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RESEARCH ARTICLEOpen Access
Identification and characterization of wheat long
non-protein coding RNAs responsive to powdery
mildew infection and heat stress by using
microarray analysis and SBS sequencing
Mingming Xin1,2†, Yu Wang1,2†, Yingyin Yao1,2†, Na Song1,2, Zhaorong Hu1,2, Dandan Qin1,2, Chaojie Xie1,2,
Huiru Peng1,2*, Zhongfu Ni1,2and Qixin Sun1,2,3*
Background: Biotic and abiotic stresses, such as powdery mildew infection and high temperature, are important
limiting factors for yield and grain quality in wheat production. Emerging evidences suggest that long non-protein
coding RNAs (npcRNAs) are developmentally regulated and play roles in development and stress responses of
plants. However, identification of long npcRNAs is limited to a few plant species, such as Arabidopsis, rice and
maize, no systematic identification of long npcRNAs and their responses to abiotic and biotic stresses is reported in
Results: In this study, by using computational analysis and experimental approach we identified 125 putative
wheat stress responsive long npcRNAs, which are not conserved among plant species. Among them, some were
precursors of small RNAs such as microRNAs and siRNAs, two long npcRNAs were identified as signal recognition
particle (SRP) 7S RNA variants, and three were characterized as U3 snoRNAs. We found that wheat long npcRNAs
showed tissue dependent expression patterns and were responsive to powdery mildew infection and heat stress.
Conclusion: Our results indicated that diverse sets of wheat long npcRNAs were responsive to powdery mildew
infection and heat stress, and could function in wheat responses to both biotic and abiotic stresses, which
provided a starting point to understand their functions and regulatory mechanisms in the future.
The developmental and physiological complexity of
eukaryotes could not be explained solely by the number
of protein-coding genes [1]. For example, the Drosophila
melanogaster genome contains only twice as many
genes as some bacterial species, although the former is
far more complex in its genome organization than the
latter. Similarly, the number of protein-coding genes in
human and nematode is extremely close. A portion of
this paradox can be resolved through alternative pre-
mRNA splicing [2]. In addition, post-translational modi-
fications can also contribute to the increased complexity
and diversity of protein species [3].
Recent studies suggest that most of the genome are
transcribed, among the transcripts only a small portion
encode for proteins, whereas a large portion of the tran-
scripts do not encode any proteins, which are generally
termed non-protein coding RNAs (npcRNA). For exam-
ple, transcriptome profiling in rice (Oryza sativa) indi-
cates that there are about 8400 putative npcRNAs,
which do not overlap with any predicted open reading
frames (ORFs) [4]. These npcRNAs are subdivided as
housekeeping npcRNAs (such as transfer and ribosomal
RNAs) and regulatory npcRNAs or riboregulators, with
the latter being further divided into short regulatory
npcRNAs (<300 bp in length, such as microRNA,
siRNA, piwi-RNA) and long regulatory npcRNAs
* Correspondence: firstname.lastname@example.com; firstname.lastname@example.com
† Contributed equally
1State Key Laboratory for Agrobiotechnology and Key Laboratory of Crop
Heterosis and Utilization (MOE) and Key Laboratory of Crop Genomics and
Genetic Improvement (MOA), Beijing Key Laboratory of Crop Genetic
Improvement, China Agricultural University, Beijing, 100094, PR China
Full list of author information is available at the end of the article
Xin et al. BMC Plant Biology 2011, 11:61
© 2011 Xin et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited.
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(>300 bp in length). With the identification of micro-
RNAs and siRNAs in diverse organisms, increasing evi-
dences indicate that these short npcRNAs play
important roles in development, responses to biotic and
abiotic stresses by cleavage of target mRNAs or by inter-
fering with translation of target genes [5-9].
Long npcRNAs are transcribed by RNA polymerase II,
polyadenylated and often spliced [10]. Studies in mice
and human suggested that at least 13% and 26% of the
unique full-length cDNAs, respectively, are thought to
be poly(A) tail-containing long npcRNAs [11-13]. Emer-
ging evidences also suggest that long npcRNAs are
developmentally regulated and responsive to external
stimuli, and play roles in development and stress
responses of plants and disease in human. For example,
some long npcRNAs are regulated in various stresses in
plants and animals [9,14-16]. In Caenorhabditis elegans,
25 npcRNAs are either over- or under-expressed under
heat shock or starvation conditions [17], while in Arabi-
dopsis, the abundance of 22 putative long npcRNAs are
regulated by phosphate starvation, salt stress or water
stress [18]. In Arabidopsis, long npcRNA, COOLAIR
(cold induced long antisense intragenic RNA), is cold-
induced FLC antisense transcripts, and has an early role
in the epigenetic silencing of FLC and to silence FLC
transcription transiently [19]. Long npcRNA HOTAIR in
human is reported to reprogram chromatin state to pro-
mote cancer metastasis [20].
Currently, two computational methods are employed
to identify long npcRNAs, genome-based and transcript-
based. Using genomic sequences, more than 200 candi-
date long npcRNAs were predicted in Escherichia coli
[21], and at least 20 long npcRNA genes have been
experimentally confirmed [22]. In Rhizobium etli, 89
candidate npcRNAs are detected by high-resolution til-
ling array, and 66 are classified as novel ones [23].
While using cDNA or EST sequences, a large number
of long npcRNAs are detected in Drosophila, mouse and
Arabidopsis [12,18,24-26].
Up to date, identification of long npcRNAs is limited
to a few plant species, such as Arabidopsis, rice and
maize. To our best knowleage, in wheat no systematic
identification of long npcRNAs is reported. Wheat (Tri-
ticum aestivum, AABBDD, 2n = 42) is the most widely
grown crop plant, occupying 17% of all the cultivated
land, provides approximately 55% of carbohydrates for
world human consumption [27], Biotic and abiotic stres-
ses are important limiting factors for yield and grain
quality in wheat production. For instance, powdery mil-
dew, caused by the obligate biotrophic fungus Blumeria
graminis f. sp. tritici (Bgt), is one of the most devastating
diseases of wheat in China and worldwide and causing
significant yield losses [28]. High temperature, often
combined with drought stress, causes yield loss and
reduces the grain quality [29]. To reduce the damages
caused by biotic and abiotic stresses, plants have evolved
sophisticated adaptive response mechanisms to repro-
gram gene expression at the transcriptional, post-
transcriptional and post-translational levels [30].
Recently, transcript profiling has been successfully
employed to determine the transcriptional responses to
powdery mildew infection and heat stress in wheat, and
the results revealed that a number of genes were signifi-
cantly induced or repressed in response to these stresses
In our previous study [33], it was demonstrated that
expression of microRNAs in wheat was regulated by
powdery mildew infection and heat stress, which stimu-
lated us to explore whether long npcRNA was also
responsive to powdery mildew infection and/or heat
stress. In this study, we performed a genome-wide in
silico screening of powdery mildew infection and heat
stress responsive wheat transcripts in order to isolate a
collection of long npcRNA genes. Combining microarray
analysis and high-throughput SBS sequencing methods,
we totally characterized 125 putative stress responsive
long npcRNAs in wheat, four of them were miRNA pre-
cursors, and one was experimentally verified by northern
blot. Wheat long npcRNAs displayed tissue-specific
expression patterns and their expression levels were
altered in response to powdery mildew infection and/or
heat stress, which suggested that at least a subset of
these newly identified wheat long npcRNAs potentially
play roles in response to biotic and/or abiotic stresses in
Identification of powdery mildew infection and heat
stress responsive long npcRNA candidates in wheat
In our previous study, a total of 9744 powdery mildew
infection and 6560 heat stress responsive transcripts
were obtained (with a fold change of at least 2) through
microarray analysis using the wheat Affymetrix Gene-
Chip®. In this study, in order to identify the putative
wheat long npcRNAs which were responsive to powdery
mildew and/or heat stress, these stress responsive tran-
scripts were used to characterize the wheat long
npcRNAs. Firstly, these transcripts were annotated by
Harvest program, and 7746 and 5754 transcripts were
identified to be protein-coding genes and therefore were
discarded in further analysis. The remaining transcripts
were then analyzed by Blastx and Blastn, 586 and 406
ESTs with no similarity to protein coding genes or
tRNA and rRNA were retained. Secondly, 125 tran-
scripts with no or short ORFs (less than 80aa) and
polyA-tails were selected as putative long npcRNAs
(Additional file 1), among which 71 were responsive to
powdery mildew infection, and 77 were responsive to
Xin et al. BMC Plant Biology 2011, 11:61
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heat stress. We found that 23 long npcRNAs responded
to both powdery mildew infection and heat stress
(designated TalnRNA). A total of 48 putative long
npcRNAs were only responsive to powdery mildew
infection (designated TapmlnRNA), and 54 were only
responsive to heat stress (designated TahlnRNA).
