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Prior to the present age it could be truly said, though one seldom had occasion for it, that there was no way to describe Charlie Callas (1924-2011). Well, it can still be said, but you don’t need to — you can just send links to film clips or GIFs and the thing is done. But no one can do what he did, so you can’t really demonstrate it or imitate it. And words fail in his case more than in most others. But I will try.
Callas was a comedian whose stand-up act involved pantomime of an extreme sort that only he could do. Yes, this was because he was enormously talented (though in a way that almost no one would actually be envious of, content to let Callas be king at whatever it was that he did), but also because, as I often say, he was “blessed with good comedy equipment”. By this, I mean, a funny face. “Ugly” is a value judgment I’m uncomfortable making, but it’s the sort of thing that was said. My mother certainly said it when he came on television: “That damn ugly Charlie Callas!”. He possessed ENORMOUS eyes that he could roll and bug and cross and sometimes half-cover with twitching eyelids as though he were having a seizure. He’d roll them into the back of his head so all you could see were the whites and he looked like a zombie. He had big furry eyebrows that he could make leap and dance up and down his forehead ONE AT A TIME. He had a huge beak of a nose that made him sort of look like a toucan. Craggy cheek bones and sunken cheeks, and a weak chin. And a mouth that was sort of an obscene gash. When I picture him, he’s usually panting like a dog, with his tongue hanging out. But it wouldn’t be just his face, but his whole (skeletal thin) body would get into whatever he was doing. And I haven’t even gotten to his sounds. He made sounds no one else on earth could make. They were the sorts of sounds that cartoonist Don Martin would spell out in Mad Magazine: “Frrrrrrp!” was a big one, or “hucketa-hucketa-hucketa!”, or “Whoa!” or a very wheezy, asthamatic sounding “HOWK!”
Of course, what I have just described is not an act. In his stand-up, he would use these skills to help him tell stories, in which he would play all the characters, illustrating what they did with sound effects, and constantly interrupting it with inexplicable tics and weird sounds. Or on variety shows, he’d be inserted into skits where this strange behavior could come into play. He first broke in the mid 1960s, and his primary medium was television. You saw him constantly on The Tonight Show with Johnny Carson, The Merv Griffin Show, The Joey Bishop Show, The Andy Williams Show, The Flip Wilson Show, The Dean Martin Show and Dean’s many celebrity roasts, The Mike Douglas Show, Dinah! (Dinah Shore’s Show), etc.
And occasionally this constant exposure and the gut-busting reaction of audiences would lead to guest spots on sit coms, or getting cast in movies. Jerry Lewis cast him in The Big Mouth (1967). Mel Brooks put him in Silent Movie (1976), High Anxiety (1978), The History of the World: Part 1 (1981), and Dracula: Dead and Loving It (1995). On rare occasions, he would even do something more “real” and “legit”. For example, he played a fairly straight, regular part as an informant on the caper show Switch (1975-78) with Robert Wagner and Eddie Albert. And he’s the voice of the dragon in Disney’s Pete’s Dragon (1977).
How does somebody like Charlie Challas (given that there is no one like Charlie Callas) happen? In his case he was born in Brooklyn, the son of a Greek-American father and a German-American mother. He started out as a drummer in big bands, playing with guys like Tommy Dorsey and Buddy Rich. This led to fooling around, which led to getting up in front of the mic. You often see him wearing a tuxedo in old clips and photos, and this is because he worked live constantly, at resorts like Las Vegas, Miami, etc. Very old school show biz.
I’ve stopped embedding or linking to youtube clips here; they all go dead. But, as a special thank you, I’ll be sending a Charlie Callas to readers who’ve given to my Patreon campaign. I had a hard time thinking what my Patreon rewards could be. This will be one — I’m sending the subscribers the kind of specially curated clips I used to post directly on Travalanche! And, believe me, a performance by Charlie Callas is not to be missed. Today is the birthday of the late Charlie Callas.


