Background
==========

Elucidating the process of adaptation and understanding its genetic basis is a major mission of modern evolutionary biology \[[@B1],[@B2]\]. Although there are a few genome-level studies of model organisms \[[@B3],[@B4]\], most previous studies have focused on a single or a few candidate genes, which limited our understanding of the molecular basis of adaptation in many non-model systems. Recent advances in DNA sequencing technology and bioinformatic tools enable us to access and analyse massive sequence data in a short time at an affordable cost, which allows large-scale comparisons at genome, exome, or transcriptome levels \[[@B5],[@B6]\]. These advances present new opportunities for examining the genetic basis of many adaptive processes.

Understanding how organisms adapt to high-elevation environments can provide important insights into the process of adaptive evolution, particularly interactions and trade-offs between genes and pathways involved in simultaneous adaptive responses to multiple environmental challenges \[[@B7]-[@B10]\]. High-elevation environments impose severe physiological challenges to organisms, particularly those related to declined levels of oxygen, low temperature, and elevated levels of ultraviolet (UV) radiation \[[@B11],[@B12]\]. Physiological variation and adaptation to high-elevation habitats are well documented in many vertebrates \[[@B13]-[@B15]\]; however, study of their genetic basis is still in its infancy. Much of the previous work has focused on a few candidate loci, such as hemoglobin related genes and mitochondrial genes \[[@B16]-[@B18]\]. Only recently, analyses of genome-wide variations among species and populations living in different altitudinal environments have begun to shed light into the genetic basis of many adaptive processes. For example, genes related to hypoxia response and energy metabolism were shown to be responsible for physiological adaptations in several high-elevation species \[[@B9],[@B10],[@B19]-[@B21]\]. All these genome-wide studies investigated endothermic species. Ectothermic species have large physiological differences and may have evolved different mechanisms for adaption to high-elevation environments \[[@B12]\]. For example, instead of maintaining high levels of metabolism for temperature stability, poikilothermic species may lower their metabolic rate to cope with low external temperatures \[[@B12]\]. Understanding the genetic basis of adaptation to high elevations in poikilothermic species will not only complement our knowledge about endothermic species, but may also reveal novel mechanisms that are applicable to species that represent a much larger portion of the tree of life.

A pair of anuran species, *Rana chensinensis* and *R. kukunoris*, provide an excellent model system to study how poikilothermic species adapt to high-altitude habitats. *Rana chensinensis* is a widespread species and primarily occurs at low elevations of 600--1300 m above sea level (a.s.l.). *Rana kukunoris* is a high-elevation species that is only found at 2800--4000 m a.s.l. \[[@B22]\]. Previous studies suggest that the two species descended from a *R. chensinensis*-like common ancestor that inhabited low elevations, and speciation of *R. kukunoris* coincided with the recent uplift of the Tibetan Plateau approximately 7.8 million years ago \[[@B23]\]. Both species are diploid with 24 pairs of chromosomes \[[@B24]\] and *R. chensiensis* has a diploid genome size of 10.05 picograms (approximately 9,800 mega base pairs) \[[@B25]\].

The objective of this study is to make a genome wide search for genes that may be involved in adaptation to high-elevation environments in these amphibians, and identify their associated functions. We sampled multiple tissues of *R. chensinensis* and *R. kukunoris* and sequenced their transcriptomes using an Illumina sequencing platform. By comparing their transcriptomes, we describe genes showing strong signals of positive selection. The functional and phenotypic outcomes of these candidate genes are further inferred by annotating to genomic resources of a variety of model vertebrate species. We report a list of candidate genes that are highly likely to be involved in high-elevation adaptation processes.

Results and discussion
======================

Illumina sequencing, *de novo* assembly, and gene annotation
------------------------------------------------------------

A total of 67,676,712 sequence reads of *R. chensinensis*, and 66,476,534 sequence reads of *R. kukunoris*, were generated by Illumina sequencing. We first filtered these reads, and removed 527,878 and 711,299 defective reads for each species, respectively.

*De novo* assembly of cleaned reads was performed using various combinations of K-mer lengths and coverage cut-off values \[[@B26],[@B27]\]. In total, 30 raw assemblies were obtained for each species and were further merged by integrating sequence overlaps and eliminating redundancies for each species. The total length of the final set of assemblies was 38.1 mega base pairs (Mb) for *R. chensinensis* and 37.6 Mb for *R. kukunoris*. For *R. chensinensis*, 41,858 transcripts were obtained with an N50 of 1,333 base pairs (bp) and a mean length of 909 bp, and for *R. kukunoris*, 39,293 transcripts were obtained with an N50 of 1,485 bp and a mean length of 956 bp. All information of data production is summarized in Table [1](#T1){ref-type="table"}, and the length distribution of all transcripts is shown in Figure [1](#F1){ref-type="fig"}.

######

**Summary of transcriptome data for*Rana chensinensis*and*R. kukunoris***

                                       ***R. chensinensis***   ***R. kukunoris***
  ------------------------------------ ----------------------- -------------------- ----------- --------
  Total number of reads                67,676,712              66,476,534
  Read length (bp)                     90                      90
  Total length of reads (bp)           6,090,904,080           5,982,888,060
  Total length of assembly (Mb)        742.5                   38.1                 751.4       37.6
  Total sequences of assembly          1,624,822               41,858               1,521,607   39,293
  N50 length of assembly (bp)          515                     1,333                586         1,485
  Mean length of assembly(bp)          456                     909                  493         956
  Median length of assembly(bp)        325                     599                  340         601
  Total number of transcripts          41,858                  39,293
  Transcript annotated                 16,738                  16,549
  Number of unique genes represented   11,179                  11,140
  Number of putative orthologs         7069

bp=base pair; Mb=mega base pairs.

![**Length distribution of transcripts in base pairs.** The numbers of transcripts are shown on top of each bar.](1471-2164-13-588-1){#F1}

We constructed a reference dataset using combined protein data from seven species that represented all major lineages of vertebrates for gene annotation. Transcripts that did not match the reference dataset were blasted against a non-redundant (NR) protein database of the GenBank. In total, 16,738 transcripts of *R. chensinensis* were annotated to 11,179 unique genes, accounting for 40.0 % of the total transcripts. Some transcripts were likely products of alternative splicing. The E-value distribution showed that 55.3% of the annotated transcripts had an E-value below 1E-50, and the similarity distribution showed that 59.2% of the annotated transcripts had a similarity greater than 60%. Similarly, 16,549 transcripts of *R. kukunoris* were annotated to 11,140 unique genes, accounting for 42.1% of the total transcripts. Furthermore, 56.6% of the annotated transcripts had an E-value below 1E-50, and 59.7% of the annotated transcripts had a similarity greater than 60% (Table [1](#T1){ref-type="table"} and Additional file [1](#S1){ref-type="supplementary-material"}).

In agreement with previous studies, we found that the quality of sequence reads and appropriate assembly methods are essential in obtaining a reliable *de novo* assembly, which is the foundation of all other analyses \[[@B26],[@B28],[@B29]\]. In particular, *de novo* assembly by exploring a wide range of K-mer and cut-off values is a powerful approach. Due to the differential gene expression, sequence coverage among transcripts tends to be uneven, which poses various challenges for *de novo* assembly \[[@B26],[@B27]\]. The use of high K-mer lengths can generate more contiguous assemblies of highly expressed transcripts. On the other hand, the use of low K-mer lengths likely helps to obtain better assemblies for under expressed genes \[[@B26]\]. Adjusting coverage cut-off value can also help to increase the assembly quality when there are similar regions among homologous transcripts \[[@B27]\]. We used five different K-mer lengths and six different coverage cut-off values. In both species, the final assembly had larger N50 and mean lengths than any of the raw assemblies generated by using a single value of both parameters (Table [1](#T1){ref-type="table"}). The final assembly also had a higher proportion of annotated transcripts (\~40%) than all raw assemblies. Moreover, a final step that maps all reads back to the assembled transcripts can also help to improve the quality of assembly. This process allows for a revision of any potential assembly errors and ascertains the consensus sequences representing the majority base calls at heterozygous sites.

Gene ontology (GO) classification
---------------------------------

Gene Ontology category was widely used to classify gene functions \[[@B30]\]. We first removed 5,559 transcripts for *R. chensinensis* and 5,409 transcripts for *R. kukunoris* that were likely generated from alternative splicing. Among the remaining 11,179 transcripts of *R. chensinensis,* which likely represented the same number of unique genes, a total of 8,781 transcripts had the GO annotation under three main divisions. Likewise, among the 11,140 annotated transcripts of *R. kukunoris*, 8,806 transcripts had the GO annotation. The GO category distributions of the transcripts for both species were highly similar ( Additional file [2](#S2){ref-type="supplementary-material"}). Within the function of "Cellular Component", 13 level-2 categories were identified, and the terms "cell", "cell part", and "organelle" were the dominant components (\>50%). Within the function of "Molecular Functions", 15 level-2 categories were identified, and the term "binding" was dominant (\>50%). Within the third function of "Biological Process", 23 level-2 categories were identified, and the terms "cellular process", "metabolic process", "biological regulation", and "pigmentation" were the main functional categories (\>50%).

Identification of putative orthologs
------------------------------------

Reciprocal best hit method was used to identify putative orthologs between the two species \[[@B31]\]. A total of 10,132 pairs of putative orthologs were identified by comparing all annotated transcripts. We then compared each pair of putative orthologs to the reference to determine their open reading frames (ORF) and eliminated the sequences with unexpected stop-codons and ambiguous alignments. After filtering the redundant orthologs that may result from alternative splicing, 7,069 pairs of unique orthologs were finally identified and used in the downstream analyses.

We also identified 8,393 pairs of possible orthologs from transcripts that could not be annotated. We were able to use EBOSS-GETORF \[[@B32]\] to predict the ORFs of 3,880 and 3,851 transcripts in *R. chensinensis* and *R. kukunoris*, respectively. Among these transcripts, 1,701 pairs were properly aligned, which were also used in the test for positive selection.

Test for positive selection
---------------------------

The Ka/Ks ratio was used to test for positive selection \[[@B33]\]. Among the 7,069 pairs of annotated orthologs, 724 pairs were identical, 2,827 pairs had only synonymous or non-synonymous substitutions, and 3,518 pairs had both types of substitutions, for which the Ka/Ks ratio were calculated. The mean values of Ka, Ks, and Ka/Ks ratio of all orthologous pairs were 0.0054, 0.0372, and 0.1452, respectively.

