Primary diagnostic cultures from patients with melioidosis demonstrate variation in colony morphology of the causative organism, Burkholderia pseudomallei. Variable morphology is associated with changes in the expression of a range of putative virulence factors. This study investigated the effect of B. pseudomallei colony variation on survival in the human macrophage cell line U937 and under laboratory conditions simulating conditions within the macrophage milieu. Isogenic colony morphology types II and III were generated from 5 parental type I B. pseudomallei isolates using nutritional limitation. Survival of types II and III were compared with type I for all assays.
Morphotype was associated with survival in the presence of H2O2 and antimicrobial peptide LL-37, but not with susceptibility to acid, acidified sodium nitrite, or resistance to lysozyme, lactoferrin, human neutrophil peptide-1 or human beta defensin-2. Incubation under anaerobic conditions was a strong driver for switching of type III to an alternative morphotype. Differences were noted in the survival and replication of the three types following uptake by human macrophages, but marked strain-to strain-variability was observed. Uptake of type III alone was associated with colony morphology switching.
Morphotype is associated with phenotypes that alter the ability of B. pseudomallei to survive in adverse environmental conditions.
Burkholderia pseudomallei is an environmental Gram-negative bacterium that causes a severe and often fatal disease called melioidosis. This is an important cause of sepsis in south-east Asia and northern Australia, a geographic distribution that mirrors the presence of B. pseudomallei in the environment . Melioidosis may develop following bacterial inoculation or inhalation and occurs most often in people with regular contact with contaminated soil and water . Clinical manifestations of melioidosis are highly variable and range from fulminant septicemia to mild localized infection. The overall mortality rate is 40% in northeast Thailand (rising to 90% in patients with severe sepsis) and 20% in northern Australia [1, 2].
A major feature of melioidosis is that bacterial eradication is difficult to achieve. Fever clearance time is often prolonged (median 8 days), antimicrobial therapy is required for 12-20 weeks, and relapse occurs in around 10% of patients despite an appropriate course of antimicrobial therapy [3, 4]. The basis for persistence in the infected human host is unknown, although several observations made to date may be relevant to the clinical behaviour of this organism [2, 5]. B. pseudomallei can resist the action of bactericidal substances including complement and antimicrobial peptides in human serum [6–8]. B. pseudomallei can also survive after uptake by a range of phagocytic and non-phagocytic cells. Macrophages have several strategies to control bacterial infection, including bacterial killing following uptake through the action of reactive oxygen and reactive nitrogen compounds, antimicrobial peptides and lysozomal enzymes. Despite this, B. pseudomallei can invade and replicate in primary human macrophages [8–10].
Bacterial survival under adverse and rapidly changing environmental conditions is likely to be facilitated by phenotypic adaptability and plasticity. A previous study conducted by us found that 8% of primary cultures of clinical samples taken from patients with melioidosis contained more than one colony morphotype on Ashdown agar. Morphotypes could switch reversibly from one to another under specific conditions, and were associated with variable expression of putative virulence determinants including biofilm and flagella . Compared with parental type I (the common 'cornflower head' morphology), isogenic type II (a small, rough colony) had increased biofilm and protease production, while isogenic type III (a large, smooth colony) was associated with increased flagella expression . In vitro models suggested that switching of morphotype impacted on intracellular replication fitness after uptake by human epithelial cell line A549 and mouse macrophage cell line J774A.1. We postulated that colony morphology switching might represent a mechanism by which B. pseudomallei can adapt within the macrophage and persist in vivo.
In this study, we investigated whether the variable phenotype associated with different morphotypes resulted in altered fitness during interactions with the human macrophage cell line U937 and after exposure to a range of laboratory conditions that simulate one or more conditions within the macrophage milieu. Isogenic morphotypes II and III generated from each parental type I of 5 B. pseudomallei strains isolated from patients or soil were used in all experiments.
Different growth rates may affect the number of intracellular bacteria following uptake by host cells. Thus, prior to observation of intracellular replication in macrophages, extracellular growth of B. pseudomallei was compared between 3 isogenic morphotypes cultured in trypticase soy broth (TSB). Using a starting inoculum of 1 × 104 CFU/ml, log and stationary phase occurred at 2 h and 12 h, respectively, for all 3 morphotypes. There was no difference in doubling time between 3 isogenic morphotypes (P = 0.14) with an average doubling time of 40.2, 39.2 and 38.3 minutes for types I, II and III, respectively.
Evaluation of the initial B. pseudomallei-macrophage cell interaction using a multiplicity of infection (MOI) of 25:1 demonstrated that 3.0% of the bacterial inoculum (range 1.2-8.0% for different isolates) was associated with macrophages at 2 h. There was no significant difference in this value between 3 isogenic morphotypes for all 5 isolates.
Following removal of extracellular bacteria and incubation for a further 2 h, 1.5% of the bacterial inoculum (range 0.4-3.4% for different isolates) was recovered. There was no significant difference in this value between 3 isogenic morphotypes for all 5 isolates.
The intracellular replication of B. pseudomallei between 4 to 8 h within macrophages is summarized in Figure 1. The replication rates for the 3 isogenic morphotypes of each strain obtained from two independent experiments were comparable (data not shown). Percent replication at 8 h was defined in relation to the 4 h time point, which was used as the reference count. Analysis of pooled data for 5 isolates demonstrated that type I had a significantly higher rate of intracellular replication than either type II or III. The mean intracellular replication of type I at 8 h was 2.0 (95%CI 1.5-2.6, P = 0.004) times higher than that of type II, and 1.9 (95%CI 1.4-2.5, P = 0.004) times higher than that of type III (Figure 1A). However, this pattern was not uniformly observed for each of the 5 isolates, as shown in Figure 1B-F. The higher replication fitness for type I based on the summary data was largely accounted for strains 164 and K96243. Other strains demonstrated a different pattern. For example, strain 153 type III had a higher intracellular replication than type I, a finding that replicates those of a previous study . The mean intracellular bacterial count also varied between individual isolates. These differences were not due to the relative sensitivities of 3 isogenic morphotypes to 250 μg/ml kanamycin, as this experimental condition removed 99.9% of extracellular bacteria independent of type for all isolates (data not shown).
Intracellular replication of 3 isogenic morphotypes of B. pseudomallei in human macrophages. Differentiated U937 cells were incubated for 2 h with B. pseudomallei at a MOI of 25:1, after which non-adherent bacteria were removed by washing and incubation for a further 2 h with kanamycin. At this 4 h time point, fresh medium containing kanamycin was added and incubation continued for a further 4 h. The bacterial count and colony morphology were enumerated at 4, 6 and 8 h by cell lysis and plating onto Ashdown agar. The data shown in Figure 1A represent mean values for each isogenic morphotype derived from 5 B. pseudomallei isolates and is expressed as the bacterial proportion at 6 and 8 h compared with the number at 4 h (which was defined as 100%). Figure 1B-1F shows the number of intracellular bacteria in CFU/ml for individual isolates. Data plots are means ± standard deviations.
To examine the effect of acid, growth of 3 isogenic morphotypes in LB at pH 4.0, 4.5, 5.0 and 7.0 was compared at each of 5 time points over 24 h of incubation. No growth difference was observed between morphotypes at any time point for pH 4.5, 5.0 or 7.0 (P > 0.10 for all time points). When cultured in LB broth at pH 4.0, all bacteria died within 12 h incubation.
The susceptibility of 3 morphotypes to ROI was initially examined on LB agar plates containing a range of H2O2 concentrations (0, 170, 310, 625, 1,250 and 2,500 μM) (data not shown). B. pseudomallei failed to grow on plates with H2O2 at a concentration higher than 625 μM, and so the percentage of viable bacteria were enumerated using agar plates with 625 μM H2O2 compared to those on plates without H2O2. This demonstrated a difference in bacterial survival between the three isogenic morphotypes (P < 0.001). Percentage survival of type I was 3.8 (95%CI 2.9-5.0, P < 0.001) times higher than that for type II, and was 5.2 (95%CI 4.0-6.8, P < 0.001) times higher than that for type III (Figure 2A).
Susceptibility of 3 isogenic morphotypes of B. pseudomallei to ROI and antimicrobial peptide LL-37. Survival was examined for 5 different B. pseudomallei isolates. (A) Percent survival in ROI was determined on LB agar plates containing 625 μM H2O2 compared to the number of bacteria on plates without H2O2. The results were obtained from 4 separate experiments. (B) Percent survival in LL-37 was determined at 6.25 μM LL-37 in 1 mM potassium phosphate buffer (PPB) pH 7.4 for 6 h. The results were obtained from 2 separated experiments. Data plots are means ± standard deviations.
Further examination was undertaken of the susceptibility of the 3 morphotypes with a range of concentrations of H2O2 in LB broth. No bacteria survived in 500 μM and 250 μM H2O2. In 125 μM H2O2, type I of all 5 isolates multiplied from 1 × 106 CFU/ml (the starting inoculum) to between 5 × 107 and 2.1 × 108 CFU/ml. By contrast, all 5 type III and 4 type II isolates (the exception being type II derived from isolate 164) obtained from the same experiment demonstrated no growth on the plates. This confirmed a higher resistance to H2O2 of parental type I compared to types II and III. A difference was also observed between three isogenic morphotypes in 62.5 μM H2O2 (P < 0.001). Bacterial growth of type I was 1.5 (95%CI 1.1-2.0, P = 0.02) times higher than that for type II, and was 2.7 (95%CI 2.0-3.7, P < 0.001) times higher than that for type III.
Susceptibility of B. pseudomallei to RNI was observed following 6 h exposure to various concentrations of NaNO2 ranging between 0.1 to 10 mM in acidified pH 5.0 in LB broth. Using a concentration of 2 mM NaNO2, the percent survival of types I, II and III were 43.8%, 43.7% and 40.1%, respectively, with no difference observed between the three morphotypes (P > 0.10).
Compared with initial inocula and untreated controls, treatment with 200 μg/ml lysozyme at pH 5.0 did not decrease the bacterial count for the 3 isogenic morphotypes of B. pseudomallei, while this concentration could reduce the number of E. coli from 4.9 × 106 CFU/ml (the starting inoculum) to 425 CFU/ml. Susceptibility was examined further in the presence of 3 mg/ml lactoferrin. A kinetic study over time demonstrated that lactoferrin alone could kill an entire E. coli inoculum of 1 × 106 CFU/ml within 3 h at pH 5.0. The same treatment did not affect the number of viable B. pseudomallei which was comparable to the inoculum and untreated control. Adding 200 μg/ml lysozyme with lactoferrin did not enhance the killing efficacy of E. coli and had no effect on B. pseudomallei.
Macrophages produce several antimicrobial peptides [12, 13]. We examined the susceptibility of isogenic morphotypes to HNP-1, HBD-2 and cathelicidin LL-37, three of the main human antimicrobial peptides. The results demonstrated that 100 μg/ml HNP-1 and 100 μg/ml HBD-2 did not reduce the bacterial count for the 3 isogenic morphotypes of any of the B. pseudomallei isolates when compared with the initial inocula and untreated controls.
In a pilot experiment with a range of LL-37 concentrations and exposure times, we found that LL-37 reduced the B. pseudomallei count at a concentration of 6.25 μM at 6 h. This condition killed 100% of a starting inoculum of 4.6 × 106 CFU/ml E. coli control and caused a 75.7 to 99.8% reduction of B. pseudomallei for different isolates. A difference in bacterial survival was observed between the three isogenic morphotypes (P < 0.001). Survival of type I was 1.5 (95%CI 1.1-2.2, P = 0.02) times higher than that for type II, but was 3.7 (95%CI 2.6-5.3, P < 0.001) times lower than that for type III (Figure 2B).