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- Open Access
Genetic diversity and population structure of genes encoding vaccine candidate antigens of Plasmodium vivax
Malaria Journal volume 11, Article number: 68 (2012)
A major concern in malaria vaccine development is genetic polymorphisms typically observed among Plasmodium isolates in different geographical areas across the world. Highly polymorphic regions have been observed in Plasmodium falciparum and Plasmodium vivax antigenic surface proteins such as Circumsporozoite protein (CSP), Duffy-binding protein (DBP), Merozoite surface protein-1 (MSP-1), Apical membrane antigen-1 (AMA-1) and Thrombospondin related anonymous protein (TRAP).
Genetic variability was assessed in important polymorphic regions of various vaccine candidate antigens in P. vivax among 106 isolates from the Amazon Region of Loreto, Peru. In addition, genetic diversity determined in Peruvian isolates was compared to population studies from various geographical locations worldwide.
The structured diversity found in P. vivax populations did not show a geographic pattern and haplotypes from all gene candidates were distributed worldwide. In addition, evidence of balancing selection was found in polymorphic regions of the trap, dbp and ama-1 genes.
It is important to have a good representation of the haplotypes circulating worldwide when implementing a vaccine, regardless of the geographic region of deployment since selective pressure plays an important role in structuring antigen diversity.
Malaria is one of the major global public health problems that affect most tropical regions of the world. Even though Plasmodium falciparum is the most virulent, it is estimated that Plasmodium vivax produces around 80 to 300 million clinical cases per year . Furthermore, there have been several reports of severe P. vivax malaria cases in the last few years [2–4]. In 2008, 560,221 malaria cases were reported in the Americas ; 74.2% of them caused by P. vivax and 25.7% by P. falciparum. About 90% of these malaria cases originated in the Amazon basin shared by Bolivia, Brazil, Colombia, Ecuador, French Guiana, Guyana, Venezuela, Suriname and Peru , whereas the other 10% was contributed by non-Amazon regions.
Developing a vaccine for P. vivax represents a major challenge especially considering the lack of in vitro cultures. Thus, current efforts focus on orthologs of P. falciparum. Over the past four decades, experiments performed in animals and human subjects have led to the development of several Plasmodium vaccine candidates. Antigenic surface proteins such as the Circumsporozoite protein (CSP), Thrombospondin related anonymous protein (TRAP), Duffy-binding protein (DBP), Merozoite surface protein-1 (MSP-1) and Apical membrane antigen-1 (AMA-1) are currently being evaluated as vaccine candidates in clinical trials [1, 6–8]. The CSP is one of the most extensively studied antigens and it is involved in the motility and invasion of the sporozoite during its entrance in the hepatocyte . Pv CSP has an immunogenic central repeat domain flanked by amino and carboxyl sequences containing highly conserved protein stretches (Regions I and II-plus) . This protein displays two major types of nonapeptide repeats: type I (VK210), which contains repeats of GDRA(A/D)GQPA or related sequences and type II (VK247), which is composed of ANGAGNQPG repeats [11, 12]. A third type of variant, called P. vivax-like has repeat units of APGANQ(E/G)GAA, identical to that described for Plasmodium simiovale[13, 14]. Currently, the P. falciparum RTS, S vaccine, which is based on CSP have showed to protect semi-immune adults  and children from P. falciparum natural infection in endemic areas [16–18]; this vaccine is being tested in Phase III clinical trials [8, 19].
Another sporozoite surface protein that stands out as a good vaccine candidate is TRAP, which is a 90 kDa protein with six distinct regions. TRAP is essential for sporozoite gliding and hepatocyte invasion [20, 21]. Genetic analysis of Pv TRAP reveals that most of the polymorphisms occur in Region II (von Willebrand factor A-domain) and III . The potential role of TRAP as a malaria vaccine candidate has been tested in mice and monkeys, inducing high levels of specific antibodies . Furthermore, a synthetic peptide-derived from a conserved region of the TRAP protein located in the N-terminal, proved to be immunogenic and provided partial protection in Aotus monkeys [1, 24].
On the other hand, the asexual P. vivax parasite antigens DBP, MSP-1 and AMA-1 have also received attention for their role in the invasion process and/or their generation of antibody response. DBP interacts with the Duffy blood group antigen of reticulocytes, MSP-1 is expressed abundantly on the merozoite surface and AMA-1 plays a role in apical reorientation of merozoites following initial attachment during invasion . Region II of DBP (DBPII) functions as a ligand domain and consists of 330 aa with 12 cysteines and numerous aromatic residues. Studies have identified the central region of the DBPII domain between cysteines 4 and 8, as the segment containing the conserved contact residues Tyr 94, Asn 95, Lys 96, Arg 103 and Ile 175 required for recognition of DARC (Duffy Antigen/Receptor for Chemokine) on human erythrocytes [25–28].
MSP-1 is part of a complex of proteins that is processed into smaller fragments by serine proteases during invasion of red blood cells . It has been suggested that fragments Pv MSP-114 and Pv MSP-120 could be included in a P. vivax multi-antigen vaccine since they contain high affinity reticulocyte binding (HARB) cluster regions and confer protection in monkeys [30–32]. AMA-1 has also been evaluated for inclusion in a multi- sub-unit vaccine for both P. falciparum and P. vivax[33, 34]. The complete AMA-1 protein contains three domains, from which a higher rate of mutations and level of diversifying selection has been shown in domain I [35, 36]. Indeed, AMA-1 is highly immunogenic, protecting against Plasmodium chabaudi infection in mice  and displaying significant antibody and T cell responses in endemic human populations [38–40].
Investigation of the sequence variation among malaria antigenic regions is necessary for the development of an effective malaria vaccine that includes haplotypes from different regions, considering that Plasmodium is highly diverse and these regions are under strong immunological pressure. In this study, important regions in Pv CSP, Pv TRAP, Pv DBP, Pv MSP-1 and Pv AMA-1 were genotyped using P. vivax Peruvian isolates collected during years 2006-2007. In addition, since natural selection plays a role in structuring diversity of these highly polymorphic antigens, the worldwide distribution of the main variants was determined by comparing the Peruvian samples with sequences previously reported from South America, Asia and Oceania.
Study area and sample collection
Blood samples from patients were collected during years 2006-2007 from endemic areas in the Peruvian Amazon region of Loreto Department using human use approved protocols (NMRCD.2005.0005). In Peru, P. vivax corresponds to more than 80% of the malaria cases and the majority of them are found in the Amazonic Department of Loreto . Fifty-seven samples were obtained from Padrecocha, in Punchana district, a village situated five km from Iquitos (Loreto's capital) and forty-nine samples were collected from a village located in the San Juan district just south of Iquitos. All samples were confirmed to be positive for P. vivax by microscopic analysis on Giemsa-stained blood smears.
PCR and DNA sequencing
DNA was purified from 200 μL of whole blood using the QIAamp DNA Blood mini kit (QIAGEN, Valencia, CA). PCR amplification was performed in a 50 μL reaction mixture containing 0.2 μM each of forward and reverse primers, 200 μM each dNTP, 0.6 units of DNA polymerase recombinant (Invitrogen, Carlsbad, CA), 3 μL of 10× PCR buffer, 1.5 mM of MgCl2 and 5 μL of DNA extracted from blood. To assess antigenic variability of Pv CSP, the Central Repetitive Region, Region I and Region II were amplified. The PCR conditions were: 5 min at 94°C; 1 min at 94°C, 1 min at 58°C, 2 min at 72°C for 35 cycles; and a final extension of 72°C for 2 min, using the primers VSC-OF (5'ATGTAGATCTGTCCAAGGCCATAAA3') and VSC-OR (5' TAATTGAATAATGCTAGGACTAACAATATG 3') . The amplification of the highly polymorphic regions II and III of Pv TRAP  was obtained using the primers Pvsspf: 5' GTCGATTTATACCTCCTAGTTGACGG 3' and Pvsspr: 5' CAGCTTGAACGTCGACCTCCGC 3' with the following conditions: 5 min at 94°C; 1 min at 94°C, 1 min at 50°C, 2 min at 72°C for 35 cycles; and a final extension of 72°C for 2 min. The amplification of the central region of the Pv DBPII domain was done using primers Liz-1 (5' TGGGGAGGAAAAAGATG 3') and Liz-2 (5' AACGGAACAAACGCATAAC 3') . The PCR conditions used were as follows: 4 min at 94°C; 1 min at 94°C, 2 min at 58°C, 2 min at 72°C for 35 cycles; and a final extension of 72°C for 3 min. Pv MSP-1 primers described by Espinosa et al. (HARB1_f: 5' GGCAACAACGATGACGAC 3', HARB1_r: 5' CTTCTCAAGTCGAGCAG 3', HARB2_f: 5' TCACTGTAAAGAAATTGCAG 3' and HARB2_r: 5' ATACATGTGTGCTCGGAG 3') were used for amplifying the two HARB containing regions: Pv MSP-120 and Pv MSP-114 kDa  with the following conditions: 5 min at 94°C; 1 min at 94°C, 1 min at 50°C, 1 min at 72°C for 35 cycles; and a final extension of 72°C for 5 min for the first region and 5 min at 94°C; 1 min at 94°C, 1 min at 45°C, 1 min at 72°C for 35 cycles; and a final extension of 72°C for 5 min for the second region. Finally, a nested PCR was used to amplify domain I of Pv AMA-1 as described by Cheng and Saul . PCR products were purified using the QIAquick PCR purifications kit (QIAGEN, Valencia, CA, USA) and sequenced using BigDye® Terminator v3.1 sequencing kit (Applied Biosystems, Foster City, CA, USA), with the primers previously mentioned and an ABI 3100 automated sequencer.
Sequence determination and data analysis
Sequences were analysed using the Sequencher™ software version 4.7 (Gene Codes Corporation, Ann Harbor, MI, USA) and sequence alignments were performed using the MEGA software version 4.1 . Comparison with previously deposited P. vivax sequences in the GenBank was performed using BLAST search. The number of haplotypes (h), haplotype diversity (hd) and nucleotide diversity (Pi) values were obtained using the DnaSP version 5.0 . The effect of natural selection was also evaluated by estimating the difference of non-synonymous and synonymous substitutions (Dn-Ds) using the Nei and Gojobori's method with the Jukes and Cantor correction. The null hypothesis of neutrality was rejected if Dn:Ds ≠ 1. Positive values for Dn-Ds indicated positive selection while negative values indicated negative selection . Standard error was calculated using bootstrap with 1,000 replications and rates of Ds and Dn were compared by the Z-test of selection using Mega version 4.1 . To test whether the haplotypes clustered according to their geographic origin, the model-based clustering algorithm implemented in the Structure 2.1 software was used . This software uses a Bayesian clustering approach to assign isolates to K populations characterized by a set of allele frequencies at each locus. The number of such populations may be either previously known or not. The program was run five times at different K values (from 2 to 10) with a burn-in period of 10,000 iterations followed by 50,000 iterations. The admixture model was used in all analyses, which allows for the presence of individuals with ancestry in two or more of the K populations. However, clustering algorithms incorporate stochastic simulation as part of the inference and, as a consequence, independent analyses of the same data may result in several distinct outcomes, even though the same initial conditions were used. To facilitate the interpretation of population-genetic clustering results, the computer program CLUMPP (Cluster Matching and Permutation Program) was used. CLUMPP strips away the 'label switching' heterogeneity so that the 'genuine multimodality' can be detected and quantified . In addition, distruct 1.1 was used to graphically display the clustering results . The genetic difference between clusters was measured by the fixation index (Fst) calculated from the haplotype frequencies (Hst) and pair-wise DNA sequence diversity (Kst*) using DnaSP version 5.0 . The nearest-neighbour statistic (Snn) was also calculated between geographic populations. Snn is a measure of how often the "nearest neighbours" of sequences are from the same locality in geographic space. This statistic is applicable when genetic data are collected on individuals sampled from two or more localities. Snn is expected to be near one when the populations at the two localities are highly differentiated and near one-half when the populations at the two localities are part of the same pan-mictic population .
A 1,100 bp PCR product corresponding to Pvcsp was amplified from 106 P. vivax samples from Peru. In addition, 326 complete CSP sequences from Brazil, Colombia, India, Iran and Korea were also included in the amino acid alignment. Sequences from Thailand, China and PNG were used to determine the percentage of strains circulating of each Pv CSP type (Table 1); however, they were not considered in the haplotype analysis since the sequences were not complete. Most of the strains circulating worldwide belonged to the VK210 type and in most population sets analysed (except Colombia and Thailand) a greater percentage of VK210 haplotypes was found. The amino acid alignment of the sequences used in this study showed 97 different VK210 haplotypes and 28 VK247 haplotypes circulating worldwide. Different haplotypes were observed in South American and Asian countries, except for one VK247 Peruvian haplotype, which was also found in Iran . Thirty-seven VK210 and 19 VK247 haplotypes were identified in South America; while in Asia, 51 and 10 haplotypes were observed from each type.
