PARADISE LOST
COMING TO LIGHT: EDWARD S. CURTIS AND THE NORTH AMERICAN INDIANS
Documentary by Anne Makepeace
Voices of Sheila Tousey, Bill Pullman
Plan B NR 85 min.
The culture and history of the North American Indian were passed down orally from generation to generation. When the Europeans came and subjugated the indigenous peoples of this continent, they did their best to eradicate the native traditions. They did it by slaughter, and they did it by enforced reeducation, sending Indian children to regimented schools to learn the white man's religions and ways, and punishing them for speaking their own languages. Native Americans were herded onto reservations, and their wardens did their best to stamp out the traditional dances and ceremonies. By the turn of the century, it was clear to anybody who cared to look that the Indian ways were headed for extinction, and that they would largely vanish without a trace.
One who cared to look was Edward S. Curtis, a society studio photographer from Seattle who developed a passionate interest in this doomed culture. He spent thirty years documenting the Indians of North America, with photographs, motion pictures, and sound recordings. He produced a stunning series of 20 volumes of photographs and text covering the native tribes. He spent all his own money and some of J.P. Morgan's, impaired his health and destroyed his marriage. For his pains, his work was soon forgotten, his collection of prints and plates sold in bulk to a Boston second-hand bookseller, where they lay collecting dust in a basement for decades until they were discovered by a clerk in the early '70s, and the Curtis revival commenced.
Anne Makepeace's engrossing documentary traces Curtis's career and shows his work, and asks contemporary Native Americans to comment on the Curtis oeuvre. There are some who resent it, who feel that the old photographs serve to arrest spirits that otherwise would have moved on. More, though, appreciate the magnificent photographs, and treasure the historical record and the link it provides with an Indian past that would otherwise be lost. Some recognize relatives, or themselves, or have even used the Curtis photographs to reconstruct traditional dances that haven't been performed in half a century or more.
In the 1920s Curtis visited Alaska's Nunivak Island, an Indian community as yet undiscovered by Christian missionaries and white settlers. He was struck by the idyllic sense of well-being and joy among the inhabitants, and it shows in the pictures he took. Within a decade Swedish missionaries had arrived, and pulled the plug on this Eden.
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I begin down the path not knowing
Silence=Death
Shakespeare Hero
Tumblr Gallery
The Random Story Machine
@DeepQuoteBot
Click Game
I Swear We Were Infinite
Story of a Single Mother
@BibleBangingBot
Sigmund Freud Has Ten Thousand Hands
Hurt
Mango Poppy
The Woman, The Rain, The Well
Rocky Horror Lost
Fashion Updatez


prepared by Abuzer Yakaryilmaz (QLatvia)
Convention: The transition probability from the 4th state to the 2nd state is presented in the 4th column and the 2nd row of the corresponsing probabilistic operator.
What is the third value of $ u = \myvector{0.1 \\ 0.3 \\ ? \\ 0} $ if it represents a probabilistic state?
What is the value of $ (x+y) $ if $ M = \mymatrix{cc}{ 0.2 & x \\ y & 0.7 } $ a probabilistic operator?
What is the value of $ (x+y+z) $ if $ M = \mymatrix{ccc}{ 0.2 & 0.3 & 0.5 \\ 0.4 & 0 & 0.1 \\ x & y & z } $ a probabilistic operator?
What is the value of $ (x+y+z) $ if $ M = \mymatrix{ccc}{ 0.8 & 0.7 & x \\ 0.2 & 0 & y \\ 0 & 0.3 & z } $ a probabilistic operator?
Which of the following probabilistic operators maps the probabilistic state $ v = \myvector{0.3 \\ 0.7} $ to the probabilistic state $ v' = \myvector{0.52 \\ 0.48} $?
We have a probabilistic system with two states. When the system is in the first state with probability $0.4$, it applies a probabilistic operator such that (i) the transition probability from the first state to the second state is $0.2$ and (ii) the transition probability from the second state to the second state is $ 0.5 $. What is the next probabilistic state?
We have a probabilistic system with two states. Starting with the initial probabilistic state $ u = \mymatrix{cc}{0.4 \\ 0.6} $, the probabilistic operator $ M = \mymatrix{cc}{0.5 & 1 \\ 0.5 & 0} $ is applied three times. What is the final probabilistic state?
We have a system composed by two probabilistic bits. The probability of being in the state $ 0 $ in the first bit is $ 0.6 $, and the same probability for the second bit is $ 0.2 $. What is the probabilistic state of the composite system?
The probabilistic state of the composite system with three bits is represented by a vector with eight entries. Which entry represents the state $ 010 $?
The probabilistic state of the composite system with three bits is represented by a vector with eight entries. Which state corresponds to the 6th entry?
We have a composite system with two probabilistic bits. Which one of the following cannot be a state of this system?
We have a composite system with two probabilistic bits. Let $ v = \dfrac{1}{7} \pstate{01} + \dfrac{2}{7} \pstate{00} + \dfrac{4}{7}\pstate{10} $ be its probabilistic state. Which one of the following represents this state?
Which of the following transitions belong to the operator $ \mymatrix{cc|cc}{ 0 & 0 & 0 & 1 \\ 0 & 0 & 1 & 0 \\ \hline 1 & 0 & 0 & 0 \\ 0 & 1 & 0 & 0 } $?
We apply a controlled-NOT operator on two probabilistic bit where the second one is the controlled bit and the first one is the target the bit. Which of the following transitions represent this operator?
We apply a controlled-NOT operator on two probabilistic bit where the second one is the controlled bit and the first one is the target the bit. Which of the following matrix represent this operator?
We have a composite system with two probabilistic bits.
- First, each bit is set to the state 0.
- Second, the fair coin operator $ \faircoin $ is applied to the first bit, and the NOT operator $ \X $ is applied to the second bit.
- Third, the CNOT operator $ \CNOT $ is applied to the composite system.
What is the final probabilistic state of the composite system?
We have a composite system with three probabilistic bits.
- First, each bit is set to the state 0.
- Second, the fair coin operator $ \faircoin $ is applied to the second bit, and the NOT operator $ \X $ is applied to the first bit.