A total of 125 (1.8%) pairs of orthologs had a Ka/Ks ratio greater than 1, indicating these genes may have experienced strong positive selection (Figure [2](#F2){ref-type="fig"}). In addition, a Ka/Ks ratio of 0.5 was also suggested as a useful cut-off value to identify genes under positive selection \[[@B34]\], and in our study, 335 (4.7%) pairs of orthologs had a Ka/Ks ratio between 0.5 and 1 (Figure [2](#F2){ref-type="fig"}). Therefore, all these 460 orthologs (Ka/Ks\>0.5) were considered as candidate genes that have likely experienced positive selection.

![**Distribution of Ka and Ks values.** Orthologous pairs with Ka/Ks ratio \>1 are above the black line; pairs with Ka/Ks ratio between 0.5-1 are between the black and grey lines.](1471-2164-13-588-2){#F2}

Among the 1,701 pairs of predicted coding sequences from un-annotated transcripts, 25 (1.5%) pairs had a Ka/Ks ratio greater than 1 and 205 (12.0%) pairs had a Ka/Ks ratio between 0.5 and 1. They were supplementary to the annotated candidate gene list for future studies.

We clustered all the unique orthologous transcripts to GO categories. Among the 7,069 annotated unique orthologs, 6,811 were assigned to 8,335 GO terms, among which 1,143 terms had at least 10 orthologs. The mean Ka/Ks ratio for each of the 1,143 terms was calculated ( Additional file [3](#S3){ref-type="supplementary-material"}), and 330 (28.9%) terms had a mean Ka/Ks ratio higher than the mean ratio of all orthologs (0.1452). The terms with the highest Ka/Ks ratio included "adult heart development", "sarcomere organization" and "condensed nuclear chromosome". Likewise, the terms with the smallest Ka/Ks ratio included "protein K63-linked ubiquitination", "protein K48-linked ubiquitination" and "ubiquitin-dependent protein catabolic process". The terms with low Ka/Ks ratio probably represent the most conservative functions, and the terms with high Ka/Ks ratio likely represent the rapidly evolving functions between the two species \[[@B10],[@B35]\].

Functional analysis for candidate genes
---------------------------------------

Among the 460 candidate genes (Ka/Ks \>0.5), 315 were annotated to GO categories (68.5%). Using the program WEGO \[[@B36]\], we compared the GO category distribution (level 3) of the candidate genes with that of the non-candidate genes (Ka/Ks \< 0.5) (Figure [3](#F3){ref-type="fig"}). Seven GO terms had a higher percentage among the candidate genes than among the non-candidate genes. Several of these terms were associated with immunity and host defense, such as "immune response" (4.4% among candidate genes vs. 4.2% among non-candidate genes), "leukocyte activation" (3.8% vs. 2.4%), and "lymphocyte costimulation" (4.8% vs. 3.8%). Immune related genes were commonly found under positive selection during speciation processes and in species that do not live at high elevations \[[@B35],[@B37]\]. Therefore, although under positive selection, these genes may not be related to adaptation to a high-elevation environment.

![**Comparison of Gene Ontology (GO) category (level 3) distribution between candidate genes and non-candidate gene.** The red and blue bars represent candidate genes (Ka/Ks ratio \>0.5) and non-candidate genes (Ka/Ks ratio \<0.5) in each GO functional category, respectively. Candidate genes have higher percentage then non-candidate gene in seven categories, which are marked with asterisks. The sum of individual percentages exceeds 100% because most genes are classified in multiple categories.](1471-2164-13-588-3){#F3}

To identify genes that may closely associate to the adaptation to high elevations, we performed a two-step approach to screen all the candidate genes according to their functions. First, we compared our candidate genes to a list proposed by Simonson *et al*. \[[@B20]\], which encompasses the functions and their associated GO categories for adaptation to high-elevation hypoxia. Two genes associated with "response to hypoxia" and two genes associated with "oxygen binding" were identified from our candidate gene list (Table [2](#T2){ref-type="table"}). Second, based on our current understanding on phenotypic adaptation to high elevations of poikilothermic species, we selected three other candidate functions, which are associated with response to ultraviolet rays \[[@B11]\], reactive oxygen species metabolism, and metabolic regulation \[[@B12]\]. Ten additional genes were identified with these candidate functions, which were in the GO categories "response to UV", "glutathione metabolic process", and "ATP catabolic process" (Table [2](#T2){ref-type="table"}).

######

List of fourteen candidate genes with related Gene Ontology (GO) categories

  **Sequence ID**   **Protein homolog**                               **Ka/Ks ratio**   **GO category**                                  **GO accession**
  ----------------- ------------------------------------------------- ----------------- ------------------------------------------------ ------------------
  Kuku_seq_14688    Glutathione s-transferase omega 2, GSTO2          1.7284            glutathione metabolic process                    GO:0006749
  Kuku_seq_38792    Sterol carrier protein 2, SCP2                    1.4657            peroxisome organization                          GO:0007031
  Kuku_seq_19028    Ubiquitin-specific protease 1, USP-1              1.2268            response to UV                                   GO:0009411
  Kuku_seq_38041    Ribosomal L1 domain containing 1,RSL1D1           1.1671            primary metabolic process                        GO:0044238
  Kuku_seq_19815    Peroxiredoxin-4, PRDX4                            1.1484            response to oxidative stress                     GO:0006979
  Kuku_seq_9362     Mitochondrial translation optimization 1, MTO-1   1.1363            mitochondrial tRNA wobble uridine modification   GO:0070899
  Kuku_seq_22135    NADPH oxidase organizer 1, NOXO1                  1.0432            NADPH oxidase complex                            GO:0043020
  Kuku_seq_29724    Aminoacylase-1, ACY1                              1.0342            protein catabolic process                        GO:0030163
  Kuku_seq_10530    DNA repair protein xrcc, XRCC                     0.6313            response to hypoxia                              GO:0001666
  Kuku_seq_19044    4-aminobutyrate aminotransferase, ABAT            0.5899            response to hypoxia                              GO:0001666
  Kuku_seq_19251    Cytochrome p450 4b1-like, CYP4B1                  0.5867            oxygen binding                                   GO:0019825
  Kuku_seq_4377     Glutathione peroxidase 4, GPX4                    0.5799            glutathione metabolic process                    GO:0006749
  Kuku_seq_29173    V-type proton ATPase subunit G 1, V-ATPase G1     0.5173            ATP catabolic process                            GO:0006200
  Kuku_seq_8042     Cytochrome p450 2f1-like, CYP2F1                  0.5062            oxygen binding                                   GO:0019825

These fourteen genes have Ka/Ks ratio greater than 0.5 and are most likely involved in the adaptation to high-elevation environments.

The two candidate genes associated with "oxygen binding", CYP2F1 and CYP4B1, belong to the cytochrome P450 superfamily (CYP). The function of CYP is to catalyze the oxidation of organic substances \[[@B38]\]. The mean Ka/Ks ratio of the term "oxygen binding" is 0.2435, much higher than the mean Ka/Ks ratio of all orthologs (0.1452), suggesting a strong overall positive selection on genes in this category. Previous studies of Tibetan people also found that several other CYP genes showed signatures of positive selection related to "oxygen binding" \[[@B20]\]. Our finding suggests a functional (phenotypic) convergent evolution between humans and *Rana kukunoris*, although the genes behind the functional adaptation are different (but belong to the same gene family).

The gene Ubiquitin-specific protease 1 (USP-1) is also under strong positive selection (Ka/Ks=1.2268), and we found a non-synonymous substitution in its His domain of N-terminal, which is likely crucial for its function \[[@B39]\]. USP-1 is a conserved gene that encode an enzyme playing an important role in DNA repair against UV damage \[[@B39],[@B40]\]. Thus, USP-1 may be especially important for amphibians to adapt to the high UV radiation environment, since their skin is completely exposed without any protection from fur or scales.

Four genes, GPX4, GSTO2, PRDX4, SCP2, which are associated with response to reactive oxygen species (ROS), have also experienced positive selection (Table [2](#T2){ref-type="table"}). The first two genes encode crucial enzymes in glutathione metabolic pathway, which prevent damage to important cellular components by ROS-like peroxides and free radicals \[[@B41]\]. The functions of the latter two genes are related to the functions of peroxisome that are also associated with response to oxidative stress \[[@B42],[@B43]\]. Bickler and Buck \[[@B12]\] proposed that the ability of poikilothermic animals to survive under low temperature and oxygen conditions depends on the ability of avoiding free-radical injury. The four genes identified here are potentially related to the adaptive process in this aspect.

We also identified five metabolism related genes that may be associated with adaptation to high elevations. The MTO-1 and V-ATPase G1 genes are related to ATP synthesis and energy metabolism in the mitochondrion \[[@B44],[@B45]\]; both functions are important in the process of high-elevation adaptation \[[@B12]\]. The ACY1, NOXO1 and RSL1D1genes are related to primary, NADPH and protein metabolism, respectively \[[@B46]-[@B48]\]. Functional changes associated with metabolic processes are likely essential for the adaptation to high-elevation environments.

A surprising absence from our candidate gene list is the hemoglobin related genes. Responses to hypoxia in endothermic animal species involve mostly hemoglobin genes and HIF-1 pathways, and we did not observe any of those genes bearing signature of positive selection. The mean Ka/Ks ratio (0.1154) of the entire term "response to hypoxia" was lower than average level (0.1452), although two genes (ABAT and XRCC) had Ka/Ks ratio greater than 0.5 but less than 1 (Table [2](#T2){ref-type="table"}). Some other genes involved in adaptation to high elevations in endothermic species also had a low Ka/Ks ratio in our comparison, for example, EPAS1 \[[@B20],[@B21]\] (Ka/Ks=0.1057), ARG2 \[[@B10]\] (Ka/Ks=0.1095), and EP300 \[[@B19]\] (Ka/Ks=0.1725). The gene EGLN1 \[[@B19]\] is an exception in this sense; it has a relatively high Ka/Ks ratio of 0.4563, which may deserve further examination. This observation is consistent with our prediction that poikilothermic species may develop different adaptive mechanisms from endotherms.

