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Identification of a Functional Homolog of the Mammalian CYP3A4 in Locusts
Line Rørbæk Olsen, Charlotte Gabel-Jensen, Peter Aadal Nielsen, Steen Honoré Hansen and Lassina Badolo
Drug Metabolism and Disposition July 2014, 42 (7) 1153-1162; DOI: https://doi.org/10.1124/dmd.114.057430
Line Rørbæk Olsen
Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark (L.R.O., C.G.-J., S.H.H.); Department of Discovery ADME, H. Lundbeck A/S, Valby, Denmark (L.B.); and EntomoPharm R&D, Lund, Sweden (P.A.N.)
Charlotte Gabel-Jensen
Peter Aadal Nielsen
Steen Honoré Hansen
Lassina Badolo
Insects have been proposed as a new tool in early drug development. It was recently demonstrated that locusts have an efflux transporter localized in the blood-brain barrier (BBB) that is functionally similar to the mammalian P-glycoprotein efflux transporter. Two insect BBB models have been put forward, an ex vivo model and an in vivo model. To use the in vivo model it is necessary to fully characterize the locust as an entire organism with regards to metabolic pathways and excretion rate. In the present study, we have characterized the locust metabolism of terfenadine, a compound that in humans is specific to the cytochrome P450 enzyme 3A4. Using high-resolution mass spectrometry coupled to ultra-high-performance liquid chromatography, we have detected metabolites identical to human metabolites of terfenadine. The formation of human metabolites in locusts was inhibited by ketoconazole, a mammalian CYP3A4 inhibitor, suggesting that the enzyme responsible for the human metabolite formation in locusts is functionally similar to human CYP3A4. Besides the human metabolites of terfenadine, additional metabolites were formed in locusts. These were tentatively identified as phosphate and glucose conjugates. In conclusion, not only may locusts be a model useful for determining BBB permeation, but possibly insects could be used in metabolism investigation. However, extensive characterization of the insect model is necessary to determine its applicability.
Today, most new drugs that fail in the clinical phase do so due to lack of desired pharmacological activity or toxicity (Hughes et al., 2011). In the development of central nervous system drugs, the blood-brain barrier (BBB) creates additional challenges, and therapeutic agents for the treatment of neurologic disorders often fail in clinical trials due to inadequate BBB permeation (Alavijeh et al., 2005; Geldenhuys et al., 2012). In vitro methods for testing BBB penetration are too simple, not integrating the complexity of the BBB, and therefore may lack important functions of the BBB (Naik and Cucullo, 2012). Preclinical in vivo models are very costly in terms of time and use of animals (rodents). Fast screening models for BBB permeation are therefore very attractive.
Insects have been suggested as novel models in drug discovery (Geldenhuys et al., 2012). It has been demonstrated that locusts, among other insects, have an efflux transporter that is functionally similar to the mammalian P-glycoprotein efflux transporter (Nielsen et al., 2011).
Currently, two insect BBB models have been reported using locusts as model insects (Nielsen et al., 2011; Andersson et al., 2013). An ex vivo model has been developed and allows application of test items directly to entire brains isolated prior to testing. Besides this model, an in vivo model has also been developed in which the test item is injected in the hemolymph and may need to pass the barriers of metabolism and excretion before reaching the brain. As such, the in vivo locust model may have more similarities to other in vivo models like rodent models. Also, the brain remains in its natural environment, with no risk of impairing the barrier function.
Despite obvious anatomic differences, there are many physiologic and biochemical similarities between insects and mammals. Essential systems such as protein synthesis and cell metabolism are not significantly different between insects and mammals (Klowden, 2007). Monooxygenases like the cytochrome P450 (P450) enzymes are found in virtually all eukaryotic organisms, including insects (Scott, 2008). This is of specific interest as some of the most important drug-metabolizing enzymes in humans are the P450 enzymes. Among mammalian P450 enzymes, the subfamilies CYP1A1/2, CYP2B6, CYP2C9/19, CYP2D6, CYP2E1, and CYP3A4/5 are the main contributors, and CYP3A4 accounts for the metabolism of almost half of all marketed drugs (Guengerich, 2008).
Most literature on insect metabolic systems concerns insecticide detoxification mechanisms and resistance development (Smith, 1962; Wilkinson and Brattsten, 1972; Feyereisen, 1999; Scott and Wen, 2001). Furthermore, most research was conducted in the 1950s and 1960s when the analytic methods were limited compared with the technologies available today.
In the present study, we have characterized the metabolism in locusts of the antihistaminic drug terfenadine, a specific human CYP3A4 substrate.
Solvents and reagents: All solvents used were at least of high-performance liquid chromatography grade. Methanol (MeOH) was purchased from BDH Prolabo (VWR, Herlev, Denmark). The following reagents were purchased from Sigma-Aldrich (St. Louis, MO): female pooled human liver microsomes (HLMs), glucose 6-phosphate, glucose-6-phosphate dehydrogenase from Baker's yeast, NADP+, formic acid, MgCl2∙6H2O, quinidine, α-naphthoflavone, sulfaphenazole, nootkatone, ketoconazole, and amitriptyline. The following were purchased from Merck (Darmstadt, Germany): K2HPO4∙3H2O, KH2PO4, ZnSO4∙7H2O, and dimethylsulfoxide (DMSO). Terfenadine was provided by NeuroSearch A/S (Ballerup, Denmark). Water for mobile phases and solutions was purified with the Direct-Q 3 UV system (Millipore, Billerica, MA).
Animals: Male desert locusts, Schistocerca gregaria (L), were obtained from a commercial breeder (Petra Aqua, Prague, Czech Republic). The locusts were housed under crowded conditions in a 12-hour light/dark cycle at 25–35°C. They had constant access to Chinese cabbage and dried wheat bran. Experiments were carried out on fed fifth instar locusts aged 3–5 weeks after final molt.
Locust experiments: Terfenadine, quinidine, sulfaphenazole, nootkatone, α-naphthoflavone, and ketoconazole were dissolved in DMSO and diluted with 0.1% lactic acid in Milli-Q water (Millipore) to the applied concentrations. Terfenadine was administered alone or in a cosolution with one of the following P450 inhibitors: ketoconazole, quinidine, sulfaphenazole, nootkatone, or α-naphthoflavone. DMSO concentrations in all solutions used for injection were 7.5% due to low solubility of some of the inhibitors. The locusts contained ∼144 μl hemolymph per g (Lee, 1961) and had an average weight of 1.59 ± 0.166 g (mean ± S.D.; n = 10), and thus the final DMSO concentration in hemolymph was expected to be <2%.
The locusts were injected with 40 μl of 500 μM terfenadine (with and without 2.5 mM inhibitor) between two terga and kept fixated at room temperature during the experiment. The effects of different P450 inhibitors were determined using a hemolymph sample 60 minutes postinjection (n = 4). If significant inhibition of terfenadine metabolism was observed, time profiles of metabolites formed were obtained from hemolymph samples over a time period of 2 hours. Hemolymph samples were taken from the locust at 1, 5, 10, 20, 30, 45, 60, and 120 minutes postadministration. Samples were collected using an end-to-end micropipette (10 μl) through a small hole punctured in the soft tissue below the head. The collected hemolymph was added to 40 μl of 2% ZnSO4 in 50% MeOH (on ice) containing amitriptyline as internal standard. After 120 minutes, the locusts were euthanized by freezing. For detection of metabolites in feces, locusts were dosed with 40 μl of 500 μM terfenadine (with or without 2.5 mM inhibitor) and left for 24 hours. Then feces were collected and weighed and metabolites were extracted with 50% MeOH-containing internal standard by ultrasonication for 30 minutes.
All samples collected were stored at −18°C prior to analyzing. After thawing, the hemolymph samples were centrifuged for 10 minutes at 15,000g at 5°C and the supernatants transferred to vials for analysis. The fecal extracts were filtered through 0.45-μm Millex-HV syringe filters (Millipore) and analyzed. Hemolymph and feces from nontreated locusts were used as controls.
HLM incubation method: HLMs (0.5 mg/ml) were preincubated at 37°C for 5 minutes in 100 mM potassium phosphate buffer (pH 7.4) with 3.3 mM MgCl2, 0.4 U/ml glucose-6-phosphate dehydrogenase, 3.3 mM glucose 6-phosphate, and 1.2 mM NADP+. After preincubation, terfenadine was added (final concentration, 4 μM). Samples (20 μl) were taken out at the same time points as for the locust experiments, and the reaction was quenched using 80 μl of 2% ZnSO4 in 50% MeOH and containing 3% formic acid (on ice). The samples were stored at −18°C.
After thawing, the samples were centrifuged for 10 minutes at 15,000g at 5°C and the supernatants analyzed with liquid chromatography–mass spectrometry (MS).
To determine pharmacokinetic parameters (Km) in locusts, five different terfenadine concentrations were used (250, 500, 1000, 1500, and 2500 μM). The concentration of DMSO was kept at 2.5% (<0.2% in hemolymph). Terfenadine solution (40 μl) was injected, and after 1 minute 20 μl of hemolymph was drawn from the locusts and added to 80 μl of 2% ZnSO4 in 50% MeOH containing internal standard. The samples were prepared in a Sirocco 96-well plate for protein precipitation (Waters, Milford, MA). The plate was centrifuged at 2000g for 4 minutes, and the filtrate was collected and analyzed.
Ultra-high-performance liquid chromatography–MS method: The chromatographic separation was performed on a Kinetex C18 (50 × 2.1 mm) column (Phenomenex, Torrance, CA) with 1.7-μm particles using a Dionex UltiMate 3000 RS ultra-high-performance liquid chromatography system (Thermo Scientific, Waltham, MA) coupled to a high-resolution MS instrument [Q Exactive Orbitrap with a HESI-II interface from ThermoFinnigan (San Jose, CA)]. Separation was performed with gradient elution using 0.1% formic acid in Milli-Q water as mobile phase A and MeOH with 0.1% formic acid as mobile phase B with a flow rate of 0.4 ml/min. The mobile phase B was kept at 5% for 0.2 minutes followed by a linear gradient of 5%–90% B over 5 minutes, 90% B for 1 minute, followed by re-equilibration for 2 minutes. The column compartment was set to 30°C, and the samples were kept at 8°C.
Detection of metabolites was performed in positive ionization mode using data-dependent tandem MS (MS2) on the top five most abundant ions in each scan with an dynamic exclusion of 5 seconds. Quantification of terfenadine was performed in targeted MS2. The lock mass used (214.0896 m/z) originated from a plasticizer, N-butyl benzenesulfonamide. MS settings were as shown in Table 1. Spray voltage was set to 3.3 kV, capillary temperature 350°C, sheath gas 30 arbitrary units, auxiliary gas 10 arbitrary units, probe heater 250°C, and S-lens RF level 50. Thermo Scientific Mass Frontier 7.0 Spectral Interpretation Software was used for identification of metabolites.
MS settings for identification of metabolites
For statistical evaluation, a Student's t test was used.
HLM Metabolites
To investigate whether metabolites obtained in the locust were similar to those obtained in humans, HLMs were incubated with the test compound and the metabolites obtained here were used as reference compounds in comparison with locust metabolites. The phase I metabolites most frequently reported in the literature are shown in Fig. 1 and were all detected in HLMs and confirmed by accurate mass and fragmentation patterns (Jurima-Romet et al., 1994; Ling et al., 1995; Rodrigues et al., 1995).
Structures of terfenadine and known metabolites formed in mammals by CYP3A4 (Jurima-Romet et al., 1994; Ling et al., 1995; Rodrigues et al., 1995).
Observed fragmentation patterns of terfenadine and metabolites are given in Table 2. Other metabolites and/or intermediates (M2, M7, and M8) were also identified in HLMs. M1, M5, and M6 were identified as azacyclonol, terfenadine alcohol, and terfenadine acid, respectively, and the structures are shown in Fig. 1.
Metabolites identified in locust feces and HLMs
Substrate Specificity in Locusts and Mammals
To determine whether there is an enzyme in locusts with the same substrate specificity as found in mammals, terfenadine was administered by intrahemolymphic injection alone or along with each of five inhibitors known to inhibit different P450 enzymes (given in parentheses) involved in drug metabolism: α-naphthoflavone (1A2), sulfaphenazole (2C9), nootkatone (2C19), quinidine (2D6), and ketoconazole (3A4). The results are shown in Fig. 2. Only coadministration with ketoconazole resulted in a significant inhibition of formation of metabolites in locusts. The terfenadine dose given to the locusts corresponds to the weight-adjusted daily oral dose in humans (120 mg ≈ 2 mg/kg) but dosed directly in the hemolymph.
Response of terfenadine and known mammalian metabolites after administration of terfenadine to locusts with and without coadministration of inhibitors. Data are means and S.E.M.; n = 4.
Elimination of terfenadine from locust hemolymph was followed over time (Fig. 3), and the area under the curve (AUC) with and without coadministration of ketoconazole was calculated. As can be seen in Fig. 3B, the AUC was significantly (1.7-fold) larger when ketoconazole was coadministered.
Elimination of terfenadine from hemolymph (A) and calculated AUCs (B). Results with (▴) and without (○) the inhibitor ketoconazole. Data are means and S.E.M.; n = 6.
Detection of P450 Metabolites in Locusts
Metabolites were detected in locust hemolymph and feces with fragmentation patterns and retention times identical to metabolites identified in HLM incubations. The metabolites were not quantified; however, the relative amounts of the different metabolites were different in hemolymph and feces. Thus, M1 and M6 were the most abundant in hemolymph, whereas M5 was more abundant in feces. M2, M3, M4, and M7 were identified only in feces. M3, M4, M9, M10, M11, and M12 were detected only in locusts.
M1, M5, M6, and M8 were all found in HLM incubations and in locust hemolymph as well as locust feces. M1, M6, and M8 increased over time when no inhibitor was coadministered, as can be seen in Fig. 4. In locust hemolymph, M5 increased rapidly in the first 5 minutes and then decreased over time. This was also seen in HLM incubations, although in the latter the decrease started after 20 minutes. The formation of all four metabolites was inhibited almost completely by coadministration of ketoconazole.
Major metabolites identical to HLM metabolites identified in locust hemolymph over time. Results with (▴) and without (○) inhibitor. Data are mean and S.E.M.; n = 6.
Detection of Non-P450 Metabolites
Non-P450 metabolites were identified based on similarities in fragmentation patterns, as seen in Table 2. Besides the six metabolites identified in HLM incubations, an additional six metabolites were detected in locusts. Only four of these metabolites were detected in hemolymph in sufficient concentrations to be measured over time (Fig. 5).
Major non-P450 metabolites identified in locust hemolymph over time. Results with (▴) and without (○) inhibitor. Data are mean and S.E.M.; n = 6.
The excreted amount of unchanged terfenadine was not significantly different with and without coadministration of ketoconazole, ranging from 3%–30% of the administered dose (data not shown). The excretion of only one metabolite was significantly increased when ketoconazole was coadministered (M10). The formation of M12 appears to be inhibited by ketoconazole; however, the excreted amount of this metabolite was not significantly different (Figs. 5 and 6). The rest of the metabolites were inhibited (M7, M8, M9, and M11) or not affected (M3). M2 and M4 were also significantly inhibited by ketaconazole; however, the responses of these metabolites were very low compared with the rest of the metabolites and therefore are not included in the figures.
MS response of terfenadine and the most abundant metabolites identified in feces collected from locusts 24 hours after injection (with and without coadministration of inhibitor). (Left) The metabolites significantly inhibited by coadministration of ketoconazole; (right) the metabolites unaffected or increased. Four locusts per cage. Data are mean and S.E.M.; n = 3. KTZ, ketoconazole.
Metabolite Identification
A spectrum of M3 (m/z 650) is shown in Fig. 7. Fragmentation of M3 is almost identical to the fragmentation of M5, suggesting that M3 is a conjugate of M5 (Δm/z = 162). M3 was identified in locust feces, and the amount excreted was not significantly changed when ketoconazole was coadministered. In hemolymph, the concentration was too low to be followed over time, suggesting that the excretion rate for this metabolite is high or that M3 is further metabolized.
Fragmentation pattern of M3 (top), with parent mass of 650.369, and M5 (bottom), with parent mass of 488.316.
As mentioned, the different metabolites were not quantified; however, based on the MS response, M9, with an accurate mass of 568.281, was the most abundant metabolite found in locust feces and hemolymph. As seen in Fig. 8, the fragmentation pattern resembles the fragmentation for terfenadine alcohol (M5). The fragment of 73 m/z is missing, and an additional fragment of 99 m/z can be seen. In Figs. 5 and 6 it appears that the formation of M9 is significantly inhibited by ketoconazole.
MS/MS spectrum of M9, with parent mass of 568.281 (top), and M5, with parent mass 488.315 (bottom).
M10 was identified as a direct conjugate of terfenadine without any prior phase I modification. According to Table 2, the mass of M10 corresponds to the m/z of terfenadine + 162 Da. The fragmentation pattern for fragments with m/z < 472 was identical to terfenadine. In Fig. 5 it can be seen that the formation rate of M10 was not affected by ketoconazole in the first 20 minutes postinjection. From 20 minutes the amount of M10 increased significantly compared with the amount in locusts receiving only terfenadine. The amount of excreted M10 was increased in locusts coadministered ketoconazole.
M11 shares some of the fragments seen for M7 and is expected to be a conjugate of M7. Fragmentation patterns were not similar regarding fragments with m/z < 450. As can be seen in Figs. 5 and 6, formation of M11 is inhibited by ketoconazole.
M12 is expected to be a direct conjugate of terfenadine. The fragment of 472 (m/z of terfenadine) is missing in the MS2 spectrum of M12, and only one water molecule is lost. Furthermore, a fragment of 98.99 m/z was observed. The formation of this metabolite in hemolymph was initially inhibited by ketoconazole (Fig. 5). No significant difference could be shown for the amount excreted in feces when the locusts were given ketoconazole (Fig. 6).
Metabolite ID
Phase I Metabolites.
The fragmentation patterns of M1, M5, and M6 are described in the literature, and these are consistent with the observed fragmentation pattern (Rodrigues et al., 1995). The fragmentation pattern of terfenadine has a characteristic loss of two consecutive water molecules from the parent mass, and the tert-butyl group results in a fragment with m/z of 57. The fragmentation patterns of M5 and M6 show that the parent masses have shifted but there is still consecutive loss of two water molecules. The primary alcohol on M5 is too stable to result in a loss of water. The tert-butyl fragment of m/z 57 has shifted according to the metabolic modification, thus confirming where oxidation has taken place. Full structural elucidation by NMR of metabolites was not attempted, and authentic standards were not available. Instead, fragmentation patterns and accurate mass were used for identification.
The MS responses of M2, M7, and M8 were minor and expected to be intermediates, as reported by Jurima-Romet et al. (1994). M2 appears to be a double-hydroxylated metabolite. Both hydroxylations occur on the tert-butyl group, as a fragment of 89.060 m/z (57 + 16 + 16) was identified. The exact mass of M7 corresponds to an aldehyde and M8 to an acid-ketone metabolite. M4 has the same accurate mass as M2 but was only detected in locust feces. As with M2, the fragmentation pattern suggests that this metabolite could be a double-hydroxylated compound; however, the fragmentation pattern was slightly different compared with M2 (Table 2).
Phase II Metabolites (Locusts Only)
Glucosylation.
The metabolites M3 and M10 were only formed in locusts and were tentatively identified as glucosides of M5 and terfenadine, respectively. The mass accuracy was better than 0.1 ppm. The glucose conjugation site of M10 could be the amine or one of the secondary alcohols. Although highly speculative, it could be suggested that the glucosylation occurs on the latter as there is only a single loss of water from M10 (634 → 616). The fragmentation pattern of M10 was identical to the fragmentation pattern of terfenadine for m/z < 472.
Regarding M3, it can be seen in Fig. 7 that there is a consecutive loss of two water molecules (650 → 632 → 614), suggesting that the glucose is conjugated to the nitrogen or the primary alcohol. The fragment with m/z 73 is formed, confirming that the hydroxylation has occurred on the tert-butyl group of terfenadine.
Phosphorylation.
M9, M11, and M12 were tentatively identified as phosphate esters of terfenadine alcohol, M7, and terfenadine, respectively. The mass accuracy was better than 0.5 ppm. Two consecutive water molecules are readily lost from M9, indicating that the phosphate ester is conjugated to the primary alcohol of M5 (Fig. 8). The fragment of 434 (M5 – 3H2O) is not seen in the MS/MS spectrum of terfenadine alcohol (M5). The fragment is, however, seen in the spectrum of M9, suggesting that the phosphate group is more readily lost by fragmentation. Also, the fragment at m/z 73 indicating a hydroxylation on the tert-butyl group is also missing, indicating that the entire phosphate group is lost. The same pattern was observed for M12, where the fragment of 472 (m/z of terfenadine) is missing in the MS/MS spectrum of M12. Only one water molecule is lost, suggesting that the phosphate forms an ester with one of the existing hydroxyl groups in the molecule. In the MS/MS spectra of all the phosphorylated metabolites a fragment of 98.99 m/z could be seen. This fragment is thought to originate from protonated phosphate (H4PO4+).
A proposed metabolic pathway of terfenadine in locusts can be seen in Fig. 9.
Proposed metabolic pathways and structures of locust metabolites.
Due to extensive phase II metabolism of both parent and phase I metabolites, a measure of the initial formation rate had to be done within 1 minute. At this time point, substantial interindividual variation was observed. Hence, estimation of pharmacokinetic parameters was not successful.
Human phase I metabolites of terfenadine were produced in HLMs and used as references to identify and characterize the metabolites in locusts. Metabolites identical to the metabolites detected in HLM incubations of terfenadine (M1, M2, M5, M6, M7, and M8) were identified in locust hemolymph and feces after administration of the drug. In humans, terfenadine is metabolized almost exclusively by CYP3A4 into the metabolites shown in Fig. 1. Human data have shown increases in terfenadine AUC, after multiple dosing, ranging from 16 to 73 times when CYP3A4 is inhibited by ketoconazole (Honig et al., 1993); however, unchanged terfenadine is normally undetectable in plasma due to extensive first-pass metabolism (Jurima-Romet et al., 1994). In in vitro assays using cryopreserved human hepatocytes, ketoconazole provides almost complete inhibition of terfenadine metabolism (Shibata et al., 2008). In HLMs, the response of azacyclonol (M1) increased over time in both HLM incubations and locust hemolymph. Terfenadine alcohol (M5) rose in the first 20 minutes and then declined, indicating that this metabolite is further converted. The same was seen in locust hemolymph, only the decline started after 5 minutes. Terfenadine acid (M6) and M8 increased after a short lag time in both HLM incubations and locust hemolymph, suggesting that M5 (terfenadine alcohol) was further converted into these metabolites. In vivo Km values for the formation of phase I metabolites are not reported, as unchanged terfenadine usually is undetectable in plasma when given in therapeutic doses (Jurima-Romet et al., 1994). In vitro data obtained using human liver microsomes resulted in Km values for azacyclonol between 0.82 and 11 μM. The Km values for hydroxy-terfenadine formation in human liver microsomes ranged between 1.8 and 18 μM (Raeissi et al., 1997; Jurima-Romet et al., 1998).
Because no in vivo data are available for comparison and because the in vitro data vary considerably, no further attempt to establish kinetic parameters for the metabolite formation was performed.
When locusts were exposed to a single dose of terfenadine combined with different P450 enzyme inhibitors, it was only ketoconazole (an inhibitor of CYP3A4) that inhibited the formation of the mammalian metabolites, indicating that the formation of these metabolites in locusts is catalyzed by an enzyme functionally related to mammalian CYP3A4. As can be seen in Fig. 3B, the AUC is significantly (1.7-fold) larger when ketoconazole is coadministered, indicating that the elimination of terfenadine is inhibited. The excreted amount of terfenadine is not changed, and compared with the inhibition observed in humans, the increase in AUC in the locusts is moderate, indicating that the metabolism of terfenadine may not be limited to a single enzyme and/or the enzyme(s) are not fully inhibited by ketoconazole. Based on the almost complete inhibition of formation of M1, M5, M6, and M8 in locusts (Fig. 4), it seems likely that alternative metabolic pathways take part in terfenadine metabolism.
M3 and M10 were tentatively identified as glucose conjugates of terfenadine alcohol (M5) and terfenadine, respectively. In hemolymph the concentration was too low to be followed over time suggesting that the excretion rate for this metabolite is high or that M3 is further metabolized. M10 increased significantly after 20 minutes from injection time compared with the amount in locusts receiving only terfenadine. The excreted amount of M3 was not significantly inhibited by ketoconazole even though the formation of M5 was significantly inhibited. M10 was increased in the presence of the CYP3A4 inhibitor. This is in agreement with the findings that other metabolic pathways are inhibited and thus that more terfenadine was available for glycosylation. However, the excreted amount of glucoside metabolites may be misleading due to glycosidase activity in the intestine (Kikal and Smith, 1959).
Previously, conjugates with amino acids, glucose, phosphate, and sulfate have been reported in insects (Myers and Smith, 1954; Smith, 1955, 1962; Cohen and Smith, 1964; Ngah and Smith, 1983). Glycosylation is mentioned a number of times in the literature as a detoxification mechanism in insects. Glycosylation can conjugate to amines and hydroxyl and carboxy groups (Smith, 1968) and has been suggested to be analogous to mammalian glucuronidation (Smith, 1962). Although glucuronic acid conjugates have been reported in insects (Terriere et al., 1961), it has later been questioned in the literature (Lowenstein, 1968). Due to inadequate identification tools available in the 1950s and 1960s, metabolites may have been wrongly identified as glucuronic acid conjugates.
It has been tested whether terfenadine and other tertiary amines could be metabolized into N-glucuronides (Luo et al., 1991). The study was conducted on healthy volunteers, and for eight of the nine compounds tested glucuronides were detected in the urine. Terfenadine was the only compound of the nine tertiary amines tested that was not metabolized by the glucuronidation pathway. Glucoside formation has been reported in humans, although the detoxification mechanism generally is considered to be characteristic in invertebrates and other lower organisms (Gessner et al., 1973). Based on the results in our study regarding substrate specificity, glucuronidation in mammals does not appear to be analogous to glycosylation in insects as no glucuronides of terfenadine have been reported. However, the two detoxification mechanisms are probably evolutionarily related (Smith, 1968).
Phosphate and sulfate metabolites have been reported in insects (Smith, 1955; Ngah and Smith, 1983). Phosphate and sulfate conjugates of the same compound differ in mass by only 10 mmu. With accurate mass techniques we were able to distinguish between these two adducts. Thus, M9, M11, and M12 were tentatively identified as phosphate conjugates of M5 (terfenadine alcohol), M7, and terfenadine, respectively. Assuming the MS responses of terfenadine and the metabolites are somewhat similar, M9 was the most abundant metabolite found in locust feces and hemolymph (Figs. 5 and 6). In Figs. 5 and 6 it appears that the formation of M9 and M11 is significantly inhibited by ketoconazole, possibly due to inhibition of the oxidation step.
The structure of M7 was not confirmed by NMR, and it is possible that the carbonyl consists of a ketone rather than an aldehyde, which is shown in Fig. 9.
The formation of M12 in hemolymph was initially inhibited by ketoconazole even though P450 is not expected to be involved and no hydroxylation is required (Fig. 5). However, the amount excreted in feces was not significantly different when the locusts were given ketoconazole (Fig. 6). Looking at Fig. 5, it appears that there is a large increase in the amount M12 between 60 and 120 minutes postinjection when ketoconazole is coadministered.
No sulfate metabolites of terfenadine were identified. Based on the present results, conjugation with phosphate and glucose seems to be a major phase II detoxification pathway in locusts. These metabolites are qualitatively different from mammalian metabolic pathways and appear to some extent to take over from P450 enzymes when these enzymes are inhibited.
Although many insecticides, such as dimethoate, have been developed due to the formation of toxic metabolites in insects but not in mammals (Smith, 1968), the phase I metabolism of terfenadine in locusts resembles mammalian phase I metabolism. It is therefore possible that the insect model could be used in phase I metabolism investigations and not only be useful for determining BBB permeation. Locusts may also be used in generating human-relevant metabolites for further identification. However, extensive characterization of the insect models is necessary to determine their applicability. Inclusion of insect experiments in early preclinical drug development could possibly reduce costs as fewer "poor" candidates would proceed to in vivo experiments in mammals. In the present study, we have shown that locusts may have an enzyme with functionality similar to mammalian CYP3A4, an important enzyme involved in the human metabolism of almost half of marketed drugs (Guengerich, 2008).
Participated in research design: Olsen, Hansen, Badolo, Gabel-Jensen.
Conducted experiments: Olsen.
Contributed new reagents or analytic tools: Hansen.
Performed data analysis: Olsen, Hansen, Badolo.
Wrote or contributed to the writing of the manuscript: Olsen, Hansen, Badolo, Gabel-Jensen, Nielsen.
Received January 31, 2014.
Accepted April 28, 2014.
This work was supported by the Danish National Advanced Technology Foundation [Grant 023-2011-3].
dx.doi.org/10.1124/dmd.114.057430.
dimethylsulfoxide
human liver microsome
cytochrome P450
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(1994) Terfenadine metabolism in human liver. In vitro inhibition by macrolide antibiotics and azole antifungals. Drug Metab Dispos 22:849–857.
Wright M, and
Neigh S
(1998) Terfenadine-antidepressant interactions: an in vitro inhibition study using human liver microsomes. Br J Clin Pharmacol 45:318–321.
Kikal T and
(1959) Comparative detoxication. 6. The metabolism of 6-amino-4-nitro-o-cresol and 4:6-dinitro-o-cresol in locusts. Biochem J 71:48–54.
Klowden MJ
(2007) Physiological Systems in Insects, Elsevier, Amsterdam.
Lee RM
(1961) The variation of blood volume with age in the desert locust (Schistocerca gregaria Forsk.). J Insect Physiol 6:36–51.
Ling KH,
Leeson GA,
Burmaster SD,
Hook RH,
Reith MK, and
Cheng LK
(1995) Metabolism of terfenadine associated with CYP3A(4) activity in human hepatic microsomes. Drug Metab Dispos 23:631–636.
Lowenstein O, editor (1968) Advances in Comparative Physiology and Biochemistry, vol 3, Academic Press, New York.
Luo H,
Hawes EM,
McKay G,
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(1991) N(+)-glucuronidation of aliphatic tertiary amines, a general phenomenon in the metabolism of H1-antihistamines in humans. Xenobiotica 21:1281–1288.
Myers CM and
(1954) Comparative detoxication. II. Glucoside formation from phenols in locusts. Biochem J 56:498–503.
Naik P and
Cucullo L
(2012) In vitro blood-brain barrier models: current and perspective technologies. J Pharm Sci 101:1337–1354.
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(1983) Acidic conjugate of phenols in insects; glucoside phosphate and glucoside sulphate derivatives. Xenobiotica 13:383–389.
Nielsen PA,
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Andersson G
(2011) Models for predicting blood-brain barrier permeation. Drug Discov Today 16:472–475.
Raeissi SD,
Guo Z,
Dobson GL,
Artursson P, and
Hidalgo IJ
(1997) Comparison of CYP3A activities in a subclone of Caco-2 cells (TC7) and human intestine. Pharm Res 14:1019–1025.
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Mulford DJ,
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Surber BW,
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Ferrero JL,
Thomas SB,
Shet MS, and
Estabrook RW
(1995) In vitro metabolism of terfenadine by a purified recombinant fusion protein containing cytochrome P4503A4 and NADPH-P450 reductase. Comparison to human liver microsomes and precision-cut liver tissue slices. Drug Metab Dispos 23:765–775.
Scott JG (2008) Insect cytochrome P450s: thinking beyond detoxification, in Recent Advances in Insect Physiology, Toxicology and Molecular Biology (Liu N ed) pp 117–124, Research Signpost, Kerala, India.
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(2001) Cytochromes P450 of insects: the tip of the iceberg. Pest Manag Sci 57:958–967.
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Takahashi H,
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Ishii Y
(2008) A novel approach to the prediction of drug-drug interactions in humans based on the serum incubation method. Drug Metab Pharmacokinet 23:328–339.
(1955) Comparative detoxication. 4. Ethereal sulphate and glucoside conjugations in insects. Biochem J 60:436–442.
(1962) Detoxication mechanisms. Annu Rev Entomol 7:465–480.
(1968) The comparative metabolism of xenobiotics. Adv Comp Physiol Biochem 3:173–232.
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(1972) Microsomal drug-metabolizing enzymes in insects. Drug Metab Rev 1:153–227.
You are going to email the following Identification of a Functional Homolog of the Mammalian CYP3A4 in Locusts
Comparison of Terfenadine Metabolism in Locusts and HLMs
Drug Metabolism and Disposition July 1, 2014, 42 (7) 1153-1162; DOI: https://doi.org/10.1124/dmd.114.057430