Among these putative long npcRNAs, the longest ORF
was 74aa, with an average of 43.5aa (Additional file 1).
In order to validate expression patterns of the long
npcRNAs in response to powdery mildew infection and/
or heat stress, expression patterns of 4 long npcRNAs,
TapmlnRNA19, TapmlnRNA30, TahlnRNA27 and
TalnRNA5, were determined by using quantitative RT-
PCR analysis. Expression levels of TapmlnRNA19 and
TapmlnRNA30 were up-regulated after powdery mildew
inoculation (Figure 1a, b), whereas expression of
TahlnRNA27 and TalnRNA5 were up-regulated after
heat stress (Figure 2a, b), which showed consistent
expression patterns with microarray analysis.
Four long npcRNA transcripts correspond to miRNA
By mapping miRNAs which were identified from our pre-
viously sequenced six small RNA libraries (S-0h, S-12h,
R-0h, R-12h, TAM-0h, TAM-1h) [33] to the complete
collection of 125 long npcRNAs, we identified that four
transcripts (TalnRNA5, TapmlnRNA8, TapmlnRNA19,
TahlnRNA27) were miRNA precursors. Prediction of the
secondary structure for the four transcripts by using the
Vienna RNA package RNAfold web interface program
showed that these four miRNA precursors had stable
hairpin structures (Additional file 2, 3, 4 and 5).
Among the four long npcRNAs, three (TalnRNA5,
TapmlnRNA19 and TapmlnRNA8) were responsive
to powdery mildew infection. Both TalnRNA5 and
TapmlnRNA19 were the precursors of miR2004, and
TapmlnRNA8 was the precursor of miR2066. It is inter-
esting to note that TapmlnRNA19 and TalnRNA5 were
up-regulated after powdery mildew infection as deter-
mined by qRT-PCR (Figure 1a, 3a), and miR2004 was
also found to be up-regulated based on the small RNA
high throughput sequencing (Figure 3b). To further
determine the expression pattern of miR2004, we per-
formed Northern blot analysis (Figure 3c) which indi-
cated that miR2004 shared similar expression pattern
with the high throughput sequencing.
The heat responsive long npcRNA TahlnRNA27 con-
tained Ta-miR2010 family sequences, and was up-
regulated in 'TAM107' (heat tolerant cultivar) 1 h after
heat treatment (Figure 2a), whereas Ta-miR2010 was
also statistically up-regulated 1 h after heat stress in the
small RNA databases of 'TAM107' in our previous study
[33]. The secondary structure and the corresponding
expression pattern indicated that TahlnRNA27 might be
the precursor of miR2010. In addition, the powdery
mildew infection responsive long npcRNA TalnRNA5
(Figure 3a) was found to be also responsive to heat
stress and the expression level was increased in 'CS' and
'TAM107' 1 h after heat stress (Figure 2b).
Characterization of putative long npcRNAs for siRNA
We found that 16 out of 71 powdery mildew responsive
long npcRNAs gave rise to small RNAs (Additional file
1), and all of them had similar expression pattern in
microarray analysis and SBS sequencing. Most of these
long npcRNAs produced more than one small RNA
family. For example, TapmlnRNA11 comprised three
small RNA family sequences and each had several mem-
bers (Figure 4). The expression level of TapmlnRNA11
in non-inoculated genotypes was quite low, but accumu-
lated to a high level after powdery mildew infection in
JD8 and JD8-Pm30 12hai (Figure 5a). Consistent with
this expression pattern, its corresponding siRNAs were
also up-regulated after powdery mildew infection (Figure
5b) in both genotypes.
For the heat stress responsive long npcRNAs, there
were nine transcripts matching the small RNAs (Addi-
tional file 1). Among them, TalnRNA21 was responsive
to both heat treated and powdery mildew inoculated
wheat leaves, however, the expression pattern was quite
Figure 1 Expression patterns of wheat long npcRNAs
TapmlnRNA19 (a) and TapmlnRNA30 (b) in response to
powdery mildew inoculation (12hai) as determined by qRT-PCR
analysis, S-0H: before Bgt inoculation in susceptible (S)
genotype, S-12H: 12 hrs after Bgt inoculation in S genotype,
R-0H: before Bgt inoculation in resistant (R) genotype, R-12H:
12 hrs after Bgt inoculation in R genotype.
Figure 2 Expression patterns of wheat long npcRNAs
TahlnRNA27 (a) and TalnRNA5 (b) in response to heat stress.
CS-0h: before heat stress treatment for heat susceptible genotype
Chinese Spring (CS), CS-1h: after 1 hour heat stress treatment, TAM-
0h: before heat stress treatment for heat tolerant genotype TAM107
(TAM), TAM-1h: after 1 hour heat stress treatment.
Xin et al. BMC Plant Biology 2011, 11:61
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different, expression of TalnRNA21 was repressed in
JD8 and JD8-Pm30 12hai (Figure 6a), but up-regulated
after heat stress in 'CS' and 'TAM107' (Figure 6b). We
also noted that TalnRNA21 accumulated to a much
higher expression level 1 h after heat treatment in heat
tolerant cultivar as compared to that in heat sensitive
cultivar (Figure 6b).
Long npcRNAs corresponding to SRP and snoRNAs
We found that 52 powdery mildew infection responsive
and 66 heat stress responsive long npcRNAs could exe-
cute their functions in the form of long molecules,
among which 21 transcripts were responsive to both
stress treatments (Additional file 1). Two transcripts,
TalnRNA9 and TalnRNA12, were identified as signal
recognition particle (SRP) 7S RNA variant 1 and 3,
respectively. It was found that the expression of
TalnRNA9 was increased in both JD8 and JD8-Pm30
genotypes 12 hours after infection (hai) (Figure 7a), but
was repressed 1 h after heat treatment in 'CS' (heat sensi-
tive cultivar) and 'TAM107' (heat tolerant cultivar) (Fig-
ure 7b). Among the 45 long npcRNAs which were only
responsive to heatstress,
TahlnRNA23 and TahlnRNA29) were characterized as
U3 snoRNAs, and their expression levels were increased
1 h after heat stress in both 'CS' and 'TAM107'(Figure 8)
Histone acetylation of TalnRNA5 and TapmlnRNA19
The histone acetylation levels of TalnRNA5 and
TapmlnRNA19 were detected using antibody H3K9 by
ChIP according to the procedure of Lawrence [34].