Low oxygen concentration may limit the intracellular growth of aerobic bacteria within the host . We examined the survival of 3 isogenic morphotypes and determined whether morphotype switching occurred in response to different oxygen concentrations during incubation on Ashdown agar at 37°C. B. pseudomallei survived in 5-15% oxygen concentration for 14 days, with an average colony count of 95% (range 72-109% for different isolates and morphotypes) compared to control plates incubated in air for 4 days (Table 1). There was no difference in the survival pattern between 3 isogenic morphotypes (P > 0.10). B. pseudomallei colonies were not visible on Ashdown agar after incubation in an anaerobic chamber for 2 weeks. The capability to recover from anaerobic conditions was observed as colonies were visible at 48 h after reincubation at 37°C in air, and colony counts were performed after incubation for 4 days. The percentage of bacteria recovered was not different between three morphotypes (P > 0.10).
The data represents the mean of 5 B. pseudomallei isolates for each morphotype. The range reflected variation of % colony count between isolates. *% Morphotype was the proportion of each morphotype on the plate. Morphotype switching was observed for type III (starting type) to either type I (isolates K96243, 164, B3 and B4) or to type II (isolate 153).
Types I and II did not demonstrate colony morphology variation over time in any of the conditions tested. Figure 3 shows the effect of various testing conditions of type III for all 5 isolates. Between 1% and 13% of colonies subcultured from 28 h TSB culture onto Ashdown agar switched to alternative types. The switching of type III appeared to be important for replication in macrophages. Following uptake, switching of type III increased over time such that by the 8 h time point, between 48-99% of the agar plate colonies (the range representing differences between isolates) had switched to type I (isolates K96243, 164, B3 and B4) or to type II (isolate 153). Morphotype switching did not increase in acid, acidified sodium nitrite, or LL-37. In contrast, morphotype switching from broth culture containing 62.5 μM H2O2 increased over time of incubation, ranging between 24-49% of the plate colonies for different isolates. Interestingly, between 15-100% of the total type III colony count switched to an alternative morphotype after recovery from anaerobic conditions. The pattern of morphotype switching in all conditions tested was specific to isolates, with four isolates switching from type III to type I (K96243, 164, B3 and B4), and one isolate switching to II (153).
Effect of seven conditions on morphotype switching of type III of 5 B. pseudomallei isolates. (i) TSB culture in air with shaking for 28 h; (ii) intracellular replication in macrophages for 8 h; (iii) exposure to 62.5 μM H2O2 in LB broth for 24 h; (iv) growth in LB broth pH 4.5 for 24 h; (v) exposure to 2 mM NaNO2 in LB broth for 6 h; (vi) exposure to 6.25 μM LL-37 in 1 mM potassium phosphate buffer (PPB) pH 7.4 for 6 h; and (vii) re-exposure to air after incubation in anaerobic chamber for 2 weeks. All experiments were performed using the experimental details described above. B. pseudomallei morphotype on Ashdown agar following incubation in air at 37°C for 4 days was defined and compared with the starting morphotype. Morphotype switching was presented as the proportion (%) of alternative types in relation to the total colonies present.
Our previous paper reported a process of B. pseudomallei colony morphology switching that occurred during human melioidosis, and in an animal model, mouse macrophage cell line J774A.1, human lung epithelial cell line A549, and under starvation conditions in vitro. In this study, we investigated whether the variable phenotype associated with different morphotypes resulted in a survival fitness or disadvantage during interactions with a human macrophage cell line U937 and after exposure to factors that simulate the macrophage milieu. Although our previous report described 7 different morphotypes from clinical isolates, the five isolates used here from 3 different clinical and 2 environmental samples were only observed to switch under nutritional limitation from parental type I to types II and III, allowing comparison of 3 isogenic morphotypes with known variable phenotype.
The initial interaction between the human macrophage cell line U937 and 3 isogenic morphotypes of B. pseudomallei was not different between the three types. Despite a comparable rate of extracellular growth between isogenic morphotypes, heterogeneity in subsequent intracellular survival/growth after this time point was observed. Type III of each isolate was inconsistently capable of multiplication after uptake by human macrophages, and was associated with a change in morphotype. This suggests that type III has a fitness disadvantage under these circumstances. A possible explanation for this is that type III does not appear to produce biofilm . A biofilm mutant demonstrated a mark reduction in intracellular survival in primary human macrophages than the wild type, suggesting that biofilm production is associated with the ability to survive in human macrophages .
Our previous study examined the survival and replication of B. pseudomallei strain 153 in the human respiratory epithelial cell line A549 and the mouse macrophage cell line J744A.1. Our finding here that type III of strain 153 had increased survival in the human macrophage cell line U937 is consistent with our previous findings for the mouse macrophage cell line J774A.1 infected with the same strain . However, the use of a wider number of strains in this study demonstrated that there was a lack of reproducibility between strains. We suggest that this is likely relate to variability in genomic content between the strains tested. Future testing strategies require the evaluation of a large numbers of strains that have undergone whole genome sequencing to facilitate statistically robust comparisons between genomic variation and phenotypic behaviour.
Several components of the innate immune system are efficient in killing organisms within human macrophages . The most important of these are the antimicrobial peptides and nitric oxide (NO), the superoxide anion (O2-), and hydrogen peroxide (H2O2), all of which are directly toxic to bacteria. Reactive oxygen species generated by the phagocyte NADPH oxidase have an essential role in the control of B. pseudomallei infection in C57BL/6 bone marrow derived macrophages . Type I of all 5 B. pseudomallei isolates tested here had the greatest resistance to H2O2, followed by types II and III, respectively, suggesting that type I has the greatest potential to scavenge or degrade H2O2 molecules. This may explain the finding that type I had the highest replication after uptake by the macrophage cell line. Type III switched to type I or II during culture in medium containing H2O2, indicated that type III had a survival disadvantage under such conditions that required switching to a more H2O2 resistant type.
One of the mechanisms by which B. pseudomallei escapes macrophage killing is by repressing inducible nitric oxide synthase (iNOS) by activating the expression of two negative regulators, a suppressor of cytokine signaling 3(SOC3) and cytokione-inducible src homology2-containing protein (CIS) . It is unknown whether there are variation between strains and isogenic morphotypes in the ability to interfere with iNOS induction. However, colony morphology differences did not influence resistance to RNI. B. pseudomallei is protected from RNI by the production of alkyl hydroperoxide reductase (AhpC) protein and depends on OxyR regulator and a compensatory KatG expression . These mechanisms may not be associated with colony morphology variability.
B. pseudomallei survive in the phagolysosome  which are acidified environments containing lysozymes, proteins and antimicrobial peptides that destroy pathogen. There was no difference in growth for the 3 isogenic morphotypes of B. pseudomallei derived from all five isolates at all pH levels tested above 4.0, but a pH of 4.0 was universally bactericidal, suggesting that morphotype switching did not provide a survival advantage against acid conditions.
All morphotypes of B. pseudomallei were highly resistance to lysozyme and lactoferrin. Lysozyme functions to dissolve cell walls of bacteria. Lactoferrin is a competitor that works by binding iron and preventing uptake by the bacteria. Common structures for resistance to these factors such as capsule and LPS  were present in all isogenic morphotypes . An alternative explanation is that B. pseudomallei may produce a morphotype-independent lysozyme inhibitor that counteracts the action of lysozyme and lactoferrin.
Antimicrobial peptides are efficient at killing a broad range of organisms. They are distributed in variety tissues, and in neutrophils and macrophages [12, 13]. All 3 isogenic B. pseudomallei morphotypes were resistant to α-defensin HNP-1 and β-defensin HBD-2, but were susceptible to LL-37. In contrast to sensitivity to H2O2, type III was more resistant than type I or II to LL-37. This ability may allow type III to survive within host cells for a limited period before successfully switching to alternative phenotypes and may provide a fitness of advantage in macrophages.
Another feature of bacterial survival during the establishment of persistent infection in the host is adaptation to hypoxia in the host microenvironment . This study demonstrated that all 3 isogenic morphotypes were able to tolerate a low oxygen concentration and anaerobic conditions for at least two weeks. Type III switching to either type I or II was observed during recovery from anaerobic incubation. The fact that types I and II were stable following anaerobic incubation suggests that they are tolerant of fluctuations in oxygen concentration.
Given the variation in the genome of different B. pseudomallei, it was not surprising to observe some variation in intracellular replication between isogenic morphotypes of different isolates. Only one strain switched from type III to II, while the other four isolates switched from type III to type I in all conditions in which a change in morphotype was observed. Analyses of 5 isolates in this study provide evidence that colony morphology variation represents heterogeneous phenotypes of B. pseudomallei with different fitness advantages to interact, survive and replicate in the presence of bactericidal substances within human macrophages.
A limitation of this study is that the experimental methods were laborious and time consuming, which restricted the number of strains we could examine. It is also unclear whether these in vitro assays using a human macrophage cell line are a good model for human infection. Further studies are required to determine the molecular mechanism of morphotype switching, and whether this is associated with persistence of B. pseudomallei in the human host.
B. pseudomallei can produce different colony morphologies in vivo and in vitro. This study has described the intracellular survival and replication of two isogenic morphotypes II and III generated from 5 different parental type I B. pseudomallei in the U937 human macrophage cell line, and has examined the survival of these isogenic morphotypes compared to the parental types in the presence of a variety of substances and under conditions which are potentially encountered within the macrophage milieu. Data for 5 isolates demonstrated that there was variability in bacterial survival and replication following uptake by human macrophages between parental type I and types II or III, as well as variability between strains. Uptake of type III alone was associated with colony morphology switching. Type I was associated with survival in the presence of H2O2. In contrast, isogenic morphotype III demonstrated higher resistance to antimicrobial peptide LL-37. Specific morphotypes were not associated with survival with susceptibility to acid, acidified sodium nitrite, or resistance to lysozyme, lactoferrin, HNP-1 or HBD-2. Incubation under anaerobic conditions was a strong driver for switching of type III to an alternative morphotype in all isolates.