- Third, the CNOT operator is applied on the second and the third bits where the second bit is the controlled bit and the third bit is the target bit.
-
Fourth, the CNOT operator is applied on the third and first bits where the third operator is the controlled bit and the first bit is the target bit.
What is the final probabilistic state of the composite system?


Zika virus (ZIKV; genus Flavivirus, family Flaviviridae) is an emerging virus of medical importance maintained in a zoonotic cycle between arboreal Aedes spp. mosquitoes and nonhuman primates in African and Asian forests. Serological evidence and virus isolations have demonstrated widespread distribution of the virus in Senegal. Several mosquito species have been found naturally infected by ZIKV but little is known about their vector competence.
We assessed the vector competence of Ae. aegypti from Kedougou and Dakar , Ae. unilineatus, Ae. vittatus and Ae. luteocephalus from Kedougou in Senegal for 6 ZIKV strains using experimental oral infection. Fully engorged female mosquitoes were maintained in an environmental chamber set at 27 ± 1 °C and 80 ± 5 % Relative humidity. At day 5, 10 and 15 days post infection (dpi), individual mosquito saliva, legs/wings and bodies were tested for the presence of ZIKV genome using real time RT-PCR to estimate the infection, dissemination, and transmission rates.
All the species tested were infected by all viral strains but only Ae. vittatus and Ae. luteocephalus were potentially capable of transmitting ZIKV after 15 dpi with 20 and 50 % of mosquitoes, respectively, delivering epidemic (HD 78788) and prototype (MR 766) ZIKV strains in saliva.
All the species tested here were susceptible to oral infection of ZIKV but only a low proportion of Ae. vittatus and Ae. luteocephalus had the viral genome in their saliva and thus the potential to transmit the virus. Further investigations are needed on the vector competence of other species associated with ZIKV for better understanding of the ecology and epidemiology of this virus in Senegal.
The online version of this article (doi:10.1186/s12879-015-1231-2) contains supplementary material, which is available to authorized users.
Zika virus (ZIKV; genus Flavivirus, family Flaviviridae) is an emerging globally mosquito-borne pathogen of growing public health importance. The virus was first isolated in 1947 from a febrile sentinel rhesus monkey and one year later from Ae. africanus in Uganda [ 1 ]. Non-human primates (NHPs) were implicated as the vertebrates hosts of ZIKV in Africa and Asia [ 2 ]. The first well-documented report of human ZIKV infection was in Uganda in 1964 when Simpson described his own occupationally acquired illness [ 3 ]. Subsequently, ZIKV has been recognized to be a cause of febrile illness in humans in Africa and Southeast Asia with symptoms including fever, headache, conjunctivitis, myalgia, rash, joint pains [ 4 – 6 ]. Serological evidence and virus isolations have demonstrated widespread distribution of the virus in Africa, the Indian subcontinent, Southeast Asia and most recently Micronesia and French Polynesia [ 3 , 7 – 11 ]. The implication of Ae. aegypti in the urban transmission of ZIKV came first from field evidence including the high prevalence of anti-ZIKV antibodies in the urban population of Nigeria [ 12 ], the coincidance of peaks of human ZIKV infections and Ae. aegypti population in Indonesia [ 7 ] and the isolation of the virus from a pool of Ae. aegypti in Malaysia [ 13 ]. This implication of Ae. aegypti was confirmed by early experimental studies wich demonstrated the competence of this species to transmit ZIKV [ 14 , 15 ]. The isolation of the virus from a pool of Ae. aegypti in Malaysia provided the first evidence of ZIKV transmission outside Africa.
ZIKV was also isolated from Ae. africanus and Ae. apicoargenteus in Uganda and the Central African Republic [ 16 , 17 ]; from Ae. luteocephalus in Nigeria in 1969 and 1972 [ 12 ]; and from Ae. vittatus, Ae. furcifer, and Ae. aegypti in Cote d’Ivoire in 1999 [ 18 ].
In Senegal, the first evidence of ZIKV circulation was the isolation of a strain from Ae. luteocephalus collected in 1968 in the Saboya forest, 187 km from Dakar, in the western part of the country [ 15 ]. One year later, the virus was isolated from Ae. luteocephalus, Ae. furcifer- taylori and An. gambiae s.l., and a human in Bandia located 65 km from Dakar. In Kedougou, Southeastern Senegal, 381 ZIKV isolates were collected as part of an entomological surveillance programme from 1972 to 2011, mainly from Ae. africanus, Ae. luteocephalus, Ae. furcifer, and Ae. taylori, 7 times from humans and twice from NHPs ( Cercopithecus aethiops, Erythrocebus patas). Serological studies in 1988 and 1990 in the area showed that 10.1 and 2.8 % of humans had Immunoglobuline M (IgM) antibodies to ZIKV [ 9 ].
Although several mosquito species have been found naturally infected by ZIKV, little is known about their vector competence. Hence, the purpose of the present study was to investigate vector competence of populations of Ae. aegypti from Dakar and Kedougou and Ae. unilineatus, Ae. vittatus, and Ae. luteocephalus from Kedougou for 6 ZIKV strains to take into account the high diversity of hosts (human, NHPs, and several mosquito species) and location from which ZIKV strains were isolated.
The protocol of this study has been approved by the Senegalese National Ethic Committee under protocol SEN29/08; 2472/MSP/DS/DER. Because this study was done in collaboration with a team from the University of Texas Medical Branch (UTMB), the UTMB Institutional Animal Care and Use Committee also approved the animal experiments under protocol 02-09-068. UTMB complies with all applicable regulatory provisions of the U.S. Department of Agriculture (USDA)-Animal Welfare Act; the National Institutes of Health (NIH), Office of Laboratory Animal Welfare-Public Health Service (PHS) Policy on Human Care and Use of Laboratory Animals; the U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Research, Teaching, and Testing developed by the Interagency Research Animal Committee (IRAC), and other federal statutes and state regulations relating to animal research. The animal care and use program at UTMB conducts reviews involving animals in accordance with the Guide for the Care and Use of Laboratory Animals (2011) published by the National Research Council.