ChIP analysis indicated that acetylation levels of
TalnRNA5 and TapmlnRNA19 in the inoculated
JD8 and JD8-Pm30 increased as compared to the non-
inoculated controls (Figure 9).
Small RNAs might influence long npcRNAs expression
Based on our analysis, two SRP 7S RNA variants
TalnRNA9 and TalnRNA12 could be regulated by 24 nt
siRNAs. There were five siRNA families complementarily
matching to the long npcRNAs, among which, three
groups (group I, group II, group III) matched both
TalnRNA9 and TalnRNA12, and other two (group IV
group V) were specific for TalnRNA9 (Additional file 6).
We designed gene specific primers (Additional file 7) and
amplified the antisense strand sequences of TalnRNA9
and TalnRNA12 (anti-TalnRNA9 and anti- TalnRNA12).
It was found that expression levels of TalnRNA9 and
TalnRNA12 were up-regulated after powdery mildew
inoculation in the two genotypes (Figure 10a), whereas
both of the antisense sequences were down-regulated
after powdery mildew inoculation in the two genotypes
(Figure 10b), and negative correlation in expression levels
was observed between sense strand and antisense strand
expression patterns in both JD8 and JD8-Pm30 (Figure
10). In addition, three long npcRNAs, TapmlnRNA11,
TapmlnRNA41 and TapmlnRNA42 also had several
group small sequences matching them, and their expres-
sion patterns could be also regulated by siRNAs.
Wheat putative long npcRNAs displayed tissue-specific
expression patterns
To investigate the expression patterns of long npcRNAs
in different wheat tissues, qRT-PCR was performed in 8
Figure 3 Expression pattern of wheat long npcRNA TalnRNA5 and its corresponding miRNA before or 12hai in both disease resistant
genotype (R) and susceptible genotype (S). (a) The expression level of TalnRNA5 as determined by qRT-PCR. (b) The expression pattern of
miR2004 based on high throughput sequencing. (c) Northern blot analysis for miR2004 expression before or 12hai in S genotype and R
Xin et al. BMC Plant Biology 2011, 11:61
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wheat tissues using gene specific primer pairs (Addi-
tional file 7), including leaf, internode, flag leaf, root,
seed, awn, young spike and glume (Figure 11).
It was found that wheat long npcRNAs displayed tis-
sue-specific expression patterns. TapmlnRNA30 was
only detected in seed, whereas TapmlnRNA19 accumu-
lated preferentially in young spike (Figure 11).
TalnRNA5 was expressed in all the tissues, but expres-
sion level was relatively higher in seed as compared to
other tissues (Figure 11). TalnRNA9 was abundantly
Figure 4 The positions of siRNAs matching to the TapmlnRNA11.
Figure 5 Expression patterns of wheat long npcRNAs and their corresponding siRNAs before or 12hai in S genotype and R genotype.
(a) The expression pattern of TapmlnRNA11 in wheat microarray analysis. (b) The abundance of corresponding siRNAs matching TapmlnRNA11
based on high-throughput sequencing.
Xin et al. BMC Plant Biology 2011, 11:61
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expressed in leaf, root and seed, no signal was detected
in other tissues (Figure 11). Interestingly, although both
TalnRNA5 and TapmlnRNA19 gave rise to miR2004,
their expression patterns were obviously different
(Figure 11). In addition, TalnRNA9 was expressed quite
differently between leaf and flag leaf, and the transcripts
accumulated predominantly in leaf (Figure 11).
Experimentally verified full length cDNA of predicted
long npcRNAs
In order to obtain the full length cDNAs corresponding
to the long npcRNAs, we performed 5'RACE for four
long npcRNAs, including TapmlnRNA26, TalnRNA21,
TahlnRNA37 and TahlnRNA47. The cDNA from young
leaf of JD8 was amplified by using gene specific primers
(Additional file 7) and sequenced. The full length
cDNAs corresponding to TapmlnRNA26, TalnRNA21,
TahlnRNA37 and TahlnRNA47 were 1599 bp, 1497 bp,
737 bp and 988 bp in length, respectively. The ORFs of
these sequences were searched by using ORF finder pro-
gram, and no ORFs longer than 80aa was found in these
full length cDNAs (Additional files 8, 9 and 10). For
example, TapmlnRNA26 contained 15 putative ORFs,
but the longest ORF was only 74aa (Figure 12).
Wheat long npcRNAs are not conserved among the plant
species and responsive to both biotic and abiotic stresses
By using combination of microarray and SBS sequen-
cing, a total of 125 putative long npcRNAs were identi-
fied in wheat leaves using strict criteria across a
collection of more than 9700 powdery mildew and 6500
heat stress responsive sequences. Our analysis could fail
to identify the bona fide long npcRNAs in wheat due to
the limited genomic information and gene annotation of
wheat, however, these 125 putative long npcRNAs con-
stituted a reliable set of wheat long npcRNAs. It must
be pointed out that, in the absence of wheat whole
genomic information and the full length sequences of
these wheat long npcRNAs, some of them might turn
out to be protein-coding RNAs when the wheat geno-
mic sequences are available. However, this study repre-
sents the first attempt to characterize the wheat long
npcRNAs and their responses to biotic and/or abiotic
stresses, which could provide a starting point for further
investigation of long npcRNAs in wheat.
As most non-protein coding RNAs were subjected to a
low degree of evolutionary constraint, we found that the
125 long npcRNAs identified in this study had no homo-
logs or significant matches out of plant, animal
and microorganism kingdoms, and were wheat specific
except for two SRP 7SRNA variants (TalnRNA9 and
TalnRNA12) and 3 U3 snoRNAs (TahlnRNA12
TahlnRNA23 and TahlnRNA29), which was in good
agreement to the previous studies in other species such
as Drosophila, Arabidopsis and mouse [12,25,26]. Also,
these long npcRNAs did not appear to form large homo-
logous family. This might suggest that during the evolu-
tion, wheat had developed a batch of specific long
npcRNAs to regulate gene expression and cell activity.
Further analysis revealed that long npcRNAs in wheat
had tissue-specific expression patterns, similar expression
patterns of long npcRNAs were also reported in other
species [24-26]. In our investigation, even in leaf and flag
leaf, TalnRNA9 was differentially expressed, which sug-
gested that long npcRNAs probably had much more pre-
cise expression regulation mechanisms. In addition,
though TalnRNA5 and TapmlnRNA19 gave rise to the
same miRNA, they displayed distinct expression patterns,
indicating that miRNA could potentially be produced by
different precursors in different wheat tissues.
SRP RNA is an exception, as it is a ribonucleoprotein
(protein-RNA complex) that recognizes and targets spe-
cific proteins to the endoplasmic reticulum in eukar-
yotes and the plasma membrane in prokaryotes.
Moreover, U3 snoRNAs predominantly found in the
nucleolus are thought to guide site-specific cleavage of
ribosomal RNA (rRNA) during pre-rRNA processing.
Therefore, they are thought to be conserved across
three kingdoms.
It was reported that increased expression of either
BC200 or an antisense transcript of the b-secretase-1
(BACE1) gene had been implicated in the progression of
Alzheimer's disease [35,36]. Ben et al [18] found that
abiotic stress altered the accumulation of 22 out of the
76 npcRNAs. These indicated that long npcRNAs had
Figure 6 The expression pattern of TalnRNA21 in response to
powdery mildew inoculation (a) and heat stress (b) based on
microarray analysis.
Figure 7 The expression patterns of TalnRNA9 in response to
powdery mildew inoculation (a) and heat stress (b) as
determined by qRT-PCR.