Five B. pseudomallei isolates were examined in this study. Isolates 153, 164 and the reference isolate K96243 were cultured from cases of human melioidosis in Thailand, and isolates B3 and B4 were cultured from uncultivated land in northeast Thailand . The colony morphology of all five parental isolates was type I, and isogenic types II and III were generated from type I of each strain using nutritional limitation . Briefly, a single colony of type I on Ashdown agar was inoculated into 3 ml of TSB and incubated at 37°C in air in static conditions for 21 days. Bacterial culture was diluted and spread plated onto Ashdown agar. Morphotypes were identified using a morphotyping algorithm . Isogenic types II and III generated from each parental type I were isolated from the plates of each strain.
Growth curves were performed for the 3 isogenic morphotypes of each of the 5 B. pseudomallei isolates. A colony of B. pseudomallei was suspended in sterile phosphate buffered saline (PBS). The bacterial suspension was adjusted to an optical density (OD) at 600 nm of 0.15 and diluted 100 times. One hundred microlitres of bacterial suspension was added to 10 ml of TSB and incubated at 37°C in air with shaking at 200 rpm for 28 h. At 2 h intervals, 100 μl of bacterial culture was removed, serially diluted 10-fold in PBS, and the bacterial count determined by plating on Ashdown agar in duplicate and performing a colony count following incubation at 37°C in air for 4 days. Doubling time was calculated.
Human monocyte-like cell line U937 (ATCC CRL-1593.2) originating from a histiocytic lymphoma was maintained in RPMI 1640 (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (PAA Laboratories), 100 units/ml of penicillin and 100 μg/ml of streptomycin (Invitrogen) and cultured at 37°C in a 5% CO2 humidified incubator . Before exposure to B. pseudomallei, 1 × 105 U937 cells per well were transferred to a 24 well-tissue culture plate (BD Falcon) and activated by the addition of 50 ng/ml of phorbol 12-myristate 13-acetate (PMA) (Sigma) over 2 days . The medium was then replaced with 1 ml of fresh medium without PMA and incubated for 1 day. The differentiated macrophage was assessed by macrophage-like morphology . Following washing 3 times with 1 ml of Hank's balance salt solution (HBSS) (Sigma), 1 ml of fresh medium was gently added to the macrophages.
The interaction assay was performed as previously described . B. pseudomallei from an overnight culture on Ashdown agar was suspended in PBS, the bacterial concentration adjusted using OD at 600 nm and then diluted in PBS and inoculated into wells containing differentiated U937 cells to obtain an MOI of approximately 25 bacteria per cell. The MOI was verified by colony counting on Ashdown agar. Infected U937 cells were incubated at 37°C in 5% CO2 for 2 h. Non-adherent bacteria were removed by washing gently 3 times with 1 ml of PBS. The U937 cells were lysed with 1 ml of 0.1% Triton X-100 (Sigma), and the cell lysates serially diluted in PBS and spread plated on Ashdown agar to obtain the bacterial count. Colony morphology was observed . The percentage of bacteria that were cell-associated was calculated by (number of associated bacteria × 100)/number of bacteria in the inoculum. The experiment was performed in duplicate for 2 independent experiments.
Intracellular survival and multiplication of B. pseudomallei in human macrophages were determined at a series of time points following the initial co-culture described above of differentiated U937 with B. pseudomallei for 2 h. Following removal of extracellular bacteria and washing 3 times with PBS, medium containing 250 μg/ml kanamycin (Invitrogen) was added and incubated for a further 2 h (4 h time point). New medium containing 20 μg/ml kanamycin was then added to inhibit overgrowth by any remaining extracellular bacteria at further time points. Intracellular bacteria were determined at 4, 6 and 8 h after initial inoculation. Infected cells were washed, lysed and plated as above. Intracellular survival and multiplication of B. pseudomallei based on counts from cell lysates were presented. Percent intracellular bacteria was calculated by (number of intracellular bacteria at 4 h) × 100/number of bacteria in the inoculum. Percent intracellular replication was calculated by (number of intracellular bacteria at 6 or 8 h × 100)/number of intracellular bacteria at 4 h. The experiment was performed in duplicate for 2 independent experiments.
B. pseudomallei from an overnight culture on Ashdown agar was suspended in PBS and adjusted using OD at 600 nm to a concentration of 1 × 106 CFU/ml in PBS. Thirty microlitres of bacterial suspension was inoculated into 3 ml of Luria-Bertani (LB) broth at a pH 4.0, 4.5 or 5.0. The broth was adjusted to acid pH with HCl. Growth in LB broth at pH 7.0 was used as a control. The culture was incubated at 37°C in air with shaking at 200 rpm. At 1, 3, 6, 12 and 24 h time intervals, the culture was aliquoted and viability and growth determined by serial dilution and plating on Ashdown agar.
The sensitivity of B. pseudomallei to reactive oxygen intermediates was determined by growth on oxidant agar plates and in broth containing H2O2. Assays on agar plates were performed as described previously , with some modifications. Briefly, an overnight culture of B. pseudomallei harvested from Ashdown agar was suspended in PBS and the bacterial concentration adjusted using OD at 600 nm. A serial dilution of the inoculum was spread plated onto Ashdown agar to confirm the bacterial count and colony morphology. Ten microlitres of serial dilutions of bacteria in PBS were spotted onto LB agar containing 0, 170, 310, 625, 1,250 and 2,500 μM H2O2. Colony counts were performed after incubation at 37°C in air for 24 h. The number of colonies on plates containing H2O2 was compared with that on control plates and presented as bacterial survival (%). The assay was performed for 4 independent experiments.
Sensitivity to killing by hydrogen peroxide was further examined in LB broth. An overnight culture of B. pseudomallei on Ashdown agar was suspended in PBS and adjusted to approximately 1 × 108 CFU/ml. Ten microlitres of bacterial suspension was added into 1 ml of LB broth containing two-fold decreasing concentrations of H2O2 ranging from 500 to 31.25 μM. The mixtures were statically incubated at 37°C in air for 24 h and then the viable count and colony morphotype were determined by serial dilution and plating on Ashdown agar. The experiment was performed for 2 independent experiments.
B. pseudomallei from an overnight culture on Ashdown agar was suspended in PBS and the bacterial concentration adjusted using OD at 600 nm. Thirty microlitres of bacterial suspension was added into 3 ml of two-fold decreasing concentrations of sodium nitrite (ranging from 10 to 0.1 mM) in LB broth at pH 5.0. The mixture was incubated at 37°C in air with shaking at 200 rpm and viable bacteria were determined at 6 h by serial dilution and plating on Ashdown agar. The number of viable bacteria in the presence of NaNO2 was compared with the number of bacteria in the inoculum and presented as bacterial survival (%). The experiment was performed in duplicate for 2 independent experiments.
B. pseudomallei cultured overnight on Ashdown agar was harvested and suspended in 10 mM Tris-HCl buffer pH 5.0 . The bacterial suspension was adjusted to a concentration of 1 × 107 CFU/ml. Fifty microlitres of bacterial suspension was added to an equal volume of 400 μg/ml chicken egg white lysozyme (48,000 U/mg protein) (Sigma) to obtain a final concentration of 200 μg/ml. The mixture was incubated at 37°C in air for 24 h, after which 10 μl of 10-fold serial dilutions were dropped on Ashdown agar. Sensitivity to lysozyme was also tested in the presence of 3 mg/ml lactoferrin (Sigma) in a separate experiment . E. coli strain HB101 was tested in parallel as a control.
B. pseudomallei was tested for resistance to HNP-1 and HBD-2 (Peptide international) as described previously , with the exception that HNP-1 was used at twice the dose. E. coli strain HB101 was tested in parallel as a control. Briefly, B. pseudomallei or E. coli strain HB101 colonies were washed and suspended in 1 mM sodium phosphate buffer pH 7.4 containing 1% TSB . The bacterial suspension was adjusted to a concentration of 1 × 107 CFU/ml. Twenty microlitres of bacterial suspension was mixed with an equal volume of 200 μg/ml HNP-1 or HBD-2 to obtain a final concentration of 100 μg/ml antimicrobial peptide and incubated at 37°C in air for 3 h. The viable bacterial count was determined by dropping a 10-fold serial dilution on Ashdown agar.
B. pseudomallei susceptibility to cathelicidin LL-37 was tested using a microdilution method . LL-37 was kindly provided by Dr. Suwimol Taweechaisupapong, Department of Oral Diagnosis, Faculty of Dentistry, Khon Kaen University and Dr. Jan G.M. Bolscher, Department of Oral Biochemistry, Van der Boechorststraat, Amsterdam, The Netherlands. A loop of bacteria was washed 3 times in 1 mM potassium phosphate buffer (PPB) pH 7.4 and suspended in the same buffer. The bacterial suspension was adjusted to a concentration of 1 × 107 CFU/ml. Fifty microlitres of suspension was added into wells containing 50 μl of a 2-fold serial dilution of human cathelicidin in PPB (to obtain a final concentration of 3.125-100 μM), The mixture was incubated at 37°C in air for 6 h and viability of bacteria was determined by plating a 10-fold serial dilution on Ashdown agar. The assay was performed in duplicate.
An overnight culture of B. pseudomallei on Ashdown agar was suspended in PBS and adjusted to a concentration of 1 × 108 CFU/ml. The bacterial suspension was 10-fold serially diluted and 100 μl spread plated on Ashdown agar to obtain approximately 100 colonies per plate. Three sets of plates were prepared per isolate and incubated separately at 37°C in 3 conditions: (i) in air for 4 days (control); (ii) in an GasPak EZ Campy Pouch System to produce an atmosphere containing approximately 5-15% oxygen (BD) for 2 weeks; or (iii) in an anaerobic jar (Oxoid) with an O2 absorber (AnaeroPack; MGC) for 2 weeks and then re-exposed to air at 37°C for 4 days. The mean colony count was determined for each morphotype from 5 B. pseudomallei isolates after incubating bacteria in air for 4 days (control). % colony count for each isolate incubated in 5-15% oxygen or in an anaerobic jar for 14 days was calculated in relation to the colony count of the control incubating bacteria in air for 4 days.
Seven conditions were observed for an effect on morphotype switching, as follows: (i) culture in TSB in air with shaking for 28 h, (ii) intracellular growth in macrophage cell line for 8 h, (iii) exposure to 62.5 μM H2O2 in LB broth for 24 h, (iv) growth in LB broth at pH 4.5 for 24 h, (v) exposure to 2 mM NaNO2 for 6 h, (vi) 6.25 μM LL-37 for 6 h, and (vii) incubation in anaerobic condition for 2 weeks and then re-exposure to air for 4 days. All experiments were performed using the experimental details described above. B. pseudomallei morphotype on Ashdown agar following incubation in air at 37°C for 4 days was defined and compared with the starting morphotype. Morphotype switching was presented as the proportion (%) of alternative types in relation to the total colonies present. Assays of resistance to HNP-1, HBD-2, lysozyme and lactoferrin employed a drop method to assess bacterial survival and colony morphology could not be accurately determined.
Statistical analysis was performed using the statistical program STATA version 10.1. Log transformation of continuous dependent variables was performed as appropriate. Nested repeated measures ANOVA was used to test continuous dependent variables between 3 isogenic morphotypes. A difference between 3 morphotypes was considered to be statistically significant when the P value was less than or equal to 0.05, after which pairwise comparisons were performed between each morphotype. All P values for pairwise analyses were corrected using the Benjamini-Hochberg method for multiple comparisons .
We are grateful to Dr. Suwimol Taweechaisupapong and Dr. Jan G.M. Bolscher for providing LL-37, to Dr. Sue Lee for statistical advice and to Mrs. Vanaporn Wuthiekanun for providing B. pseudomallei isolates. We thank staff at the Mahidol-Oxford Tropical Medicine Research Unit for their assistance and support. S.T was supported by a Siriraj Graduate Thesis Scholarship, Thailand. N.C. was supported by a Wellcome Trust Career Development award in Public Health and Tropical Medicine, UK, and a Thailand Research Fund award, Thailand.
ST carried out the experiments and data analysis. AT isolated and maintained isogenic morphotypes. DL participated in statistical analysis. SK and ND provided materials and intellectual comments. SJP participated in the design of the study, and assisted in the writing of the manuscript. NC participated in the design of the study, data analysis and coordination and writing of the manuscript. All authors read and approved the final manuscript.