Table 1 describes the characteristics and geographic origin of the populations of Ae. aegypti, Ae. unilineatus, Ae. vittatus, and Ae. luteocephalus tested in this study. These species were chosen taken into account their abundance, anthrophophilic behaviour and association to ZIKV in the field [ 19 ]. For each population, several breeding habitats were prospected and collected eggs reared in the laboratory. Females F0 were fed several times on guinea pigs to obtain F1 generation eggs. These eggs were hatched and the larvae reared to F1 adults used in this study. This F1 generation were maintained exclusively with a 10 % sucrose solution at 27 °C, 80 % relative humidity (RH), 16:8 h (L:D) photoperiod.
Mosquito species tested for vector competence for Zika virus
| Species | Source a | Habitat | Generation |
|---|---|---|---|
| Ae. aegypti | Dakar | Domestic | F
1
|
| Kedougou | Sylvatic | F
1
|
|
| Ae. unilineatus | Kedougou | Sylvatic | F
1
|
| Ae. vittatus | Kedougou | Sylvatic | F
1
|
| Ae. luteocephalus | Kedougou | Sylvatic | F
1
|
aAll mosquitoes were collected during 2012
Hosts origin, year of collection and passage histories of the six ZIKV strains used in this study are presented in Table 2 . To prepare viral stocks, ZIKV isolates were amplified in Ae. albopictus C6/36 cells for a week at 27 °C and infection progression was monitored using indirect immunofluorescence assay. Supernatant fluids were collected and viral titers estimated by serial 10-fold dilutions on Vero cells [Plaque forming unit (PFU)/ml] as previously described by De Madrid and Porterfield [ 20 ]. Each virus stock was divided into 500 μl aliquots and stored at - 80 °C until use.
Zika virus strains used for this study
| ZIKV strains | Host origin | Year of collection | Location | Passage history a |
|---|---|---|---|---|
| ArD 128000 | Mosquito ( Ae. luteocephalus) | Oct. 11 th 1997 | Kedougou (Senegal) | 6 |
| ArD 132912 | Mosquito ( Ae. dalzieli) | Nov. 20 th 1998 | Kedougou (Senegal) | 4 |
| ArD 157995 | Mosquito ( Ae. dalzieli) | Nov. 17 th 2001 | Kedougou (Senegal) | 6 |
| ArD 165522 | Mosquito ( Ae. vittatus) | Oct. 21 st 2002 | Kedougou (Senegal) | 5 |
| HD 78788 | Human blood | Feb. 14 th 1991 | Dakar (Senegal) | Unknown |
| Ref. (MR 766) | Monkey | Apr. 18 th 1947 | Zika Forest (Uganda) | 20 |
aPassages were conducted with Aedes ( Stegomyia) pseudoscutellaris 61 cells (AP-61)
Mosquito infection has been performed according to procedures already described [ 21 ]. Briefly, one-week-old females of each mosquito species that have never taken blood meal were starved for 24 h before the infectious meal. These females were allowed to feed through a chicken skin membrane by the artificial feeding method described by Rutledge et al. [ 22 ]. The infectious meal consisted of two-thirds washed rabbit erythrocytes mixed with sucrose and fetal bovine serum (FBS) and one-third viral suspension. Adenosine 5’-triphosphate (ATP) was added at a final concentration of 5 × 10 −3 M as phagostimulant. For each infection experiment, a sample of the virus-blood suspension was taken at the end of the mosquitoes feeding and stored at - 80 °C for titration as previously described [ 20 ]. After 30 min of exposure, Mosquitoes were cold anaesthetized and sorted according to their stomach repletion. Fully engorged specimens were preserved and maintained at 27 °C, 80 % RH, 16:8 h (L:D) photoperiod and fed on 10 % sucrose for extrinsic incubation of the virus.
At 5, 10, and 15 dpi, samples of mosquitoes were collected randomly, cold anesthesized, and their legs and wings removed and transferred individually into separate tubes. The proboscis of each mosquito was then inserted into a capillary tube containing 1 μL of FBS for salivation for up to 30 min. After salivation, each mosquito body (whole body except legs and wings) and saliva sample was placed in a separate tube. Aedes luteocephalus was tested only at 15 dpi because this species is difficult to rear, feed and maintain in the laboratory, so a small sample was available.
Mosquito bodies, and legs/wings were triturated using cold pestles in 500 μl of L-15 medium (GibcoBRL, Grand Island, NY, USA). After trituration, pools were centrifuged at 7500 rpm for 10 min at 4 °C. For each sample, 100 μl of supernatant were used for RNA extraction with the QiaAmp Viral RNA Extraction Kit (Qiagen, Heiden, Germany) according to the manufacturer’s protocol with slight modification [ 23 ]. The RNA was amplified using a real-time RT-PCR assay and an ABI Prism 7500 SDS Real-Time apparatus (Applied Biosystems, Foster City, CA) using the QuantiTect kit (Qiagen, Hilden, Germany). The 25 μl reaction volume contained 5 μl of extracted RNA, 10 μl of buffer (2x QuantiTect Probe), 6.8 μl of RNase free water, 1.25 μl of each primer, 0.5 μl of probe, and 0.2 μl of enzymes. The primers and probe sequences were described by Faye et al. [ 23 ].
Only RT-PCR was used to detect ZIKV because the objective of this study is only to show the competence of the vector. Thus, if we are able to show that the virus reached the saliva, it implies that the vector is competent. In our previous experiences with other viruses (West Nile, Usutu), we have noticed that RT-PCR and infectious viral particles are generally very consistent and concordant in their conclusions and trends [ 24 , 25 ]. Such a trend has also been confirmed on C6/36 cells for Chikungunya virus by Chen et al. [ 26 ].
Infection (number of positive bodies/total number of mosquitoes tested), dissemination (number of infected legs & wings/total number of infected bodies), and transmission (number of positive saliva/number of infected legs & wings) rates were calculated for each species on each dpi. Rates were compared using Fisher’s exact test. The associations between viral titers and infection rates were assessed by Spearman’s rank-order correlation test. Differences were considered statistically significant at p < 0.05. Statistical tests were performed using R v. 3.0.1 (R Foundation for Statistical Computing, Vienna, Austria).