Xin et al. BMC Plant Biology 2011, 11:61
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been linked to biotic and/or abiotic stresses, though in
most instances, evidence had relied on differences in
transcript expression levels between treated and non-
treated samples. Our analysis added further evidence for
the responsiveness of long npcRNAs to both biotic and/
or abiotic stresses, since 71 wheat long npcRNAs were
responsive in defense against powdery mildew infection,
and 77 were responsive to heat stress.
Some of the wheat long npcRNAs are small RNA
Study showed that miR675 was derived from the long
npcRNA H19 which was endogenously expressed in
human keratinocytes and neonatal mice [37], and .
npcRNA78 gene contained the miR162 sequence in an
alternative intron and corresponded to the MIR162a locus
[24]. The 'BIC' noncoding RNA that served as the precur-
sor for miR155 was also readily detectable in vivo as full-
length transcripts [38]. In our identified wheat long
npcRNAs, four transcripts (TalnRNA5, TapmlnRNA8,
TapmlnRNA19, and TahlnRNA27) were characterized as
putative miRNA precursors. Among them, TapmlnRNA8,
TapmlnRNA19 were specific to powdery mildew infection,
while TahlnRNA27 was only responsive to heat stress.
Increasing evidence indicated that miRNAs played impor-
tant roles in plant responses to biotic stresses [9,39,40].
After powdery mildew infection, TalnRNA5 and
TapmlnRNA19 were up-regulated 12hai in JD8 and JD8-
Pm30 genotypes, and their corresponding miR2004 was
also increased in abundance, which strongly indicated that
these two long npcRNAs were processed to miRNAs to
regulate wheat response to powdery mildew infection.
However, as there were no significant expression differ-
ences between the NILs JD8 and JD8-Pm30, we speculated
that these two long npcRNAs functioned as basal defense.
To further confirm this hypothesis, TalnRNA5,
TapmlnRNA19, TalnRNA9 and TapmlnRNA30 were ana-
lyzed using qRT-PCR in 12 hrs after-touched JD8 and
JD8-Pm30 as well as their controls, and their expression
level had no differences between two treatments (data not
show), which suggested that the expression alteration were
caused by powdery mildew infection, not by touching.
In addition, we revealed that 26 wheat long npcRNAs
produced siRNAs and 97 sequences could function in
the form of long molecules involved in wheat resistance
to powdery mildew infection and/or heat stressed. The
collection of long npcRNAs offered candidates for
further analysis of this kind of npcRNAs, which gained
increasing attention in recent years [9,41,42].
Our analysis revealed that two SRP 7S RNA variants
(TalnRNA9 and TalnRNA12) as well as TapmlnRNA11,
TapmlnRNA41 and TapmlnRNA42 could be regulated
by siRNAs. Coram et al [43] reported that the antisense
strands of probe sets Ta.21480 and Ta.24771 (namely,
Figure 8 The expression patterns of TahlnRNA12, TahlnRNA23 and TahlnRNA29 1 h after heat stress in heat sensitive genotype ('CS')
and heat tolerant genotype ('TAM107') based on microarray analysis.
Figure 9 The H3K9 acetylation levels of TalnRNA5 and
TapmlnRNA19 in S and R genotypes before or 12 hrs after
powdery mildew inoculation as determined by qRT-PCR.
Xin et al. BMC Plant Biology 2011, 11:61
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Figure 11 Expression patterns of TalnRNA5, TapmlnRNA19, TalnRNA9 and TapmlnRNA30 in eight tissues as determined by qRT-PCR.
Figure 10 Expression patterns of sense and antisense sequences for TalnRNA9 and TalnRNA12 before or 12 hrs after Bgt inoculation
in S genotype and R genotype. (a) Expression patterns of sense sequences revealed by microarray analysis. (b) Expression patterns of antisense
sequences as determined by qRT-PCR.
Xin et al. BMC Plant Biology 2011, 11:61
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TalnRNA9 and TalnRNA12) were expressed using
wheat Affymetrix genome array, which was in good
agreement with our experimental results. And interest-
ingly, our analysis also shown that the expression pat-
terns of antisense had negative correlations with sense
sequence for both TalnRNA9 and TalnRNA12, which
strongly indicated that the siRNAs generated from anti-
sense strands might regulate expression of their corre-
sponding sense strands. Collectively, this study indicated
that expression of wheat long npcRNAs might be regu-
lated by other non-protein coding RNAs, as was the
case for Xist gene [44].
In summary, by using computational analysis and experi-
mental approach, for the first time, we identified 125
putative wheat long npcRNAs. These identified wheat
long npcRNAs were not conserved among plant species,
and some of them were small RNA precursors. Wheat
long npcRNAs showed a tissue dependent expression
patterns and their expressions were responsive to pow-
dery mildew infection and/or heat stress, suggesting that
they could play roles in development and regulation of
biotic and/or abiotic stresses. Our analysis also indicated
that expressions of some wheat long npcRNAs could be
regulated by small RNAs and through histone acetylation,
but this need further investigation. The identification and
expression analysis of wheat long npcRNAs in this study
would provide a starting point to understand their func-
tions and regulatory mechanisms in the future.
Plant materials
Seeds of powdery mildew susceptible wheat cultivar 'JD8'
(designated as S) and its near isogenic line carrying a pow-
dery mildew resistance gene Pm30 (designated as R) were
planted in 8-10 cm diameter pots. Seedlings were
artificially inoculated when the first leaf was fully
expanded, with a local prevalent Blumeria. graminis f. sp.
tritici isolate E09. Inoculation was performed by dusting
or brushing conidia from neighboring sporulating suscep-
tible seedlings onto the test seedlings. Leaf samples were
collected from both genotype at 0 and 12 hrs post inocula-
tion (designated as S-0 h, S-12 h, R-0 h, R-12 h), respec-
tively, and frozen in liquid nitrogen and used for RNA
For heat stress, heat tolerant genotype 'TAM107' was
used in this study. Seeds were surface-sterilized in 1%
sodium hypochlorite for 15 min, rinsed in distilled
water, and soaked in dark overnight at room tempera-
ture. The germinated seeds were transferred into the
pots (25 seedlings per pot) containing vermiculite. The
treatments were carried out as described by Qin et al
[32]. Leaves were collected at 0 and 1 hour after heat
treatment (designated TAM-0 h and TAM-1 h) At the
end of heat treatments, the leaves were frozen in the
liquid nitrogen immediately, and then stored at -80°C
for further use.
Microarray analysis
Total RNA was extracted using Trizol reagent (Invitro-
gen) following the manufacture's recommendations.
Briefly, mRNA was enriched from 80~90 μg total RNA
using the RNeasy Plant Mini Kit (QIAGEN) according
to the protocol, and was subsequently reverse-
transcribed to double stranded cDNA using the Gene-
Chip®Two-Cycle cDNA Synthesis Kit. The biotin
labeled cRNA was made using the GeneChip®IVT
Labeling Kit (Affymetrix, CA, USA). Twenty micrograms
of cRNA samples were fragmented and hybridized for
16 h at 45°C to the Affymetrix Wheat Genome Array
(Santa Clara, CA, USA). After washing using the Gene-
chip®Fluidics Station 450, arrays were scanned using
the Genechip®3000 Scanner that is located in
Figure 12 The 15 short possible open reading frames (ORFs) positioned in TapmlnRNA26.
Xin et al. BMC Plant Biology 2011, 11:61
Page 9 of 13
Page 10
Bioinformatics Center at China Agriculture University
(NCBI accession Number: GSE27339 http://www.ncbi.
Small RNA sequencing
Small RNA libraries (S-0 h, S-12 h, R-0 h, R-12 h,
TAM-0 h, TAM-1 h) preparation and sequencing were
performed with Solexa sequencing technology (BGI,
Shenzhen, China) as described by Sunkar et al [45].