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Q: How do you find height of mesh object? I need a way to find height of object(.x). Is there simple way without using some complex math, E.g con or sin.

A:
Is there simple way without using some complex math

For each vertex, store the min and max value of the dimension that represents height (For example, y). If this is still "complex math", then you can use D3DXComputeBoundingBox.

redpajama_set_name: RedPajamaStackExchange


Nirvana Venture Advisors, the early-stage VC fund of the Patni family, has acquired 10 per cent stake in Games2win from a clutch of VC and PE firms, as well as from employee shareholders, for an undisclosed amount.
Early investors Nexus India Pvt Ltd and ICICI Venture part-exited the company by selling their shares to the new investor. However, lead investors Clearstone Venture Partners and SVB Holdings have not divested their shares in this transaction.
As part of the transaction, Amit Patni, co-founder of Nirvana Venture Advisors, will join the board of Games2win.
Alok Kejriwal, CEO and co-founder of Games2win, said, "We have very aggressive expansion plans for Games2win. Given that we have sufficient capital for growth, Nirvana invested in the company by providing partial exits to the early shareholders and key employees."
According to Kejriwal, the company's games have had a reasonable success on the Android platform and the mobile apps have crossed the 10 million download mark. The games have registered over 100 million game plays in just one month. "We are now set to create a large, global, consumer-revenue centric, social mobile gaming play," he added.
The online games company released its first mobile game Makeover Madness in 2009 and since then, has released close to 40 apps. One of the most successful games is a parking game called Parking Frenzy 2.0 (Android), which has been downloaded more than 3.6 million times. Other popular games include Super Mom (iOS), Makeover Madness (iOS), Volcano Flight (iOS) and Best Friends Forever (iOS).
As of now, the company owns more than 600 proprietary games and has over 15 million players each month on its network.
Rajan Mehra, MD of Nirvana Venture Advisors, said, "We are excited by what Games2win is creating and this investment allows us to align with the company at the right stage and also participate in its future success."
Games2Win has recently roped in Tirath Kamdar who has joined its board of advisors.
The company had raised $6 million in a series B funding from Clearstone Venture Partners and Silicon Valley Bank (SVB India Capital Partners) in March 2011.
"We have supported Alok across various firms in the 2win Group and we look forward to having Nirvana as a partner in the new growth phase of Games2Win," said Sandeep Singhal, MD of Nexus Venture Partners.
Is it purely secondary share sale by early investors or includes fresh share issue?
We didn't actually raise money or receive any funds; it's only the shares of the company that exchanged hands.
What does the transaction signify?
Today, there are many companies in the country that are ready to list but are not able to do it because of the market conditions.
Many people have asked me "what happens to people who are investors in shares of companies that are unlisted (like Games2Win)?" I have always answered this question by saying if you create value proposition and a great company, people will still be interested in your shares even if you are not listed. This development signifies that.
What are your investment and expansion plans?
We plan to delve completely into casual social games on mobile phones and as far as generating revenues are concerned, we will be focused on sale of virtual goods and not advertisements. Additionally, very soon a senior US game designer will join us. Our investments will primarily be in people (getting new talent).
Have you launched any new product?
We recently launched our latest product called 'Appucino', which is a social gaming platform.
Like any other entrepreneur; our end game is to make a massive exit. As far as when can we see it happening, is not clear at this point of time.