Simple Network Management Protocol i.e. SNMP is a simple request/response protocol in which SNMP manager communicates with SNMP agents/managed devices using SNMP PDU's (Packet Data Unit). Security is an important aspect wrt. SNMP as the pdus that are being broadcasted contain critical information about the managed devices.
Think of a scenario when someone gets access to this critical information and reboot the managed devices without the knowledge of network administrator or network operator. This can result in loss of service and hence loss of business. There can be many such scenarios where in invalid commands can be issued to the managed devices. Thus security should not be overlooked and modification of contained information or disclosure of information should not be allowed.
Different versions of SNMP i.e. SNMPv1, SNMPv2 and SNMPv3 implement security in different ways and as obvious, each version provides improvements over it's predecessors. Here is a brief overview of how security is implemented in each of the SNMP versions.
Only form of security used in SNMPv1 is community names. Community names are similar to passwords. The concept behind using community name is that all the managed devices in an SNMP network which are being managed by a network manager are considered to be in a community and a specific name i.e. community name can be assigned to it. While creating the SNMP PDU, this community name is set in the message header and any message received with the wrong value of the community name would be rejected by the receiver.
The community name set in the message header is in the form of plain text. Thus, it is very easy to fiddle with this data by eavesdropper.
In SNMPv2 all the information in pdu which contains source IP address, community name and other object instances/values except the destination address is encrypted. The encryption is done using DES. This is called as Party Based Security Model and is also known as SNMPv2p.
The SNMP agent can decode the encrypted data in the pdu and use the accepted IP address and accepted community name to validate the request.
SNMPv3 provides the latest architecture for SNMP security. USM i.e. User Based Security Model and VACM i.e. View Based Access Control Model are part of SNMPv3. USM is based on the user's access to a specific machine whereas VACM is based on user's access to a view that contains specific MIB objects. Unless specified, default model in SNMPv3 is USM.
Thus all the versions of SNMP provides security at some level. Hope you find the information presented here useful. Please let us know for any queries, feedback or suggestions.
I went through most of the articles in this website. They are so superb and great thing is each and every thing is explained in very simple terms. Thank you very much for sharing the knowledge,Keep it up!
I'd appreciate if you please explain the operation of USM/VACM in SNMPv3 in the same way yiu have explained existing articles. Thank you so much in Advance!!
I went through most of the topics in this tutorial, it was so useful. It helps newbie to understand SNMP protocol better.