Automated base calling of the raw sequence and vector
removal were performed with PHRED and CROSS
MATCH programs [46,47]. Trimmed 3' and 5' adapters
sequences, removed RNAs less than 17 nt and polyA,
only sequences longer than 17 nt with a unique ID were
Figure 13 Schematic representation of computational method for long npcRNA identification.
Xin et al. BMC Plant Biology 2011, 11:61
Page 10 of 13
Page 11
used for further analysis. We calculated sequencing fre-
quency of each small RNA sequence, the number of
reads for each sequence reflecting relative abundance
(NCBI accession Number: GSE27339, http://www.ncbi.
Computational methods for long npcRNA identification
Firstly, powdery mildew infection and heat stress
responsive transcripts were annotated by Harvest pro-
gram, and protein-coding genes are discarded, the
reminding sequences were analyzed by Blastx and
Blastn, and ESTs with no similarity to protein coding
genes or tRNA and rRNA were retained. Secondly, we
screened the reminding ESTs, and the sequences with
polyA-tail were selected. Thirdly, we predicted the long-
est ORF of these transcripts using ORF finder, and the
sequences with no or short ORFs (less than 80aa) were
retained for further analysis (Figure 13).
Chromatin-Immunoprecipitation (ChIP) assays
ChIP was modified from published protocol [34]. Approxi-
mately 1 g of leaves was used for each ChIP assay. The
fresh tissues were subjected to vacuum infiltration in for-
maldehyde (1%) solution for cross-linking the chromatin
proteins to DNA. Chromatin was extracted and sonicated
for 4 × 10 sec pulses, 40% duty cycle and 20% power with
chilling on ice for 1 min after each pulse. The average size
of the resulted DNA fragments ranged between 0.2~2.0-
kb, centering around 500 bp. An aliquot of chromatin
solution (1/10 of total volume) was used to determine the
DNA fragment sizes and serve as input control. The
remaining chromatin solution was diluted 10-fold and
divided into two aliquots. One aliquot was incubated by
adding 10 μl of antibodies (anti-acetyl-histone H3K9,
Upstate Biotechnology, NY). The other aliquot was incu-
bated without antibodies (mock). After incubation at 4°C
with rotation for overnight, the solution was added to
40 μl of protein A agarose and incubated for another
1 hours. The immunocomplexes were eluted and cross-
links were reversed by incubation at 65°C for 15 min.
Residual protein was degraded by proteinase K and DNA
was extracted and dissolved in 50 μl of ddH2O.
Quantitative Real-Time PCR (qRT-PCR) Analysis
qRT-PCR was performed using the ChIP DNA or cDNA
samples in a 10-μl mixture containing 1× LightCycler-
FastStart DNA master SYBR Green I. According to
MIQE guidelines, qRT-PCR was performed as follows:
initial denaturation for 10 min at 95°C, followed by 40
cycles of 30 s at 95°C, 45 s at 55 to 60°C, 10 s at 72°C,
and 72°C for 5 min as the last step. The threshold cycles
(Ct) of each test target were averaged for triplicate reac-
tions and the values were normalized according to the
Ct of the control products (Ta-actin).
Additional material
Additional file 1: Wheat long npcRNAs responsive to powdery
mildew infection and/or heat stress. The table includes lnpcRNAs' ID,
probe set ID, the longest ORF, the number of putative ORFs and
corresponding siRNAs.
Additional file 2: The hairpin structure of putative TahlnRNA27. The
figure shows the secondary structure of putative wheat long npcRNA
TahlnRNA27 by using the Vienna RNA package RNAfold web interface
program, the perfect hairpin structure indicates that it might give rise to
Additional file 3: The hairpin structure of putative TalnRNA5. The
figure shows the secondary structure of putative wheat long npcRNA
TahlnRNA5 by using the Vienna RNA package RNAfold web interface
program, the perfect hairpin structure indicates that it might give rise to
Additional file 4: The hairpin structure of putative TalnpmRNA8. The
figure shows the secondary structure of putative wheat long npcRNA
TahlnRNA8 by using the Vienna RNA package RNAfold web interface
program, the perfect hairpin structure indicates that it might give rise to
Additional file 5: The hairpin structure of putative TalnpmRNA19.
The figure shows the secondary structure of putative wheat long
npcRNA TahlnRNA19 by using the Vienna RNA package RNAfold web
interface program, the perfect hairpin structure indicates that it might
give rise to miRNA.
Additional file 6: Categories of siRNAs corresponding to SRP1 and
SRP3 7S RNA variants and sequences of SRP1 and SRP2
corresponding siRNAs. (a) The siRNAs corresponding to SRP1 and SRP3
7S RNA variants are categorized to 5 groups according to their locations,
most members of group I, II, and III match both TalnRNA9 and
TalnRNA12, and other two (group IV group V) are specific for TalnRNA9.
(b) The table includes sequences of of SRP1 and SRP3 corresponding
Additional file 7: Primer sequences for 5'RACE and real time PCR.
The table displays the sequences of primers used for both 5'RACE and
real time PCR
Additional file 8: The short possible ORFs in TalnRNA21. The figure
displays all the possible ORFs in the full length cDNA of TalnRNA21, and
none of them are longer than 80aa.
Additional file 9: The short possible ORFs in TahlnRNA37. The figure
displays all the possible ORFs in the full length cDNA of TalnRNA37, and
none of them are longer than 80aa.
Additional file 10: The short possible ORFs in TahlnRNA47. The
figure displays all the possible ORFs in the full length cDNA of
TalnRNA47, and none of them are longer than 80aa.
This work was financially supported by National Basic Research Program of
China (2007CB109000), 863 Project of China (2007AA10Z138, 2006AA10A104)
and National Natural Science Foundation of China (30871529, 30871528).
Author details
1State Key Laboratory for Agrobiotechnology and Key Laboratory of Crop
Heterosis and Utilization (MOE) and Key Laboratory of Crop Genomics and
Genetic Improvement (MOA), Beijing Key Laboratory of Crop Genetic
Improvement, China Agricultural University, Beijing, 100094, PR China.
2National Plant Gene Research Centre (Beijing), Beijing 100094, PR China.
3Department of Plant Genetics & Breeding, China Agricultural University,
Yuanmingyuan Xi Road No. 2, Haidian District, Beijing, 100193, PR China.
Authors' contributions
MX, YW, YY and DQ carried out the microarray analysis and small RNA
sequencing, participated in the long npcRNA identification, and draft the
manuscript. MX and ZH carried out ChIP analysis, NS, CX and HP carried out
Xin et al. BMC Plant Biology 2011, 11:61
Page 11 of 13
Page 12
the qRT-PCR analysis. ZN and QS carried out the design of the study and
finish the manuscript. All authors read and approved the final manuscript.
Received: 21 October 2010 Accepted: 7 April 2011
Published: 7 April 2011
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the rice genome. PLoS One 2007, 2:e294.
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Cite this article as: Xin et al.: Identification and characterization of
wheat long non-protein coding RNAs responsive to powdery mildew
infection and heat stress by using microarray analysis and SBS
sequencing. BMC Plant Biology 2011 11:61.