After exposure to virus titers ranging from 2.7 × 10 6 to 4 × 10 7 PFU/ml (Additional file 1: Table S1), overall, 111 (50.2 %) of the 221 Ae. aegypti from Dakar, 216 (57.6 %) of the 375 Ae. aegypti from Kedougou, 56 (18.7 %) of the 300 Ae. unilineatus, 37 (14.4 %) of the 256 Ae. vittatus and 45 (75.0 %) of the 60 Ae. luteocephalus tested were infected by the six ZIKV (Fig. 1). No correlation was found between viral titer and mosquito infection rates at 5, 10 and 15 dpi (spearman test; rho > - 0.18, p > 0.95). Infection rates varied significantly between strain for each species tested at 5, 10 and 15 dpi ( p < 0.05). Highest infection rates were generally observed at 10 dpi for Ae. aegypti from Dakar (3/6 viral strains), Ae. aegypti from Kedougou (4/6 viral strains) and Ae. vittatus (3/6 viral strains) and 15 dpi for Ae. unilineatus (4/6 viral strains). Aedes aegypti, Ae. unilineatus, and Ae. vittatus populations showed a decreasing trend in infection rates either from 5 to 10 dpi or from 10 to 15 dpi in 19 of the 24 infection assays performed.
Infection, dissemination and transmission rates at different incubation period for different mosquito species orally exposed with six zika virus strains
The overall trend has been the low dissemination in the populations of Ae. aegypti from Dakar (6.3 % of the 111 specimen tested), Ae. aegypti from Kedougou (5.6 % of the 216 specimen tested) and Ae. unilineatus (5.3 % of the 56 specimen tested) and a relatively high dissemination rates in Ae. vittatus (27.0 % of the 37 specimen tested) and Ae. luteocephalus (42.2 % of the 45 specimen tested). Only Ae. unilineatus disseminated one viral strain at 5 dpi. Dissemination rates varied between 0 and 10 % for the populations of Ae. aegypti and beteween 0 and 100 % for Ae. vittatus at 10 dpi. At 15 dpi, these dissemination rates varied between 0 and 50 % for the populations of Ae. aegypti and beteween 0 and 100 % for Ae. vittatus and Ae. luteocephalus. Dissemination rates were statistically comparable for all species at 10 and 15 dpi ( p > 0.09) except Ae. luteocephalus at 15 dpi ( p = 0.03). The 2 populations of Ae. aegypti and Ae. luteocephalus disseminated 4 of the 6 ZIKV strains tested, Ae. vittatus 3 strains and Ae. unilineatus only 2 strains.
Taken into account all the six viral strains, a total of 7, 12, 3, 10 and 27 saliva of Ae. aegypti from Dakar, Ae. aegypti from Kedougou, Ae. unilineatus, Ae. vittatus and Ae. luteocephalus, respectivly were tested for the presence of ZIKV RNA. Only Ae. vittatus and Ae. luteocephalus transmitted ZIKV strains HD 78788 (20 %) and MR 766 (50 %) respectively at 15 dpi indicated by the presence of the viral genome in the mosquito saliva. These transmission rates were comparable ( p = 1).
Only RT-PCR was used to detect ZIKV because the objective of this study is only to show the competence of the vector. Thus, if we are able to show that the virus reached the saliva, it implies that the vector is competent. Mean Ct values decreased gradually from 5 to 15 dpi for most infected species and ZIKV strain association suggesting that the mean amount of virus in each mosquito has increased and thus the RT-PCR was able to detect the ZIKV amplification (Additional file 1: Table S1).
All mosquito species tested were susceptible to ZIKV infection with infection rates varying according to viral strains and extrinsic incubation periods. In some cases, these rates were as high as those obtained with Ae. aegypti and Ae. albopictus populations from Singapore [ 27 ].
Our results also revealed globally in Ae. aegypti, Ae. unilineatus, and Ae. vittatus populations a decreasing trend in infection rates either from 5 to 10 dpi or from 10 to 15 dpi. Indeed, in 24 infection assays performed, 17 showed this profile. Similar profile were also observed in Ae. aegypti populations from Singapore between 6 and 7 dpi and those of Ae. albopictus from 6 to 10 dpi [ 27 , 28 ]. This decrease following high infection rates obtained at 5 or 10 dpi suggest that ZIKV infection could induce an immune response that can lead to replication of the virus at undetectable levels.
The main findings of our study was the low dissemination and lack of transmission of the population of Ae. aegypti tested here. This result was unexpected taken into account very high transmission rates obtained in previous studies with this species in Senegal and Asia. Indeed, a previous study had shown that Ae. aegypti from Kebemer (a locality near Dakar, Senegal), were competent to ZIKV with a transmission rate of 88 % at 7 dpi [ 15 ], reaching as high as 95 % at 21 dpi. However, intrathoracic inoculation and transmission to newborn mice used in that study were different from our methods. With other viruses, intrathoracic inoculation is well known to bypass the midgut infection barrier, leading to a shorter period of extrinsic incubation, a direct exposure of the virus to the salivary glands, and thereafter generally more efficient infection and transmission [ 29 ].
The recent studies conducted on Ae. aegypti populations from Singapore using an oral infection methods similar to our method have generated high salivary glands infection rates of 62 and 100 % at 5 and 10 dpi, respectivly [ 27 ].
The low or lack of transmission we observed could not be attributed to the transmission method used (salivation on capillary tubes) which efficacy has been proven. Indeed, this system has been recently used to demonstrate vector competence of Ae. albopictus for the same ZIKV [ 28 ]; but also with various other mosquito species tested for various viruses [ 21 , 30 ]. In addition, a comparative study has shown that Ae. albopictus and Ae. taeniorhynchus transmit better with the method of capillary tubes than with the use of an animal for transmission monitoring [ 31 ]. Because the experiments were carried out at 27 °C, the average temperature in Senegal during ZIKV transmission, the temperature may not be impacting our transmission results. Also the titers of the ZIKV we used could not be responsible because a virus suspension at a final concentration of 7.0 log
The difference between the transmission rates observed in this study and the others could be explained by genetic variability between Ae. aegypti populations from different geographical origin. The impact of this variability in vector competence results was demonstrated for several vector/virus associations [ 33 , 34 ]. Although the lack of transmission by Ae. aegypti populations was not expected, this is consistant with the low number of ZIKV strains isolated from this species in West Africa. Indeed only 2 strains of ZIKV were isolated from this species in Senegal, one strain in Burkina Faso and one in Ivory Coast [ 35 , 36 ].