Not gonna lie...I've taken a liking to caribou's lately. Don't @ me. They cute!
To my friends on snouts.online and all my other friends and followers here, please, read this thread.
I'm always around if another on the spectrum wants to talk.
I am smol fennec fren. Who wants to hold and snuggle me in their lap?
I'm in a big cuddle mood. Who wants to snuggle?
I hate this time of year sometimes....it just reminds me that I'm alone, like always. Everyone always asks me if I got my shopping done but I have no one in my life to shop for..... it just beings me down.
Aaaa this so adorable! I love this piece! I love it when snow lands on my snoot.
They need to bring goku and vegeta or goku and freeza to the smash roster!
Oh wow! I just realized I never did an #introduction! So here's mine!
You guys can call me Nitro! My main two sona's are Nitro, an Arctic Fennec and Blaze, a wolf right now but I am thinking about changing him to a Shadow Wolf, who I may switch to from time to time.
I'm 22, and live in Coventry. RI. I'm pansexual, although I can be very gay xD. I play a lot of video games and listen to music in my spare time when I'm not at work!
Thinking about butts. I just want to play with them and kiss them. I love butts.
Tmi Tuesday! Ask me anything!
I honestly might knock out early tonight. Got nothing special going on anyway.
Ugh. I can't sleep. I love being a switch but I'm feeling really subby tonight. All I can think about is I want someone to dominate me.
My cat is doing a heckin blep and I'm dying of cuteness!