In Peru, 23 different VK210 subtypes and three different VK247 subtypes were found (Figure 1). Pv CSP displayed a different pattern of a nine amino acid oligopeptide repetitions depending on the subtype. The VK210 subtypes contained the repeat region G(D/N)RA(A/D/G/V)G(Q/H)PA either 16, 17, 18 or 19 times, while VK247 subtypes contained the repeated region ANGA(G/D)(D/N)QPG, 17 or 18 times. Region I of Pv CSP showed a polymorphism in position 3 (L/V), whereas the 18 amino acid long linear peptide EWTPCSVTCGVGVRVRRR, located on Region II [53, 54], was highly conserved. Pv CSP analysis of Peruvian isolates also revealed high variability of VK210 and VK247 haplotypes compared to other populations from South America [55, 56]. Region II-plus, which contains dominant B-cell epitopes [57, 1] and block hepatocyte invasion by sporozoites , was highly conserved in Peruvian isolates, with the same amino acid sequence as the Sal I strain.
A total of 104 sequences of regions II and III of Pvtrap were obtained from the Peruvian isolates. In addition, 63 Thailand sequences from GenBank were included in the amino acid alignment. The N-terminal region (209-256aa), which has been proved to be immunogenic and to provide partial protection in Aotus monkeys  was highly conserved with only three amino acid substitutions in positions 216 (R/K), 219 (H/Q) and 229 (G/K). In general, more diversity was found in Thailand (h = 34, π = 0.007 ± 0.00029 and Hd = 0.961 ± 0.01) (Table 2) compared to Peru (h = 7, π = 0.00235 ± 0.00012 and Hd = 0.641 ± 0.026) and according to the Dn-Ds test, regions II and III of Pvtrap were under positive selection (Table 3).
A 1,150 bp DNA fragment of Pvdbp was amplified from all 106 Peruvian isolates and 601 P. vivax sequences from Brazil, Colombia, India, Sri Lanka, Thailand, Korea, Iran and Papua New Guinea were also included in the analysis. The alignment of 510 bp from region II of dbp showed 150 different haplotypes, from which 39 were found in South America, 42 in Asia, 56 in Oceania and 13 were found in South America and Asia. In the Peruvian isolates 26 different haplotypes were found with genetic diversity values of π = 0.008 ± 0.00048 and Hd = 0.903 ± 0.014. In general, the average genetic and haplotype diversity in South America and Asia was very similar (Table 2). On the other hand, the analysis of DNA sequence polymorphisms showed a significant difference between synonymous and non-synonymous substitutions, indicating balancing selection in dbp II (Table 3). In addition, several substitutions in the positions underlined were found in DBPII regions corresponding to peptides H1: F HR DITFRKLYLKRKL, H3: DEK AQ QRR KQWWN ES K an 1653: RD YVS ELPTEVQ KLKEK C, previously reported as immunogenic [27, 59].
According to the amino acid alignment of PvMSP-120 and PvMSP-114, a total of 28 haplotypes were found worldwide, 11 in South America, 14 in Asia and three in both continents. When comparing genetic diversity between populations, Peru (π = 0.00286 ± 0.00029, Hd = 0.665 ± 0.041) and Brazil (π = 0.00293 ± 0.00061, Hd = 0.75 ± 0.139) showed the highest levels of nucleotide and haplotype diversity, while in Korea, Thailand and Turkey these HARBs were highly conserved. In general, values of genetic diversity for Pvmsp-1 were low when compared with the other genes and no clear signature of selection was found according to the Dn-Ds test (Table 3).
The polymorphic region of Pvama-1 (Domain I of protein) was amplified and sequenced from 98 of the 106 Peruvian isolates. A total of 774 Pvama-1 partial sequences were used in the analysis, including samples from Brazil, Venezuela, India, Sri Lanka, China, Korea, Myanmar, Philippines, Thailand, Papua New Guinea and Solomon Islands. A total of 168 haplotypes were identified; 135 were exclusively found in Asia, nine in Oceania and 12 in South America. In addition, seven haplotypes were shared in America and Asia and five in Asia and Oceania. According to the analysis of polymorphisms, the dN-dS test indicated balancing selection in this domain (Table 3) with several haplotypes in low frequency. In addition, values of nucleotide diversity were very similar between geographic regions and populations (Table 2).
Population genetic analysis
When analysing Plasmodium isolates, samples are usually grouped by geographic location; nevertheless, this approach assumes that geographic barriers or distance are the underlying factors that limit gene flow. It also assumes that genetic exchange within each population geographically defined is random. To better estimate the distribution of haplotypes of different P. vivax vaccine candidate antigens, a Bayesian clustering algorithm  was used to sub-group samples according to related SNPs haplotypes. The Structure model assumes that loci are independent within populations; however, this assumption is likely to be violated for sequence data, or data from non-recombining regions. It is a matter of debate whether it is valid to group sequences containing potentially linked polymorphisms in a single gene under balancing selection . The main cost of the dependence within regions will be that Structure underestimates the uncertainty in the assignment of particular individuals. However, if there is enough independence across regions than LD within regions, then Structure may actually perform reasonably well . Moreover, our main goal is not to describe the demographic history of the population but to get a graphically representation of the vaccine candidate's haplotype distribution. Genetic differentiation between clusters was calculated using Fst from haplotype-frequencies and pairwise DNA sequence diversity. The haplotype frequency-based statistics are more sensitive for small sample sizes, while the sequence-based statistic is a more sensitive method for detecting population structure in highly polymorphic loci [61, 62].
The results obtained with Pv TRAP, Pv DBP and Pv AMA-1 showed three different clusters. Significant differentiation was observed in haplotype and pair-wise comparisons between all clusters according to the Fst values (Table 4), except for Pvmsp-1, from which a significant number of clusters it was not possible to identify. Although admixed haplotypes were observed in Pvdbp and Pvama-1, the estimate of K = 3 best explained the distribution of haplotypes from all geographic populations; besides, the defined sub-groups were not geographically or even regionally restricted (Figure 2). Snn values between the different geographic populations were also calculated for each of the antigens. We observed that for most of the comparisons in DBP, AMA-1 (Table 5) and TRAP (Snn = 0.86803*), Snn values were near one, which indicates that the localities are highly differentiated. However, close geographic populations such as Sri Lanka and India in South Asia and Peru and Brazil in South America showed almost not genetic differentiation (Snn = 0.58, p < 0.01 and Snn = 0.65, p < 0.01). Thus, relative low Snn values were found between Brazil, India and Sri Lanka, since there are shared haplotypes circulating in South America and South Asia. Moreover, previous work also indicated low genetic differentiation using Fst analysis between dbpII sequences from Brazil and Iran, India or Sri Lanka . On the other hand, at least seven of the pairwise comparisons made using MSP-1 resulted in non-significant values.
When designing an efficient anti-malarial vaccine, worldwide information of the circulating antigenic variants is necessary for the formulation of a polyvalent vaccine, which would be effective in different regions. Because of the complex parasite life cycle and the co-existence of P. falciparum and P. vivax infections in most endemic areas, it is accepted that a multi-antigen and multi-species vaccine should be developed for most endemic regions . In P. falciparum, the majority of haplotypes upon which current vaccines are based have been found to be present at extremely low frequencies in the global parasite population, except for lsa-1 and msp-1 haplotypes . Since immune selection might be playing a role in structuring the diversity of highly polymorphic antigens, population genetic studies in endemic regions are essential for the next generation of vaccines. In this study, the information obtained from the Peruvian isolates contribute to get a better understanding of the haplotypes circulating in the Amazon basin and to compare these results with other previously reported from different parts of the world.
For the analysis of Peruvian isolates, samples form Padreococha and San Juan were considered as a single geographic population since these areas are relatively closed and they present similar haplotype distribution in all antigens with little or none genetic differentiation between them. In general, data from a two-year period (2006-2007) in Peru revealed high nucleotide and haplotype diversity compared to other geographic populations included in the final analysis. These results suggest that the extent of allelic diversity in this region is determined by complex demographic processes that include heterogeneity of transmission intensities in time and space, as well as the number of recurrent infections that are common in the region, allowing for a high turnover of parasite genotypes . The presence of asymptomatic cases  participating in the infection process might also be contributing to the persistence of genetic variation since they are a "reservoir" of untreated patients. In addition, we need to take in consideration possible P. vivax multi clonal infections that might be difficult to observe by direct sequencing. This limitation could potentially affect the overall estimates of genetic diversity and population structure; however, for the goal of this study, the data collected is useful to obtain a broad picture of how these vaccine candidate haplotypes are worldwide distributed. If a fine scale study is considered in which the goal is to understand the population structure of Plasmodium in a specific geographic area then, unlinked microsatellite loci should be used to get a better idea of the parasite's population history.
When considering worldwide data, genes encoding non-merozoite antigens in P. falciparum showed variation in diversity . In P. vivax, the same pattern was observed using CSP and TRAP. Although, limited population samples were available for TRAP sequences. On the other hand, the genes encoding merozoite antigens in P. falciparum and P. vivax showed different patterns. P. falciparum genes encoding AMA-1, EBA-175 and MSP-1, MSP-3 and MSP-4 showed similar levels of diversity among countries and regions . In P. vivax, genes dbpII and ama-1 also showed this trend; however, msp-1 exhibited very low but different polymorphism and haplotype diversity values among regions and countries. These results support the hypothesis that maintaining the structural conformation of the HARBs regions (MSP-120 and MSP-114) is important for recognition and binding of reticulocytes . The limited variation of amino acid substitutions in these fragments makes them strong candidates to be tested in immunological studies. Also, TRAP sequences showed low levels of nucleotide and haplotype diversity with a highly conserved region between positions 209 to 256, previously reported to be immunogenic.
Contrary to P. falciparum, a largely structured diversity in all P. vivax populations was not found on the basis of geography. The clusters determined by Structure were found in most countries except in those with small number of samples, suggesting that haplotypes corresponding to those clusters were underrepresented. Moreover, a high level of admixture in all countries was observed, which could be explained by natural and high fluctuations over time, as a result of frequency-dependent selection or the discrepancy due to the sampling used , since the worldwide sequences obtained from GenBank came from different time points and locations. On the other hand, the lack of geographic clustering could be explained by balancing selection acting on the genes studied and as a result high worldwide antigen diversity was observed not only in Asia but also in South America.
In conclusion, it would be ideal to have a good representation of the haplotypes circulating worldwide when implementing a vaccine regardless of the geographic region of deployment. Considering that selective pressure plays an important role in structuring the diversity of antigens, it cannot be assumed that a single strain could accurately represent the haplotypes circulating worldwide and as a consequence be effective in all endemic regions. In addition, Plasmodium population definition should be revised since parasite sub-groups are not necessarily geographically restricted and treatment or vaccination in a region cannot be considered as if it was isolated but a dynamic map needs to be acknowledged. Thus, important epidemiological units should be established according to the parasite's gene flow, which would lead to a more effective malaria control strategy.
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Several authors of this manuscript are employees of the US Government. This work was prepared as part of their duties. Title 17 U.S.C. § 105 provides that 'Copyright protection under this title is not available for any work of the United States Government.' Title 17 U.S.C. § 101 defines US Government work as a work prepared by a military service member or employee of the US Government as part of that person's official duties. The views expressed in this article are those of the authors only and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the US Government. This research was supported in part by the grant R01GM80586 from the National Institute of Health to AE.
The authors declare that they have no competing interests.
SC conducted the molecular genetic studies, performed the genetic diversity experiments, analysed the data and wrote the first draft of the manuscript. LT performed part of the genetic diversity experiments. AE, CL, SD contributed to the write up and reviewed the final draft. DB designed the project, supervised and directed the research team and contributed to the writing and editing of the manuscript. All authors read and approved the final manuscript.
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Chenet, S.M., Tapia, L.L., Escalante, A.A. et al. Genetic diversity and population structure of genes encoding vaccine candidate antigens of Plasmodium vivax. Malar J 11, 68 (2012). https://doi.org/10.1186/1475-2875-11-68
- Plasmodium vivax
- Vaccine candidates