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text:
Feb. 23 2007
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subset: commoncrawl


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Pound's Cavalcanti: |
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Message board »Message Board home »Sign-in or register to get started
Online now: 1 member: CRB6970; 48 anonymousDiscussion: FELIZ NAVIDAD Y PROSPERO A~O NUEVO
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NPY and galanin in a hibernator: hypothalamic gene expression and effects on feeding. | |
MedLine Citation:
PMID: 7693306 Owner: NLM Status: MEDLINE |
Neuropeptides such as neuropeptide Y (NPY) and galanin may play a role in regulating the pronounced seasonal changes in food intake shown by golden-mantled ground squirrels (Spermophilus saturatus). We used in situ hybridization histochemistry to localize the expression of NPY and galanin mRNA in the hypothalamus of normally feeding animals. NPY mRNA was abundantly expressed in the arcuate nucleus, while galanin mRNA was concentrated in both the arcuate nucleus and the dorsomedial nuclei. When NPY (0.1, 0.5, 2, and 8 micrograms) or galanin (0.1, 0.5, 2, and 8 micrograms) were injected into the third cerebral ventricle, food intake was significantly and dose-dependently increased over the subsequent 30 min. NPY stimulated significant increases in food intake for up to 2 h whereas galanin's effect did not extend beyond 30 min. Our results suggest that hibernating and nonhibernating rodents share common neural substrates for the regulation of food intake. Seasonal modulation of these neural pathways may contribute to annual cycles of food intake in hibernating mammals. |
T Boswell; R D Richardson; M W Schwartz; D A D'Alessio; S C Woods; A J Sipols; D G Baskin; G J Kenagy |
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Publication Detail:
Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
Title: Brain research bulletin Volume: 32 ISSN: 0361-9230 ISO Abbreviation: Brain Res. Bull. Publication Date: 1993 |
Date Detail:
Created Date: 1993-12-03 Completed Date: 1993-12-03 Revised Date: 2007-11-14 |
Medline Journal Info:
Nlm Unique ID: 7605818 Medline TA: Brain Res Bull Country: UNITED STATES |
Other Details:
Languages: eng Pagination: 379-84 Citation Subset: IM |
Department of Zoology, University of Washington, Seattle 98195. |
Export Citation:
APA/MLA Format Download EndNote Download BibTex |
MeSH Terms | |
Analysis of Variance Animals Arcuate Nucleus / metabolism Base Sequence Cerebral Ventricles / drug effects, physiology* Dorsomedial Hypothalamic Nucleus / metabolism Eating / physiology* Feeding Behavior / drug effects* Galanin Gene Expression* Hibernation / physiology* Hypothalamus / cytology, metabolism* In Situ Hybridization Molecular Sequence Data Neuropeptide Y / administration & dosage, biosynthesis*, pharmacology* Neuropeptides / biosynthesis* Oligonucleotide Probes Peptide Biosynthesis* RNA, Messenger / biosynthesis Sciuridae / physiology* |
Grant Support | |
Chemical | |
Reg. No./Substance:
0/Neuropeptide Y; 0/Neuropeptides; 0/Oligonucleotide Probes; 0/RNA, Messenger; 88813-36-9/Galanin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Afferent projections to the reticular thalamic nucleus from the globus pallidus and the substantia n...
Next Document: Muscarinic agonists have no measurable effect on cGMP formation of rat pinealocytes.


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Contact and additional information for this author • All NBER papers and publications • NBER Working Papers only
Published: Self-Reported Disability and Reference Groups, Arthur van Soest, Tatiana Andreyeva, Arie Kapteyn, James P. Smith. in Investigations in the Economics of Aging, Wise. 2012
Published: Disability Insurance and Labor Market Exit Routes of Older Workers in the Netherlands, Klaas de Vos, Arie Kapteyn, Adriaan Kalwij. in Social Security Programs and Retirement around the World: Historical Trends in Mortality and Health, Employment, and Disability Insurance Participation and Reforms, Wise. 2012
5. Brown, J.R., Kapteyn, A., and Mitchell, O., "Framing and Claiming: How Information-Framing Affects Social Security Claiming Behavior", The Journal of Risk and Insurance, 2014, forthcoming .
Published: Work Disability, Work, and Justification Bias in Europe and the United States , Arie Kapteyn, James P. Smith, Arthur van Soest. in Explorations in the Economics of Aging, Wise. 2011
Published: " The Effects of Lottery Prizes on Winners and Their Neighbors : Evidence from the Dutch Postcode Lottery" (with Peter Koorem a n, Adriaan Soetevent, and A rie Kapteyn) American Economic Review 101 (5) (August 2011). pp 2226 - 2247
Published: Labor Market Status and Transitions during the Pre-Retirement Years: Learning from International Differences, Arie Kapteyn, James P. Smith, Arthur van Soest, James Banks. in Research Findings in the Economics of Aging, Wise. 2010
Published: James Banks & Arie Kapteyn & James P. Smith & Arthur van Soest, 2009. "Work Disability is a Pain in the ****, Especially in England, the Netherlands, and the United States," NBER Chapters, in: Health at Older Ages: The Causes and Consequences of Declining Disability among the Elderly National Bureau of Economic Research, Inc.
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Map/Directions: 1050 Massachusetts Ave., Cambridge, MA 02138
ph: 617-868-3900 email: email@example.com
ph: 617-868-3900 email: firstname.lastname@example.com


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Key things you need to know aboutDr. Shugart:
Dr. Shugart's Experience
Research training, expertise and qualifications
Dr. Shugart's Specialties
- Orthopedic Surgery
- View all 2 specialties and 1 board certification
Procedures Dr. Shugart Performs
- Disc Replacement
- Dressing and-or Debridement of Wound, Infection, or Burn (incl. Negative Pressure Wound Therapy)
- See all 15 procedures
Conditions Dr. Shugart Treats
- Adhesive Capsulitis
- Bursitis
- See all 49 conditions
More About Dr. Shugart's Background
Hospital Quality
The right hospital is as important as the right doctor
- Orthopedic Hospital of the Lutheran Health Network
- 7952 West Jefferson Boulevard
- Fort Wayne, IN 46804
- Lutheran Hospital of Indiana
- 7950 West Jefferson Boulevard
- Fort Wayne, IN 46804
9 Facts About Hospital Quality
Patients' risk of dying during a hospital stay for heart bypass surgery could be 85.2% lower, on average, at a hospital rated 5 stars compared to a hospital rated 1 star for that procedure. Choosing the right hospital is critical to your health. View Slideshow
Patient Satisfaction
Patients' feedback on their experience with Dr. Shugart
Likelihood of recommending Dr. Shugart to family and friends is 3.8 out of 5
See More Survey Results
8 Facts About Patient Satisfaction Surveys
Patient satisfaction surveys provide a standardized way of reporting about healthcare experiences. But there can be more than meets the eye when reviewing scores. Here's what you should know
Appointments & Offices
Check insurance plans, locations and make an appointment
Accepting new patients?
- Yes
- Fort Wayne Orthopedics LLC
- Fort Wayne Office
- 7601 W Jefferson Blvd
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7 Tips to Prepare for Your Appointment
The key to a successful appointment is communication. To get the most out of your visit, prepare ahead of time so you can lead the conversation with your doctor. These 7 tips will get you ready


text:
Pulmonary arteriovenous fistula showing a fall in shunt fraction during exercise. | |
MedLine Citation:
PMID: 6705589 Owner: NLM Status: MEDLINE |
A 23-year-old man with pulmonary arteriovenous fistulas of the right middle lobe is described. During the incremental exercise test, the shunt fraction dropped from 19 percent to 12 percent as the cardiac output increased. We discuss the mechanism of this fall in shunt fraction in this patient during exercise. |
H Maeda; Y Monden; K Nakahara; S Miyoshi; Y Kawashima |
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Publication Detail:
Type: Case Reports; Journal Article |
Journal Detail:
Title: Chest Volume: 85 ISSN: 0012-3692 ISO Abbreviation: Chest Publication Date: 1984 Apr |
Date Detail:
Created Date: 1984-05-07 Completed Date: 1984-05-07 Revised Date: 2004-11-17 |
Medline Journal Info:
Nlm Unique ID: 0231335 Medline TA: Chest Country: UNITED STATES |
Other Details:
Languages: eng Pagination: 575-7 Citation Subset: AIM; IM |
Export Citation:
APA/MLA Format Download EndNote Download BibTex |
MeSH Terms | |
Adult Arteriovenous Malformations / physiopathology* Cardiac Output Exercise Test Humans Male Oxygen / blood Pulmonary Artery* Pulmonary Veins* |
Chemical | |
Reg. No./Substance:
7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Metastatic renal cell carcinoma simulating sarcoidosis. Analysis of 12 patients with bilateral hilar...