The low competence of Ae. vittatus and Ae. luteocephalus is discordant with the high abundance of these species and their frequent association with ZIKV in the field [ 19 , 35 , 36 ]. Indeed, Ae. vittatus, with 22.98 % of the fauna, was the most abundant mosquito species collected by human landing catch in the Kedougou area between June 2009 and December 2010. This species has beeen found associated with ZIKV in Ivory Cost (2 strains) and Senegal (15 isolates at least) in West Africa. Ae. luteocephalus is generally less abundant than Ae. vittatus but more ZIKV was isolated in this species in Burkina Faso (40 strains), Ivory Cost (48 strains) and Senegal (92 strains).
However, an Ae. aegypti population with poor vector competence but high density have been shown to be the principal vector of a Yellow Fever outbreak in Nigeria [ 37 ].
The low infection and dissemination rates and the lack of transmission by Ae. unilineatus are in agreement with the single isolation of ZIKV from this species in nature and its low representation from human landing catch collection [ 38 ].
Our results are discordant with the pattern of ZIKV transmission in Southeastern Senegal. Indeed ZIKV has the highest frequency of detections among arboviruses found in this area. Its amplifications have been detected during 20 of the 34 years of monitoring that took place between 1972 and 2005 there. Furthermore, the virus has been isolated continuously every year from 1984 to 1994 [ 35 ]. Our low transmission rate are difficult to reconcile with continuous ZIKV transmission, and suggest the involvement of other vectors or other mechanisms of maintenance and transmission. The involvment of other vectors is supported by the fact that lesser number of strains were isolated from the tested species compared to Ae. furcifer and Ae. taylori [ 35 , 36 ]. Vector competence of these species should be investigated to better understand ZIKV epidemiology and transmission in Senegal. Our results suggest that the species tested here are probably less implicated in the regular ZIKV amplifications in Senegal. Other mechanisms of maintenance and transmission probably include the vertical and/or venereal transmission of the virus supported by its detection in several pools of male Ae. furcifer in nature [ 35 , 36 ]. This phenomenon, already demonstrated in nature has never been proven experimentally and therefore needs further investigation.
All the populations of Ae. aegypti, Ae. unilineatus, Ae. vittatus, and Ae. luteocephalus tested here were susceptible to oral infection of ZIKV but only a low proportion of Ae. vittatus and Ae. luteocephalus had the viral genome in their saliva and thus the potential to transmit the virus. Vector competence and vertical transmission studies involving other species more often associated with ZIKV should be undertaken to better understand the ecology and epidemiology of this arbovirus of growing medical interest.
Ct (threshold cycle) mean values for each mosquito species body (the whole body except legs and wings) and Zika virus strain at different days post-infection (dpi). (DOC 73 kb)
Aedes
Adenosine 5’-triphosphate
days post-infection
Fetal bovine serum
Immunoglobuline M
Non-human primates
Plaque forming unit
Relative humidity
Ribonucleic Acid
Reverse transcription-polymerase chain reaction
50 % tissue culture infectious dose
Zika virus
Competing interests
The authors declare that they have no competing interests.
Authors’ contributions
CTD, DD, AAS, SCW and MD conceived and designed the study. CTD, AG and DD collected the mosquito populations. CTD, AG, OumF, AAS and OusF performed the laboratory experiments. CTD, DD, and MD analyzed the data and drafted the manuscript. CTD, DD, YB, ID, OumF, AAS, OusF and MD critically revised the manuscript. All authors read and approved the final manuscript.
This work was supported by grant from the US National Institute of Allergy and Infectious Diseases [Grant R01AI069145] of the National Institutes of Health. The authors gratefully thank Amadou Thiaw, Abdou K. Bodian for their technical assistance during fieldwork and mosquito rearing.


TOYS A film review by Frank Maloney Copyright 1992 Frank Maloney
TOYS is a film directed by Barry Levinson, written by Valerie Curtin and Barry Levinson. It stars Robin Williams, Michael Gambon, Robin Wright, Joan Cusak, LL Cool J, with Donald O'Connor. It is rated PG-13, for mild profanity and mature humor.
TOYS was a disappointment. I had hoped for something a lot more energetic, if nothing else. There is no plot, merely a skeleton on which to hang a dazzling visual experience. Michael Gambon and Robin Williams were both too restrained, too whimsical. They were both outshone by Joan Cusak, the only member of the cast who really got the idea of what they should have been doing. In fact, the rapper LL Cool J put in a more interesting performance, even given his inexperience as an actor.
Barry Levinson is to blame, of that there can be no doubt. It was his idea nearly 15 years ago. He and his former writing partner Valerie Curtin wrote the script then; their collaborations include BEST FRIENDS and JUSTICE FOR ALL, both inferior to his solo writing work (DINER, TIN MEN) or to the films he only directed (RAIN MAN, BUGSY). Perhaps, he should have asked Terry Gilliam to be his script doctor; it definitely lacks (and needs) that kind of dark imagination. Levinson is more interested in making a couple of rather obvious points than in telling a story. For example, he rathers earnestly pursues the idea that weapons are toys for the bloody-minded, that we can choose whether we want our toys to be life-affirming or lethal. He gives us some wonderful visual effects, including a very pointed and chilling scene of generals qua skeletons. But, are we to suppose that Barry Levinson thinks that this is some special vision, some unique insight, vouchsafed only to him that he can squander so much talent to so little purpose?
If you want to immerse yourself in Robin Williams' genius for improvisation, go back to ALADDIN or rent Levinson's GOOD MORNING, VIETNAM. You get only a few really memorable Williamsesque moments in TOYS and long stretches where he has all too little to do. Williams gives his all to promoting the film for Levinson and indeed the trailer, shot in a Spokane wheat field, is vastly better than the movie it advertises.