domain: medical


text:
Every couple years, the University of New Mexico duck pond is drained by UNM’s Facilities Management department. The drainage makes the pond a cleaner and healthier environment for the fish, ducks and turtles that reside in the center of campus.
“We take all the turtles and fish to one of the golf courses while it’s emptied, then bring them back once the cleaning is done,, but there’s still a few of them — turtles and ducks — around here. They hang out, you know?” said Dwayne Jones, UNM’s water systems master technician.
Cleaning the pond is a diligent process that takes various steps. First the pond is drained completely, then Facilities Management cleans out the grime and takes it to an offsite landfill. The project is led by Richard Schorr, head of groundskeeping at UNM.
“With all the duck feces and animal droppings, it just builds up a lot of sludge over time,” Jones said.
The cleaning started on July 7, followed by the refilling of the pond on July 11. While no courses are currently taking place in-person due to the COVID-19 pandemic, some students still live close enough to campus to witness the wildlife walking around.
Elizabeth Dooner was walking her dog through the campus when she noticed three turtles roaming around.
“My dog began to sniff the area where the turtles were,” Dooner said. “Then I looked around and saw three different ones. I didn’t know they were draining the duck pond this summer.”
Now, the duck pond is restored with clearer waters and healthy animals to enjoy the new environment.
Lauren McDonald is a freelance reporter at the Daily Lobo. She can be contacted at email@example.com or on Twitter @old_mcdonald25

domain: news


Wilson close to signing for Peterborough - reports
PUBLISHED: 14:38 26 June 2012 | UPDATED: 14:38 26 June 2012
Winger on the move according to reports coming out of south London
Lawrie Wilson is close to signing for Peterborough United, according to reports.
The winger has interested The Posh as well as Charlton Athletic and Millwall this past year, and reports coming out of south London suggest Charlton have missed out on his signature as he is close to signing for United.
If you value what this story gives you, please consider supporting the Comet. Click the link in the yellow box below for details.