Oakland Park & Rec Holiday Tournament — Registration open for boys and girls, grades 6-8. Tournament runs Dec. 20-21 at Skyline High for boys and Oakland Tech High School for girls. $200. For boys contact Chris Kyriacou 510-577-9100ext 292, for girls Curtis Cooper 510-414-3570.


text:
Succubus (eBook XIII)
Succubus Reissue eBook Cover,
written by Miss Irene Clearmont
Miss Irene Clearmont
December 2, 2013 (Original)
June 28, 2016 (Reissue)
B01H89RWKQ
Succubus Original eBook Cover, written by Miss Irene Clearmont
Succubus Alternative eBook Cover, written by Miss Irene Clearmont
Succubus is an eBook written by Miss Irene Clearmont. In this work the character Elspeth is a Succubus and the character Jason is an Incubus.
Title: Succubus
Author: Miss Irene Clearmont
Published By: FDC
ASIN: B01H89RWKQ
Publishing Date: December 2, 2013 (Original), June 28, 2016 (Reissue)
Erotic horror and female domination, in both modern and classical form, is the order of the day in this horrific tale of men under female control from the ever inventive and diabolical Miss Irene Clearmont.
With "Succubus", the glorious Miss Irene gives us the tale of a beauty with roots dating back to the time of Carthage – and a need to control, dominate and destroy her male victims that is equally as old.
Male victims whose essence she takes during sex ensures she maintains her youth and beauty.
The following review was originally published by Tera on her Blog, A Succubi's Tale on June 28, 2015
The world contains mysteries and questions beyond understanding. The path of Elspeth is one that has been touched by her for centuries uncountable. A need within growing stronger, a means being searched for. But along with that comes another searching for something else which will draw them both and other together into fates unknown.
Simply the most amazing piece of Succubus literature that I have read this year. The detail in the story, the presence of the characters is just exquisite. The work is not a simple piece of erotica by any means, there is so much more to be seen within the pages. It is complex, gripping, and once begun simply cannot be put down.
The main character, Elspeth, passes along a journey from the far past to the present day and as that unfolds history is seen through her eyes in one moment, but at the next, her own need and desire pushes at the limits. The story tells of her struggle with herself on the one moment, but in the next her deeply rooted need to dominate, control, and survive.
The myths of Succubi are mixed with that of Incubi and the Lamia among others. There are some wonderfully complex myths that come into play over the course of the story and this brings Elspeth's journey into sharpen focus.
Elspeth herself changes with time, becoming kind of a chameleon as need be in order to survive. Along the way we see her encounter her prey and then see what her belief within drives her to do. There is a good deal of dominance in her character which I thought was well told in the work. She does have needs as a Succubus, which cause problems for herself and the ends to many others, but this isn't done haphazardly or at a whim. There is purpose, direction, and reason to a point in her actions which did hold the story at the fore of the work. There is some lovely heat in the erotica, a bit of Succubus mind control appears here and there which I thought was well done also.
The writing is solid, there are no moments of being taken out of the story, or a misstep in a word or phrase being out of place. Every action matters. Every event in important. Every character, no matter how small their role, drives the story onwards and that tells of someone that knows their craft, why they are writing the work and where they want to be when it is over.
There is an Incubus that appears in the work as well, and it is an interesting exercise to compare him and Elspeth's natures. Both are complex, both are predators, but more importantly, it is the difference in their attitudes, what they desire most of all that separates them.
The work is told in three arcs, all interconnected and I found that in being written this way there were moments that connected back and forth which added to the overall understanding of Elspeth which I appreciated. To see how events swirled around her, what the consequences were at later points along the story was breathtaking at times.
The work ends on a twist that is quite unexpected, but in doing so leaves behind many questions and thoughts to wonder over. In doing so the work leaves a lingering impression that is wonderfully done. The strength of the work comes in the characters, their lives and the choices they make. Simply wonderfully told and more.
Five out of five pitchforks.
Highly recommended. While a bit less romantic than I normally prefer, that is well overshadowed by the strength of the characters, the power of the story, and more.
My thanks again to the author for sharing this work with me and satisfying my curiosity. Well worth the read by any measure and I wish the author all of the best in her future writings…
The author Miss Irene Clearmont's website
This work available for purchase on the author's website
Retrieved from "http://succubus.net/wiki/index.php?title=Succubus_(eBook_XIII)&oldid=48083"