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Home » Kristian Mercado's "Pa'lante" Takes Top Music Video Award at SXSW
Kristian Mercado's "Pa'lante" Takes Top Music Video Award at SXSW
NEW YORK—April 2, 2019— Pa'lante, director Krisitan Mercado's riveting short film exploring the struggles of working-class Puerto Ricans in the aftermath of Hurricane Maria, won the Jury Award for Music Videos at the recently concluded SXSW Film Festival in Austin. Based on the song of the same name by the band Hurray for the Riff Raff, Pa'lante was a decided underdog as one of 20 finalists for the award, beating out films from several chart-topping artists, including Childish Gambino's This is America, Anderson .Paak's Bubblin' and Kaskade's Tight.
It was a huge win, not only for Mercado, whose acceptance speech was the emotional highlight of the night, but also for the broader Puerto Rican and Latin communities. It underscored the power of cinema as a tool for social change. "In a world where honesty is rare, we took a chance to share our pain," says Mercado. "SXSW has shown that the future of cinema is now. Cinema provides a platform for people on the fringe, people who have been marginalized and oppressed. SXSW heard the voices of the people of Puerto Rico and provided them a forum to tell their story to the world."
Mercado, whose own family suffered dislocation and loss in the storm, had a visceral, emotional reaction when first presented with the song Pa'lante(from a Spanish phrase meaning "forward") by Hurray for the Riff Raff singer Alynda Segarra. "Alynda was looking for a director of Puerto Rican descent to help tell her story," he recalls. "When we spoke on the phone, we connected immediately. I understood her perspective, because the hurricane also affected me and my family deeply."
The director adds that he hopes the film's success at SXSW will draw attention to the continued plight of Puerto Rico and open doors for other Latino filmmakers. "I'm grateful for the opportunity to tell the story of the people of Puerto Rico, who have not only lost their homes, but have also suffered a loss of dignity," he says. "For a director like me, who does not come from means, winning this award is very significant. I hope it will inspire others to step forward and share their stories."
Starring Mela Murder (The Florida Project) as Milagros and Kareem Savingnon (The Girl Is in Trouble) as Manuel, Pa'lante was shot over five days in Puerto Rico. Although it was months after the hurricane, working conditions were still challenging as electricity was spotty and debris had yet to be cleared. "It was a very scrappy production," Mercado says. "We shot in a lot of places that were devastated, including my grandparents' neighborhood. It was odd to be in a place that was so familiar, yet so changed."
Editorial for the film was done by Michael Reuter. Post-production audio finishing was completed at Sound Lounge in New York. Mercado teamed with Mixer/Sound Designer Pete Crimi in finalizing the film's soundtrack. Crimi created an environment through sound for Pa'lante's opening and closing narrative sequences that was authentic to the island. He also blended subtle effects with the music in the body of the piece to help tie it to the visuals. "The life of Puerto Rico comes through in its sounds…the roosters, the wind, the palm trees, the ocean," observes Mercado. "Pete helped flesh out that soundscape. He captured the texture of that world."
Crimi says it was an honor to work on a project that sheds light on a story of the strength of the human spirit, noting, "All of us at Sound Lounge are grateful for opportunities to use our skills to build awareness for important issues. Films like this deserve to be awarded and celebrated."
If you'd like to see Pa'lante, click herehttps://vimeo.com/266748395
Pa'lante Credits
Director: Kristian Mercado Figueroa
Talent: Mela Murder (Milagros), Kareem Savinon (Manuel), Amethyst (Baby Linda), Kylie B (Maria).
DOP: Rudolf Costin
Edit: EDITBAR. Michael Reuter, Editor.
Story: Kristian Mercado Figueroa.
Director Rep: Reprobates, Doug Klinger.
Production: BabyPanther, Andrew Nisinson and Avtar Khalsa, Executive Producers; Yanira Figueroa Velázquez, Matthew Glueckert and Luis E. Mendez; Producers; Tiffany Stephens, Assistant Director/Story Supervisor; Luis Mendez, Locations; Christina Morales, Casting Director; Daniel Maldonado, Driver; Nora Cruz, Event Organizer; Balois Cora, Emily Iraola, Francheska Figueroa and Maria Justiniano, Additional Locations; Felipe Figueroa Jr., Puerto Rico Prep; Peter Roman; DIT; Tony Colon, Additional Housing; Jose Chamorro, Hispanic Federation; Club Raul Reyes, El Nuevo Caribe Democratic; Alex Mercado and Jabreel Shakir, Pas; Dennis Febo, Street Casting; Ali, Abel, Street Casting; Paige Campbell, Hair and Makeup; Ramona Todoca, Art Director; Ionut Radulescu, Type Design.
Post: The Mill. Josh Bohoskey, Colorist.
Sound: Sound Lounge, Pete Crimi, Sound Designer, Mixer
Rentals: Flug. Kristine Guzman.
Aerials: Drone Dogs. Su-Jeng Sang, Drone Operator; Gilberto Cruz, Drone Gimbal Operator.
Thanks: Comunidad La Perla, Case De Poeta Luis Pales Matos, Guayama, Gaby's Restaurant, Arroyo, Residencial Las Margaritas, Municipio De Arroyo, Barrio Yaurel, Municipio De Maunabo, Municipio De Patillas, Municipio De San Juan
Special Thanks: Jose Enrique Segarra, The Community of Spanish Harlem El Barrio NY, The City of Newark NJ, The Young Lords Party, The Figueroa Family, Ineabell Figueroa, Yanira Figueroa Velázquez, Eveny Padilla, Juanita Velázquez, Jennifer R. Colón, Yolanda Figueroa, The Sand and Sea Restaurant In Cayey Rey and Irelsa, La Perla y su bella gente,
Yashira en Impactó la Perla con Coco y la comunidad, Don Benny el Frutero, Milk Burger & Erik Mayor.
Filmed on Location, New York, New Jersey, Puerto Rico
About Sound Lounge
Sound Lounge is an audio post-production facility, providing services for TV and radio commercials, feature films, television series, digital campaigns, gaming and other emerging media. Based in Manhattan, Sound Lounge is artist-owned and operated. Follow on Facebook, Twitter, LinkedIn and Instagram or visit http://www.soundlounge.com for the latest Sound Lounge news.
www.soundlounge.com
Taylor Maggard
taylor@soundlounge.com
Community & Partner Links
How Sony's New Virtual Sound Technology Can Change How We Hear Films
Kami Asgar and Jessica Parks are post-production heavyweights who work with major studios, namely Sony. As a sound designer (Asgar) and as a post executive (Parks), their collective resume touches on everything from Apocalypto to Grandma's Boy to Venom.
Parks has recently shifted her focus from supervisor to hands-on sound design, and we talk about how it's never too late to pivot on your career path and find the thing you love doing wherever you are in life.
Click on this link to read the rest of the article on No Film School's site.
NJ – Governor Murphy signs $14B Incentive Program Bill – the NJ Economic Recovery Act of 2020
Film tax credits — amending existing programs to include provisions for so-called New Jersey film partners and New Jersey film-lease partners and allowing an additional $200 million of tax credits annually over 13 years.
Click this link if you want to read the full article on the Lexology site. http://bit.ly/35NtDx6
Film Commish announces date for production restart
In her December 18, 2020 news update, MOME Commissioner Anne del Castillo announced that the Film Office is now accepting permit applications for production activity that begins on July 27th.
She also announced awards now (Awkwafina) and more. To read all of the Film Commish's bloggy sort of news column, click here.
The coronavirus relief package that Congressional leaders agreed to this week includes grant money that many small proprietors described as a last hope for survival.
For the music venue owners, theater producers and cultural institutions that have suffered through the pandemic with no business, the coronavirus relief package that Congress passed on Monday night offers the prospect of aid at last.
To read the full article on The New York Times' site, click here.
If you want to start production, here's the latest news from the Mayor's Office
Phase 4 production guidance is available on the Film Permit website. All production activity, whether it requires a Film Permit or not, must comply with New York Forward Industry Guidance.
For more information see, please refer to the State Department of Health's Interim Guidance for Media Production During the COVID-19 Public Health Emergency. Please review the guidelines and FAQ before submitting permit applications. The Film Office is operating remotely, so please allow additional time for Film Permit processing.
The above paragraphs contain links to the various FAQ – just mouse over the relevant words.
Nikon to Stop Making Cameras in Japan
Nikon has fallen on hard times as of late as its camera sales have cratered, and now there's a new indicator of how dire its financial situation is: the company is reportedly pulling the plug on making cameras in Japan after over 70 years of doing so.
To read the full article on Petapixel's site, click here.
NVIDIA Uses AI to Slash Bandwidth on Video Calls
NVIDIA Research has invented a way to use AI to dramatically reduce video call bandwidth while simultaneously improving quality
What the researchers have achieved has remarkable results: by replacing the traditional h.264 video codec with a neural network, they have managed to reduce the required bandwidth for a video call by an order of magnitude. In one example, the required data rate fell from 97.28 KB/frame to a measly 0.1165 KB/frame – a reduction to 0.1% of required bandwidth.
To read the rest of this article on Petapixel, click this link.
Union Health Plan Dodges Film Workers' Suit Over Virus Relief
Law360 (October 9, 2020, 5:22 PM EDT) — The Motion Picture Industry Health Plan's board can't be sued under ERISA for allegedly flouting its duties when it relaxed plan rules in response to COVID-19, a California federal judge has ruled, nixing a proposed class action filed by two cinematographers who still couldn't qualify for benefits.
In an order entered Thursday, U.S. District Judge R. Gary Klausner granted the board of directors' motion to dismiss Greg Endries and Dee Nichols' Employee Retirement Income Security Act suit accusing board members of breaching their duty to treat all plan participants fairly.
Endries and Nichols, members of Local 600 of the International Cinematographers Guild, said in July that the board left them and others "out in the cold" in its attempts to address the problems COVID-19 caused for plan participants.
But Judge Klausner agreed with the board's contention that the case, which alleged a fiduciary breach, should be tossed because plan administrators don't act as fiduciaries when they amend health care plans.
Read the full article on the Law360 site by clicking here.
Russo Brothers Received Close to $50 Million From Saudi Bank
Anthony Russo and Joseph Russo photographed at the PMC Studio in Los Angeles for the Variety Playback Podcast.
The Russo brothers, directors of the all-time top grossing film "Avengers: Endgame," quietly secured a roughly $50 million cash infusion for their production company AGBO from Saudi Arabia earlier this year, multiple sources tell Variety.
In a deal brokered and closed at the beginning of the pandemic, the Russos received the investment from an undisclosed Saudi bank in exchange for a minority stake in the brothers' Los Angeles-based shop.
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The #RNCinCLE hashtag has exploded on Twitter, as everyone from pundits and politicians to citizen viewers have been weighing in on the Republican National Convention. Day One of the convention was full of excitement; we saw a long-anticipated floor fight sparked by the Never Trump movement, which made a last-ditch effort to change the nomination rules in a way that unbinds them from the results of the primaries —an effort that, once again and for the last time, failed. We also heard a bunch of super intense speeches in honor of the night's theme, "Make America Safe Again." There was no shortage of material to discuss, and Twitter users made use of the platforms' emojis during the convention to not only tell, but show others how they felt about the proceedings.
American pride reigned supreme among emoji-using RNC tweeters, according to Twitter's internal data; the American flag emoji scored the number one slot for Monday. Second place went to a likely less supportive emoji — the "laughing so hard I'm crying" emoji. Given the general dearth of light spirits and joking at Monday's event (save a few pantsuit cracks at Hillary Clinton's expense), Twitter users probably weren't laughing at something meant to be funny. In third place, we have the double exclamation mark emoji — not surprising, given the number of surprises Donald Trump has brought to the race.
In fourth place, we have an interesting choice: the finger point. This could be in reference to Trump's emphatic manner of pointing at his wife Melania as he introduced her before her keynote speech.
The remainder of common convention feels expressed via emoji on Twitter Monday indicate positive vibes. In fifth we have the heart, followed by a steam locomotive — possibly indicating that the right was hopping on the "Trump Train" the day preceding his official nomination. Next, we have the basic thumbs up, a classic expression of approval, though it can be used sarcastically.
Coming in eighth place, there's the flame, which can be used in a couple different ways. Some tweeters may have found aspects of the convention to be "lit"; but others probably used it during the walk-out of several Never Trump delegates following the fight over rules changes, to indicate that this thing was burning to the ground.
Clapping hands and a proud gold trophy finish of the top-10 emoji list. Emotions around the 2016 Republican primary and the convention have run high, and emoji use is keeping pace.


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Sierra (spanisch für "Säge" oder "Bergkette") steht für:

 Sierra (Geographie), Gebirge
 Sierra (Film), US-amerikanischer Spielfilm (1950)
 Sierra (Fernsehserie), US-amerikanische Fernsehserie
 Sierra (Supercomputer), US-amerikanischer Supercomputer
 La Sierra, kolumbianisch-US-amerikanischer Dokumentarfilm (2005)
 Sierra Entertainment, US-amerikanischer Computerspielhersteller
 Sierra Island, Insel der Südlichen Shetlandinseln, Antarktis
 Sierra-Klasse, Klasse russischer Atom-U-Boote
 Sierra Tequila, deutsche Spirituosenmarke
 Sierra Wireless, kanadischer Hersteller von Mobilfunkgeräten
 Beechcraft Sierra, Leichtflugzeug, siehe Beechcraft Musketeer
 Ford Sierra, Pkw-Modell von Ford
 GMC Sierra, Pick-up-Modell von GMC
 macOS Sierra, Version von macOS
 Monteverdi Sierra, Pkw-Modell von Monteverdi
 Sierra Kidd, Rapper
 S als Buchstabier-Ansagewort im ICAO-Alphabet ("NATO-Buchstabieralphabet")

Sierra ist der Familienname folgender Personen:
 Alfredo Castro Sierra (1942/1943–2010), spanischer Mediziner
 Álvaro Sierra (* 1967), kolumbianischer Radrennfahrer
 Arlenis Sierra (* 1992), kubanische Radrennfahrerin
 Bianca Sierra (* 1992), mexikanisch-US-amerikanische Fußballspielerin
 Carlos Osoro Sierra (* 1945), spanischer Geistlicher, Erzbischof von Madrid und Kardinal
 Carmen Martínez Sierra (1904–2012), spanische Sängerin (Mezzosopran) und Schauspielerin
 Elias Sierra (* 2001), belgischer Fußballspieler
 Enrique Sierra (1957–2012), spanischer Musiker
 Gregory Sierra (1937–2021), US-amerikanischer Schauspieler
 Ismael Rueda Sierra (* 1950), kolumbianischer Geistlicher, Erzbischof von Bucaramanga
 Iván De La Sierra (* 1986), uruguayischer Fußballspieler
 Javier Sierra (* 1971), spanischer Journalist
 Jordi Sierra i Fabra (* 1947), spanischer Autor