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Boba Fett: Pursuit hits stores this December and poses some very interesting ideas that might be answered finally as well as possibly in Episode III itself. A lot of fans are intrigued by the relationship Boba Fett has between Mace Windu and would love to see a fight sequence between Samuel L. Jackson’s character and the mysterious bounty hunter still coping with loss.
According to sources producer of the Star Wars prequels, Rick McCallum has hinted at this actually appearing in the film, “Well, remember what Obi-Wan learned on Kamino. Jango didn’t want Boba to be artificially aged like the other clones, but whose to say that Boba wouldn’t do it himself, in order to become strong enough to seek his revenge.” George Lucas chimed in once before about Mace Windu’s fate in Episode III stating about Samuel L. Jackson “…He’s looking forward to the fact that he knows that at the end of the Clone War he gets killed so he gets hopefully a great death scene.”
“Young Boba Fett is quickly making a name for himself as one of the most capable bounty hunters in the galaxy. Having survived an encounter with the deadly General Grievous, there’s one bounty he wants more than any other: Mace Windu, the Jedi who killed his father.
“When fate and Anakin Skywalker bring Boba Fett to Coruscant, Boba plunges into the city-planet’s underworld and arms himself for the greatest confrontation in his young life.
“It’s bounty hunter vs. Jedi Master as Boba Fett seizes his one and only chance at vengeance, in the dramatic conclusion to the popular series. Pursuit is the sixth novel in the series, a Clone Wars novel by Elizabeth Hand. It features cover art by Louise Bova, and is scheduled for publication in December, 2004 by Scholastic Inc.”
According to a transcript from the Official Star Wars web site representative, “Scholastic will end the Boba Fett series with book six, Pursuit. It’s mostly how Boba Fett ends up where he is in Episode IV.”

domain: literature


text:
Zombie Kid Creepy Crawlies
by CL Cook
Searching for creepy crawly bugs is one of Zombie Kid's favorite things to do. One day he has to share his favorite hobby with his zombie sister. Learn with Zombie Kid the importance of patience and being kind to your siblings as he teaches his sister how to be an expert bug catcher. This book is a great addition to any kid's collection!

domain: library


text:
With a stay at Zhengzhou Galaxy Hotel in Zhengzhou (Jinshui District), you'll be close to Century Amusement Park. This hotel is within the vicinity of Erqi Pagoda and Renmin Park.
Stay in one of 160 guestrooms featuring flat-screen televisions. Complimentary wired Internet access is available to keep you connected. Conveniences include phones and desks.
Make use of convenient amenities such as concierge services, babysitting/childcare, and a hair salon.
Enjoy a meal at a restaurant, or stay in and take advantage of the hotel's room service (during limited hours).
Business, Other Amenities
Featured amenities include a business center, limo/town car service, and dry cleaning/laundry services. Event facilities at this hotel consist of a conference center and meeting rooms.

domain: travel


text:
We are creatures of habit. Despite the fact that many of us claim that we like ‘taking risks’, we rarely do. We do what we always do. Rarely do we ever ‘rock the boat’.
We have become a society that doesn’t do politics. We talk about it – lots! However, people’s participation in democratic activities is on the decrease – not only voting, but also campaigning/fighting for what we believe is right.
Alinsky argued, in his Rules for Radicals, that Community Organisers need to stay within the experience of our communities. Ironically, he argued this point to ensure that people do get out of their comfort zones and act more politically than they would otherwise do.
The world as it is is a world where the powerful aim to keep things as they are. It works for them. Why change things that work?
To build a world as it should be, it therefore takes people who are willing to subvert the status quo. Change rarely happens without tension.
In our schools, things are very prescribed: there are frameworks (Ofsted, the National Curriculum, assessment frameworks and their connected progress measures), there are structures, and these are often resting on rules set by people whose self-interest is not to question the world as it is. Remember, the world as it is works for them, so why change it?
I’ve argued before that teachers can easily become tactical disruptors. Building on their experience of teaching the skills required for children to succeed within our current system (e.g. reading, mathematics, writing, etc.), elements of disruptions can easily be added to the mix.
In our school, we increasingly – increasingly as this is still work-in-progress – start with the following postulate: it is our duty to make the world a better place through everything we teach. What we teach in terms of knowledge and skills, consequently, has to revolve around that. If not, we often tell prospective families, we’ve missed the trick.
There is nothing radical as such in our approach. Simply, we start with what Simon Sinek talks about in his Start with Why. Why do we do what we do? What’s the point?
Our point is that it is politics which makes the world go round. It is people getting involved and taking ownership of what they want society to look like that make our lives more meaningful. The choice to be a shaper of society – as opposed to a receiver – is exactly that: a choice.
School leaders interested in change need to not shy away from politics. As Cicero wrote, ‘Citizens are made, not born’. Democracy is learned by doing, good practice and eventually ‘informed consent’. And that’s tricky, because we’re told to remain neutral. Neutrality paralyses us and we therefore never reach ‘informed consent’. Doing so is exactly why our world either stagnates or derails in ways which, personally, bother us.
How to reconcile this tension between the need to be neutral/impartial and the need to realise that politics isn’t neutral is a key question we need to ask ourselves – openly.
Politics, as people mostly define it, is mostly partisan. And that’s where we often get stuck. We have to take sides and that makes us biased, something many frown upon within schools. But politics doesn’t have to be – and, in fact – shouldn’t be partisan. Politics should simply be about ‘informed consent’, reached through dialogue and the need to establish what works for me, beyond me. In the words of Rabbi Hillel: If I am not for myself, who am I? When I am only for myself, what am I? And if not now, when?
Veteran Community Organiser Mike Gecan recently argued, in an op-ed published in the New York Daily News, that we need to be much more fluid about our political views. Having to stick to a ‘party line’ isn’t healthy.
So, taking this back to experiences we, as teachers, are very familiar with, let’s do something straightforward: let’s talk about what we believe is right in terms of the challenges our communities face and let’s take on those who have the power to affect the change we seek and convince them that they need to implement the changes we believe are right.
Take the issue of Pupil Premium funds being used by some private sector companies to the disadvantage of vulnerable families: the fact that some companies don’t give students in receipt of the Pupil Premium the change they’re entitled to when they purchase meals (charging them the full equivalent of their Pupil Premium allowance, without giving them their change back when using less than said allowance) is wrong. Taking on those private companies and/or the Local Authorities that use them doesn’t make us partisan or anti-capitalist: it simply makes us justice seekers.
Let’s open up a wider dialogue between teachers, students, and families within our schools so we establish what our views are and let’s make our teaching clearer about what we stand for and how our curricula can meet the wider needs of the society we have the power to shape. Let’s not shy away from saying what we think, as school communities. And more than that, let’s act on what we believe is right for, as we’ve said before, it is not hope that leads to action but, rather it is action that leads to hope.

domain: sociology


About eight months after securing the city's approval for a 29-story condominium project in the Gold Coast, a Chicago developer might not build the tower after all.
Lexington Homes hired the Chicago office of HFF to sell the development site at 1022-1028 N. Dearborn Parkway, on a parking lot north of the Ruth Page Center for the Arts. A Lexington venture bought the property only in July, a few months after the City Council signed off on a zoning change to allow the 50-unit project.
Lexington is still weighing its options for the 11,281-square-foot property and could still develop the building if HFF finds a joint venture partner willing to invest in the project on attractive terms, said HFF Managing Director Danny Kaufman.
“It could be a sale of the site,” he said. “It could also be a capital raise.”
Lexington Co-Principal Jeff Benach declined to comment.
Amid a resurgent condo market, especially at the top end, demand for development sites is strong these days. That's especially true for parcels like Lexington's that fall under the city's older affordable-housing regulations. The city approved tougher rules last year that will increase development costs, making the grandfathered properties more attractive to developers.
The Lexington venture paid $16.8 million for the Gold Coast site in July, a price that reflects the higher value based on the property's new zoning. Considering how much the downtown condo market has risen since Lexington signed its contract to buy the site, it's likely that the firm could sell the property for more than it paid, said Susan Tjarksen, principal and managing broker at KIG, the Chicago-based brokerage that sold the site to Lexington for the Ruth Page Foundation.
Lexington also bought air rights over the existing low-rise buildings to the south to protect views in that direction, according to an HFF flier.