redpajama_set_name: RedPajamaCommonCrawl


Watching and Listening – An Alternative to Direct Questioning?
In the ea rly 90's, the 'tagline' of a leading Japanese agency's brochure was 'the key to understanding was to read between the lines of what was not said'. Two decades later, Shobha Prasad re-visited the issue focusing on the layering and subtleties of Asian languages in 'Listening to the Sounds of Silence' at ESOMAR's APAC Conference.
As Asia grows more healthy, wealthy, and wired, superficially its consumers resemble their Western counterparts. Sometimes even, having leapfrogged technological lifecycles, they appear more advanced. Nevertheless, although these new trappings bring an almost 'stateless' vocabulary to the world's languages, the fundamental challenges of interpreting local cultural nuance and international comparisons remain.
The Myth of Market Research's Failure.
I am getting increasingly angry about the number of posts, books, You Tube Videos and articles – often by market researchers themselves – that imply "conventional Market Research" is a failure.
Here's a good example, 'futurist' Patrick Dixon talking about why market research is "often wrong": http://tinyurl.com/25kp34z .
An assertion that "traditional MR" misses out on some extraordinarily key factor in understanding consumers, be it an inability to capture emotion, or failure to understand the role of Social Media or whatever uber-trend the author is fascinated by.
Avoiding Extinction – Reinventing the Survey.
Are Surveys Heading For Extinction? The Approaching Storm.
In the next few years there will be a significant lift in the use of traditional "fact gathering", ad hoc surveys. This will be driven by demand for data in growth economies and in sectors where currency services and data-bases are still in their infancy. At the same time, in developed markets, as the recession recedes and firms start to generate cash they will realise a need to reacquaint themselves with consumers whose lives and values have been transformed by recent events. This Indian Summer of the traditional survey will not last, however, and firms that get fooled by the burst of prosperity and fail to prepare (by investing in R&D, software, and training) for a new world will ultimately fail.
Are Surveys Headed For Extinction?
Every year an increasing number of surveys are being carried out in more and more countries. Only a decade or so ago many people lived out their lives without a single interviewer asking their opinion on the government of the day or paper towel brands. In major cities at least, it's a rare person who can say that today. Yet, these staples of our industry are increasingly under threat, partly by new alternatives, partly by social changes and possibly, in largest part, by our collective inattention as an industry. Time, in my view, to rethink what a survey is good for and reinvent what we do with them.


text:
LOCAL SENIOR FOOTY
Blumers Lawyers CRC Round 13 Preview
crrl.com.au
Fri 12 Jul 2019, 01:54 PM
The Southside Showdown takes centre stage this weekend as the Woden Valley Rams host the Tuggeranong Bushrangers in the Match of the Round.
Woden Valley Rams v Tuggeranong Bushrangers
The Woden Valley Rams will be looking to stay within the top four as they host league leaders the Tuggeranong Bushrangers.
The Rams suffered a big loss last weekend after going down to the Kangaroos, who now sit one point behind them on the ladder.
The Bushrangers further increased their lead at the top of the table after narrowly defeating the Bulls 26-24.
The league leaders sit comfortably on top of the Blumers Lawyers CRC Ladder, three points ahead of the chasing pack.
Yass Magpies v Belconnen United Sharks
The Yass Magpies will be looking to make it two wins in a row and get their season back on track when they host the Belconnen United Sharks.
The Magpies came from 12-points behind last weekend against the Queanbeyan Blues to win 36-32 and claim their second win of the season.
The Sharks will be hoping to return to winning ways after going down to the Bulldogs 34-6.
The away side sit three wins outside the top four, so a Round 13 victory would be crucial to help keep their finals hopes alive.
Queanbeyan Blues v Gungahlin Bulls
The Queanbeyan Blues will be hoping to regain their winning mentality when they face the Gungahlin Bulls.
The Blues suffered a shock four-point loss to last placed Magpies despite leading at the break 16-4.
The Bulls also head into the Round 13 clash of the back of a defeat, narrowly going down to league leaders the Bushrangers.
The Bulls sit eighth on eight points and will be hoping a victory can keep their finals hopes alive with only just five rounds left.
Goulburn Workers Bulldogs v West Belconnen Warriors
Goulburn Workers Bulldogs will be looking to close the gap at the top of the table when they face the West Belconnen Warriors.
The Bulldogs enter the clash off the back of a huge 34-6 win over the Sharks with the Cornish brothers Nick and Mitch control the game.
The Warriors head into the game raring to go off the back of a bye, hoping to remain in close nit with the top four.
The Warriors sit in sixth with 14 points, with a win in Round 13 putting them on level points with majority of the top four.


text:
package gtd.util;

public class MarkableForwardSplittingLink<E>{
	public final ArrayList<MarkableForwardSplittingLink<E>> nexts;

	public final E element;

	public boolean mark;

	public MarkableForwardSplittingLink(E element){
		super();

		this.nexts = new ArrayList<MarkableForwardSplittingLink<E>>();

		this.element = element;
	}

	public void addLink(MarkableForwardSplittingLink<E> next){
		nexts.add(next);
	}

	public void setMark(){
		mark = true;
	}
}

redpajama_set_name: RedPajamaGithub


We've been installing tile bathrooms, tile showers and serving other high end tiles needs to Douglas County and the Denver metro area for some time. "Brunetti Tile" - We are a Tile installation company with over 11 years experience in working with, setting or repairing custom tile. We can custom build our customers anything from hand-made mortar beds, shower benches and damns to tile 100% custom built tile counter-tops or tables. We work with tile from mosaic and field tiles to large 24" or bigger tile. "Brunetti Tile" provides these services to both residential and commercial customers. Tile Company in or near the DTC Colorado. We can provide help, sales and service to Russian speaking clients if needed (no not Italian, weird I know). A tile shower should be fun, unique and relaxing! Work with a tile shower contractor in Castle rock colorado you can trust! Shall a customer just need a large SF area of floor tiles laid down as quickly (but professionally) as possible, well we can do that too! We do not sub out our clients work. We meet with our customers, we provide the shower bathroom tile estimate, and WE show up to do the work. Contact Brunetti tile today for a free estimate on a tile bathroom, tile mortar bed or damn, tile shower, tile sauna or bathroom remodel that needs tile.
Will we install tile or repair it for you!
We love to provide free onsite tile estimates and answer any tile questions you may have if your anywhere around the Denver Tech Center (DTC) to Douglas County. Brunetti Tile also services Lone Tree and Parker. Free Tile Estimates for the DTC, online or on site!


text:
<?php
// Check for empty fields
if(empty($_POST['name'])      ||
   empty($_POST['email'])     ||
   empty($_POST['phone'])     ||
   empty($_POST['message'])   ||
   !filter_var($_POST['email'],FILTER_VALIDATE_EMAIL))
   {
   echo "No arguments Provided!";
   return false;
   }

$name = strip_tags(htmlspecialchars($_POST['name']));
$email_address = strip_tags(htmlspecialchars($_POST['email']));
$phone = strip_tags(htmlspecialchars($_POST['phone']));
$message = strip_tags(htmlspecialchars($_POST['message']));

// Create the email and send the message
$to = 'administracion@briccancun.org'; // Add your email address inbetween the '' replacing yourname@yourdomain.com - This is where the form will send a message to.
$email_subject = "Formulario de Contacto:  $name";
$email_body = "Recibiste un nuevo mensaje del formulario de contacto del sitio web.\n\n"."Nombre: $name\n\nEmail: $email_address\n\nTelefono: $phone\n\nMensaje:\n$message";
$headers  = 'MIME-Version: 1.0' . "\r\n";
$headers .= 'Content-type: text/plain; charset=UTF-8' . "\r\n";
$headers .= "From: noreply@briccancun.org" . "\r\n"; // This is the email address the generated message will be from. We recommend using something like noreply@yourdomain.com.
$headers .= "Reply-To: $email_address";
mail($to,$email_subject,$email_body,$headers);

// Email to Person
$título = 'Gracias por su interés';
// mensaje
$mensaje = '

            Brigada de Rescate Internacional Cancún, A.C.

Gracias por su interés.

            Se ha enviado su consulta a la persona correspondiente; se pondrán en contacto con usted lo antes posible.

';
// Para enviar un correo HTML, debe establecerse la cabecera Content-type
$cabeceras  = 'MIME-Version: 1.0' . "\r\n";
$cabeceras .= 'Content-type: text/html; charset=iso-8859-1' . "\r\n";
$cabeceras .= 'From: noreply@briccancun.org';
// Enviarlo
//mail('conomia_alfredo@hotmail.com', $título, $mensaje, $cabeceras);
mail($email_address, $título, $mensaje, $cabeceras);

return true;
?>


WAIR appeared in the early 00's with indie pop and indie rock music. Various hosts were heard on the air, including "Ravi" and "Ira Glass", though the main host seemed to be Robert J. Yardbrown.
This page was last modified on 25 February 2011, at 22:31.


Huma is pleased to welcome Lisl George, Administrative Officer, responsible for financial and human resource management. Lisl has worked in research administration at UCT for 13 years and has been in office management for more than 20 years. She has a business administration qualification from the University of Cape Town's Graduate School of Business.


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These machines are built in-house at the Genelite Brisbane workshop using quality heavy-duty Australian-made anti-vibration mounts and solid heavy-duty powder-coated frames.
These Yanmar engines have a highly precise and efficient FIE direct injection system, making use of every droplet of fuel, and they offer superb lifetime value. They are powerful and compact, making them suitable for a variety of mobile uses.
This model is a 'mine spec' model which has many additional features suitable for mining applications.
The Genelite range are Power You Can Rely ON!
Be the first to review "5/6.5kVA Yanmar GYD5000E Mine Spec" Click here to cancel reply.


