Vanity Fair) drags him to NYC and gives him a lucrative contract for what are really very unlikely and mostly unexplained reasons. Pegg takes a while to adjust to kissing celebrity ass rather than kicking it, but eventually he succumbs to the lures of the fast life.
How to Lose Friends is probably annoying and tedious to those who have actually read Young's memoir; I haven't, so I found it a serviceable comedy along very obvious lines. It resolutely stays as far away from new ground as it possibly can, but it's pleasant enough as it goes along, and Pegg's character is a lot of fun as long as he keeps his British rough edges (which isn't nearly long enough). Dunst is cute and lovable; Fox is scorching and yet clueless; and Bridges good as the grumpy soft-living boss whom Pegg, in the end, can't follow. It's not nearly as smart and interesting a movie as it should have been, but it's not a disaster, just a disappointment.
I may have to read the book now.
Monday, August 17, 2009
Movie Log: How to Lose Friends & Alienate People
Your Hornswoggler is Andrew Wheeler Released into the wild 8/17/2009 12:30:00 PM
|Jerked Knees:
Recurring Motifs: Movie Log


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Foundation for Tomorrow's Schools want to build football ground for Naxxar Lions
FTS has referred directly to Naxxar Lions FC's lack of "adequate sporting facilities that cater for today's needs and expectations for a football club"
The authority responsible for the construction of Maltese schools is seeking a planning permit to build a football ground and an underground car park for 212 cars in Naxxar.
The Foundation for Tomorrow's Schools has picked an 8,830 square metre site within the precincts of the Giovanni Curmi Higher Secondary and MCAST complex.
The foundation intends making the ground available for the wider Naxxar community including its football club, which had previously expressed an interest in developing a full-blown stadium and sports hotel in Gharghur.
While the degraded land earmarked for the sports facility is deemed to have no particular environmental value, the Environment and Resources Authority has objected to an access road that would pass through undeveloped land outside building zones.
An underpass is now being considered instead of a surface road.
The new ground will face the villas along Triq Giorgio Locano in Iklin but the access will be through the school complex itself, which already includes two smaller football grounds and various other sports facilities.
The FTS's own project description referred directly to Naxxar Lions FC's lack of "adequate sporting facilities that cater for today's needs and expectations for a football club".
But so far no formal agreements have been signed with any club or nursery. "Actual operational and lease arrangements with stakeholders, including the local community such as the football nursery, still need to be worked out," a spokesperson for FTS told MaltaToday.
The construction of the football ground is considered by the FTS as an important aspect of its strategy for integrating school facilities in the wider community, so that buildings and equipment are made available to the community during the evenings and in other non-teaching hours.
"In this way greater use is made of these resources and the relationship between the educational authorities, schools, students and other stakeholders are further strengthened," FTS said.
The ERA has warned that building a surface road would "formalise the undeveloped peripheries and commit additional land" and questioned why the whole access road cannot be accommodated through the school grounds. The FTS is now revisiting its designs to include the construction of an underpass and avoid the take up of ODZ land.
The application for the new football ground in the school premises dates back to September 2017.
Last year MaltaToday had revealed that two letters of intent for a Naxxar Lions stadium in Gharghur were issued right in the middle of the 2017 election campaign – one by the parliamentary secretariat for sports, and one by the Lands Authority. The project for a stadium for Naxxar Lions FC and a hotel, attracted the interest of Syrian-Canadian investor Yahya Kirdi with a €10 million commercialisation project.
But tangible progress on Kirdi's investment for Naxxar, having been installed as president of the club, has so far not materialised.
Kirdi was in the past associated with a bid for Liverpool, and later in 2017, reportedly fronting a bid to acquire the smaller club Carlisle United. In those negotiations he was represented by Nick McCreery, a north east-based agent and son of the former Carlisle boss David McCreery – a friend of former Naxxar president and benefactor Michael Zammit Tabona.


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Average rate
Amber D.
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- Joined October 2017
Let me just start with, I LOVE Programming. For me it's about figuring out the solution to the puzzle and an insatiable curiosity.
- Las Vegas, NV
Amber D.
Las Vegas, NV
Taking ideas from concept to production and watching them come to life is my passion. I have spent the last 10 years learning every new framework and library I could get my hands on. I love working with JavaScript and all of it's wonderful possibilities.
Work history
Oxford Digital
Senior JavaScript Developer
Develop applications using multiple JavaScript technologies for several clients in different industries. Skills: React, Redux, AngularJS, Angular4, MEAN, Node, Express, Mongo, Ramda, RxJs, D3, ThreeJS, SVG Animation, CSS3, SASS, LESS, jQuery
Senior Development Consultant
Develop applications for several clients in the casino sector including custom game and management software as well as hardware driver development for integration with web-based applications deployed on self-serve kiosks.
Senior Responsive UI/UX Developer
Managed the Git repository for the project with multiple developers committing from across the globe and developed several internal tools to enhance the employee network and asset search capabilities
AgFirst Farm Credit
Senior Development Consultant
Developed the Get Farm Credit app and several association websites. Streamlined their process for Kentico development. Worked with designers to overcome Kentico restrictions and built websites on the Kentico CMS framework in an easy to update format.
Premier Inc
Senior Development Consultant
Lead the front end development of several pharmaceutical processing applications. Collaborated with executives and managers to decide and evaluate requirements for the applications; then directing the design team on how best to implement the requirements.
Ally Financial
Senior Application Developer
Developed the responsive and mobile web redesign version of their main online banking portal using CSS3, SASS, HTML5, JavaScript, jQuery, Handlebars, and Ember.js. Also worked on the print features, desktop dashboard redesign, and defect remediation.
Intel Corp
Graphics Architecture Intern
Built web based reporting applications that could gather information and provide reports on the benchmarking software, worldwide network of servers and current build. Multilevel reporting with file extraction, filterable results & notification system.
AvailabilityNot seeking work
- Available during business hours, evenings, and weekends
- Can work 50hrs per week
- Can commit to 1+ years


text:
Hi Gents
I recently received a letter of refusal after submitting my application in October 2008. The reasons given was that i "did not provide adequate reasons to prove that the need exists to posses the particular firearm and that the need cannot be satisfied by any other means than possessing the said said firearm" !!!!!!!!!!!!
also i did not convince the registrar that the needs exists that necessitate the possession of the firearm!!!!!!
now i would like to appeal by submitting a comprehensive motivation as requested by them.
My question is: What are the main pointers that should or should not be in a motivation letter with regards to new firearm application :?:
I have tried the seach option but came up with nothing concrete :? any assistance is greatly appreciated. Also does the make and model of the firearm have anything to do with the application :?: i bought a CZ-75 9mm. I know its old but like a toyota its a die hard and i got it for a good price....


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How do I generate a random number between 0 and some integer n?
Created May 4, 2012
John ZukowskiThis would return a value from 0 (inclusive) to maxvalue (exclusive).
While you can use Math.random() to generate random numbers, the Random class in the util package is actually a little easier to work with if you don't just want a number between 0 and 1.
First you would need to create an instance of Random somewhere, like your constructor or the init() method of an applet:
Random random = new Random();Then, whenever you need a number, you ask it:
int pick = random.nextInt(maxvalue);


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