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domain: finance


text: Protein phosphatase 5 (Ppp5) is one of several proteins that bind to the Hsp90 chaperone via a tetratricopeptide repeat (TPR) domain. We report the solution structure of a complex of the TPR domain of Ppp5 with the C-terminal pentapeptide of Hsp90. This structure has the "two-carboxylate clamp" mechanism of peptide binding first seen in the Hop-TPR domain complexes with Hsp90 and Hsp70 peptides. However, NMR data reveal that the Ppp5 clamp is highly dynamic, and that there are multiple modes of peptide binding and mobility throughout the complex. Although this interaction is of very high affinity, relatively few persistent contacts are found between the peptide and the Ppp5-TPR domain, thus explaining its promiscuity in binding both Hsp70 and Hsp90 in vivo. We consider the possible implications of this dynamic structure for the mechanism of relief of autoinhibition in Ppp5 and for the mechanisms of TPR-mediated recognition of Hsp90 by other proteins.


Many foreign companies have considered relocating factories to Vietnam after the COVID-19 epidemic
According to JLL, Vietnam has been a promising destination since the wave of factory relocation from China began. Manufacturing industries can easily switch to Vietnam after the COVID-19 epidemic.
The research agency pointed out that, when factories were forced to close during the COVID-19 outbreak in China, many large companies had to plan to overcome the production restriction period.
A number of multinational companies have been planning to expand operations in Vietnam since last year, in order to reduce the pressure of new tariff barriers on goods exported from China to the United States and seeking markets to replace when prices increase.
Data from the U.S. Census Bureau shows that U.S. merchandise imports from Vietnam in 2019 increased by 35.6% over the same period, in contrast to a 16.2% decline in imports from China. This year's data will be affected by the impact of COVID-19 on global supply chains, but production trends will shift from China to countries in Southeast Asia.
“Vietnam has been a promising destination since the wave of factory relocation from China began. Although the COVID-19 epidemic is causing certain difficulties for decisions and relocation activities, investors are still confident to raise land prices in the first quarter of 2020 as this is a long-term investment trend ” Mr Stephen Wyatt, General Director of JLL Vietnam emphasized.
According to the JLL quarter 1/2020 report, the North attracts the majority of large corporations who want to diversify their product portfolio next to their facilities in China, with well-developed infrastructure and have a location near to China.
The average land price reached 99 USD / m2 / rental cycle, up 6.5% over the same period last year. Available factories - a favourite of small and medium-sized businesses - remain stable at rents ranging from the US $ 4.0-5.0 / sqm/month, and are already full.
In the South, JLL recorded an increase in the number of land lease requirements and developers became more confident in increasing the land rent. The average land price in the first quarter of 2020 reached US $ 101 / m2 / rental cycle, up 12.2% over the same period. However, the development of logistics/infrastructure does not correspond to this increase in land prices because the development process for significant changes in infrastructure is still slow, so potential investors have to start looking for other alternatives. Available factories in the South have a rent ranging from 3.5-5.0 USD / m2 / month, a slight increase in Binh Duong, Ho Chi Minh City, Long An and maintain a stable level in the remaining provinces.
According to this unit, under the impact of COVID-19, the suspense of lease agreements and new needs will become more and more evident if the situation does not improve soon. However, the market will recover and grow rapidly after the epidemic is regulated. The disruption of the global supply chain due to the impact of the disease has made businesses realize the urgency of diversifying their production and avoiding being dependent on one country.
COVID-19 could be the new catalyst to promote production shifts, after being promoted by trade tensions last year. Therefore, Southeast Asia in general and Vietnam, in particular, will be more attractive to businesses in the future.
Meanwhile, China focuses on developing value-added industries. It is home to the world's leading companies for solar cells, 5G networks, artificial intelligence and battery manufacturing. The main reason is that these businesses produce high-value products, creating high tax revenues for the government. In addition, low-value manufacturing industries often cause more pollution, while China is expecting to improve the environment in urban areas. Switching to cleaner production with less space will also free up land for re-planning.
However, according to JLL, not all manufacturing sectors can easily move to Vietnam. The salary of workers in China is three times higher than in Vietnam, but the skill level of workers here is also higher. The size of China cannot be replicated: the number of migrant industrial workers in China is even higher than Vietnam's population. Moreover, a large volume of manufactured goods is for the Chinese domestic market.
“In the long run, many businesses are likely to change their production plans to ensure supply chain continuity and minimize the risk of similar shocks in the future. Along with initiatives to improve sustainable performance and limit the environmental impact of manufacturing operations, retailers can choose to produce and buy products from the domestic market ”said JLL representative.
If you are looking for offshore partners to develop outsourcing in Vietnam, besides the companies that have been in Vietnam for many years, Hachinet is also a place to experience with the following characteristics.
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Recently, 80% of Hachinet’s clients are Japanese software companies, our engineers not only have high technical performance and professional qualifications but also are fluent in Japanese and able to communicate directly with customers.
Please contact us via email: firstname.lastname@example.org


text:
During her concert at the Cape Town Stadium, Lady Gaga called a lucky few concertgoers onto the stage. One fan got the chance to sing along with Gaga on an acoustic version of Born This Way.
24.com publishes all comments posted on articles provided that they adhere to our Comments Policy. Should you wish to report a comment for editorial review, please do so by clicking the 'Report Comment' button to the right of each comment.
Data released by Minnesota authorities provide some new details about more than 40 calls for service involving Prince's Paisley Park complex. Read More »
The Rolling Stones, Bob Dylan, Paul McCartney, Neil Young, Roger Waters and The Who - the most celebrated acts in rock 'n roll - will perform at a mega festival in October. Read More »
Cape Town show pics.
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domain: entertainment