Future directions
-----------------

Our study demonstrated that comparing sequences of orthologous genes between species by *de novo* transcriptome sequencing offers a fast and cost-effective approach to detect positive selection and to understand adaptive evolution at the genomic level. Further tests are needed to validate these candidate genes that we have identified. Population-level analysis can be applied to these candidate genes to examine if signals of positive selection on these genes are also evident at the population level. Detailed functional research is also needed to illustrate how the positively selected genes promote the fitness of poikilothermic species under the high-elevation environment. Finally, to understand the general genetic mechanisms of adaptation to high elevations of poikilothermic animals, it is necessary to examine other poikilothermic species from various groups. For example, hemoglobin genes, which were thought to play an important role in adaptation to high elevations in endothermic species, were absent from our candidate gene list and the importance of their role remained largely unknown in other poikilothermic species \[[@B17]\]. Studies on a wide range of species are necessary to understand parallel and convergent adaptive evolution to high elevations between poikilotherms and endotherms.

Better and more elaborative tests for positive selection should be explored in order to improve their sensitivity. Ka/Ks ratio can only serve as a preliminary approximate estimator. Functional analysis with pre-defined "potential functions for adaptation", or candidate genes, largely limits the scope of discovery. Only with tests that are solely based on genetic data at a genome-wide scope, it is possible to establish a new 'genotype to phenotype' approach to the study of adaptation, which will complement the traditional 'phenotype to genotype' approach. With this new approach, we can take the great advantage of rapidly accumulating genomic data to reveal novel phenotypes of adaptation.

Conclusions
===========

By comparing the transcriptomes of two ranid species, one from high and one from low altitudes, we identified 125 protein-coding genes that have experienced strong positive selection (Ka/Ks\>1), and 335 genes that may also bear signatures of positive selection (1≥Ka/Ks\>0.5). Among them, 14 genes are most likely involved in the adaptive process to high-elevation environments, particularly genes associated with oxygen binding (CYP4B1and CYP2F1), response to UV (USP-1), and repair of free radical injuries (GSTO2, GPX4, PRDX4, and SCP2). Furthermore, we found evidence for convergent evolution between poikilothermic and endothermic species; genes from the CYP superfamily are under positive selection in both humans and amphibians. Yet, we also observed differences between them; hemoglobin related genes are commonly involved in high-elevation adaptation in endotherms, but are absent from our candidate gene list. The genes that display signatures of positive selection will serve as a baseline for further investigations that aim to understand high-elevation adaptation at both molecular and phenotypic levels.

Methods
=======

Sample collection
-----------------

To include as many expressed genes as possible, we sampled five different types of tissues from six individuals of each species, including liver, heart, skeletal muscle, testicle/ootheca from two males and two females, and the bodies of two tadpoles. Samples of *R. chensinensis* were collected in April, 2011, from the Yaodu Township, Qingchuan County, located on northern Sichuan Basin, China (105.45°E, 32.79°N) with an altitude of 604 m above sea level (a.s.l.). Samples of *R. kukunoris* were collected in April, 2011, from the Zoige County, situated on the eastern margin of the Tibetan Plateau, China (102.90°E, 33.58°N) with an altitude of 3,358 m a.s.l. Sample locations are depicted on a map in Additional file [4](#S4){ref-type="supplementary-material"}. Tissue samples for RNA extraction were stored in RNALater (QIAGEN) immediately following euthanasia and dissection.

cDNA library construction and Illumina sequencing
-------------------------------------------------

RNA extraction, cDNA library construction, and sequencing were performed by BGI (Shenzhen, China). The entire process followed a standardized procedure and was monitored by BGI's Quality Control System. For each species, total RNA was extracted using SV Total RNA Isolation System (Promega) from each tissue sample independently and then mixed with approximately the same quantity. RNAs with poly (A) tail were purified from total RNA by oligonucleotide (dT) magnetic beads and interrupted into short sequences. Subsequently, cDNAs were synthesized and purified with PCR extraction kit (QiaQuick). The cDNA library had an insert size of 200--700 base pair (bp), and paired-end of the library was sequenced with a read length of 90 bp using the Illumina HiSeq™ 2000 sequencing platform. Output of image data from the sequencer was transformed by base calling into raw sequence data, which formed the raw reads and then stored in the FastQ format.

Data filtration and *de novo* assembly
--------------------------------------

We first cleaned the raw sequence reads by removing the exact duplicates from both sequencing directions. We further trimmed reads by removing adapter sequences, reads with unknown bases call (N) more than 5%, low complexity, and low quality (\<Q20) using Blat \[[@B49]\]. We also removed reads likely derived from contaminants by comparing with the human and *Escherichia coli* genomes using Bowtie \[[@B50]\].

*De novo* assembly of cleaned reads was performed using a combination of various multiple K-mer lengths and coverage cut-off values \[[@B26],[@B27]\]. Using VELVET \[[@B51]\] and OASES \[[@B52]\], five different K-mer lengths (21, 31, 41, 51, and 61) and six coverage cut-off values (2, 3, 6, 10, 15, and 20) were used to produce a total of 30 raw assemblies for each transcriptome. We then used CD-HIT-EST \[[@B53]\] and CAP3 \[[@B54]\] to produce a final assembly by integrating sequence overlaps and eliminating redundancies. All reads were mapped back to the assembled contigs using SOAP-ALIGNER \[[@B55]\] and SNPs were identified using SOAP-SNP \[[@B56]\]. For all variable sites, the base call with the most mapped reads was selected as the consensus using an in-house Python script.

Gene annotation
---------------

Due to the lack of genomic resources from any *Rana* species, we combined protein data from seven species covering all major vertebrate groups to create a reference dataset for gene annotation, which includes *Anolis carolinensis*, *Danio rerio*, *Gallus gallus*, *Homo sapiens*, *Mus musculus*, *Oryzias latipes*, and *Xenopus tropicalis*\[[@B57]\]. All protein data were downloaded from the Ensembl database \[[@B58]\]. Transcripts were annotated to the reference dataset based on the BLAST similarity using blastx with an E-value cut-off of 1E-5. Transcripts that did not match to any gene in the reference dataset were searched against the non-redundant (NR) protein database of GenBank.

Gene ontology classification
----------------------------

Gene Ontology \[[@B30]\] categories were used to classify the functions of the transcripts. In order to exclude the interference from alternative splicing of transcripts, we first clustered the transcripts that matched to the same reference gene; then we removed the redundant transcripts and only reserved the longest transcript for each cluster to represent the unique gene. GO classification was performed following the Blast2GO pipelines \[[@B59]\], based on a standard cut-off E-value of 1E-3 and a maximum of 20 hits of the BLAST process, and the default parameters of mapping and annotation procedure.

Identification of putative orthologs
------------------------------------

A reciprocal best hit method was used to identify putative orthologs between the two species with tblastx of BLAST-2.2.26+ and a bit-score threshold of 300 \[[@B31]\]. Subsequently, sequences in each orthologous pair were compared to the combined protein reference to determine their open reading frames (ORF) by blastx. The coding region of both sequences was then extracted and aligned by MUSCLE \[[@B60]\]. Aligned sequences with unexpected stop-codons, ambiguous alignments, and/or shorter than 200 bp were discarded from further analysis. A manual check was also conducted to correct potential errors. All transcripts, including products of potential alternative splicing, were blasted to maximize the chance of finding matches between the two species. If any orthologs that matched to the same reference gene had overlapping annotated region, we removed the shorter sequences to ensure that all orthologous pairs represented unique genes. For contigs that could not be annotated, we also applied the same method to identify orthologous pairs, then used EMBOSS-GETORF to predict the ORF for each sequence \[[@B32]\]. We adopted a strict criterion that only reserved the predicted coding sequence longer than 300 bp of each sequence. The resulting sequence pairs were analysed following the same approach described above.

Test for positive selections
----------------------------

The ratio of the number of nonsynonymous substitutions per nonsynonymous site (Ka) to the number of synonymous substitutions per synonymous site (Ks) was used to test for positive selection. A Ka/Ks ratio less than one indicates purifying selection, while a Ka/Ks ratio greater than one provides evidence for positive selection \[[@B33]\]. For its simplicity and applicability, the ratio has been widely used in genome wide comparative studies, despite its limitations. Yang and Bielawski \[[@B33]\] provided a thorough evaluation of the application of Ka/Ks for testing selection. We employed the software KaKs_CALCULATOR \[[@B61]\] to estimate the Ka, Ks and Ka/Ks ratio of each orthologous pair, and the YN model \[[@B62]\] was used. The sequences with Ks \>0.1 were excluded to avoid potential paralogs \[[@B63]\]. Based on the GO classification, the unique orthologs were clustered into different functional terms and the mean Ka/Ks ratio for each term was also calculated.

Functional analysis for candidate genes
---------------------------------------

The orthologs that had Ka/Ks ratio above 0.5 were considered as candidate genes \[[@B34]\]. To detect which functions were more likely under positive selection, we compared the GO distribution (level 3) of the candidate genes with distribution of the non-candidate genes (Ka/Ks \< 0.5) by the program WEGO \[[@B36]\]. Furthermore, we performed a two-step approach to identify candidate genes that might have been involved in facilitating the adaptation process to high-elevation environments. First, we examined the list of a priori functional candidate genes and the associated GO categories made by Simonson *et al*. \[[@B20]\], which represents the functions for adaptation to high elevation hypoxia and was used to screen candidate genes identified from a Tibetan Chinese population \[[@B20]\]. Second, based on previous studies on poikilotherms, we also studied other GO categories and selected the terms potentially associated with response to ultraviolet rays (UV), reactive oxygen species metabolism and metabolic regulation, which may represent other functions that have likely facilitated the adaptation to high elevations \[[@B11],[@B12],[@B64]\].

Availability of supporting data
-------------------------------

The data supporting the results of this article are available in the NCBI Sequence Read Archive (SRA) repository \[Accession Number: SRA060325\].

Abbreviations
=============

ACY1: Aminoacylase-1; ARG2: Arginase, type II; bp: Base pair; CYP2F1: Cytochrome p450 2f1-like; CYP4B1: Cytochrome p450 4b1-like; CYP: Cytochrome P450 superfamily; EGLN1: Egl nine homolog 1; EP300: E1A binding protein p300; EPAS1: Endothelial PAS domain-containing protein 1; GO: Gene Ontology; GPX4: Glutathione peroxidase 4; GSTO2: Glutathione s-transferase omega 2; Mb: Mega base pairs; MTO-1: Mitochondrial translation optimization 1; NOXO1: NADPH oxidase organizer 1; ORF: Open reading frame; PRDX4: Peroxiredoxin-4; ROS: Reactive oxygen species; RSL1D1: Ribosomal L1 domain containing 1; SCP2: Sterol carrier protein 2; USP-1: Ubiquitin-specific protease 1; UV: Ultraviolet; V-ATPase G1: V-type proton ATPase subunit G 1; XRCC: DNA repair protein xrcc.