 Juan Hernández Sierra (* 1969), kubanischer Boxer

 Liza Del Sierra (* 1985), französische Pornodarstellerin
 Manuel Sierra (* 1981), mexikanischer Eishockeyspieler
 Nadine Sierra (* 1988), US-amerikanische Sängerin (Sopran)
 Roberto Sierra (* 1953), puerto-ricanischer Komponist

 Sebastián Sierra, uruguayischer Tischtennisspieler
 Segundo García de la Sierra y Méndez (1908–1998), spanischer Erzbischof
 Sergio Sierra Bernal (* 1950), mexikanischer Diplomat
 Solana Sierra (* 2004), argentinische Tennisspielerin
 Stella Sierra (1917–1997), panamaische Dichterin
 Terencio Sierra (1839–1907), honduranischer Politiker, Präsident 1901 bis 1903
 Valentín de la Sierra (1898–1926), mexikanischer Cristero
 Verena Sierra (* 1978), deutsche Radiomoderatorin und Redakteurin
 Vicente Horacio Saeteros Sierra (* 1968), ecuadorianischer Geistlicher, Bischof von Machala

Siehe auch:
 Serra

redpajama_set_name: RedPajamaWikipedia


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One of the crown jewels of complexity theory is Valiant's 1979 theorem that computing the permanent of an n*n matrix is #P-hard. Here we show that, by using the model of linear-optical quantum computing—and in particular, a universality theorem due to Knill, Laflamme, and Milburn—one can give a different and arguably more intuitive proof of this theorem.
For decades, Harvard's Leslie Valiant has obviously deserved a Turing Award—and today, the ACM most excellently announced its agreement with the obvious. I have little to add to the prize citation (see also Lance's post): from launching new fields whose reach extends beyond theory (PAC-learning), to proving epochal results (#P-completeness of the permanent), to asking hugely influential questions (permanent vs. determinant), Valiant has been a creative powerhouse of theoretical computer science for longer than I've been alive.
One thing the prize citation doesn't mention is that Valiant is now the third Turing Award winner (after Andy Yao and Len Adleman) to have made a major contribution to quantum computing theory. Valiant's 2001 paper Quantum Computers that can be Simulated Classically in Polynomial Time introduced the beautiful computational model that computer scientists now know as "matchgates," and that physicists know as "noninteracting fermions." It still amazes that Valiant proposed this model for purely mathematical reasons—hitting physical relevance straight between the eyes despite (as far as I can tell) not having that target anywhere in his sights.
Valiant's work has shown that, even if our universe hadn't been made of bosons and fermions, theoretical computer scientists would have had compelling reasons of their own to invent those particles or something equivalent to them—and furthermore, that at least one theoretical computer scientist would have had the imagination to do so.
Certainly Valiant has had a huge influence on me, both through his work and as someone who made time to talk to me as an obscure grad student a decade ago. Three of my papers—The Learnability of Quantum States, A Full Characterization of Quantum Advice, and The Computational Complexity of Linear Optics—would collapse entirely without Valiant-laid foundations.
This entry was posted on Wednesday, March 9th, 2011 at 5:17 pm	and is filed under Announcements, Complexity, CS/Physics Deathmatch, Quantum. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.
Valiant is one of the men who I would love to treat to a meal as an obscure grad student (whenever some stubborn CS department finally decides I deserve an acceptance rather than a rejection letter).
Scott, this might be a good time to broach the subject of whether awards like the Turing (e.g., the Nobel, Fields medal, etc.) should even exist. What legitimate purpose do they actually serve? Surely, Valiant (and every other award winner) has received plenty of recognition before getting the prize; if he hadn't there's almost no way Valiant could have won to begin with. Given that odds are you will one day win the Turing (assuming you keep on as you have been) would you feel cheated if the prize were abolished before you had a realistic chance to win it? Shouldn't the energy and the money go to a better purpose (that is, one which helps directly advance science or philanthropy)?
Why don't you think awards directly advance science? I'm not a scientist or professional mathematician, but awards do many things, including: incentivize, reward, publicize, and glorify (in a good way) progress, whether it's scientific or of another variety. I can tell you from personal experience that there are few things worse or more demoralizing than a community (e.g., a workplace) that doesn't celebrate its successes adequately. And besides, reading about great computer scientists and mathematicians being recognized for their work is inspiring to many people.
Will: Within the theory community, Oded Goldreich is well-known for sharing your anti-award sentiments.
On the other hand, I also find it grossly unfair that (with a few exceptions, like Einstein and Hawking) the world's greatest scientists probably get less than 1% of the fame, glory, etc. of a mediocre NFL player. And that, in turn, affects what kids dream about being when they grow up, and probably even the relative social status of nerds vs. other kids. To whatever tiny extent the Nobel Prize, Fields Medal, and Turing Award rectify that situation, I find myself supporting their existence despite my reservations about their fairness.
Oded's essay was not familiar to me, and thank you for providing a link to it.
On a wide city, the colored splendor, galaxies of light.
now there is no escape.
It seems to me that those mathematicians, scientists, engineers (and physicians, artists, politicians, teachers, …) who are most worthy of our appreciation, include in particular those whose work helps us to an appreciation that the space inside our global purse-seine, is (hopefully) more resource-rich, and also vastly more beautiful, than is apparent at first sight.
Note however that the very few scientists who do get really famous get fame for centuries, whereas any NFL player will only be famous for a few decades at most.
By the way, while we're at famous scientists, the readers of this blog may want to look at "http://www2.stetson.edu/~efriedma/periodictable/" which has a bibliography of 111 famous mathematicians.
Giving enormous respect to Valiant, and especially to the work on permanent vs. determinant. But let me take the opportunity to put forth a contrarian viewpoint on the matchgates work (again, with absolutely the highest respect for everything else!): to a physicist, it really is just non-interacting fermions! IMO, when I see people in CS talk about how holographic algorithms might be a key to understanding deep problem in complexity theory, etc…, I really think they miss the point. There are multiple instances in physics where one can make enormous progress on a particular point, and yet a minor modification of the model makes that machinery much, much less effective (compare the power of CFT in 2d vs CFT in 3d), and noninteracting fermions are one example. Further, physicists already did invent the noninteracting fermion model multiple times, at least once for essentially the same reasons as Valiant, without any motivation by actual fermions. Namely, Onsager's solution of the 2d Ising model is now usually presented in a much simpler way using noninteracting fermions than its original more complicated construction. Bruria Kaufman was responsible for that breakthrough, but she didn't, as far as I understand it, immediately leap to noninteracting fermions using physical knowledge. Rather, she found a way of using spinor representations of the orthogonal group, and then turned that into fermions, essentially following the math. Then, this was re-invented in the non-interacting fermion mapping of counting dimer covers by Kasteleyn and company (interestingly, while they did it directly, one can find frustrated Ising models on the dual lattice which also count the dimer covers, so given the fermion mapping of the Ising model one can also derive the fermion counting of dimers this way…I don't know when this was realized, whether it was before or after). Just to add an interesting complexity theoretic remark: one interesting outcome of this work has been the ability to count coverings on nonplanar graphs at a complexity which scales polynomially on the graph size but exponentially in the genus. Anyway, again, huge respect to Valiant, but I think many physicists, if given the paper to referee, would have pointed out references in the earlier literature, and only accepted it in a journal which was not top rank. IMO, this was an area where CS didn't know the literature. Hope this isn't considered offensive!
Matt: Thanks for the interesting perspective!
Indeed, I strongly disagree with the view Valiant has sometimes expressed in talks that there might well be an "accidental algorithm" proving NC = P#P. My own view on holographic algorithms is much closer to yours—that what they reveal are surprising weaknesses in very specific models.
While I agree that physicists had been interested in noninteracting fermion systems for some time (to put it mildly), I'm curious: can you give me an earlier source than Valiant for the claim that such systems can be simulated in P?
People can get confused about the difference between real and imaginary time, yes (QMC pretty much works well for imaginary time and can in some cases be analytically continued to real time, but the continuation is often very hard to do without encountering numerical instability). Not sure why you say it's different than the observation that fermions=determinant, as if the Hamiltonian is quadratic then any expectation value which is a product of fermion operators can be calculated via Wick's theorem as a determinant (going to the Heisenberg representation). That also gives you the ability to sample, if desired with a little more work, but that is a separate discussion.
Agreed that matchgates are an example of particular weaknesses in certain models. I see the theory of NP-completeness (or any other theory which works using reductions from one problem to another) as basically a way of saying that "this problem almost certainly does not have a very large weakness, as if it did then so would all these other problems", which is indeed opposite to the viewpoint that one might find a weakness in a large class of models.
Interestingly, Onsager's work was not actually the first example of solving an integrable model. Bethe beforehand had solved the Heisenberg spin-1/2 chain (although many steps were done later by others to further develop the Bethe ansatz, and I don't know much about the history or actual results). And it wasn't the last example: there are many interesting models with names like Baxter, Lieb, etc… which have been exactly solved. So, of course one can ask whether these solutions give rise to nontrivial algorithms, just as Onsager's work gives rise to certain polynomial time algorithms for computing partition functions. Alas, it seems that (as far as I can tell from talking to my colleagues who actually understand integrable systems in 2d), these models are only exactly solvable with uniform interactions (or perhaps interactions which vary in space in certain particularly simple ways), not with arbitrary spatially varying interactions, so they cannot be viewed in the same way as corresponding to a network of gates where each gate can be chosen arbitrarily so long as it belongs to a particular class. But perhaps there are some examples where these tools do give rise to new algorithms that I just don't know about yet.
I agree with Lagrange that 1 is a prime number.
Raoul Ohio, mathematical definitions can change over time.
There are various concrete reasons why the current definition is the way it is. Yet, in the end it is: the general theory, essentially inexistant at Lagrange's time, is 'nicer' that way, as I believe somebody explained you recently.
My understanding of holographic algorithms is limited but still I feel Valiant's question makes sense.
After all these (allow me the word crazy) looking algorithms can solve problems minor variations of which are #P-Complete. Typically, we come very close to solving the actual #P-Complete problem and hit a dead end when we discover that no matchgate can realize the said signature.
But still, from what I have gathered so far, I guess the theory of signatures of these matchgates is not yet complete and certainly, IMHO it can be a fruitful avenue for research.
So my question is – why is the theory community not very excited about the theory of realizable signatures? Moreover, what is wrong with Valiant's belief that any proof of P \neq NP may need to explain the unsolvability of holographic algorithms using this approach.
Sure, minor variants of matchgates are #P complete. But, 2-SAT is in P (actually, can be done in linear time, if you are allowed to manipulate integers in O(1) time), while MAX-2-SAT is a "minor variation" and is NP-complete. And it is "surprising" that 2-SAT is in P since if you just tried every possible solution it would take exponential time. Similarly, Dijkstra's shortest path algorithm is a fast way to find the shortest path between 2 points (again, "surprising" given that trying every path takes exponential time). But if we make the "minor variation" that you look for the shortest path going to every point once, we are at travelling salesman. So, it is actually a very common phenomenon that if you solve a problem by expoloiting structure, then any minor variant of that structure can kill your solution. The same happens all the time in physics, too, where a minor change of an exactly solvable problem can be hopeless. So, matchgates (ahem, "Onsager/Kaufman/Kasteleyn/Fisher solution of dimer covering and Ising model") are just another case of this, most likely.
I agree any correct proof of P \neq NP would need to avoid proving that the problems solved by matchgates are hard. Similarly, a correct proof of P \neq NP can't prove that 2-SAT is hard (and it seems like a lot of proofs fail this test). But again, this is just another case of the fact that there are lots of fast, clever algorithms and proving that a problem is hard is a hard thing to do.
Thanks for sharing your thoughts. I guess, I see your point. However, I still feel that holographic algorithms are kinda cool maybe because of the surprising way in which they work (and I would also accept that them being cool is not the point you have dissension with. You do not buy into them being any helpful as far as NP-Hard problems are concerned and I see why).
Use matchgates with appropriate signatures, invoke Holant theorem and game over.
They (Aluffi and Marcolli) bound the algorithmic complexity of the set of real zeroes of the partition function of a Potts model on a graph (and show that this complexity increases exponentially with the size of the graph), by using high-powered motivic mathematics (recently employed in the formal theory of QFT renormalization) to compute a topological property of the set of zeroes. This will then have implications for understanding phase transitions in these Potts models (but they don't say exactly what, e.g. they don't say that calculating the phase diagram will also be exponentially hard).
In the present context, the paper caught my eye because the partition function of the Potts model is the Tutte polynomial, and this is one of those functions which is #P-hard to compute except for a few special cases, like Onsager's algorithm. Also because the paper is employing very advanced algebraic concepts. There may be a "motivic complexity theory" waiting to be discovered, somewhere above and beyond Mulmuley's "geometric complexity theory".
Hey Scott, that new paper of yours isn't half bad. But what I'm wondering about is why you say "we" instead of "I" when you are the sole author. Isn't that convention a little out-dated? Forget modesty. Take the glory that is yours by right.
I am from Electrical Engineering. So I do not know much about the jargon. BUt I do know permanent is hard to calculate. Supposing if someone finds a way to calculate permanent efficiently, does that mean P=NP? What are the consequences of such a result to complexity theory and cryptography?
And ofcourse to quantum computing. If you can give some toy examples to each case it would be great:)!!
That is what I dont get. Permanent of an all one square matrix of size n is n!. Say someone computes this in polynomial time which is n^k time. Then how can one factor a number that has 100-300 zeros. Wouldn't that be still 10^100k time?
KnowledgeSeeker: No. If you had a fast algorithm for the permanent, then you could use it to get a fast algorithm for factoring also! That statement is the end result of combining two reductions: from factoring to satisfiability, and from satisfiability to the permanent. You shouldn't expect to understand it right away (that's what makes the #P-completeness of the permanent a great result, and not just some trivial observation).
If you want to understand why the permanent is #P-complete (and what that means), then google for course lecture notes, read the Wikipedia page, read the book Computational Complexity by Christos Papadimitriou, or (for an alternative, quantum computing proof) try my recent paper.
Thank you very much. I will read the most accessible material that you have quoted given my limited knowledge in the field.
I was looking at your paper. It makes more sense now with the introduction you have given:)!
BTW someone(a math prof) told me that if RSA is broken, then people may go for ECC. So does it mean, permanent is easy does not imply ECC is easy? In that case one has ECC to fall back on correct?
No, if permanent is easy then every public-key cryptosystem is breakable. Do you see the logical flaw in what you wrote? Breaking RSA wouldn't necessarily imply a break of ECC, but breaking RSA by computing the permanent would certainly imply a break of ECC.
Thank you. It is very enlightening. So if permanent is easy then no one-way functions exist.
So is the converse known?
For example if someone breaks RSA(through factoring) or discrete log would that provide evidence of permanent is easy?
I had the following question as well in mind. I found permanent is easy to approximate. Does it provide any help in providing an easier way with one-way functions such as multiplication by two primes?