text:
On The Power of Vulnerability, Dr. Brown offers an invitation and a promise - that when we dare to drop the armor that protects us from feeling vulnerable, we open ourselves to the experiences that bring purpose and meaning to our lives. Here she dispels the cultural myth that vulnerability is weakness and reveals that it is, in truth, our most accurate measure of courage.
"The audio makes all the difference."
The physics of vulnerability is simple: If we are brave enough often enough, we will fall. The author of the number-one New York Times best sellers Daring Greatly and The Gifts of Imperfection tells us what it takes to get back up and how owning our stories of disappointment, failure, and heartbreak gives us the power to write a daring new ending. Struggle, Brené Brown writes, can be our greatest call to courage and rising strong our clearest path to deeper meaning, wisdom, and hope.
"Learn to live your life with compassion, integrity and authenticity."
An accessible and practical toolkit that teams and companies in all industries can use to increase their customer base and market share, this book walks listeners through the process of creating and executing their own custom-made growth hacking strategy. It is a must listen for any marketer, entrepreneur, innovator or manager looking to replace wasteful big bets and "spaghetti-on-the-wall" approaches with more consistent, replicable, cost-effective, and data-driven results.
"A must read book if u wanna grow ur business"
On The Gifts of Imperfect Parenting, Dr. Brené Brown invites us on a journey to transform the lives of parents and children alike. Drawing on her 12 years of research on vulnerability, courage, worthiness, and shame, she presents ten guideposts to creating what she describes as "wholehearted" families where each of us can continually learn and grow as we reach our full potential.
"A very good refresher, but not her best book."
What does it take to be secure in our sense of belonging and self-worth? We may hustle to attain this security through achievements, meeting expectations, or repeating affirmations to ourselves - but Dr. Brené Brown's research has shown there is ultimately one obstacle to our sense of worthiness. “Shame is the barrier,” she teaches, “and building shame resilience is how we overcome it.”
"Incredibly helpful book"
Ten-year-old Annemarie Johansen and her best friend Ellen Rosen often think of life before the war. It's now 1943, and their life in Copenhagen is filled with school, food shortages, and the Nazi soldiers marching through town. When the Jews of Denmark are "relocated," Ellen moves in with the Johansens and pretends to be one of the family. Soon Annemarie is asked to go on a dangerous mission to save Ellen's life.
"People of all ages will enjoy this book."
Don't just let influence just happen to you. Instead, take charge of your life by grasping the science behind how influence works and by strengthening your own skills at using it to your advantage. In this dynamic 12-lecture series, you'll discover how to tap into the hidden powers of influence - and use these powers to enhance your life in ways you never thought possible. Using clear and accessible language, Professor Brown teaches you how and why influence works.
The Art of War is the most famous and longest-lasting book on the practice of war. The book does not mince words: war is about winning; it is not a gentleman's game; it is about using every means at one's disposal to tip the odds in one's favor. Of the centuries, this book has been used by military figures, war departments, business people, educators, and politicians, to name a few. This is the original work without additional editorial comment.
"Good presentation for Audio"
It was the night before his wedding, 15 years ago, when the nightmare began for Jay Crawford - locked up for a crime he never committed. Now he's escaped prison and wants nothing more than to clear his name and protect his family. To get justice,he'll need the help of the three best friends who have always had his back - Wil, Kyle, and Allan. But a man on the run requires absolute trust...and Jay may just be setting himself up for the ultimate betrayal.
"Good Listen but inconsistent"
Study the science of achievement through the wisdom of the world's leading motivational speaker, Les Brown. In this collection of live keynote speeches, Les Brown's straight-from-the-heart approach will change your belief in what you can achieve. This popular collection of audio recordings includes Les Brown's most powerful speeches. You'll have unlimited access to his timeless insights and discover how to live the life of your dreams. You have greatness within you; it merely takes persistence to let it unfold.
"Not really a collection"
Bobby Brown has been one of the most compelling American artists of the past 30 years, a magnetic and talented figure who successfully crossed over many musical genres, including R&B and hip-hop as well as the mainstream. In the late 1980s, the former front man of New Edition had a wildly successful solo career - especially with the launch of Don't Be Cruel - garnering multiple hits on the Billboard top 10 list as well as several Grammy, American Music, and Soul Train awards.
Rosie Daniels leaves her husband, Norman, after 14 years in an abusive marriage. She is determined to lose herself in a place where he won't find her. She'll worry about all the rest later. Alone in a strange city, she begins to make a new life, and good things finally start to happen. Meeting Bill is one, and getting an apartment is another. Still, it's hard for Rosie not to keep looking over her shoulder, and with good reason. Norman is a cop, with the instincts of a predator.
"Completely brought me in"
All of us have what it takes to make it in today's competitive and changing world. You may have doubted your ability to get the job done in the past, but something else may have been getting in the way of your success. The key to ultimate achievement has very little to do with your education or skill level. The secret to accomplishing great things is simple: Find and pursue the kind of work you are meant to do - your "purpose."
"Have you ever given a book a standing ovation?"
This audiobook uses a flash card system to teach new and renewing drivers what they need to know in order to pass the CDL as well as be better drivers. Listeners can use this to study while driving or at home. The book was suggested by a long-distance driver, Nathan Huckaby, who needed an audiobook to help him refresh his memory for his latest renewal test.
This second volume in William Manchester's three-volume biography of Winston Churchill challenges the assumption that Churchill's finest hour was as a wartime leader. During the years 1932-1940, he was tested as few men are. Pursued by creditors (at one point he had to put up his home for sale), he remained solvent only by writing an extraordinary number of books and magazine articles.
"Senseless change of narrators"
This audiobook was created based on Michael Porter's landmark book Competitive Strategy. This was Mr. Porter's synopsis of his book for the Harvard Business Review.
"Tough to listen to this one"
Eleven-year-old Reuben spends his days exploring, hiding, and practicing parkour among the abandoned buildings of the Lower Downs as a way to escape the rough times that have befallen him and his mom - but his discovery of an extraordinary antique pocket watch changes everything. When Reuben finds that the watch has the power to turn him invisible, he's propelled on the adventure of a lifetime. Now Reuben is being pursued by a group of dangerous men called the Directions, and someone - or something - ominously called The Smoke.
"Great book and amazing story"
When most of us think about artificial intelligence, our minds go straight to cyborgs, robots, and sci-fi thrillers where machines take over the world. But the truth is that artificial intelligence is already among us. It exists in our smartphones, fitness trackers, and refrigerators that tell us when the milk will expire. In some ways the future people dreamed of at the World's Fair in the 1960s is already here. We're teaching our machines how to think like humans, and they're learning at an incredible rate.
"Shallow, disjoint, disappointing"
The book you can't afford NOT to listen to. It is already affecting your life... And it WILL affect your future! Is it possible that there exists a three-thousand-year-old mystery that... Has been determining the course of your life without your knowing it? Foretells current events before they happen? Revealed the dates and the hours of the greatest crashes in Wall Street history before they happened? And much more....
"Why is this narrated by Optimus Prime??"
Graciousness is practicing the arts of kindness, thoughtfulness, good manners, humanity, and basic decency. As a bonus, it makes your life and even the world (or at least the room you're in) much lovelier. It's not about memorizing every rule of traditional etiquette to a T (though there is so much there to explore!) or spending all your time maintaining a spotless house. It's about approaching the world with compassion, conviction, self-confidence, and wholeheartedness, whether you're at the Met Gala or saying good morning to a convenience store clerk.