Of the other performers, Joan Cusak is the most memorable in the role of Williams' sister. She is otherwordly, fragile, the last word in gentle space cadets; when her big denouement arrives, we actually can believe it as earned and established both by the script and by her performance. It is one of the few bright spots in the film. Michael Gambon, who was so loathsome in THE COOK, THE THIEF, HIS WIFE, AND HER LOVER, is mostly addled here. His evilness become really clear only too late in the film, a villain who consults his invalid father and calls him Daddy; it does not suffice. Robin Wright as the love interest is supposed to be a female version of Williams, but she fails to convince. LL Cool J is blunt and slightly amateurish, but earnest and direct in the way of gifted amateurs. He plays Gambon's son and the facts of skin color and lineage are entertainingly ignored. Finally, Donald O'Connor puts in a charming cameo as the dying owner of the Zevo toy company.
But the real and only star of TOYS is the visual effects, the sets, the production design, the computer animation, and the wonderful, wonderful toys. Everything that TOYS lacks in the way of story it possesses in abundance in visuals. It is this aspect of the film, a la Tim Burton, that gives it any chance at all of surviving as something like the masterpiece that Levinson clearly had in mind for himself. And there is something about TOYS that lingers in the mind. I woke up this morning thinking about it, having dreamed about it during the night.
However, I really cannot recommend TOYS to most of you. If you have a strong taste for Robin Williams, if you must see every movie that Barry Levinson makes, if your primary interest is in the visual effects, then you will go to TOYS. A lot of non-specialists, however, will stay away, unless they find themselves with no other movie and the possibility of a discount ticket.
-- Frank Richard Aloysius Jude Maloney .
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Figure 55: Johann van der Schijff. Cyborg - 16, 1993.
[ Return to: Part 1 - 3.Artistic Feedback; 3.1.Electronic Media ]
A large surveillance antenna, turning pointlessly, seeking humans that are no longer there, forms the central motif of this sculpture. This element of communication hardware is situated on top of a camouflaged aeroplane, which has an absurd out-stretched hand forming the one wing. This hybrid form suggests a combined fighting machine and victim. The symbol of flight is contradicted by the landing wheel that slots into the stable, square box that forms the base of the sculpture. [Figure 55]
Figure 56: Globo Cop. Cover of Time Magazine.
(Time 1991, April 1)
This work stems from the arms race and subsequent New World Order (NWO) that developed after the cold war. The NWO is envisaged as a 'benign' international entente on deterring and disciplining aggressors as a replacement for the cold war's grand alliances. Washington made it clear that it saw such a system as a way of institutionalising the spirit of collective law enforcement evident in U.N. Security Council verdicts against Iraq. However those people who remained dubious about the Gulf War and Somalia suspect 'Globo Cop' of bolder designs. 'The NWO carries the ring of a metallic tread, the flash of an electronic eye and the wind from a sudden whirl of high-tech weapons. It conjures up an amplified voice advising overly independent countries to face the wall and assume the position.' (Walsh 1991: 19) [Figures 56-57]
Figure 57: An Airborne Warning and Control System (AWACS) aircraft.
(Bonds 1983: 141)
The sculpture speaks of a post-apocalyptic world where the technology that humans have created, has destroyed life, and machines are left to communicate and control each other in senseless feedback loops. The technique and materials of construction were inspired by the planes made by black children, from cans and detritus found in townships. [Figure 58]
Figure 58: Titus Moteyane. Pan Am 747, 1983.
(Younge 1988: 41)
Figure 59: Johann van der Schijff. Cyborg - 16, 1993.
Mild-steel sheet metal; enamel paint;
found mechanical parts; electric motor.
H: 1660 mm W: 2700 mm L: 2940 mm
[ Return to: Part 2 - 1.Working Method ]
Figure 60: Johann van der Schijff. Drawing no. 1, Cyborg - 16, 1993.
Figure 61: Johann van der Schijff. Drawing no. 2, Cyborg - 16, 1993.
Figure 62: Johann van der Schijff. Drawing no. 3, Cyborg - 16, 1993.
Figure 63: Johann van der Schijff. Drawing no. 4, Cyborg - 16, 1993.
Figure 64: Johann van der Schijff. Drawing no. 5, Cyborg - 16, 1993.
Figure 65: Johann van der Schijff. Drawing no. 6, Cyborg - 16, 1993.


{% extends "app/layout.html" %} {% block content %}
Load, Scrape, Load the internal and external Datasets
Load data from an external source. This can be an excel file, but also anotehr database.
Re-create a Index for certain web-site(s). Re-creating an index on the Elastic Search Engine means that the existing index, with all of its content, is first deleted. Next the Index is created with a field for each facet that is defined for the connected site(s).


Manages the Game.
More...
#include <game.h>
|
| static Game * | Instance () |
| | Retrieves a pointer to the Game instance. More...
|
| |
| static void | LuaRegisterFunctions (lua_State *L) |
| | Registers this objects functions. More...
|
| |
| static int | LuaUpdate (lua_State *L) |
| | A placeholder update function. More...
|
| |
| static int | LuaDraw (lua_State *L) |
| | A placeholder draw function. More...
|
| |
| static int | LuaRender (lua_State *L) |
| | Renders all rendertargets with given camera. More...
|
| |
| template<typename T > |
| static int | LuaConstructor (lua_State *L) |
| | The LuaConstructor used by any userdata which is "new"-able. More...
|
| |
| template<typename T > |
| static int | LuaDestructor (lua_State *L) |
| | The LuaDestructor used by any userdata which is "new"-able and also needs to be garbage collected. More...
|
| |
| template<typename T > |
| static int | LuaToString (lua_State *L) |
| | The LuaToString function that is set in the metatables of generated objects. More...
|
| |
| template<typename T > |
| static int | LuaIndex (lua_State *L) |
| | The LuaIndex metamethod. More...
|
| |
| static void | LuaRegisterFunctions (lua_State *L) |
| | Registers functions of this class. More...
|
| |
Manages the Game.
- Author
- Riko Ophorst
| Game * blowbox::Game::Instance |
( |
| ) |
|
|
static |
Retrieves a pointer to the Game instance.
- Returns
- A pointer to the Game's instance
| int blowbox::Game::LuaDraw |
( |
lua_State * |
L | ) |
|
|
static |
A placeholder draw function.