text:
Home Activism Accountability Aviation companies blame FBI, CIA and terrorists for 9/11
Aviation companies blame FBI, CIA and terrorists for 9/11
NEW YORK: Aviation companies sued by the families of Sept. 11 victims for failing to safeguard air travel are in turn blaming federal investigators — arguing the Federal Aviation Administration was not alerted that al-Qaida was poised to launch terrorist attacks.
In court documents filed this week in U.S. District Court in Manhattan, aviation companies are seeking to force five FBI employees to provide testimony that may help defend against claims the companies share blame in the attacks.
"The aviation parties anticipate that the FBI witnesses' testimony will demonstrate that the FBI had information before Sept. 11 indicating that al-Qaida may have been about to launch terrorist attacks on civil aviation, which it did not timely pass along to the Federal Aviation Administration," lawyers wrote.
The airlines and aviation companies are defending themselves against lawsuits seeking billions of dollars in damages for injuries, fatalities, property damage and business losses related to the Sept. 11, 2001, terrorist attack.
The companies in turn filed separate lawsuits against the CIA and the FBI last August to force terrorism investigators to tell whether the aviation industry was to blame for the Sept. 11 attacks.
The latest documents filed by the airlines, airport authorities, security companies and an aircraft manufacturer argue that if the FAA had known about an FBI investigation of Zacarias Moussaoui weeks before the Sept. 11 attacks, it could have amended security measures to guard against the type of terrorist attack Moussaoui was planning.
The aviation defendants said the FBI has refused to permit even a single deposition, although the agency does not deny that five potential witnesses in the case have already testified and made other public statements before the 9/11 Commission, the Moussaoui trial jury and the media.
In the lawsuit against the CIA, companies including American Airlines Inc., United Airlines Inc., US Airways Group Inc., Delta Air Lines Inc., Continental Airlines Inc. and The Boeing Co. are seeking to interview the deputy chief of the CIA's bin Laden unit in 2001, and an FBI special agent assigned to the unit at that time. The names of both are secret.
The lawyers said testimony from both agencies was critical to their defense.
"After weighing this evidence together with the criminal acts of the terrorists, the jury would be entitled to conclude that any act or omission by the aviation parties was so dwarfed by the other causal factors of the attacks that the aviation parties' conduct was not a substantial cause of the plaintiffs' injuries," the lawyers wrote.
U.S. Attorney Michael Garcia said last year that the request for depositions with FBI agents was rejected because the aviation companies didn't explain how FBI information before the attacks would relate to their defense efforts.
Garcia said much of the information was protected from disclosure, "because it involves classified national security information or matters protected by the law enforcement investigative privilege."
He also said it would be "extremely difficult and burdensome" to separate the classified information from the non-classified information and risk that some classified materials may be inadvertently disclosed.
Yusill Scribner, a spokeswoman for government lawyers in the case, said there was no comment Wednesday.
Source URL: http://www.iht.com/articles/ap/2008/04/30/america/Sept-11-Lawsuits.php
Related article: Of course, this reports omits the action taken by the FAA prior to the attacks to warn airports in the spring of 2001 that if "the intent of the hijacker is not to exchange hostages for prisoners, but to commit suicide in a spectacular explosion, a domestic hijacking would probably be preferable.".
Zacarias Moussaoui
Previous articleRoots of surveillance standoff go back decades
Next articleIt's Not About Revenge, This Is About Justice–Lorie van Auken
FBI agent sought CIA help to investigate Sarasota Saudis who fled U.S. before 9/11
Barr, Grenell block 9/11 families access to evidence with broad, 'unprecedented' claims of secrecy
9/11 Families, Experts Mount Unprecedented Challenge to NIST in New Filing


Astronomy is a subject in science. Now in the modern world of computers, people can create pictures by completely different programs which can be really captivating. In accordance with future technology information nature will be fully protected from air pollution utilizing these new technologies as it won't produce any deadly or hazardous chemical solvents or gasoline products into the air.
In my observation it is most of the time the successful one that is artistic, not a selected kind of particular person. Digital artists already in contact with technology are conscious of the methods which can be in trend and use them to create their artwork work and promote them on-line.
Although one part of technology tends to dominate a nation's actions at a specific time, different phases will probably be usually practiced at the similar time. Technology opens completely different paths for the artists to enter into for a great incomes. This ensures that after graduation they are going to be capable of use the technology in a piece setting, which can put them forward of somebody who didn't have entry to a selected technology or software in their very own college setting.


text:
Middle East World
Saudi girl facing death for non-violent protest (V)
By Reuters News Service October 25, 2018 October 25, 2018 0166
Saudi Arabia must immediately and unconditionally release all women it has detained for campaigning for human rights, officials mandated by the United Nations said.
Saudi authorities have detained more than a dozen women's rights activists since May. Most campaigned for the right to drive – which was granted in June – and an end to the kingdom's male guardianship system, which requires women to obtain the consent of a male relative for major decisions.
This statement, from experts who report to the U.N. Human Rights Council, called for the release of six women.
They include one, Israa Al-Ghomgham, who is facing possible execution, a threat the experts called "reprehensible".
They said they were in touch with Saudi authorities.
The Saudi mission in Geneva did not immediately respond to a Reuters request for comment.
Israa Al-Ghomgham is a 29-year-old Saudi woman who is facing the death penalty for her peaceful protesting. Here are all the shocking details before her sentencing on October 28th.
picks4Saudi Arabia
Finance ministry decision to hire consultants directly was unlawful, AG says (updt)
Hemp grower remanded for growing THC-rich plants (Updated)
Lovers in exile in Cyprus
Kato Amiandos house damaged by fire, no injuries
Police warn over icy roads, help eight drivers (updated)


text:
Mangum Manor Park
☏ Useful Numbers
This is an informational page for Realtors.
Mangum Manor is a civic club with voluntary annual dues of $15 per year per household.
There are no transfer fees.
Mangum Manor does have deed restrictions which may be viewed in the Deed Restrictions section of this website. Mangum Manor deed restrictions are also on file with Harris County. To obtain a copy, please contact the Harris County Clerk's Office at (713) 755-6405 during business hours. If you wish, a copy may be obtained in person at the Harris County Clerk's Office located downtown in the Harris County Civil Courthouse, 201 Caroline, 3rd Floor, Monday through Friday from 8:00 a.m. to 4:30 p.m.
Mangum Manor is divided into three sections, which each have a distinct set of deed restrictions. The sections are delineated in the Maps section of this website.
Buyers, Builders, Developers, etc. are expected to follow the law with respect to City of Houston noise ordinances, construction codes, trash and refuse laws, etc.
Mangum Manor has opted in to the city ordinance which prohibits parking vehicles on yards.
Parking in a manner which blocks a sidewalk is prohibited by city ordinance.
Trash collection day is every Thursday and trash may be placed out no earlier than 6 PM the preceding day by city ordinance.
Mangum Manor was a pioneer in the City of Houston single-stream recycling program. Recylcing is collected the first and third Thursday of each month and may be placed out no earlier than 6 PM the preceding day by city ordinance. It is also against the law to remove items from a recycling container not your own.
Heavy Trash(Tree/Junk Waste) is the fourth Tuesday of each month and may be placed out no earlier than 6 PM the preceding Friday by city ordinance.
This page was created to help assist and inform realtors and developers with working in Mangum Manor and answer some of the most often asked questions. Mangum Manor is a community of both newcomers as well as original residents living here since the neighborhood was first built in 1955 and everything in between.
If you have additional questions, please feel free to email them to contact@mangummanor.org
© 2020 Mangum Manor Civic Club
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I offer customised tuition on either a one-to-one basis or for small groups (up to a maximum of 6). Sessions can be from one hour to a whole day (or more) and can be field-based photography sessions or indoor technical tuition, or even a mixture of the two.
Modern cameras and computer software can be daunting in their complexity, and many people need a little help to get the most from their photography.
Please contact me for more details (including rates), or to discuss your requirements.


Chat bots are web applications which use artificial intelligence to carry out believable conversations with human users. I have always found chat bots interesting, and decided to create my own using Java. The chat bot I created was very simple and did not work terribly well, but gave reasonable responses to common conversations. While it could in some sense learn, the bot I made had no understanding of the English language and was effectively hardwired to give certain responses. In this article I will briefly outline how it worked. It is interesting how simple it can be and still give sensible responses. Below is a conversation I had with the chat bot. The percentages correspond to how certain the bot is that his replies make sense.
A chat bot is not directly programmed to talk in and understand English. It would be almost impossible to do that. Instead, the bot is created in a way where it can learn to converse with humans. Therefore it begins not really being able to do anything until it learns from examples. The set of examples that the bot is given to learn from is called the training set. The training set for a chat bot would probably be a set of conversations that demonstrate sensible responses to questions the chat bot is likely to be asked.
My chat bot stored a set of pieces of dialog in a file on the computer. Specifically it stored a set of statements and responses to the statements. For example, a statement might be "How are you?" and the response might be "Good." I called each such pair of sentences a dialog fragment. A set of dialog fragments can tell the computer what to say in many situations. For example, if someone says "What is your name?" to the bot, it can look up the dialog fragment that contains "What is your name?" and will know that "My name is ChatBot" is an appropriate response. The bot can learn how to respond to different phrases by being told appropriate responses, and can save them as dialog fragments. The bot could even learn new dialog fragments on the fly.
There are a few small optimizations that can make this simple technique work reasonably well. I will outline exactly how the bot I created works. After having saved a large sample of dialog fragments that are reflective of common conversation, the chat bot is ready to talk to an actual user. The user says some statement to the bot. The bot reads the statement and breaks it up into separate words. The bot then searches all of the dialog fragments it has saved with similar initial statements. It determines which fragment contains an initial statement that is most similar to the one the user entered (has most words in common). Finally, the bot prints the response it has associated with this statement out.
It is also worth noting that the percent similarity between the initial statement in the fragment and the user's statement can be used to gauge how confident the bot is that it has a sensible answer. The confidence is less than 50% or so the bot probably does not know what to say, and can say something else like "I don't know what you mean."
Using the technique I described the chat bot can produce reasonable responses for very common statements like "How are you?" I hope you found it interesting to know how you can do it! Chat bots can of course be much more sophisticated than this and demonstrate a much deeper understanding of the conversation. My chat bot did not do any "natural language processing", so was not very well equipped to understand statements. I did find it fun to make though. I have not trained my chat bot to respond to many different things. It also does not format it's output properly, and seems to have a few bugs. It certainly isn't finished. However, if you are interested you can download the source code here and try talking to it. It is just a console application. Maybe in the future I will improve the bot more, give it a GUI, and train it to respond to more things. If and when the program is more capable I will put it under the projects section of my website.