Competing interests
===================

The authors declare that they have no competing interests.

Authors' contributions
======================

WY carried out most of the data analyses and drafted the manuscript. YQ collected the specimens. KB participated in data analysis and writing. JF conceived the project and finalized the manuscript. All authors contributed to the project design and manuscript preparation. All authors have read and approved the final manuscript.

Authors' information
====================

WY is a Ph.D. candidate and this project is part of his dissertation work. He is mostly interested in bioinformatics and related genomic work. YQ is a herpetologist and mostly interested in high elevation herpeto-fauna. KB is a post-doctoral fellow who is interested in detecting genome-wide signatures of natural selection using museum historical specimens. JF is an evolutionary biologist particularly interested in evolutionary phenomena around the Tibet Plateau.

Supplementary Material
======================

###### Additional file 1

**Characteristics of gene annotation of assembled transcripts against the reference dataset.** A. E-value distribution of blastx hits for each transcript with a cut-off E-value of 1E-5. B. Similarity distribution of blastx hits for each transcripts.

######

Click here for file

###### Additional file 2

**Distribution of Gene Ontology (GO) categories (level 2) of transcripts for*Rana chensinensis*and*R. kukunoris.*** The GO functional annotations are summarized in three main categories: cellular component, molecular function and biological process. The red bars and blue bars represent *R. chensinensis* and *R. kukunoris*, respectively. (PDF 240 kb)

######

Click here for file

###### Additional file 3

**Average Ka/Ks ratio for each GO term.** Only terms that have more than 10 orthologs are presented.

######

Click here for file

###### Additional file 4

**Sampling sites and altitude distribution.** Samples of *Rana chensinensis* were collected from Yaodu Township (105.45°E, 32.79°N) with an altitude of 604 m above sea level. Samples of *R. kukunoris* were collected from Zoige County (102.90°E, 33.58°N) with an altitude of 3,358 m above sea level.

######

Click here for file

Acknowledgements
================

We would like to thank D. Hu for field assistance. J. P. Bogart, Q. Dai, B. Lu, T. F. Mitterboeck, and C. Rochus provided many valuable comments on an early version of this manuscript. This project is funded by a Chinese Academy of Sciences Knowledge Innovation Program grant (Y1C2021203) and an NSERC discovery grant (Canada) to JF.


Studies of the tumorigenic effect of two goitrogens.
The nature of tumors appearing in the thyroid gland and in the lungs of mice fed two standard goitrogenic drugs (MTU and MII) has been studied. These tumors have been considered malignant on the basis of their morphological appearance and their occurrence in abnormal locations. Some investigatiors, however, have questioned that they are actually malignant. The present results indicate that these tumors are most likely not malignant even if it is shown that the pulmonary nodules are of thyroid origin. The thyroid adenomas disappear once the goitrogen is withdrawn, but thryoid enlargement pesists, and event 6 months after discontinuation of the goitrogenic treatment , pulmonary nodules are still produced. Evidence is presented that these nodules are emoli from hyperplastic thyroid tissue and not tumors.


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[Oral medicine 2. Treatment of dry mouth].
Treatment of dry mouth starts with determining the salivary gland function by measuring the unstimulated and stimulated flow rate.Treatment depends on these measurements. Iffunctioning salivary gland tissues with saliva producing potential are present, stimulation of the salivary glands by gum chewing or sucking sugar-free sweets is recommended. Salivary gland stimulation may also be achieved using medications, acupuncture and electrostimulation. If stimulation is insufficient, moistening the oral mucosa with a sip ofwater regularly during the day is one of the easiest and most effective methods of easing dry mouth. In addition, the use of saliva replacement therapy might be beneficial, but only with proper instruction. With regard to the caries risk in dentate patients, a fluoride rinse or gel should be prescribed.


It was a deadly weekend around the world, with significant terrorist attacks in Bangladesh and Iraq, and close on the heels of the attack on Istanbul's airport.

And it's been less than nine months since ISIS-backed terrorists mounted a deadly attack in Paris — and little more than three months since the attack on Brussels airport.

But there have been more than 1,000 terror attacks since the November 13 attacks in Paris. Factor out the ongoing conflicts in Afghanistan, Iraq, Libya and Syria and you still have more than 100 attacks in that time.

Some, like the attack in San Bernardino, California, or at the Pulse nightclub in Orlando were certainly well-reported. An attack in Ivory Coast in March also was widely covered. As was a June attack in Mogadishu.

But other attacks have flown under the radar. Here are some of the most significant terrorist attacks in the past eight months. Attacks in the conflict zones in Iraq, Afghanistan, Syria and Libya are not included, despite being increasingly violent.

Bamako hotel attack: November 20, 2015

A group of al-Qaeda-linked militants took 170 people hostage, ultimately killing 20, during a mass shooting at a Radisson Blu hotel in Bamako, the capital of Mali in West Africa.

Malian commadoes ultimately raided the hotel and freed the hostages, killing the two assailants. Another seven people were wounded.

Tunis bus attack: November 24, 2015

A bus loaded with Tunisian presidential guards was struck by a suicide bomber linked to the so-called Islamic State.

The lone bomber killed 13 people in what was the third significant attack attributed to ISIS in Tunisia in 2015.

San Bernardino attack: December 2, 2015

Candles are seen at a pop-up memorial in San Bernardino, California, December 4, 2015, following Wednesday's attack. Credit: Mario Anzuoni/Reuters

In the attack, 14 people were killed and another 22 were seriously injured.

A manhunt ensued and the two perpetrators, Syed Rizwan Farook and Tashfeen Malik were killed in a shootout with police. The FBI said the two shooters were "inspired" by ISIS, but conducted the attacks on their own.

Bring new voices into public media.

Istanbul bombing: January 12, 2016

A suicide bombing in a popular central square in Istanbul, Turkey's largest city, left 13 people dead and 14 people injured. All of the dead were foreigners.

The perpetrator was identified as Nabil Fadli, an ISIS follower from Syria.

Jakarta attacks: January 14, 2016

A series of suicide bombings and shootings in Indonesia's capital left eight people dead, including four assailants. Another 24 people were wounded.

ISIS claimed responsibility for the attack, which was conducted by an unknown number of assailants. Though four assailants were killed, as many as 10 others may have been involved.

Splendid Hotel attack: January 15, 2016

French Prime Minister Manuel Valls stands in front of Cappuccino cafe, the site of an al-Qaeda attack that killed 30 in January in Ouagadougou, Burkina Faso, February 20, 2016. Credit: Nabila El Hadad/Reuters

Government forces mounted a raid and freed more than 170 hostages. Four assailants were killed and another two or three escaped.

Ankara bombing: February 17, 2016

Kurdish freedom fighters attacked a convoy of buses killing military personnel and civilians during evening rush hour. At least 29 people were killed and another 60 people were injured.

The attack, which was carried out by a car bomb, was targeted at military forces, the Kurdistan Freedom Falcons said. One perpetrator died in the attacks.

Mogadishu hotel attack: February 26, 2016

A group of militants linked to al-Shabbab killed at least 15 people and left others wounded after a suicide bomber detonated a bomb at the gate of the SYL hotel in Mogadishu.

Four gunmen forced their way past the first security barrier before being killed in a shootout with guards manning the hotel's last security line. More than a dozen people were wounded.

Grand-Bassam resort shootings: March 13, 2016

A police cordon is seen while Ivorian police prepare to inspect the area of the hotel Etoile du Sud following an attack by gunmen from al Qaeda's North African branch, in Grand Bassam, Ivory Coast, March 14, 2016. Credit: Luc Gnago/Reuters

The hotel is popular with ex-pats in Ivory Coast. At least three assailants involved in the attack were killed and another as many as three escaped.

Ankara bombing: March 13, 2016

A second attack in Ankara this year saw the deaths of 37 people and another 127 people injured. As with the first attack, the Kurdistan Freedom Falcons claimed responsibility for the assault.

The perpetrator was killed in the attack, which targeted a transportation hub. The Kurdish group said civilians weren't its main target, but more civilian casualties were inevitable.

Istanbul bombing: March 19, 2016

At least four people were killed by a suicide bomber that has rocked the residents of Istanbul to their core. Another 36 people were wounded by the attack on Istanbul's main shopping street.

About one-third of the injured were foreign tourists. Thus far, no one has claimed responsibility.

Yemen suicide bombing: May 15, 2016

An ISIS suicide bombing and attack killed 47 people in the Yemeni port of Mukalla. The attack targeted police cadets, killing at least 41 of them.

Another 60 people were wounded.

Assault on Ambassador Hotel: June 2, 2016

At least 16 people were killed and dozens more wounded when Al Shabab attacked the Ambassador Hotel in Mogadishu, Somalia. Armed men stormed the hotel before being repelled by armed guards and security agents.

A second assault in Mogadishu in June had a similar death toll.

Aktobe shootings: June 5-10, 2016

Twenty-five people died, 18 of them terrorists, in a series of shootings in Aktobe, Kazakhstan. Seven civilians were killed and another 40-plus were wounded.

The attack was blamed on Islamic radicals, some of whom were believed to have return from fighting for ISIS in Syria.

Pulse nightclub shooting: June 12, 2016

A lone gunman walked into a nightclub in Orlando popular with the LGBT community where he shot and killed 49 people and wounded another 53 before being killed himself by law enforcement.

The gunman, Omar Mateen, proclaimed his loyalty to ISIS.

Istanbul airport suicide bombing: June 28, 2016

A trio of suicide attacks at the heavily guarded Istanbul Ataturk International Airport left 45 people dead and more than 200 people wounded. All three attackers were also killed, either in the blasts or shot by law enforcement.

No one has claimed responsibility for the attack — but it has been blamed on ISIS.

Dhaka attack: July 1, 2016

At least 22 people, plus six attackers, were killed after a group linked to ISIS assaulted and took hostages in the diplomatic neighborhood of Gulshan, in Dhaka, Bangaldesh.