- Parameters
-
| [in] | L | (lua_State*) the lua state |
| void blowbox::Game::LuaRegisterFunctions |
( |
lua_State * |
L | ) |
|
|
static |
Registers this objects functions.
- Parameters
-
| [in] | L | (lua_State*) the lua state |
| int blowbox::Game::LuaRender |
( |
lua_State * |
L | ) |
|
|
static |
Renders all rendertargets with given camera.
- Parameters
-
| [in] | L | (lua_State*) the lua state |
| int blowbox::Game::LuaUpdate |
( |
lua_State * |
L | ) |
|
|
static |
A placeholder update function.
- Parameters
-
| [in] | L | (lua_State*) the lua state |
| void blowbox::Game::Reload |
( |
const std::string & |
path | ) |
|
Invokes the reload callback in lua.
- Parameters
-
| [in] | path | (const std::string&) the path to the file that was reloaded |
The documentation for this class was generated from the following files:
- C:/Users/Riko pc/Documents/Visual Studio 2013/Projects/blowbox/Blowbox/game.h
- C:/Users/Riko pc/Documents/Visual Studio 2013/Projects/blowbox/Blowbox/game.cc


Programming is a tool I plan to use for health data analytics and informatics, to improve efficiency for doctors and patients.
Prior Education and Work
- Doctorate in Podiatric Medicine
- Board-accredited Foot and Ankle Surgeon
- Prior Technical Trainer for Biologic startup
- Taught scientists at major pharma how to use software for high throughput chemical testing
- Prior Reseach Study Coordinator
- University of Washington School of Public Health
- Ran complex multi-day multi-datapoint study
- Managed exposing subjects with asthma to diesel exhuast with varied particulate matter loads
- Prior Systems Administrator
- F5 Networks
- Managed 200+ server farm with multiple platforms
- Managed and implemented redundant data server sites throughout country
Only those who risk going too far can possibly find out how far they can go.


Macht Schnell
The Dark Side of the Performance Moon
Performance Golden Rule
Make Less Requests for Smaller Content
- Global styles, not per-page styles
- Lazy-load images
- Lazy-load javascript
- Concatenate css, javacsript
- Load using sharded, cookieless domains
The Speed of Light
299,792,458 m / s
From EC2 in Virginia
- 15ms to San Francisco
- 37ms to Beijing
But Wait, I Don't Have a Cable From VA to My Computer
Oh. Right.
Packet Loss
Another round-trip if anything is corrupted along the
way
distance * speed of light gives us two things to change
Lower the distance
Increase the speed of light
Wormholes
Are Prohibitively Expensive
Move The Content Closer
(Edge Caching)
(Akamai, ChinaCache)
Edge Caching
Local servers keep content cached, so users can load
from somewhere close instead of across the world.
- Images
- CSS
- Javascript
- Pages that don't refresh often
TCP (how the internet works)
Imagine a train.
TCP Connection
-
Someone telegrams airbnb.com to say a train is coming
-
airbnb.com sends a telegram back to say "okay"
-
airbnb sets up a train track connection
-
the train cars start small and get larger as they arrive
successfully, until they get too big and start to fail
TCP Connection
You must establish a new connection for every request.
SPDY (how the better internet works)
Instead of new connections, SPDY reuses the existing connection
and makes parallel requests.
SPDY, Also.
- Header compression (you only send cookies, etc once)
- Always SSL (yay security)
- Less packet loss, better socket utilization
If You Have a SPDY Connection
- No more domain sharding (a0-a3.muscache.com)
- No need to concatenate scripts (except legacy support)
- It's currently enabled for our static assets
Overview
- Cheat spacetime by using edge caches
- Lower time spent on data transfers by re-using connections
- Any Questions?


First, "Wong Tai Sin", Wong Tai Sin enshrined in the temple, is said to have a request for help. The side of the Kowloon Wall is modeled after the Forbidden City of Beijing. Second, Ocean Park, Hong Kong Ocean Park is divided into: Waterfront Paradise is divided into three theme parks-Hawaii World, Asian Animal World and Dream Water. The peak park is divided into five theme parks: Ocean World, Rapids World, Tropical Rainforest and Ice World. Go. Third, Rainbow Village can be said to be the C position in the estate. "Lonely Planet" is also the third place to list it as "Hong Kong's Top Ten Ins Hotspots." The roads in the four villages are named after the seven colors of the rainbow, just like the pink Pantone card board-the sky is rainbow color, and the person is the youngest.


Unexpected system error.
Please report to your local system administration, referring to error ID [errorID]
The application must be stopped to avoid inconsistencies


clustifyr
?Single cell transcriptomes are difficult to annotate without extensive knowledge of the underlying biology of the system in question. Even with this knowledge, accurate identification can be challenging due to the lack of detectable expression of common marker genes defined by bulk RNA-seq, flow cytometry, other single cell RNA-seq platforms, etc.
clustifyr
solves this problem by providing functions to automatically annotate single cells or clusters using bulk RNA-seq data or marker gene lists (ranked or unranked). Additional functions allow for exploratory analysis of calculated similarities between single cell RNA-seq datasets and reference data.
In this example, we take a 10x Genomics 3’ scRNA-seq dataset from peripheral blood mononuclear cells (PBMCs) and annotate the cell clusters (identified using Seurat
) using bulk RNA-seq data from isolated PBMC cell populations. The cell types in this dataset have been previously annotated, so this is a good dataset to use for testing the accuracy of clustifyr
.
To identify cell types, the clustifyr()
function requires several inputs:
Using the plot_tsne()
function, we can plot the known identities of each cluster, which were stored in the “classified” column of the meta.data table.
library(clustifyr)
library(ggplot2)
library(cowplot)
# Matrix of normalized single-cell RNA-seq counts
full_pbmc_matrix <- clustifyrdata::pbmc_matrix
# meta.data table containing cluster assignments for each cell
# The table that we are using also contains the known cell identities in the "classified" column
full_pbmc_meta <- clustifyrdata::pbmc_meta
# Create tSNE showing known cell identities for each cluster
plot_tsne(
data = full_pbmc_meta, # meta.data table containing UMAP or tSNE data
feature = "classified", # name of column in meta.data to color cells by
do_label = TRUE, # should the feature label be shown on each cluster?
do_legend = FALSE # should the legend be shown?