text:
So, a lot of people are very competitive in everything they try. They want to have the best business if they get into it, they want to be the best at their job when they get it, etc. If you are that sort of person and you look for excellence in your game but do not want to commit professionally to this then it is going to take an upwards of 5-10 years to play at some decent level like club level or USTA sponsored tournaments.
Tennis racquets come in a very wide variety of price ranges, to say the least. You may want to set yourself a budget to help narrow down your choices because you will have so many of them. Do we find a correlation between the price of a tennis racquet and the quality of it? The answer here is absolutely but you have to be careful here too.In general, the higher the price a tennis racquet is the better quality that tennis racquet is because it most likely much innovative design features built into it. Be careful here too, though. Because things such as the tennis racquet brand or a tennis racquet that is designed by a famous player may up the price of a tennis racquet without it making the tennis racquet better than some lower priced models.
A game consists of a sequence of points played with the same player serving, and is won by the first side to have won at least four points with a margin of two points or more over their opponent. Normally the server's score is always called first and the receiver's score second. Score calling in tennis is unusual in that (except in tie-breaks) each point has a corresponding call that is different from its point value. The current point score is announced orally before each point by the judge, or by the server if there is no judge.
The best marks for this racquet were awarded to the services on which the racquet split the air with power on contact to allow them to reach their targets. However, the EZONE 98 (305 g) will not suit everyone. Some players will enjoy the more damped feel of the DR, while others will prefer the firmer, more powerful response offered by this new version.
So, if you are reading this one, you are probably a parent who sees some talent in their child and wants them to go pro when they grow up. It is essential for your child to start Tennis at a very young age, like close to 5-6 years. What this does is make the game instilled in their muscle memory forever. Tennis becomes a habit for them and it becomes second nature.
Baseline Battler: This is top spin guru Rafael Nadal's racket of choice. It has a very stiff body that allows for better ball feel on each hit and can help improve control for those looking to tighten up their game. Aero is still using their wing shaft design in conjunction with an 11.2oz weight for fast speed both from the back court and for quick exchanges at the net. It has a 100 square-inch head with a nice, big sweet spot which makes is good for players of any experience level. While you'll be able to move the ball with precision, you can also really flatten it out. Even from deep in the back court you can rack up some fairly vicious speed to cut your opponent off at the knees. [Purchase: $194+]
During the 19th century the Royal Tennis Court played a variety of different roles. It served as a theatre storeroom, a workshop and storage space, and a studio for the painters Antoine-Jean Gros and then Horace Vernet. It was listed as a national monument in 1848, then became a games room again under the Second Empire. Approaching the centenary of the oath, under the Third Republic, architect Edmond Guillaume was invited to restore it and convert it into a museum of the French Revolution. The architect used original engravings to reconstitute the spirit of the room, which had been modified and had deteriorated over the course of the century. The museum housed the statue of Bailly, busts of the most important signatories of the oath, and a monumental canvas by Luc-Olivier Merson, based on Jacques-Louis David's preparatory drawing. The museum was inaugurated on 20 June 1883 in the presence of Jules Ferry.
If you are beginner starting out you might have been scared by all the different complexities of the game and might have felt overwhelmed by them. You might be wondering that it takes a very very long time to achieve some sort of proficient level in this game. You will be surprised to know that tennis is a very awesome game and if you keep your level of play consistent and employ the right technique, you will be able to play at a pretty good level in no time at all compared to other sports.
^ Lorde (2013). Lyrical Influences (VEVO LIFT): Brought to You By McDonald's (video). VEVO/YouTube. Event occurs at 2:25. Retrieved 22 November 2013. When I wrote "Tennis Court", I was looking at a lot of photography by this guy called Gregory Crewdson who, um, makes these huge, super intricate, really beautiful photographs. They kind of just, like, depict human life, and there is this an emptiness to them and there is a suburbia to them which I find really compelling. And so I was definitely thinking of that visually.
He said there was no way he could ever have been a pro player, and that he knew that in his early teens — for one thing, there wasn't the money to get him to tournaments beyond those near his home. He plays competitively now one night a week, which is all his schedule allows, hitting with guys his age who had played serious college tennis and, in some cases, joined the low rungs of the pro tour for a year or two.
In the US, "Tennis Court" was released as a 7-inch vinyl single on 27 August 2013.[14] Lava and Republic Records initially planned to service "Tennis Court" to US modern rock radio on 11 March 2014 and contemporary hit radio (CHR) on 8 April 2014 as the album's third US airplay single, following "Royals" and "Team".[15] Republic cancelled the scheduled release in favour of "Glory and Gore", which impacted modern rock radio on 11 March.[16][17] The label also planned to service "Glory and Gore" to CHR, but subsequently called of its release and serviced "Tennis Court" to CHR as originally planned.[18] "Tennis Court" ultimately impacted hot adult contemporary radio and CHR on 21 and 22 April 2014, respectively.[19][20] It was released to UK radio on 12 May 2014.[21]
A tennis court is the venue where the sport of tennis is played. It is a firm rectangular surface with a low net stretched across the center. The same surface can be used to play both doubles and singles matches. A variety of surfaces can be used to create a tennis court, each with its own characteristics which affect the playing style of the game.
Consider a player who wins six games in each of two sets, all by a score of game–30. The winner has scored 4×12 = 48 points and the loser 2×12 = 24. Suppose also that the loser wins four games in each set, all by a score of game-love. The loser has scored 4×8 = 32 points and the winner zero in those games. The final score is a win by 6–4, 6–4; total points 48–56.
Net Worth: The Radical Pro was what was sitting in Andy Murray's hand when he rocked Wimbeldon, so it damn sure deserves a place in your bag. At 11.5oz it is a nice mid-weight racket, though thanks to the way its balanced it moves like a much lighter piece of equipment. On power swings you are going to notice some sluggishness, but barely enough to make a difference. This is definitely a control players racket with its 98 square-inch head which allows for decent serve speed but sluggers won't get the crushing power they desire. For smooth operation when making a net play, it works wonders and allows for sniper-level ball placement. For pure power it's not a winner. [Purchase: $190]
Well, as you learn to play tennis, you need to know how to continue the play. If the serve is successful, the play may continue with a variety of shots. Often, play will continue with groundstrokes. Groundstrokes are shots taken after the ball has bounced once. These can be hit with a forehand or a backhand stroke. The forehand stroke is made with the face of the racket, the palm of your hand facing the ball. The backhand stroke is made with the reverse side of the racket, with the palm of your hand facing away from the ball. To successfully learn to play tennis, you'll need to learn to hit forehand shots; and to successfully learn to play tennis, you'll need to learn to hit backhand shots.
When playing a match, it is usually best to report each score out loud with one's opponent to avoid conflicts. During a game, the server has the responsibility to announce the game score before serving. This is done by announcing the server's score first. If, for example, the server loses the first three points of his or her service game, he or she would say "love–40". This is to be done every time. After a set is complete, the server, before serving for the first game of the next set, announces the set scores so far completed in the match, stating his or her own scores first. If the server has won the first two sets and is beginning the third, he or she would say, "two–love, new set." If the server had lost the first two sets, he or she would say, "love–two, new set." Finally, after the completion of the match, either player, when asked the score, announces his or her own scores first.
Outside Australasia, the single achieved modest commercial success. It peaked at number 78 on the Canadian Hot 100 and remained for five weeks on the chart.[61] The track earned platinum certification from Music Canada, which denotes shipments of 80,000 copies in the country.[62] The single had the similar position on the UK Singles Chart and was certified silver by the British Phonographic Industry for track-equivalent sales of 200,000 units, based on sales and streams.[63][64] It also entered the top 100 of the German Singles Chart, where it reached number 83.[65] Despite not entering the record chart of Norway, the song was certified platinum by IFPI Norway for exceeding sales of 10,000 units in the country.[66] In the US, "Tennis Court" debuted at number 71, which was also its highest placement, on the Billboard Hot 100 chart dated 19 October 2013.[67] The single spent 18 weeks on the chart and had sold 355,000 copies in the US by April 2014.[68][18] It also reached number nine on the Hot Rock Songs, a Billboard auxiliary chart.[69]
As it happens, I was reading a ruefully captivating new memoir called "Swimming Studies," by a onetime contender for the Canadian Olympic team, Leanne Shapton, which explores how growing up a competitive swimmer formed her habits of heart and mind. Years later, her daily rhythms and life choices, her nightly dreams, her understanding of duration, pleasure, pain and reward, remain informed by her hours in the training pool. Swimming strokes carved the contours of her inner life, and it's not at all clear she is thankful for that. Of course, Andre Agassi, in his memoir, writes of how the aloneness of singles tennis — the very thing that imparted to Kirill a kind of Emersonian self-reliance, as he understands it — just enlarged his loneliness. You never know.
Learn to play doubles tennis. Doubles tennis has two players on each side instead of one. You'll use the wider court dimensions, but the rest of the scoring and rules remain the same. The big challenge for doubles tennis beginners is to learn how to interact with a teammate. Ask other tennis-playing friends to teach you the best strategies for doubles tennis.[23]
Advantage sets sometimes continue much longer than tie-break sets. The 2010 Wimbledon first-round match between John Isner and Nicolas Mahut, which is the longest professional tennis match in history, notably ended with Isner winning the fifth set by 70–68. The match lasted in total 11 hours and five minutes, with the fifth set alone lasting eight hours, 11 minutes. Whoever wins by a margin of two wins the set, but this could take a very long time to finish.
By 1975, aluminum construction improvements allowed for the introduction of the first American "oversized" racket, which was manufactured by Weed. Prince popularized the oversize racket, which had a head size of approximately 110 square inches (710 cm2). Howard Head was able to obtain a broad patent for Prince, despite the prior art of the Bentley Fortissimo (the first oversize, made in Germany of fiberglass) and the Weed. The patent was rejected by Germany but approved in the USA. The popularity of the Prince aluminum oversize had the side effect of popularizing rackets having other non-standard head sizes such as mid-size 85–90 square inches (550–580 cm2) and mid-plus sizes 95–98 square inches (610–630 cm2). Fairly quickly, midsize frames began to become the most-used frames in the pro tours. Martina Navratilova popularized the midsize graphite racket, with her wins using the Yonex R-7, the first midsize graphite racket made by Yonex. Nearly at the same time, however, she said the "jumbo" rackets (midsize included) should be removed from the sport for making it easier. She said she would use them only because other players could, as they were tournament-legal. Fewer players chose to use oversize rackets, and some switched to midplus frames after their earliest career for more control. Fiberglass frames also had a brief period of limited popularity, making fewer inroads among top players than aluminum. Also, the earliest composites, such as the Head Competition series, used by Arthur Ashe, were made without graphite. These were more flexible than a typical early graphite composite but stiffer than wood, fiberglass, and aluminum.
If I had come too late to tennis to get very good, or for the game to play a role in who I am, there are experiences and sensations it has been providing me while I'm on the court that are new and surprising. I feel helpless when I tire and lose focus — focus has never been an issue for me. I get angry with myself at times while playing, Andy Murray angry, though I have never gone so far as to repeatedly slap my forehead with my palm. I think you have to be really good to get away with that.
Regardless of the number of games and sets in a match, players must continuously rotate sides when the total number of games in a set is an odd number. For instance, if the total game score in a set is 3-2, simply add 3 + 2 = 5. Since 5 is an odd number, the players rotate sides prior to starting the next game. If the total game score in a set is 5-1, simply add 5 + 1 = 6. Since 6 is an even number, the players do not rotate sides.
This is Kirill's twice-a-week way of reminding me, however inadvertently, that I started playing tennis too late. Footwork is key to tennis, and taking short strides as you are approaching the ball sets in place the underpinnings of everything that might lead to a good shot. Short steps allow one to be neither too close to nor too far astride the ball. But short steps are not what you learn when you spend your childhood playing football and baseball and basketball.
Before and during the open era, Rod Laver remains the only male player in history to have won the calendar year Grand Slam twice in 1962 and 1969 [101] and also the calendar year Professional Grand Slam in 1967.[102] More recently Björn Borg and Pete Sampras were regarded by many of their contemporaries as among the greatest ever. Andre Agassi, the first of two male players in history to have achieved a Career Golden Slam in singles tennis (followed by Rafael Nadal), has been called the best service returner in the history of the game.[103][104][105][106] He is the first man to win grand slams on all modern surfaces (previous holders of all grand slam tournaments played in an era of grass and clay only), and is regarded by a number of critics and fellow players to be among the greatest players of all time.[103][107][108] However it must be noted that both Rod Laver and Ken Rosewall also won major Pro Slam tournaments on all three surfaces (grass, clay, wood) Rosewall in 1963 and Laver in 1967.[109]
In 1926, promoter C. C. Pyle established the first professional tennis tour with a group of American and French tennis players playing exhibition matches to paying audiences.[26][37] The most notable of these early professionals were the American Vinnie Richards and the Frenchwoman Suzanne Lenglen.[26][38] Once a player turned pro he or she was no longer permitted to compete in the major (amateur) tournaments.[26]