Among the dead were 20 civilians and two police officers. One gunman was captured and 13 hostages were rescued.


Perceptions of shared decision making among health care professionals.
Scholars have progressively promoted shared decision making (SDM) as an optimal model of treatment decision making in clinical practice. Nevertheless, it is unclear whether health care professionals (a) understand SDM well, (b) believe that SDM is helpful in their daily practice, and (c) are willing to practice SDM during their daily activities. These are crucial research topics; however, such research is still limited. The aim of this study was to apply the knowledge-attitude-behavior (KAB) model to probe health care professionals' perceptions of SDM. A questionnaire was delivered to health care professionals working in various hospitals in southern Taiwan from 9 November 2018 to 8 January 2019. In addition to KAB constructs, this study explored the barriers to SDM practice and willingness to practice SDM among health care professionals. Predictive variables were subjected to multiple linear regression analysis to investigate health care professionals' views of SDM. Valid respondents numbered 400, including physicians, pharmacists, nurses, and other health care professionals. The characteristics of these health care professionals significantly affected the mean scores of the KAB model. A correlation analyses indicated that the KAB constructs were positively correlated with each other. The top three barriers reported were lack of time (57.50%), lack of knowledge (38.25%), and difficulty of developing patient decision aids (37.75%). Respondents who were willing to practice SDM opined that SDM can provide the best health care for patients (81.62%), can maintain and improve individual clinical expertise (77.38%), and can meet patient and social expectations (48.40%). Continuous emphasis on education regarding SDM and continuous promotion of a positive attitude of SDM acceptance can influence the behaviour of practicing SDM among health care professionals. Further study is required to assess the SDM practices of various health care professionals in different settings.


Q:

how to fix error 403 when update new tweet to twitter using xcode?

Request DBE044D8-3C3D-4122-95FF-F0E2063FB9DF failed with error: Error Domain=HTTP Code=403 "The operation couldn’t be completed. (HTTP error 403.)"

that error log is showed like that when I want to update new tweet, I am using this code
- (void) requestSucceeded: (NSString *) requestIdentifier {
    CLog(@"Request %@ succeeded", requestIdentifier);
}

- (void) requestFailed: (NSString *) requestIdentifier withError: (NSError *) error {
    CLog(@"Request %@ failed with error: %@", requestIdentifier, error);
}

-(void)twit:(NSString *)text{
    CLog(@"test to tweet");
    [_engine sendUpdate:@"hallo"];
}

- (IBAction)twitter:(id)sender {
    NSString * user=[[NSUserDefaults standardUserDefaults] objectForKey:@"username"];
    if(![user isNotEmpty]){
        _engine=nil;
    }

    if(!_engine){
        CLog(@"engine create");
        _engine = [[[SA_OAuthTwitterEngine alloc] initOAuthWithDelegate:self] autorelease];
        _engine.consumerKey= kOAuthConsumerKey;
        _engine.consumerSecret = kOAuthConsumerSecret;
    }

    if(![_engine isAuthorized]){
        UIViewController *controller = [SA_OAuthTwitterController controllerToEnterCredentialsWithTwitterEngine:_engine delegate:self];

        if (controller){
            [self presentModalViewController:controller animated:YES];
            [[BNUtilitiesQuick FrontPageofBadger] presentModalViewController:controller animated:YES];
        }
    }
}

#pragma mark SA_OAuthTwitterEngineDelegate
- (void) storeCachedTwitterOAuthData:(NSString *) data forUsername:(NSString *) username {
    NSUserDefaults          *defaults = [NSUserDefaults standardUserDefaults];

    [defaults setObject:data forKey:@"authData"];
    [defaults setObject:username forKey:@"username"];

    [defaults synchronize];
    CLog(@"username:%@", username);
}

- (NSString *) cachedTwitterOAuthDataForUsername:(NSString *) username {
    return [[NSUserDefaults standardUserDefaults] objectForKey:@"authData"];
}

- (void) OAuthTwitterControllerFailed:(SA_OAuthTwitterController *) controller{
    CLog(@"failed");
}
- (void) OAuthTwitterControllerCanceled:(SA_OAuthTwitterController *) controller{
    CLog(@"cancel");
}

anyone get that error too? and how to fix this problem?

A:

I think this problem occur because your twitter API may be deprecated also closed by twitter....
use latest twitter API or also inBuilt Twitter Framework which work in iOS 5 or Later.
its very useful and also very easy..
:)


Organization of the exoerythrocytic stage of the rodent malaria parasite Plasmodium berghei. A cytochemical study.
With the osmium tetroxide-zinc iodide impregnation technique the visualization of the internal organization of the exoerythrocytic form of the rodent malaria parasite Plasmodium berghei was improved. Osmium impregnation leached the ground matrix of the parasite thereby displaying a system of intermediate-sized filaments. Because microtubules are only present as part of the mitotic spindle and as remnants of the sporozoite cytoskeleton, the observed intermediate-sized filaments comprise most of the cytoskeletal organization of the liver stage malaria parasite.


A file server is a computer that provides file service relating to the organization of information on persistent storage devices, such memories, tapes or disks. The file server or filer may be embodied as a storage system including a storage operating system that implements a file system to logically organize the information as a hierarchical structure of directories and files on, e.g., the disks. Each “on-disk” file may be implemented as set of data structures, e.g., disk blocks, configured to store information, such as the actual data for the file. A directory, on the other hand, may be implemented as a specially formatted file in which information about other files and directories are stored.
One type of file system is a write-anywhere file system that does not overwrite data on disks. If a data block on disk is retrieved (read) from disk into memory and “dirtied” with new data, the data block is stored (written) to a new location on disk to thereby optimize write performance. A write-anywhere file system may initially assume an optimal layout such that the data is substantially contiguously arranged on disks. The optimal disk layout results in efficient access operations, particularly for sequential read operations, directed to the disks. An example of a write-anywhere file system that is configured to operate on a storage system, such as a filer, is the Write Anywhere File Layout (WAFL™) file system available from Network Appliance, Inc., Sunnyvale, Calif. The WAFL file system is implemented as a microkernel within an overall protocol stack of the filer and associated disk storage.
The disk storage is typically implemented as one or more storage “volumes” that comprise a cluster of physical storage devices (disks), defining an overall logical arrangement of disk space. Each volume is generally associated with its own file system. In the WAFL file system, a special directory, called a “qtree”, may be created that has the properties of a logical sub-volume within the namespace of a physical volume. Each file system object (file or directory) is associated with one and only one qtree, and quotas, security properties and other items can be assigned on a per-qtree basis. Each volume has its own file system identifier (ID) and each qtree within a volume has its own qtree ID.
A difficult and time-consuming issue in managing a file server or filer is copying data, e.g., a data set, from an original filer (“primary filer”) to another filer (“backup filer”) to protect from data loss/corruption due to primary filer failure. As used herein, a data set is defined as one or more storage units, such as volumes or qtrees, that when combined represent data being served, migrated, and protected against disaster. A primary filer is a physical filer that is serving a client's data set and a backup filer is a different physical filer functioning as a target of a data set being served from the primary filer. The backup filer may also serve its own independent data set.
One way to copy a data set onto a backup filer to ensure against total primary filer failure is to physically move the storage disks from the primary filer to the backup filer. Another approach is to generate copies of the data set on backup tapes for use with the backup filer. However, these approaches can be slow and labor-intensive processes to manage. Yet another approach is to replicate a primary filer data set at the backup filer using a conventional storage replication technique, such as remote synchronous mirroring. In a remote synchronous replication environment, a mirrored data set must be transferred over a computer network and stored (“written”) to the backup filer before the write operation completes on the primary filer. Although this technique assures a high level of data currency for the backup filer, it also introduces data latency issues as a result of, among other things, performance delays inherent to remote synchronous mirroring.
The copied, mirrored data set can be used in a variety of useful ways, e.g., as local read-only copies of data that can be accessed quickly, or as an online consistent backup copy of data that can be used in the event of a failure of the original data set. However, the mirrored data set must typically be restored to the primary filer before it becomes available because the operating environment, e.g., operating system, security domain, etc., of the backup filer may be different from that of the primary filer. Thus to effectively use a mirrored data set, that data set usually must be returned to the filer of origin after the primary filer has been repaired or rebuilt.
A known Remote Disaster Facility (RDF) system available from Compaq Computer (Tandem Dictionary) maintains synchronized and consistent local and remote copies of a data set based on transfer of audit information. Briefly, the RDF system comprises a transaction-processing database on a local computer that utilizes audit information to monitor changes to the local data set. The audit information is transferred to a backup computer of the RDF system and applied to the remote copy of the data set. The backup computer can then take over a function from the local computer in the face of a disaster. However, transfer of the audit information is implemented at a “middle” software layer using, e.g., a Transaction Monitoring Facility (TMF), that resides between an application layer and a file system layer. Use of the TMF consumes substantial overhead, without providing general file system recovery. Moreover, clients must be notified of new location of the data set in the event of takeover by the backup computer.


In this training video watch Lightwave Instructor Adam Gibson build a Sci-Fi Style Corridor/Hallway similar to many that have been created for big screen sci-fi action films.

Learn how to take simple shapes and turn them into highly detailed geometry. Learn how to make your polygons glow to be used a light sources. A great video for the beginner 3D modeler and those who wish to learn a few more new modeling tricks.

Table of Contents for Sci-Fi Modeling (Volume #1)

Chapter 1- Intro

Chapter 2- Creating the Base of the Corridor

Chapter 3- Modeling the Grill Style Floor (Part One)

Chapter 4- Modeling the Grill Style Floor (Part Two)

Chapter 5- Boolean Substract Grill Ends

Chapter 6- Reduce Grill Polygons

Chapter 7- Mirroring Grill

Chapter 8- Mirroring Grill to Match Corridor

Chapter 9- Naming Surfaces and Statistics Panel

Chapter 10- Creating Support Structures

Chapter 11- Detail Support Structures Pt 1

Chapter 12- Detail Support Structures Pt 2

Chapter 13- Mirror Support Structures

Chapter 14- Create Caution Bars

Chapter 15- Create Side Lights

Chapter 16- Ceiling Panels (Part One)

Chapter 17- Ceiling Panels (Part Two)

Chapter 18- Cloning Ceiling Panels

Chapter 19- Ceiling Pipes

Chapter 20- Long Pipe and Supports

Chapter 21- Long Pipe Rail Extrusion for Curving

Chapter 22- Wall Panel Boxes (Part One)

Chapter 23- Wall Panel Boxes (Part Two)

Chapter 24- Ceiling Lights

Chapter 25- Wall Panel Detailing (Part One)

Chapter 26- Wall Panel Detailing (Part Two)

Chapter 27- Wall Panel Detailing (Part Three)

Chapter 28- Wall Panel Detailing (Part Four)

Chapter 29- Mirroring Wall Panel Detail (Part One)

Chapter 30- Mirroring Wall Panel Detail (Part Two)

Chapter 31- Level 01 Text Boolean

Chapter 32- Electrical Cables for Boxes

Chapter 33- Lower Wall Panels

Chapter 34- Lower Wall Panels and Adding a Texture

Chapter 35- Texturing Mapping

Chapter 36- Test Renders (Part One)

Chapter 37- Test Renders (Part Two)

Running Time: 8 hrs. 10 mins.