)
The clustifyr()
function returns a matrix of correlation coefficients for each cell type and cluster, with the rownames corresponding to the cluster number.
# Calculate correlation coefficients for each cluster (spearman by default)
vargenes <- pbmc_vargenes[1:500]
res <- clustify(
input = full_pbmc_matrix, # matrix of normalized single-cell RNA-seq counts
metadata = full_pbmc_meta, # meta.data table containing cell clusters
ref_mat = pbmc_bulk_matrix, # matrix containing bulk RNA-seq data for each cell type
query_genes = vargenes, # list of highly varible genes identified with Seurat
cluster_col = "seurat_clusters" # name of column in meta.data containing cell clusters
)
# Tidy up column names
colnames(res) <- gsub("primary human ", "", colnames(res))
# Peek at correlation matrix
res[1:5, 1:5]
#> B cells rep 1 myeloid DC rep 1 monocytes rep 1 neutrophils rep 1
#> 0 0.4765782 0.4530271 0.4669347 0.3601846
#> 1 0.4789201 0.4578896 0.4653634 0.3632720
#> 2 0.2586354 0.6328819 0.7125717 0.5247699
#> 3 0.6485402 0.4648393 0.4593464 0.3072537
#> 4 0.4102300 0.4071095 0.4080622 0.3214890
#> NK cells rep 1
#> 0 0.5368206
#> 1 0.5463310
#> 2 0.3373901
#> 3 0.4403688
#> 4 0.6370834
To visualize the clustifyr()
results we can use the plot_cor_heatmap()
function to plot the correlation coefficients for each cluster and each cell type.
clustifyr
also provides functions to overlay correlation coefficients on pre-calculated tSNE embeddings (or those from any other dimensionality reduction method).
# Overlay correlation coefficients on UMAPs for the first two cell types
plot_cor(
cor_matrix = res, # matrix of correlation coefficients from clustifyr()
metadata = full_pbmc_meta, # meta.data table containing UMAP or tSNE data
data_to_plot = colnames(res)[1:2], # name of cell type(s) to plot correlation coefficients
cluster_col = "seurat_clusters" # name of column in meta.data containing cell clusters
)
#> [[1]]
#>
#> [[2]]
The plot_best_call()
function can be used to label each cluster with the cell type that gives the highest corelation coefficient. The plots below show that the highest correlations between the bulk RNA-seq data and the 10x Genomics scRNA-seq dataset are restricted to the correct cell clusters.
# Label clusters with clustifyr cell identities
clustifyr_types <- plot_best_call(
cor_matrix = res, # matrix of correlation coefficients from clustifyr()
metadata = full_pbmc_meta, # meta.data table containing UMAP or tSNE data
do_label = TRUE, # should the feature label be shown on each cluster?
do_legend = FALSE, # should the legend be shown?
cluster_col = "seurat_clusters"
) +
ggtitle("clustifyr cell types")
# Compare clustifyr results with known cell identities
known_types <- plot_tsne(
data = full_pbmc_meta, # meta.data table containing UMAP or tSNE data
feature = "classified", # name of column in meta.data to color clusters by
do_label = TRUE, # should the feature label be shown on each cluster?
do_legend = FALSE # should the legend be shown?
) +
ggtitle("Known cell types")
plot_grid(known_types, clustifyr_types)
Correlation coefficients can also be calculated on a per cell basis by setting per_cell = TRUE
.
# Calculate correlation coefficients for each cell
cell_res <- clustify(
input = full_pbmc_matrix, # matrix of normalized single-cell RNA-seq counts
ref_mat = pbmc_bulk_matrix, # matrix containing bulk RNA-seq data for each cell type
query_genes = pbmc_vargenes, # list of highly varible genes identified with Seurat
per_cell = TRUE
)
# Overlay correlation coefficients on UMAPs
plot_cor(
cor_matrix = cell_res, # matrix of correlation coefficients from clustifyr()
metadata = full_pbmc_meta, # meta.data table containing UMAP or tSNE data
data_to_plot = colnames(cell_res)[c(1, 5)] # name of cell type(s) to plot correlation coefficient
)
#> [[1]]
#>
#> [[2]]
The clustify_lists()
function allows cell types to be assigned based on known marker genes. This function requires a table containing markers for each cell type of interest. Cell types can be assigned using several different statistical tests including, hypergeometric, Jaccard, Spearman, and GSEA.
# Take a peek at marker gene table
cbmc_m
#> CD4 T CD8 T Memory CD4 T CD14+ Mono Naive CD4 T NK B CD16+ Mono
#> 1 ITM2A CD8B ADCY2 S100A8 CDHR3 GNLY IGHM CDKN1C
#> 2 TXNIP CD8A PTGDR2 S100A9 DICER1-AS1 NKG7 CD79A HES4
#> 3 AES S100B CD200R1 LYZ RAD9A CST7 MS4A1 CKB
#> CD34+ Eryth Mk DC pDCs
#> 1 SPINK2 HBM PF4 ENHO LILRA4
#> 2 C1QTNF4 AHSP SDPR CD1E TPM2
#> 3 KIAA0125 CA1 TUBB1 NDRG2 SCT
# Available metrics include: "hyper", "jaccard", "spearman", "gsea"
list_res <- clustify_lists(
input = full_pbmc_matrix, # matrix of normalized single-cell RNA-seq counts
cluster_info = full_pbmc_meta, # meta.data table containing cell clusters
cluster_col = "seurat_clusters", # name of column in meta.data containing cell clusters
marker = cbmc_m, # list of known marker genes
metric = "hyper" # test to use for assigning cell types
)
# view as heatmap, or plot_best_call
plot_cor_heatmap(
cor_matrix = list_res, # matrix of correlation coefficients from clustify_lists()
col = not_pretty_palette, # colors to use for heatmap
cluster_rows = FALSE, # cluster by row?
cluster_columns = FALSE, # cluster by column?
legend_title = "-log10(padj)" # title of heatmap legend
)