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A quest to understand how memory works: interview with neuroscientist Eric R. Kandel
At 82, the Nobel Prize-winning neuroscientist Dr. Eric R. Kandel is still constantly coming up with new ideas for research.
This winter, he has been working on a project that he hopes will lead to a new class of drugs for treating schizophrenia. Last year he collaborated, for the first time, with Denise B. Kandel — his fellow Columbia University research scientist and wife of 55 years — investigating the biological links between cigarette and cocaine addiction. And this month his newest book, "The Age of Insight: The Quest to Understand the Unconscious in Art, Mind and Brain, From Vienna 1900 to the Present," is to be released by Random House.
A condensed and edited version of our two interviews follows. As in his new book, the conversation begins with memories of Vienna, his birthplace.
How old were you when the Nazis marched into Vienna?
I was 8 ½. Immediately, we saw that our lives were in danger. We were completely abandoned by our non-Jewish friends and neighbors. No one spoke to me in school. One boy walked up to me and said, "My father said I'm not to speak to you anymore." When we went to the park, we were roughed up. Then, on Nov. 9, 1938, Kristallnacht, we were booted out of our apartment, which was looted. We knew we had to get out.
Fortunately, my mother had the foresight to apply for visas to the United States earlier. For more than a year, we waited in the terror of Vienna for our immigration quota number to come up. When it finally did, my older brother, Ludwig, and I made the Atlantic crossing alone. Our parents came later. On the trip, it's amazing how unfrightened I was, considering that even before the Nazis, I was an apprehensive child. You rise to the occasion....
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- Historians named to the 2015 class of the American Academy of Arts and Sciences


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Quick Links |
Information for: For the Media: |
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The Summer Fast Track Institute is an intensive three week academic-skill building program offered to help students achieve"college ready" status and register for college level courses. Students testing into one or more developmental courses and meeting program criteria are invited to participate.
Participants receive the following incentives:
* Weekly stipends to cover food & transportation
* Daily instruction in the necessary area and "College Success Skills" classes aimed at preparing students for on-time graduation
* Access to online tutorials
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* An opportunity to explore the campus and make new friends prior to the start of classes
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Page maintained by University College
Last updated or reviewed on 3/19/15


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There is not much to tell about me, but my name is Jacob Everhart. I like that name when I study more on Jacob in the Bible. But any way Christ has been my everything from birth, and he is still making me into what he wants me to become in Him.
Please add a name and description in order to save your playlist to your user profile.


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Again sorry pal, This is what Wallace said "Ball when it came out of the hands kind of looked like it was going to the right so I turned that way. then it was going back to the left so I turned around and tried to find the ball I just didn't make the play. " He clearly stated he misjudged the direction of the ball when it came out of Tannehills hands.
Still with all that said and done the ball still went through Wallaces hands. That is all on Wallace.


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Rep. Bob Franks
Former Representative from New Jersey's 7th District
Elected Positions
Dates | Title | State / District |
Representative | New Jersey's 7th District |
See Also: Bioguide
Sponsorship Analysis
Our unique analysis of the bills Franks sponsored and cosponsored provides insight into his position in the House of Representatives.
Each dot in the chart below is a member of the House of Representatives. The dots are positioned horizontally according to our progressive—conservative ideology score and vertically according to our leadership score (leaders toward the top). Franks is shown as a purple triangle. (analysis methodology)
Bills Sponsored
Some of Franks's most recently sponsored bills include...
- H.Res. 623 (106th): Regarding the adoption of Resolution 1322 by the Security Council of the ...
- H.R. 5252 (106th): Release Our Reserve Now Act
- H.R. 5227 (106th): 21st Century Community Learning Centers Act
- H.R. 4849 (106th): Pipeline Safety Improvement Act of 2000
- H.R. 4519 (106th): Baylee's Law
- H.R. 4433 (106th): Charitable Contribution Tax Relief Act of 2000
- H.R. 4076 (106th): Cybermolesters Enforcement Act of 2000
View All » (including bills from previous years)
Voting Record
From Jan 1993 to Dec 2000, Franks missed 245 of 4,863 roll call votes, which is 5.0%. This is worse than the median of 3.0% among the lifetime records of representatives serving in Dec 2000. The chart below reports missed votes over time.
Primary Sources
The information on this page is originally sourced from a variety of materials, including:
- The House and Senate websites, for committee membership and voting records
- Congressional Biographical Directory for elected positions
- Congressional Pictorial Directory for the photo
- THOMAS, for sponsored bills


text:
It took many years of intense work
and development to make it: The first 100% biodegradable, high-performance baby
wipe. Naty by Nature Babycare Baby Wipes perform as well as the best
?traditional? wipes, and are more gentle to your baby.
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Aqua (Water), Glycerin, Benzyl Alcohol, Sodium Benzoate, Propylene Glycol, Potassium Sorbate, Dehydroacetic Acid, Decyl Glucoside, Citric Acid, Chamomilla Recutita Flower Extract, Aloe Barbadensis Leaf Juice.


text:
SAN RAFAEL, Calif. — Serial murder suspect Joseph Naso delivered an hours-long personal history Monday replete with childhood photos, as he launched his defense, denied the decades-old slayings of four women and claimed he is not the "monster" prosecutors have made him out to be.
The 79-year-old defendant's opening statement came after prosecutors spent the morning showing the jury graphic images of the four women's bodies discovered in Northern California, leading some on the panel to wipe tears from their eyes.
Naso, wearing a dark suit and spectacles, rose after Marin County prosecutor Rosemary Slote had called him a "serial rapist and murderer" and said he was anxious to tell his side.
"I've been waiting two years and two months for this day, to tell my side of the story," Naso told the panel.
"I'm not the monster they say killed these women," he said. "I don't kill people and there's no evidence of that in my writings and photography."
Naso, has pleaded not guilty to four counts of first-degree murder with special circumstances for the slayings of four women — all prostitutes with matching initials: 18-year-old Roxene Roggasch in 1977; 22-year-old Carmen Colon in 1978; 38-year-old Pamela Parsons in 1993; and 31-year-old Tracy Tafoya in 1994.
Whether the "double initials" in each victim's name was a coincidence or a plan, investigators have not said. Prosecutors are seeking the death penalty.
Prosecutors say Naso drugged and photographed his unconscious victims, then strangled them and dumped their naked bodies in rural areas.
Slote read from sections of a diary found at Naso's home that detailed rapes of women.
In a 1961 entry, the journal describes a man picking a girl up and raping her in a car in the Berkeley Hills. Other entries made vague references to victims in this case.
Naso was arrested at the time on suspicion of assault, but he said he was never charged. Prosecutors say the woman named in that entry will testify about the incident.
Authorities around the country have also looked at Naso as a suspect in cold cases.
Marin County prosecutors have built a significant case against Naso.
Investigators discovered DNA matching Naso's profile on at least one victim, Roggasch, and a partial DNA match from material collected from under the fingernails of Colon.
Naso said he plans to challenge that, saying the DNA is inconclusive.