Level: Beginner to Intermediate

Screen Resolution: 1440 x 900 pixels

Format: Quicktime Video (.mov)

In this training video watch Lightwave Instructor Adam Gibson build a Sci-Fi Style Corridor/Hallway similar to many that have been created for big screen sci-fi action films.Learn how to take simple shapes and turn them into highly detailed geometry.

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Q:

Wordpress PHP title creation - Solve errors on 'child theme'

I have a wordpress theme running the "static" side of my website (www.url.com/home), and I'm working outside of it for the App (www.url.com/app), but I'm using the wordpress header and footer in the App so as to mantain the look and feel.
The problem is that all pages in the /app directory are coded as a 404 Error. I can somewhat solve this using js to change the title of the page once the page loads, but that isn't a good solution (all search engines ignore those pages).
The wordpress part defines the title like this:
<?php
if ( is_plugin_inactive('wordpress-seo/wp-seo.php') ) {
    bloginfo( 'name' );
}
?>
<?php
    wp_title("|", true);
?>

I'm not very good with PHP. Can anyone offer a solution so that the title is set normally within the wordpress environment, but instead of automatically tagging the website as an error it will check if the url address starts with "url.com/app" and if it does, it will just title the page something ("Web app", for example).
Thanks, and sorry if the question is lame but I'm really new with PHP.

A:

The correct way is to add a filter into your functions.php file within your theme. So:
wp-content/themes/[YOUR THEME]/functions.php
Add this to the end and then just change the "Your Page Title Here" string. It will add a filter and change any pages that include "/app" in the URI.
function set_title_if_slash_app( $title ){

    $full_url = $_SERVER[HTTP_HOST].$_SERVER[REQUEST_URI];

    if (strpos($full_url,"/app") !== false) {
        return "Your Page Title Here";
    }

    return __( 'Home', 'theme_domain' ) . ' | ' . get_bloginfo( 'description' );
}

add_filter( 'wp_title', 'set_title_if_slash_app');


Transforming growth factor-beta1 codon 10 polymorphism is associated with acute GVHD after allogenic BMT in Iranian population.
Certain cytokine genotypes are associated with acute graft versus host disease (aGVHD) after bone marrow transplantation (BMT). The present study aimed to determine existing association between TGF-beta1 codon 10 polymorphism and aGVHD after HLA-identical sibling BMT in the Iranian population. In a retrospective case-control study, 168 subjects including 84 Iranian HLA-identical sibling BMT donor/recipient pairs were recruited. All of the patients were affected by hematological malignancies (AML=39, ALL=23 and CML=22). PCR-SSP method was performed to determine TGF-beta1 codon 10 T/C polymorphism genotypes. The frequency of TGF-beta1 codon 10 TT, TC and CC genotypes among all subjects were 26.8%, 33.3% and 39.9% respectively. Recipients with the T allele developed aGVHD significantly less than those without the T allele (odds ratio =0.334, P=0.026). Genetic background of TGF-beta1 may be involved as a protective factor in the development of aGVHD in HLA-matched sibling BMT in Iranian population. Moreover, this finding may indicate that the genetic markers in Iranians are, at least to some extent, linked to distinct genetic event from Japanese.


Q:

Windows zip error: Windows cannot complete the extraction. The destination file could not be created

I have a strange error in Windows 7 when I want to execute/open a file inside ZIP archive.
I have two files:
File1.dwg
File2.dwg

The archive is not corrupted ( I checked with 7zip utility )

When I double click on the any file inside ZIP (I opened ZIP with Windows Explorer), the error occured:

Ok. If I open zip file with 7-zip or WinRAR every file could be opened(executed) without error and the AutoCAD is opened to see the dwg content.
Why in Windows ZIP ?
What points have I to see to manage/remove this error ?
Thank you.
UPDATE
If I click Extract all from Windows ZIP, the error occured as unspecified error:

A:

That error message can be a bit misleading.  If the file in the zip you are opening is password protected that could be the problem. Windows can't handle AES-256 encryption which comes up as the default in the 7zip UI when password protecting files through the UI.  If you change to ZipCrypto, then windows should be able to read it after prompting for the password.

A:

It looks like Windows is having a hard time creating the temporary file for you to view (it extracts it into temp when you double-click it, then opens it for viewing). Perhaps clear out your temp folder (there are tools online for this) and try again to view the files.
Out of interest, though, can you extract the files normally (via the "Extract all" command) specifying a path (which obviously isn't temp)?


Sex-determining region Y box 4 is a transforming oncogene in human prostate cancer cells.
Prostate cancer is the most commonly diagnosed noncutaneous neoplasm and second most common cause of cancer-related mortality in western men. To investigate the mechanisms of prostate cancer development and progression, we did expression profiling of human prostate cancer and benign tissues. We show that the SOX4 is overexpressed in prostate tumor samples compared with benign tissues by microarray analysis, real-time PCR, and immunohistochemistry. We also show that SOX4 expression is highly correlated with Gleason score at the mRNA and protein level using tissue microarrays. Genes affected by SOX4 expression were also identified, including BCL10, CSF1, and NcoA4/ARA70. TLE-1 and BBC3/PUMA were identified as direct targets of SOX4. Silencing of SOX4 by small interfering RNA transfection induced apoptosis of prostate cancer cells, suggesting that SOX4 could be a therapeutic target for prostate cancer. Stable transfection of SOX4 into nontransformed prostate cells enabled colony formation in soft agar, suggesting that, in the proper cellular context, SOX4 can be a transforming oncogene.


Leaving French Assets to Step-Children

Charlotte Macdonald, an associate solicitor in Stone King’s international and cross-border team, sets out a number of scenarios that a British owner of French property may encounter.

Over a 12-part series of articles Charlotte will cover topics such as how to buy your property as a couple (jointly or not), the different legal implications for a married couple and a civil partnership, and what happens to the property on your death, including who it will pass to and what succession tax consequences there might be.

Scenario Two – My spouse and I both have children from our previous marriages. We want to make sure that our children receive the same amount after we die.

Beth and Barry Brown are English. They moved to Annecy in France eight years ago following their retirement.

They have been married for 20 years and each have a child from a previous marriage but have no children together.

When Beth and Barry moved to France they sold their home in England and purchased a property overlooking Lac d’Annecy. They bought this property jointly (50% each) and moved all their assets to France.

Following a health scare they decided to make wills. They wish to leave all their assets to one another and then to their children equally. As explained in our previous article, because Beth and Barry are English, they can elect for English law to apply to the succession of their property in France. This means they can leave their assets to whomever they choose in their wills.

However, the couple have heard that doing this might be very tax inefficient.

Differences between French and UK inheritance tax

In the UK, each individual’s estate has an inheritance tax free allowance on death, which is currently £325,000. Anything over that tax free allowance (subject to exemptions and allowances) is taxed at the flat rate of 40%. This tax rate does not change if you leave your assets to your children, step-children or friends.

In contrast, in France it is the individual beneficiary (the person receiving an inheritance) who has an inheritance tax free allowance, and they pay inheritance tax at a rate dependent upon their relationship with the deceased. The closer the relationship, the greater the tax free allowance and the lower the rate of tax that is paid.

For example, a child inheriting from a parent will receive the first €100,000 free of inheritance tax. Anything they inherit over this amount will be taxed on a sliding scale between 5% and 45%. A step-child is classed as an unrelated beneficiary. This means they have a very small tax free allowance of €1,594 and pay inheritance tax at the flat rate of 60% on anything they inherit over this amount.

The effect of the EU succession regulation on taxation

Although the EU succession regulation gives foreign nationals the option to avoid French succession law, it does not change the inheritance taxation position in France. This means if you pass away owning property in France, it will be assessed for tax by the French tax authorities.

What does this mean for Beth and Barry?

If Beth dies before Barry and leaves all her assets to him, he will inherit them free of inheritance tax, because they are married. If Barry then leaves all their assets equally to his child and to Beth’s child, the children will be taxed as follows:

Barry’s child €100,000 tax free. Balance taxed between 5% – 45%.

Beth’s child €1,594 tax free. Balance taxed at a flat rate of 60%.

This is not what the couple wished, as Beth’s child will receive substantially less than Barry’s child after tax.

To avoid this as far as possible, Beth and Barry should obtain professional advice before making their wills. They may decide it is appropriate for each to leave some of their assets directly to their own child, or they may decide a French ownership structure such as an usufruit is appropriate, to try and reduce the French inheritance tax.

The next scenario in the series will look at issues that can arise when you do not want your children to inherit your assets.

For more information please contact Charlotte Macdonald, Dan Harris or Raquel Ugalde at Stone King LLP either by calling +44(0)1225 337599, or by emailing: international@stoneking.co.uk

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Victoria Park, Widnes

Victoria Park is the premier urban park in Widnes, Cheshire, England. It is managed by Halton Borough Council.

History

The park was opened in 1900 to celebrate the Diamond Jubilee of Queen Victoria. It is on the former Appleton House estate and was created by the local council with funds collected via a public subscription.

Facilities
There are formal gardens, a lake, a fountain, a skate park, climbing boulder, bandstand, cafe, children's play area, tennis courts and a butterfly house.

Landmarks

There is a memorial fountain in honour of William Ewart Gladstone. A milestone marker with plaque commemorates the last effective Zeppelin air raid of World War I in England. Five Zeppelins dropped bombs in Widnes, Ince and Wigan. There is a statue of Sgt. Thomas Mottershead V.C., DCM (1892 -1917) who was born in Widnes and was awarded a Victoria Cross and Distinguished Conduct Medal in World War I. There is a combined memorial headstone for Mottershead and two other recipients of the V.C. from Halton: Thomas Wilkinson and Thomas Alfred Jones.