text:
Samini Features In Official Theme Song Of AFCON 2021
Ghanaian reggae/dancehall artistes, Samini has made his country part after joining an exclusive list of musicians to feature in the African Cup of Nations (AFCON) theme song ahead of the AFCON to be held in Cameroon.
Samini joined other musicians from across the continent to come up with a track deserving to be used as a theme song for the biggest continental tournament in Africa.
After many trails, with three songs submitted prior to the release of the official theme song, "African Smile", with "African Smile" getting the nod with 86.32 votes, knocking out singer Grace Bethel's narrowly by 1.61 votes.
Salatiel, Timi Dakolo, Assia, Hakim, Jaylann, Rayen Youssef, Tukkiman, Imilo Iechanceux, and Mylmo are among the other African singers featured in the 5-minute song.
The African Cup of Nations is set to kick off on January 16 and will come to an end on February 6, 2022 and will showcase some of Africa's very best players from across Europe and the continent.
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text:
Is Woody Harrelson Living or Dead?
Has American actor (1961-) Woody Harrelson died? Or is he still alive?
American actor (1961-)
Woody Harrelson is ...
Born 23 Jul 1961 in Midland
Woody Harrelson is dead
Actor Woody Harrelson talks to the Austin media before a screening of his new movie, LBJ, at the LBJ Presidential Library. On Saturday evening October 22, 2016, the LBJ Presidential Library held a sneak peek of Rob Reiner's new film LBJ, starring Woody Harrelson as the 36th president. The film, which premiered at the Toronto International Film Festival in September, chronicles the life and times of Lyndon Johnson who would inherit the presidency at one of the most fraught moments in American history. Following the screening, director Rob Reiner, actor Woody Harrelson, and writer Joey Hartstone joined LBJ Library Director Mark Updegrove on stage for a conversation about the film.
About Woody Harrelson
Woodrow Tracy Harrelson born on 23 July 1961 in Midland to Charles Harrelson. He is a citizen of United States of America. Woody Harrelson was educated at Hanover College and Lebanon High School.
He is an American actor (1961-) known for his work on Indecent Proposal, Natural Born Killers, The People vs. Larry Flynt, No Country for Old Men, The Messenger, Zombieland and The Hunger Games.
He has one sibling Brett Harrelson.
Woody Harrelson has worked as a television actor, film actor, playwright, stage actor, producer, theatre director, film director, screenwriter, film producer, voice actor and environmentalist. He has been a member of Sigma Chi. He has been nominated for an Academy Award for Best Supporting Actor and Academy Award for Best Actor. He has has won Independent Spirit Award for Best Supporting Male and Primetime Emmy Award for Outstanding Supporting Actor in a Comedy Series.
He is also known as Woodrow Tracy Harrelson.
Woody Harrelson has worked on the following films:
2012 film by Gary Ross
2018 American epic space opera film in the Star Wars franchise directed by Ron Howard
2007 film directed by Ethan and Joel Coen
The Hunger Games: Mockingjay – Part 2
2015 American epic dystopian science fiction war film
2018 film by Ruben Fleischer
2021 film by Andy Serkis
2014 film by Francis Lawrence
War for the Planet of the Apes
2017 science fiction film directed by Matt Reeves
2017 film by Martin McDonagh
2009 film by Roland Emmerich
1998 film by Terrence Malick
Now You See Me 2
2016 film by Jon M.
2011 film by Will Gluck
Natural Born Killers
1994 film by Oliver Stone
2019 film directed by Roland Emmerich
A Scanner Darkly
2006 film by Richard Linklater, Tommy Pallotta, Steven Soderbergh
2013 film by Louis Leterrier
The Edge of Seventeen
2016 film by Kelly Fremon Craig
2010 film by Guy Moshe
The Highwaymen
2019 film directed by John Lee Hancock
Seven Psychopaths
American crime-drama heist film
Seven Pounds
2008 film by Gabriele Muccino
Out of the Furnace
2013 film by Scott Cooper
Austin Powers: The Spy Who Shagged Me
1999 film by Jay Roach
2005 film by Niki Caro
2021 action film
2017 American drama film directed by Destin Daniel Cretton
2008 film by Kent Alterman
The People vs. Larry Flynt
1996 film by Miloš Forman
Indecent Proposal
1993 film by Adrian Lyne
1997 film by Barry Levinson
2003 film by Peter Segal
2011 drama film
Money Train
1995 film by Joseph Ruben
2006 film by Robert Altman
2009 film by Peter Stebbings
White Men Can't Jump
1992 American sports comedy film
2017 film by Woody Harrelson
1988 film by John Hughes
1996 film by Peter Farrelly, Bobby Farrelly
The Man from Toronto
2022 film by Patrick Hughes
2004 film by Spike Lee
2009 war drama film
After the Sunset
2004 crime comedy movie
1991 film by Michael Caton-Jones
Battle in Seattle
2007 film by Stuart Townsend
Transsiberian
2008 film by Brad Anderson
EDtv
1999 film by Ron Howard
2007 film by Bill Guttentag
2008 film by Bill Maher
By Way of Helena
2015 film by Kieran Darcy-Smith
Welcome to Sarajevo
1997 film by Michael Winterbottom
L.A. Story
1991 film by Mick Jackson
2002 film by Gavin Grazer
2008 American romantic comedy-drama
I'll Do Anything
1994 film by James L. Brooks
2017 film by Craig Johnson
The Hi-Lo Country
1996 film by Michael Cimino
Triangle of Sadness
Film by Ruben Östlund
The Grand
2007 film by Zak Penn
Play It to the Bone
1999 film by Ron Shelton
2005 film by Mark Mylod
The Prize Winner of Defiance, Ohio
2005 film by Jane Anderson
True Believer
2007 film by Paul Schrader
1986 film by Michael Ritchie
The Cowboy Way
1998 film by Volker Schlöndorff
Surfer, Dude
Welcome to Hollywood
2000 film by Adam Rifkin
2003 film by Ron Mann
Ted & Venus
1991 film by Bud Cort
Milos Forman – What Doesn't Kill You
2016 film by Miloslav Šmídmajer
Kiss the Ground
documentary on soil health
Woody Harrelson has worked on the following shows:
American late-night live television sketch comedy and variety show
US sitcom (1993–2004)
Walt Disney anthology television series
anthology television series
Live in Front of a Studio Audience: Norman Lear's All in the Family and The Jeffersons
live performance of ''All in the Family'' and ''The Jeffersons''
1994 American television sitcom
Woody Harrelson has been celebrated for these works:


text:
from __future__ import annotations

import copy
import unittest
from unittest import mock

from google.api_core.gapic_v1.method import DEFAULT
from google.cloud.automl_v1beta1 import BatchPredictResult, Dataset, Model, PredictResponse

from airflow.providers.google.cloud.hooks.automl import CloudAutoMLHook
from airflow.providers.google.cloud.operators.automl import (
    AutoMLBatchPredictOperator,
    AutoMLCreateDatasetOperator,
    AutoMLDeleteDatasetOperator,
    AutoMLDeleteModelOperator,
    AutoMLDeployModelOperator,
    AutoMLGetModelOperator,
    AutoMLImportDataOperator,
    AutoMLListDatasetOperator,
    AutoMLPredictOperator,
    AutoMLTablesListColumnSpecsOperator,
    AutoMLTablesListTableSpecsOperator,
    AutoMLTablesUpdateDatasetOperator,
    AutoMLTrainModelOperator,
)

CREDENTIALS = "test-creds"
TASK_ID = "test-automl-hook"
GCP_PROJECT_ID = "test-project"
GCP_LOCATION = "test-location"
MODEL_NAME = "test_model"
MODEL_ID = "TBL9195602771183665152"
DATASET_ID = "TBL123456789"
MODEL = {
    "display_name": MODEL_NAME,
    "dataset_id": DATASET_ID,
    "tables_model_metadata": {"train_budget_milli_node_hours": 1000},
}

LOCATION_PATH = f"projects/{GCP_PROJECT_ID}/locations/{GCP_LOCATION}"
MODEL_PATH = f"projects/{GCP_PROJECT_ID}/locations/{GCP_LOCATION}/models/{MODEL_ID}"
DATASET_PATH = f"projects/{GCP_PROJECT_ID}/locations/{GCP_LOCATION}/datasets/{DATASET_ID}"

INPUT_CONFIG = {"input": "value"}
OUTPUT_CONFIG = {"output": "value"}
PAYLOAD = {"test": "payload"}
DATASET = {"dataset_id": "data"}
MASK = {"field": "mask"}

extract_object_id = CloudAutoMLHook.extract_object_id

class TestAutoMLTrainModelOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.AutoMLTrainModelOperator.xcom_push")
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook, mock_xcom):
        mock_hook.return_value.create_model.return_value.result.return_value = Model(name=MODEL_PATH)
        mock_hook.return_value.extract_object_id = extract_object_id
        op = AutoMLTrainModelOperator(
            model=MODEL,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.create_model.assert_called_once_with(
            model=MODEL,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
            metadata=(),
        )
        mock_xcom.assert_called_once_with(None, key="model_id", value=MODEL_ID)

class TestAutoMLBatchPredictOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        mock_hook.return_value.batch_predict.return_value.result.return_value = BatchPredictResult()
        mock_hook.return_value.extract_object_id = extract_object_id

        op = AutoMLBatchPredictOperator(
            model_id=MODEL_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            input_config=INPUT_CONFIG,
            output_config=OUTPUT_CONFIG,
            task_id=TASK_ID,
            prediction_params={},
        )
        op.execute(context=None)
        mock_hook.return_value.batch_predict.assert_called_once_with(
            input_config=INPUT_CONFIG,
            location=GCP_LOCATION,
            metadata=(),
            model_id=MODEL_ID,
            output_config=OUTPUT_CONFIG,
            params={},
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLPredictOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        mock_hook.return_value.predict.return_value = PredictResponse()

        op = AutoMLPredictOperator(
            model_id=MODEL_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            payload=PAYLOAD,
            task_id=TASK_ID,
            operation_params={"TEST_KEY": "TEST_VALUE"},
        )
        op.execute(context=None)
        mock_hook.return_value.predict.assert_called_once_with(
            location=GCP_LOCATION,
            metadata=(),
            model_id=MODEL_ID,
            params={"TEST_KEY": "TEST_VALUE"},
            payload=PAYLOAD,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLCreateImportOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.AutoMLCreateDatasetOperator.xcom_push")
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook, mock_xcom):
        mock_hook.return_value.create_dataset.return_value = Dataset(name=DATASET_PATH)
        mock_hook.return_value.extract_object_id = extract_object_id

        op = AutoMLCreateDatasetOperator(
            dataset=DATASET,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.create_dataset.assert_called_once_with(
            dataset=DATASET,
            location=GCP_LOCATION,
            metadata=(),
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )
        mock_xcom.assert_called_once_with(None, key="dataset_id", value=DATASET_ID)

class TestAutoMLListColumnsSpecsOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        table_spec = "table_spec_id"
        filter_ = "filter"
        page_size = 42

        op = AutoMLTablesListColumnSpecsOperator(
            dataset_id=DATASET_ID,
            table_spec_id=table_spec,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            field_mask=MASK,
            filter_=filter_,
            page_size=page_size,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.list_column_specs.assert_called_once_with(
            dataset_id=DATASET_ID,
            field_mask=MASK,
            filter_=filter_,
            location=GCP_LOCATION,
            metadata=(),
            page_size=page_size,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            table_spec_id=table_spec,
            timeout=None,
        )

class TestAutoMLUpdateDatasetOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        mock_hook.return_value.update_dataset.return_value = Dataset(name=DATASET_PATH)

        dataset = copy.deepcopy(DATASET)
        dataset["name"] = DATASET_ID

        op = AutoMLTablesUpdateDatasetOperator(
            dataset=dataset,
            update_mask=MASK,
            location=GCP_LOCATION,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.update_dataset.assert_called_once_with(
            dataset=dataset,
            metadata=(),
            retry=DEFAULT,
            timeout=None,
            update_mask=MASK,
        )

class TestAutoMLGetModelOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        mock_hook.return_value.get_model.return_value = Model(name=MODEL_PATH)
        mock_hook.return_value.extract_object_id = extract_object_id

        op = AutoMLGetModelOperator(
            model_id=MODEL_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.get_model.assert_called_once_with(
            location=GCP_LOCATION,
            metadata=(),
            model_id=MODEL_ID,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLDeleteModelOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        op = AutoMLDeleteModelOperator(
            model_id=MODEL_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.delete_model.assert_called_once_with(
            location=GCP_LOCATION,
            metadata=(),
            model_id=MODEL_ID,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLDeployModelOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        image_detection_metadata = {}
        op = AutoMLDeployModelOperator(
            model_id=MODEL_ID,
            image_detection_metadata=image_detection_metadata,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.deploy_model.assert_called_once_with(
            image_detection_metadata={},
            location=GCP_LOCATION,
            metadata=(),
            model_id=MODEL_ID,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLDatasetImportOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        op = AutoMLImportDataOperator(
            dataset_id=DATASET_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            input_config=INPUT_CONFIG,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.import_data.assert_called_once_with(
            input_config=INPUT_CONFIG,
            location=GCP_LOCATION,
            metadata=(),
            dataset_id=DATASET_ID,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLTablesListTableSpecsOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        filter_ = "filter"
        page_size = 42

        op = AutoMLTablesListTableSpecsOperator(
            dataset_id=DATASET_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            filter_=filter_,
            page_size=page_size,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.list_table_specs.assert_called_once_with(
            dataset_id=DATASET_ID,
            filter_=filter_,
            location=GCP_LOCATION,
            metadata=(),
            page_size=page_size,
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

class TestAutoMLDatasetListOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.AutoMLListDatasetOperator.xcom_push")
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook, mock_xcom):
        op = AutoMLListDatasetOperator(location=GCP_LOCATION, project_id=GCP_PROJECT_ID, task_id=TASK_ID)
        op.execute(context=None)
        mock_hook.return_value.list_datasets.assert_called_once_with(
            location=GCP_LOCATION,
            metadata=(),
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )
        mock_xcom.assert_called_once_with(None, key="dataset_id_list", value=[])

class TestAutoMLDatasetDeleteOperator(unittest.TestCase):
    @mock.patch("airflow.providers.google.cloud.operators.automl.CloudAutoMLHook")
    def test_execute(self, mock_hook):
        op = AutoMLDeleteDatasetOperator(
            dataset_id=DATASET_ID,
            location=GCP_LOCATION,
            project_id=GCP_PROJECT_ID,
            task_id=TASK_ID,
        )
        op.execute(context=None)
        mock_hook.return_value.delete_dataset.assert_called_once_with(
            location=GCP_LOCATION,
            dataset_id=DATASET_ID,
            metadata=(),
            project_id=GCP_PROJECT_ID,
            retry=DEFAULT,
            timeout=None,
        )

redpajama_set_name: RedPajamaGithub


Warkworth Training Centre is Dynamic Martial Arts second full time centre.
After three years of operating out of halls and schools the strength of the small but successful club lead to a new full time venue created.
Warkworth Training Centre builds on our successful Kickboxing program which has been taught in Rodney for over a decade.
Why train in Martial Arts?
Represent New Zealand at the largest world combat event.
To train at this club, contact Dave Sawyer on 0212147204.


Traveling on a budget? Hostels are an affordable alternative to expensive hotels. The Avenues Hostel is a Salt Lake Hostel with dorm style and private rooms. One of the great advantages of a hostel is that you get to meet fellow travelers on your vacation. Of course, that is also one of the big disadvantages. Your dorm mate might end up being some sort of know-it-all site of the day reviewer who thinks he's some sort of hot stuff, but isn't.
As I know people in Salt Lake. I have not stayed in this particular hostel. I wish the site had pictures. Hostels do very in quality.