We all know that performing aerobics and other workouts will make our physique fitter and healthier. Flutter Kick: Legs are prolonged straight back, in keeping with your physique, as you kick them up and down. Now that you have the basics down, it's time to improve the intensity of your swimming routine. Swim 75 yds., then swim 75 yds., respiratory each 5 strokes; relaxation 15 seconds.
Adding coaching aids to enhance your swim when fatigued helps promote great technique being carried over into your subsequent session. And because water is sort of 800 occasions denser than air, each kick, push, and pull is sort of a mini resistance workout in your entire body—particularly your core, hips, arms, shoulders, and glutes.
Listed below are 40 swimming workouts for sprinters, distance swimmers, butterfliers, IM'ers, and everybody in between courtesy of a number of the high programs, swimmers and coaches on this planet. Hit the pool to scorch fat and enhance your stamina with this 650-yard exercise.
From distance to dash , newbies to triathletes-take a look at our ever-growing assortment of swimming sets. Distances are based on a 25-yard pool (one length); to do these routines, you should really feel comfy swimming at least 100 yards without stopping.
John Mora, writer of Triathlon 101," means that new swimmers take a beginner's swim class to study the fundamental fundamentals of swimming. For these nicely versed in IM and trying to improve their distance per stroke ratio, get able to work.


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Border Patrol agents in Arizona discovered two large groups of Central American migrants, 306 people in all, this week near Lukeville in southern Arizona.
The group included families, pregnant women and children primarily from Guatemala, El Salvador and Honduras, according to a U.S. Customs and Border Protection said in a statement.
Agents found the first group of 242 people Wednesday while patrolling the border about 15 miles west of Lukeville, the agency said. On Thursday, agents patrolling on the Tohono O'odham reservation discovered 64 more people. Agents took the migrants to Tucson for further processing and medical evaluation, the agency said.
Officials also identified nine children in the group ages of 1 to 13 with flu-like symptoms, Customs and Border Protection said. They were taken to a Tucson hospital.
Border Patrol agents in Arizona have found numerous groups of Central American migrants traveling into the state from Mexico since summer. The incidents preceded the movement of large border-bound caravans.
Migrants say policy keeping asylum-seekers in Mexico won't stop more from coming
As ICE releases migrant families, churches and businesses step up to help
What will $5 billion in border wall funding build? 215 miles of fence, feds now say


text:
package org.springframework.boot.actuate.autoconfigure;

import java.lang.annotation.Annotation;

import org.springframework.boot.autoconfigure.condition.ConditionMessage;
import org.springframework.boot.autoconfigure.condition.ConditionOutcome;
import org.springframework.boot.autoconfigure.condition.SpringBootCondition;
import org.springframework.context.annotation.ConditionContext;
import org.springframework.core.annotation.AnnotationAttributes;
import org.springframework.core.env.Environment;
import org.springframework.core.type.AnnotatedTypeMetadata;

/**
 * Base endpoint element condition. An element can be disabled globally via the
 * {@code defaults} name or individually via the name of the element.
 *
 * @author Stephane Nicoll
 * @author Madhura Bhave
 * @since 2.0.0
 */
public abstract class OnEndpointElementCondition extends SpringBootCondition {

	private final String prefix;

	private final Class<? extends Annotation> annotationType;

	protected OnEndpointElementCondition(String prefix, Class<? extends Annotation> annotationType) {
		this.prefix = prefix;
		this.annotationType = annotationType;
	}

	@Override
	public ConditionOutcome getMatchOutcome(ConditionContext context, AnnotatedTypeMetadata metadata) {
		AnnotationAttributes annotationAttributes = AnnotationAttributes
				.fromMap(metadata.getAnnotationAttributes(this.annotationType.getName()));
		String endpointName = annotationAttributes.getString("value");
		ConditionOutcome outcome = getEndpointOutcome(context, endpointName);
		if (outcome != null) {
			return outcome;
		}
		return getDefaultOutcome(context, annotationAttributes);
	}

	protected ConditionOutcome getEndpointOutcome(ConditionContext context, String endpointName) {
		Environment environment = context.getEnvironment();
		String enabledProperty = this.prefix + endpointName + ".enabled";
		if (environment.containsProperty(enabledProperty)) {
			boolean match = environment.getProperty(enabledProperty, Boolean.class, true);
			return new ConditionOutcome(match, ConditionMessage.forCondition(this.annotationType)
					.because(this.prefix + endpointName + ".enabled is " + match));
		}
		return null;
	}

	/**
	 * Return the default outcome that should be used if not property is set. By default
	 * this method will use the {@code <prefix>.defaults.enabled} property, matching if it
	 * is {@code true} or if it is not configured.
	 * @param context the condition context
	 * @param annotationAttributes the annotation attributes
	 * @return the default outcome
	 * @since 2.6.0
	 */
	protected ConditionOutcome getDefaultOutcome(ConditionContext context, AnnotationAttributes annotationAttributes) {
		return getDefaultEndpointsOutcome(context);
	}

	/**
	 * Return the default outcome that should be used.
	 * @param context the condition context
	 * @return the default outcome
	 * @deprecated since 2.6.0 for removal in 2.8.0 in favor of
	 * {@link #getDefaultOutcome(ConditionContext, AnnotationAttributes)}
	 */
	@Deprecated
	protected ConditionOutcome getDefaultEndpointsOutcome(ConditionContext context) {
		boolean match = Boolean
				.parseBoolean(context.getEnvironment().getProperty(this.prefix + "defaults.enabled", "true"));
		return new ConditionOutcome(match, ConditionMessage.forCondition(this.annotationType)
				.because(this.prefix + "defaults.enabled is considered " + match));
	}

}


We've received a Voltera One PCB printer and reflow oven in recent weeks. We'll have it on site to showcase a few things about PCB design and production.
Everything about printed circuit board (PCB) printing, reflow soldering and CAD you always or never wanted to know. Want an animated light me up nifty 9V PCB powered by a Signetics NE555 (most popular integrated circuit ever manufactured) timer, oscillator, and flip-flop IC chip? Come to check out our work and watch the machine print traces and reflow paste to PCB substrates.
This page was last modified on 22 December 2016, at 23:20.


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Microsoft's Edge adds an automatic description to images that don't have it to help those who don't see it
Microsoft estimates that more than half of the images circulating on the Internet have no captions, which is irrelevant information for those who can see the content, but critical for giving oral context to those who cannot see or who have a reduced level of vision.
To minimize the impact of this lack of information, the company has updated Edge with a new feature, which will automatically create a description for images that don't have it. This textual description will now be available to be read by screen readers, who until now the only information they could return, when a description was not detailed by the person who created the image, was a message that the chart has no legend.
Now, "when a screen reader encounters an image without a caption, that image can be automatically processed by machine learning algorithms to describe the image in words and capture any text it may contain," Microsoft says. .
The company admits that "the algorithms aren't perfect and the quality of the description will vary, but for those using screen readers, having some description of an image will be much better than having no context at all." at all", defends Travis Leithead, product manager of the Microsoft Edge browser, quoted in a note on the site. On the other hand, and since it uses artificial intelligence, the functionality will improve over time, as the task is repeated and the diversity of the images analyzed increases.
The captioning work will be done by a computer vision API on the Azure platform, which will process the information from each image to create alternative text, which will be associated with the features of the image. This written account of visual content will be available in Portuguese, English, Spanish, Japanese and Simplified Chinese.
The update takes effect immediately for all desktop versions of Edge, i.e. Windows, Mac, and Linux. It can be enabled in the accessibility options of browser settings and used with any screen reader. Anyone using the browser on Android or iOS will not be able, at least for now, to take advantage of the novelty. Also excluded from this effort are images that the API identifies as sexual, obscene, or decorative, as well as very large or very small images (less than 50×50 pixels).
Jornal Médico – Raul Marques Pereira: Pain in patients at the end of life – approach and treatment "is one of the many leaflets aimed, in particular at CSPs" launched by MGF.dor
Microsoft and Sony plan to monetize free games with ads
Gamers' most anticipated game is coming to Xbox Game Pass this week
Forget the computer. That's all you need to work… – C Studio
Scientists simulate what a 'twin' planet of Earth would look like in Alpha Centauri
TP-Link Tapo C320WS Outdoor CCTV Camera
Samsung Galaxy Watch 5 Battery Capacity Leaks
V Rising sold half a million copies in three days after release
Egg study says dinosaurs were in decline when asteroid hit
Back Then, 5th Edition PlayStation Talents Winner Receives Trailer
Tuning launches the Tesla Model S Plaid. It already exceeds 322 km/h – Observer
Assassin's Creed Liberation will no longer be playable on Steam