War memorial pillar
The park contains a Grade II listed war memorial pillar. Incorporating detailed carvings and standing more than 16m high, the memorial is a striking and prominent architectural building. The pillar was designed by Harold E Davies and unveiled by the 17th Earl of Derby in a ceremony on 28 September 1921 after a civic parade from Widnes town hall. The memorial cost £6000 and was paid for by public subscription. The sculptural additions were undertaken by the national artist Herbert Tyson Smith.

Events

Vintage Steam Rally
Each year the park hosts North West England's largest Vintage Steam Rally, featuring a vintage fair, and vintage steam machinery.

Parkrun
A 5 km parkrun takes place every Saturday morning beginning at 9am.

See also
List of parks and open spaces in Cheshire

References

Category:Parks and open spaces in Cheshire
Category:Widnes


// -------------------------------------------------------------------------------------------------
// Copyright (c) Microsoft Corporation. All rights reserved.
// Licensed under the MIT License (MIT). See LICENSE in the repo root for license information.
// -------------------------------------------------------------------------------------------------

namespace Microsoft.Health.Fhir.CosmosDb.Features.Storage
{
    /// <summary>
    /// Cosmos DB customer-managed key (CMK) sub status values
    /// </summary>
    public enum KnownCosmosDbCmkSubStatusValue
    {
        // Customer-Managed Key (CMK) values
        AadClientCredentialsGrantFailure = 4000,
        AadServiceUnavailable = 4001,
        KeyVaultAuthenticationFailure = 4002,
        KeyVaultKeyNotFound = 4003,
        KeyVaultServiceUnavailable = 4004,
        KeyVaultWrapUnwrapFailure = 4005,
        InvalidKeyVaultKeyUri = 4006,
        InvalidInputBytes = 4007,
        KeyVaultInternalServerError = 4008,
        KeyVaultDnsNotResolved = 4009,
    }
}


Apple is 'dragging its feet' by not matching rivals such as Microsoft and Google in barring access to sites secured with suspect DigiNotar certificates

A security researcher criticized Apple for what he called "foot dragging" over the DigiNotar certificate fiasco, and urged the company to quickly update Mac OS X to protect users.

"We're looking at some very serious issues [about trust on the Web] and it doesn't help matters when Apple is dragging its feet," said Paul Henry, a security and forensics analyst with Arizona-based Lumension.

Unlike Microsoft, which updated Windows Tuesday to block all SSL (secure socket layer) certificates issued by DigiNotar, Apple has not updated Mac OS X to do the same.

DigiNotar, one of hundreds of firms authorized to issue digital certificates that authenticate a website's identity, admitted on Aug. 30 that its servers were compromised weeks earlier. A report made public Monday said that hackers had acquired 531 certificates, including many used by the Dutch government, and that DigiNotar was unaware of the intrusion for weeks.

Because almost all the people who were routed to a site secured with one of the stolen certificates were from Iran, many experts suspect that the DigiNotar hack was sponsored or encouraged by the Iranian government, which could use them to spy on its citizens.

Microsoft isn't the only software maker to block all DigiNotar certificates: Google, Mozilla, and Opera have also issued new versions of their browsers -- Chrome, Firefox, and Opera -- to completely, or in Opera's case, partially prevent users from reaching websites secured with a DigiNotar certificate.

Users of Safari on Mac OS X, however, remain at risk to possible "man-in-the-middle" attacks based on the fraudulently obtained certificates.

And that rubbed Henry the wrong way. "Mac users have been left hanging in the wind," Henry said.

Because Safari relies on the underlying operating system to tell it which certificates have been revoked or banned entirely, Apple must update Mac OS X. The Windows edition of Safari, which has a negligible share of the browser market, taps Windows' certificate list: That version is safe to use once Microsoft's Tuesday patch is applied.

Henry admitted he wasn't surprised by the fact that Apple was odd-man out.

"No, I'm not, not after it took them a month to respond to the Comodo issue," Henry said, referring to a smaller-scale hack of another certificate authority last March.

Then, Apple updated Mac OS X on April 14, nearly a month after Comodo discovered the intrusion and began notifying browser makers. Microsoft had patched Windows to block the stolen Comodo certificates on March 23, while Google and Mozilla had even earlier.

The DigiNotar hack, however, was larger -- criminals acquired only nine certificates from Comodo -- and more serious in that the Dutch government relied on DigiNotar to secure its websites.

And then there are the boasts made by an unidentified hacker who has claimed responsibility for both the Comodo and DigiNotar breaches.

On Monday, that hacker -- known only as "Comodohacker" -- also asserted that he had penetrated four other CAs, or certificate authorities, including GlobalSign, a U.S.-based CA much more widely used than DigiNotar.

Tuesday, GlobalSign suspended certificate sales and said it had launched an investigation into Comodohacker's claims. A day later the New Hampshire company said it had hired Fox-IT, the forensics firm that is still digging into the DigiNotar hack for the Dutch government.

If Comodohacker's claims are accurate, and there are other CAs besides DigiNotar that have issued unauthorized SSL certificates, Apple's sluggishness is even more inexcusable, Henry said.

"We may be looking at the tip of the iceberg," said Henry. "If there are four other CAs [involved] as this guy claims, we could be in a hell of a mess in the next few months. We'll be playing catch-up, but that seems to be more difficult for Apple than for Microsoft, or Google and Mozilla."

But Henry didn't limit his criticism to Apple.

"What about smartphones ?" he asked. "I'm not aware of a single carrier or vendor that has pushed out a browser update for this."

Neither Google nor Apple have said anything about plans for updating Android or iOS to follow the road taken by most desktop browsers. The companies declined to comment Tuesday for a story written by IDG News Service reporter Robert McMillan. (Like Computerworld, IDG News is part of IDG.)

According to Web metrics company Net Applications, about three-out-of-four Mac OS X users run Safari as their primary browser.


Rosa 'Apricot Nectar'

Rosa 'Apricot Nectar'  is a hybrid tea rose cultivar, bred by Eugene Boerner before 1964. The plant is considered very easy to grow and is well-known for its attractive pale apricot pink color. It was named an All-America Rose Selections (AARS) winner in 1966.

Description
'Apricot Nectar' is a medium, upright shrub, 3 to 4 ft (90—121 cm) in height with a 3 to 4 ft (90—121 cm) spread. Blooms are  4.5 in (11.43 cm) in diameter, with 26 to 40 petals. Flowers have a high-centered, double bloom form, and are borne in large clusters of 3—11.  The flowers are a pale apricot-pink color with darker backs and yellow at the center. The entire flower fades to light yellow as it ages. The flower is exceptionally attractive in evening light, and produces the best flowers in warm, dry climates. The rose has a strong and fruity fragrance and glossy, dark green foliage. It is very disease resistant and blooms in flushes  throughout its growing season. The plants does well in USDA zone 6 and warmer.

Child plants
'Apricot Nectar' was used to hybridize one child plant, the hybrid tea rose Rosa 'Serendipity'. The new cultivar is yellow in color with apricot hues, fading to cream as the flowers mature. The cultivar was developed by Dr. Griffith Buck in  1978 by crossing ('Western Sun' x 'Carefree Beauty')  and ('Apricot Nectar' and 'Prairie Princess').

Awards
 All-America Rose Selections (AARS) winner, USA, (1966)

See also
Garden roses
Rose Hall of Fame
List of Award of Garden Merit roses

Notes

References

Apricot Nectar


Maternal effects of regular salicylate ingestion in pregnancy.
6.6% of all Australian patients attending an antenatal clinic were found to be taking salicylate preparations regularly, most commonly in powder form and almost always as self-medication. In this group of salicylate takers there was an increased incidence of anaemia, anti-partum and post-partum haemorrhage, prolonged gestation, complicated deliveries, and perinatal mortality compared with non-takers. These findings suggest that regular salicylate consumption has detrimental effects in pregnancy. Routine antenatal urinary screening for salicylate in pregnancy is recommended.


Theoretical study of the 1:1 and 2:1 (homo- and heterochiral) complexes of XOOX' (X, X' = H, CH3) with lithium cation.
A theoretical study of the 1:1 and 2:1 complexes of XOOX' (X, X' = H, CH(3)) with the lithium cation has been carried out by means of ab initio computational methods up to the MP2/aug-cc-pVTZ level. The optical rotatory power and NMR parameters (absolute chemical shielding and indirect coupling constants) have been calculated. In addition, the racemization barriers within the complexes formed have been evaluated. Special attention has been paid concerning the differences between the 2:1 homo- and heterochiral complexes.


Germany's Justice Ministry says former chief federal prosecutor Harald Range, who was fired in 2015 over a controversial investigation into two journalists, has died at age 70.

The ministry confirmed media reports Thursday that Range died of a heart attack while out at a cafe in Karlsruhe on Wednesday evening.

Then-Justice Minister Heiko Maas, now Germany's foreign minister, dismissed Range over his decision to open an investigation into two journalists from the website Netzpolitik.org who had reported that Germany's domestic spy agency planned to expand surveillance of online communication.

The treason probe, later dropped, was widely criticized and regarded as an embarrassment to the government, which was forced to defend Germany's commitment to protecting press freedom.

Range accused Maas of political interference in the case.


[Phacoemulsification and pseudoexfoliative syndrome].
To compare the results and the complications of cataract surgery by phacoemulsification in eyes with pseudoexfoliation syndrome (PEX) and without pseudoexfoliation syndrome. A retrospective clinical study (January 2001 - January 2004; mean follow-up 21.5 months) of 116 cases divided in two groups: group A (55 eyes) with senile cataract and pseudoexfoliation syndrome and group B (61 eyes) with senile cataract but without pseudoexfoliation syndrome. The eyes were without another associated ophthalmological diseases and the phacoemulsifications were performed by the same surgeon. Intraoperative observations included anterior chamber depth, pupil size after maximal mydriasis, zonular dehiscence, capsule tear, vitreous loss and postoperatively: best corrected visual acuity, intraocular pressure (IOP), aqueous flare response and IOL stability were evaluated at 1 day, 1 week, 1 month and until 3 years. The Student-t test was used for statistical analyses. There were determined significant correlations between pupil size, anterior chamber depth and the rate of complications in the group A. The visual acuities were similar in both groups. IOP reduction was superior in the eyes with PEX as compared with control group, but without statistically significantly difference. In group A, there were a non-statistically significantly percentage of IOL dislocation as compared with group B. In the eyes with PEX, it is necessary to utilize IOLs with bigger optic size. Phacoemulsification represents a safe method in cataract surgery in eyes with pseudoexfoliation syndrome in the presence of careful surgical technique: maximal mydriasis, circular continuous curvilinear capsulorhexis, adequate hydrodissection. On the long run, the postoperative results were comparable between eyes with PEX and the control group (without PEX).