text:
Reviews of 'Ohana Dinner
Family Style brought to your table
AllEars Review: http://land.allears.net/blogs/jackspence/2009/10/ohana_1.html
Reviewer: SCOTTMOP Review Date: 08/27/2018
Rating: (10) Would Recommend:
I think this is the best dinner value at Disney. The amount of food you get. You get wings, noodles, veggies, etc. Start with delicious bread. The meat is yummy. Chicken, shrimp and steak. I prefer the shrimp and steak.if you don't like how something is cooked just tell your server and they will make sure you get what you want. We got some steak that was a little rare and told our server who then brought steak that was cooked right. don't be afraid to ask. THe bread pudding for dessert is to die for. They also make some of the best alcoholic and non alcoholic drinks here. so good we ate here twice on our trip and both times were great
Pros:delicious food and atmosphere
Reviewer: Disneydad7506 Review Date: 08/01/2018
Rating: (6) Would Recommend:
This was our sixth time having dinner at Ohana and as always the food was excellent. It was a Thursday evening and we only had to wait 10 minutes for our table. The service that night was a big disappointment. Our waiters were rushing us through the meal and were not very attentive. The appetizers and salad came out together and within a few minutes the grilled meats very being put on our plates. We asked the server politely to wait on the meats, but he just put them on our plates anyway. This all happened before they even took our drink order. When we finally got our drinks we were being told our dessert was on the way! I did inform one of the managers that we were disappointed in the service that evening and her response was that they were extremely busy and she was even having to clean the tables herself! Hopefully this was just a off night for the staff at Ohana.
Cons:Rushed, poor service
Reviewer: Dsnygrl Review Date: 07/06/2018
Rating: (8) Would Recommend:
Excellent service and entertainment while we ate dinner. Servers did rush a little bit as all the food courses were brought to the table without allowing time to enjoy each course. The Pot Stickers are the best I have ever had and the wings are great. The salad dressing is light and slightly sweet without being over powering. Three type of meat are served, chicken, top sirloin and peel and eat shrimp in shell, all grilled over an open fire pit. The chicken was tasty and not too dry. The sirloin was hit or miss depending on its placement on the skewer. the shrimp were my favorite. The highlight of dinner was the banana foster bread pudding. Make sure you save lots of room for this. Our server was very attentive and brought out seconds and then thirds of the dessert as the portion they bring out is really better suited for a single serving and we were a party of three. maybe they figure by the time we got to dessert we would be full? I highly recommend this restaurant for dinner!
Pros:Location and service
Reviewer: KenHutch Review Date: 06/25/2018
Rating: (10) Would Recommend:
How much does my family like this place? We eat dinner there twice every trip and 3 out of 5 trips we have had breakfast as well. This time the service was great and so was the food. I was surprised that the grilled chicken was the big winner in my family this time. The 1st time our waiter was great, he brought out 2 trays of desert and when one of my daughter maternal great aunts grabbed one and said MINE in a loud voice he just turned around and brought a 3rd tray without being asked.
Pros:Food and great atmosphere
Reviewer: TiggerTrigger Review Date: 03/15/2018
Rating: (5) Would Recommend:
We have been to 'Ohana for dinner multiple times. We go in expecting a long wait to be seated, but a fun and relaxing dining experience with great food to follow. The food this time was still good...well, except for a change in the dumplings (not sure if they changed a recipe or vendor, but the were not what they used to be)...but everything else with the meal was bizarre.
We had an 8:20pm ADR. We checked in at 8:05pm and were seated at 8:10pm. That has NEVER happened. We have always waited 30-40 minutes to be seated for dinner at 'Ohana. So, we knew something had to be up.
Our server was very friendly, but I found it odd that our appetizers were being placed on the table right as he was taking our drink orders. Just a couple of minutes later, he came by with the meat and was forcing it upon us. We asked him to wait, but in a sort of fun be part of the party 'Ohana style way...he insisted. Not only did he give us meat at that point, he loaded it onto our plates on top of the appetizers we literally just served ourselves. To give you some perspective, he unloaded a completely full skewer of shrimp onto my 11 year old's plate. Every time he came by, he loaded more food on top of food.
The meal was so rushed and poorly paced that we were being moved onto dessert about 30 min after being seated. They were also finally getting me my lapu lapu that I'd ordered at the beginning of the meal at this time. The drink was also poorly mixed.
By the time we were finishing up our bread pudding and paying out the check, we'd been there for about 45 minutes. We lingered a little longer so I could try and have a little more of my drink, but we were done at some point between the 50-55 minute mark.
Our server was kind enough to give us the passholder discount even if we're not passholders, so that was a plus. Still, we all kind of stood there looking at each other like "what just happened?" I just can't believe how rushed it was...and how that kind of pacing just destroys the experience.
I think we'd still go back to 'Ohana at some point, but I really hope Disney quits this nonsense and goes back to pushing for more normal meal pacing.
Pros:The Majority of the Food
Reviewer: ginny3 Review Date: 02/06/2018
Rating: (10) Would Recommend:
For 3 months I attempted to snag an ADR at Ohana with no luck.
When we checked in to our resort I asked Concierge to check for me for any day of our stay and he was able to get one for us albeit a bit late in the evening. Early in the morning of our ADR I had them check again and we were able to move the time to a reasonable hour.
Awesome food and atmosphere. We were very lucky to be seated at a window table that had a fantastic view of MK fireworks.
Pros:Fireworks viewing a plus
Reviewer: jeconk Review Date: 01/08/2018
Rating: (9) Would Recommend:
After hearing many others rave about this restaurant, we decided to give it a try for dinner. We choose dinner because we are not really into character meals and don't have any young children.
Not being a big fan of Polynesian food, I was somewhat nervous about eating here but boy was I wrong! The food is wonderful and very good even for a picky eater like me.
We were fortunate to get a table by the window facing the pool and Magic Kingdom just as the sun was going down so we got to see the lights coming on at the park. After we were seated, our waitress, Houda, came over. She explained how dinner works at Ohana and took our drink orders. She was very personable and we enjoyed her service.
Everything (except the meat) is served family style with serving plates placed on the table so each person could take as much or as little of each item as they liked. Houda told us that if we wanted more of any one item to let her know. The first course was a tossed salad with "Lilikoi" dressing. The dressing tasted pretty much like italian. Typical salad....don't fill up on this course because the best is yet to come...
Next we were brought a plate that had chicken wings, terriyaki noodles, pork dumplings and stir fry vegetables. There was more than enough of each item on this plate for the four of us!! (in fact, we couldn't eat it all.). Then came the main course(s). Houda came first with two skewers of chicken and put some on each persons plate....right off the skewer! The chicken was very moist, had a good flavor and was tender. Next, our waitresses came with two skewers of steak and did the same thing. The steak had an ok flavor (chicken was more flavorful) but was tender. Finally, she arrived with two skewers of grilled shrimp. Since I don't like shrimp, I skipped this course (everyone else just gave me some of their steak or chicken....although the waitress said she could bring more of any meat we wanted.) Those who ate the shrimp said it too was flavorful and tasty. There was more than enough meat for the four of us (and of course we were asked if we wanted more...)
Finally came the dessert....make sure you save room for this....Ohana Bread Pudding. I am not a fan of bread pudding but this is possibly the best bread pudding that I have ever had. Houda poured the warm sauce over it right at our table when she served it. (it was served family style like everything else). This was a wonderful ending to the meal (and the part we almost asked for more of....lol.).
While this is not a character breakfast, there was entertainment during the meal. A Hawaiian gentleman sang traditional songs and even taught a class in the hula for anyone brave enough to go up. Entertainment was a nice addition and not over the top.
The cost of this meal can be high for a large family but after all it is Disney World! This restaurant is definately one of the must do meals for those visiting Disney and is worth the cost.
Pros:Great food served family style so each person can pick and choose and eat as much or as little as they want. Dessert is out of this world!
Cons:Must make reservations far in advance.
Reviewer: Luvtheparks Review Date: 11/13/2017
Rating: (3) Would Recommend:
Was looking forward to a much hyped dinner here. Started with Cousin somebody (so loud in restaurantt didn't catch her name) telling us to take our time, she's there all night - that is, as long as you don't mind being rushed through dinner. Salad arrived then family style apps (pretty good) then they brought the chicken (thighs on a skewer) slimy and my least favorite type of chicken. Then came the peel and eat rubber shrimp followed by the VERY rare chewy skewered beef. Honestly, the dessert was the best part of the meal (which we ate with no coffee or addition beverages offered) Not sure why this place has such a great reputation and a loyal following. Perhaps just an off night? Definitely not worth the price - this was a one and done.
Pros:Trying to think of any
Cons:Wait time, food, waitstaff
Reviewer: MichelleR Review Date: 10/07/2017
Rating: (9) Would Recommend:
We love this restaurant! We had stumbled upon this one trip and couldn’t wait to go back and take our family. We were thrilled when we went back for our vacation with our extended family and children and everyone loved it. It is a new restaurant to add to our list of favorites. Food delicious, great quality, friendly staff and atmosphere. Drinks were all on point. The only thing that we had a slight issue with is we asked for some additional noodles and dumplings and this took a very very long time. Service this night was a little slow (our kids wanted to get back into the park and were getting antsy) but other than that we loved it and can’t wait to go back!
Pros:Delicious tasting food, friendly staff, portion size
Reviewer: Fitzboyzmom Review Date: 08/31/2017
Rating: (9) Would Recommend:
Dinner was great! Lots and lots of good food. I would say that they should change out the shrimp with something else. It was not that great and very hard to eat. My son loves shrimp and the server gave him lots, which was nice, but he ate one...I did as well. Other than that, we really enjoyed the dinner and would definitely come back.

domain: food