2059 Baboquivari

2059 Baboquivari, provisional designation , is an asteroid classified as near-Earth object of the Amor group, approximately 1.9 kilometers in diameter. Discovered by the Indiana Asteroid Program in 1963, it was later named after the Baboquivari Mountains in Arizona, United States.

Discovery and recovery

Baboquivari is one of the lowest numbered near-Earth asteroids as it was already discovered on 16 October 1963. The discovery observation was made by the Indiana Asteroid Program at Goethe Link Observatory near Brooklyn, Indiana, in the United States. Three months later, it became a lost asteroid until June 1976, when it was recovered by the Steward Observatory's 90-inch Bok Telescope at Kitt Peak National Observatory located in the Sonoran Desert of Arizona.

Classification and orbit

Baboquivari is an Amor asteroid – a subgroup of near-Earth asteroids that approach the orbit of Earth from beyond, but do not cross it. It orbits the Sun at a distance of 1.2–4.1 AU once every 4 years and 4 months (1,577 days). Its orbit has an eccentricity of 0.53 and an inclination of 11° with respect to the ecliptic.
The body's observation arc begins at the discovering observatory, 10 days after its official discovery observation.

Close approaches

The asteroid has an Earth minimum orbit intersection distance of , which corresponds to 98.8 lunar distances. It approached the Earth at a similar distance on 20 October 1963, shortly after its discovery. The eccentric asteroid is also a Mars-crosser and approached Jupiter at a distance of about 1.4 AU on 20 April 1970.

Physical characteristics

Little is known about Baboquivaris physical characteristics. Its spectral type has never been determined.

Diameter and albedo

It is classified as a near-Earth object larger than one kilometer in diameter by the Minor Planet Center ("1+ KM"). A generic magnitude-to-diameter conversion gives a diameter of 1.9 kilometers, based on the body's absolute magnitude of 16.0 and an assumed standard albedo for stony S-type asteroids (Baboquivari would still measure 1.3 kilometers in diameter, if it had a higher albedo of 0.4, typically seen among bright members of the Hungaria family).

Rotation period

As of 2017, no rotational lightcurve of Baboquivari has been obtained from photometric observations. The asteroid's rotation period, poles and shape remain unknown.

Naming

This minor planet was named after the main-peak of the Baboquivari Mountains, a sacred location in the mythology of the Papago Indian Tribe. The Observatories of the Association of Universities for Research in Astronomy (AURA) are located on the Baboquivari land, just a few kilometers south of Kitt Peak. The approved naming citation was published by the Minor Planet Center on 1 December 1979 ().

References

External links
 Asteroid Lightcurve Database (LCDB), query form (info)
 Dictionary of Minor Planet Names, Google books
 Asteroids and comets rotation curves, CdR – Observatoire de Genève, Raoul Behrend
 Discovery Circumstances: Numbered Minor Planets (1)-(5000) – Minor Planet Center

002059
002059
Category:Minor planets named for places
Category:Named minor planets
19631016


Looking for the agent: an investigation into consciousness of action and self-consciousness in schizophrenic patients.
The abilities to attribute an action to its proper agent and to understand its meaning when it is produced by someone else are basic aspects of human social communication. Several psychiatric syndromes, such as schizophrenia, seem to lead to a dysfunction of the awareness of one's own action as well as of recognition of actions performed by others. Such syndromes offer a framework for studying the determinants of agency, the ability to correctly attribute actions to their veridical source. Thirty normal subjects and 30 schizophrenic patients with and without hallucinations and/or delusional experiences were required to execute simple finger and wrist movements, without direct visual control of their hand. The image of either their own hand or an alien hand executing the same or a different movement was presented on a TV-screen in real time. The task for the subjects was to discriminate whether the hand presented on the screen was their own or not. Hallucinating and deluded schizophrenic patients were more impaired in discriminating their own hand from the alien one than the non-hallucinating ones, and tended to misattribute the alien hand to themselves. Results are discussed according to a model of action control. A tentative description of the mechanisms leading to action consciousness is proposed.


/*
 * Licensed to the Apache Software Foundation (ASF) under one
 * or more contributor license agreements.  See the NOTICE file
 * distributed with this work for additional information
 * regarding copyright ownership.  The ASF licenses this file
 * to you under the Apache License, Version 2.0 (the
 * "License"); you may not use this file except in compliance
 * with the License.  You may obtain a copy of the License at
 *
 *  http://www.apache.org/licenses/LICENSE-2.0
 *
 * Unless required by applicable law or agreed to in writing,
 * software distributed under the License is distributed on an
 * "AS IS" BASIS, WITHOUT WARRANTIES OR CONDITIONS OF ANY
 * KIND, either express or implied.  See the License for the
 * specific language governing permissions and limitations
 * under the License.
 */
package org.apache.struts.beanvalidation.constraints;

import javax.validation.groups.Default;
import java.lang.annotation.ElementType;
import java.lang.annotation.Retention;
import java.lang.annotation.RetentionPolicy;
import java.lang.annotation.Target;

/**
 * Validation with Group Constrains on execution method (Action)
 *

 * Example, Validate with on special group
 *

 *

 *
 *  {@literal @}ValidationGroup(CarChecks.class)
 *  {@literal @}Action...
 *

 *

 *

 * Example, Validate with severals special group
 *

 *

 *  {@literal @}ValidationGroup(Default.class, CarChecks.class, DriverChecks.class)
 *  {@literal @}Action...
 *

 */
@Target({ElementType.METHOD})
@Retention(RetentionPolicy.RUNTIME)
public @interface ValidationGroup {

    Class<?>[] value() default {Default.class};
}


The difference between Hall and Gray's bills and the bills that have come before?

The two men chair their respective Governmental Organization committees, which oversee gambling in the State.

"A Framework That Can Be National Model"

Can California set bar for America?

While still a "placeholder" bill the prominence and influence of the two politicans does make it likely these new bills will be the forebearers of intrastate online poker in California in some form.

2016 is the likely adoption date of some form of the bills but several key obstacles remain with all of the legislation in play including bill AB 9, introduced by Mike Gatto last year, and AB 167 introduced by Reggie Jones Sawyer last month.

A "bad actor" clause in AB 9, ostensibly preventing PokerStars from returning to the US market, has been an issue although a new agreement between Caesars, PokerStars and the support of several local tribes has reduced that concern.

Opposition from race tracks in the state is also a concern. Hall and Gray, however, say it's time for all parties to get on the same page:

“The issue of iPoker in California has historically been divisive; dealing legislators, the governor and the public a folding hand,” said Senator Hall and Assemblymember Gray. “It is time to work together, stop bluffing and take control of this issue."

“This iPoker proposal must include a strong regulatory framework that protects Californians,” Gray added. “We need to crack down on illegal online gaming and replace it with a safe and responsible entertainment option for adults, which includes safeguards against compulsive and underage gambling, money laundering, fraud, and identity theft.”

“We intend to develop a framework in California that can be a national model,” said Senator Hall.

PokerListings.com is the world's largest and most trusted online poker guide, offering the best online poker bonus deals guaranteed, over $1m in exclusive freerolls every year and the most free poker content available on the Web.


Q:

Could Removing A Nintendo Game Boy Game While The Console Is Powered-On Do Any Damage?

I've looked all over the Internet to try to find an answer to this, but I can't. I want to know if removing a Nintendo Game Boy game while the console is powered on will do any damage, like rendering the cart and/or console unusable.
I don't think it will cause any damage, seeing as most games have volatile RAM and should go back to normal when inserted into the Game Boy again and powered on, but I'd like an actual source.
Also, I understand that with a standard Game Boy and game, removing the game while the console is on is impossible, due a plastic arm sliding over to cover the game when the power switch is moved. This is assuming I'm using another Game Boy product that doesn't have that feature, such as the Game Boy Color, or an original model modded to remove the arm.

A:

What's up?
No, not beyond save data corruption.
In fact, the Game Boy's big brother console, the Nintendo 64 had games that planned on using live cart swapping as a feature, to link games (most famously with Banjo Kazooie/Banjo Tooie), but it was scrapped due to other concerns (essentially ram being too volatile for it to work reliably).
There is one caveat: Poorly designed cartridges could be knocked out of commission by corrupted save data alone.
Note also: All of this applies to the OG Game Boy, the Color, the Pocket, the Advance and consoles NES, SNES and N64. From DS onwards, the system is designed specifically to detect cartridge removal and handle it gracefully; and the GC/Wii (U) don't use cartridges anymore.

So... why?
The question is, of course, why do these systems tolerate this at all? The basic reason is that in straightforward terms: There's nothing there to damage. The consoles mentioned above are, in essence, just a circuit board with a processor on it, some ROM chips that provide libraries, some volatile RAM, I/O, and a power supply. There's no OS or persistent storage or really anything else. When a cartridge is plugged in, it's plugged into what essentially works as an expansion slot on this board, providing the bulk of the actual executable code in ROM, as well as anything else the cartridge might need (such as flash or battery RAM memory for save files).
Most of the systems have physical barriers to removing the cartridge (usually the power button engages a lever that inserts itself into a groove in the cartridge, preventing it from being removed (or inserted) while the power is on).
The first console Nintendo shipped without a physical barrier like this was the Game Boy Color, which allows a cartridge to be hot swapped; as far as I recall it detects this situation and gracefully shuts down when this happens. This is likely a feature that wasn't present in the earliest Game Boys or home consoles due to the physical barriers simply being easier to implement; but the proliferation of different-shaped Game Boy cartridges made the existing safeguards ineffectual or counter-productive, and the console was made to detect the situation and handle it instead.
In any case, the simplicity of design means that there's not really anything for pulling out a cartridge to damage in any meaningful way; which is a premium design feature in a product designed to be roughed around by kids.