Also discovered were photographs — including images of at least one of the victims in the case — of women who appeared dead or unconscious and what prosecutors called a "rape journal" during a search of Naso's Reno, Nev., house.
Naso characterized the photographs as his art and said all of his "models" were willing participants.
He showed the jury on Monday dozens of photographs he took of weddings, landscapes and family pictures along with what he called "glamour" or "cheesecake" photographs of nude women. He said he never forced any of them to do anything.


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Belle Epoque
Simple luxury at its best, this Euro sham is rendered in an incredibly soft 420-thread count Egyptian cotton in a warm taupe hue.
Elegantly running the gamut from clean, crisp contemporary design to classic traditional styles, Belle Epoque offers superlative materials, super-luxurious quality and beautiful design. Each piece is subjected to rigorous testing, so you can rest easy that your beautiful bedroom investment will also be an enduring one.


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Goal-directed Resuscitative Interventions During Pediatric Inter-facility Transport (The GRIPIT Trial)
Recruitment status was Recruiting
- Full Text View
- Tabular View
- No Study Results Posted
- Disclaimer
- How to Read a Study Record
Recent advances in critical care medicine have dramatically improved morbidity and mortality of critical illness. Goal-directed therapy protocols have been instrumental in this change. Goal-directed therapy standardizes the rapid delivery of definitive care in illnesses such as SIRS (Systemic Inflammatory Response Syndrome) and head trauma. Although this treatment approach has been shown to improve clinical outcomes, it has not been widely adopted outside academic medical centers. Further improvement in outcomes of critical illness is likely if goal-directed therapy is utilized early in the course of care. To facilitate this early adoption, goal-directed therapeutic protocols should be developed and implemented by specialized pediatric transport teams. The investigators hypothesize that the institution of goal-directed therapy during pediatric interfacility transport will improve the outcomes of critically ill children.
The GRIPIT Trial (Goal-directed Resuscitative Interventions during Pediatric Inter-facility Transport) will compare outcomes of pediatric SIRS patients before and after the implementation of a goal-directed therapeutic protocol during transport. This will be the first test of goal-directed therapy in the transport environment. Data will be collected on pediatric SIRS patients transported by the Angel One Transport Team at Arkansas Children's Hospital before and after protocol implementation. Outcome measures will include length of hospital stay, length of intensive care unit (ICU) stay, incidence of multiple organ dysfunction syndrome (MODS), and required therapeutic interventions during ICU stay (TISS-28 scores). In addition, NIRS (Near-Infrared Spectroscopy) monitoring will be used as a cerebral and somatic oxygenation trend monitor, to determine its effectiveness as a resuscitation guide for pediatric SIRS during transport. NIRS trends are useful as a surrogate marker for systemic venous saturations, known to decrease with severe SIRS.
Condition | Intervention |
SIRS (Systemic Inflammatory Response Syndrome( |
Procedure: Goal-directed Therapeutic Protocol |
Study Type: | Interventional |
Study Design: | Allocation: Non-Randomized Intervention Model: Single Group Assignment Masking: Open Label Primary Purpose: Treatment |
Official Title: | Goal-directed Resuscitative Interventions During Pediatric Inter-facility Transport (The GRIPIT Trial) |
- Incidence of Multiple Organ Dysfunction [ Time Frame: ICU Admission to ICU Discharge ] [ Designated as safety issue: No ]Computed utilizing PELOD scores.
- Required ICU Interventions [ Time Frame: ICU Admission to ICU Discharge ] [ Designated as safety issue: No ]Utilizing TISS-28 Scores
- Hospital Length of Stay [ Time Frame: Hospital Admission to Hospital Discharge ] [ Designated as safety issue: No ]Subjects will be followed from hospital admission to hospital discharge.
- Changes in cerebral and somatic oxygenation [ Time Frame: During Inter-facility Transport (Average) ] [ Designated as safety issue: No ]Utilizing NIRS Technology
Estimated Enrollment: | 200 |
Study Start Date: | October 2010 |
Estimated Study Completion Date: | September 2012 |
Estimated Primary Completion Date: | September 2012 (Final data collection date for primary outcome measure) |
Procedure: Goal-directed Therapeutic Protocol
Ages Eligible for Study: | 1 Month to 17 Years |
Genders Eligible for Study: | Both |
Accepts Healthy Volunteers: | No |
Inclusion Criteria:
- Pediatric patients transported to Arkansas Children's Hospital by the Angel One Transport Team
- Age Group: 1 month - 17 years
- Systemic Inflammatory Response Syndrome (Age-specific SIRS criteria)
Exclusion Criteria:
- Known or suspected congenital heart disease
- Known or suspected cardiomyopathy or myocarditis
- Diabetic Ketoacidosis (DKA) as SIRS-inducing illness
- Traumatic Brain Injury (TBI) as SIRS-inducing illness
- Burn Injury as SIRS-inducing illness
- Patients with known or suspected active hemorrhage
Please refer to this study by its ClinicalTrials.gov identifier: NCT01293500
Contact: Michael H Stroud, MD | 501-364-1861 | email@example.com |
United States, Arkansas | |
Arkansas Children's Hospital | Recruiting |
Little rock, Arkansas, United States, 72202 | |
Contact: Michael H Stroud, MD 501-364-1861 firstname.lastname@example.com | |
Principal Investigator: Michael H Stroud, MD |
Principal Investigator: | Michael H Stroud, MD | Arkansas Children's Hospital Research Institute |
No publications provided
Responsible Party: | Arkansas Children's Hospital Research Institute |
ClinicalTrials.gov Identifier: | NCT01293500 History of Changes |
Other Study ID Numbers: | 1R21HD060171-01A2 |
Study First Received: | October 22, 2010 |
Last Updated: | January 18, 2012 |
Health Authority: | United States: Federal Government |
Keywords provided by Arkansas Children's Hospital Research Institute:
SIRS shock sepsis pediatric |
inter-facility transport specialized transport NIRS monitoring |
Additional relevant MeSH terms:
Systemic Inflammatory Response Syndrome Inflammation Pathologic Processes Shock |
ClinicalTrials.gov processed this record on April 26, 2015


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We've heard your feedback and are continuing to build a better Slickdeals. Click here to check out the updated site.
I managed to enter my billing address incorrectly and I have not been able to put in an order for the past 2 days and no reply to any of my inquiries. How did you get in touch with anyone, is there a number I can call? I fear I will miss this promo because noone is getting to my emails in time
check the status of your order by phone by calling 425.861.9100 |
playseat arrived today, assembled.
Fanatec deals are far and few in between so I just bought the XL pack http://us.fanatec.com/index.php?r...u you save a little bit. I decided not to go for the elite one after hearing about issues and shoddy customer service. Not something you want to deal with when you spend 550 on a wheel.... Also to put the shifter in the proper place u need to buy the playseat shifter and an adapter from germany for the fanatec shifter. It's a pity u have to spend that $33 extra |
Got my setup installed. I can say that this setup is many times better than my old wheel stand. Seating position is a big deal. And mostly finally playing forza with a wheel is amazing
One thing is that while these come with the adapter plate to work with the Fanatec CSR Wheels, the screws included do not work as they are too short (20mm) I just picked up 2 30mm m6 screws at home depot and that fixed the problem. Also I have some concern with the pedal mount with the CSR Elite Pedals. They only mount 2 holes instead of all 4. This leaves some flex in the middle which looks dangerous over time. I wish they had an adapter plate or something. |
So how does everybody find driving with a pole between their legs? I don't mean- well, you know what I mean.
I don't usually pay attention while driving a real car but I do now, and I noticed my legs are often very close together, especially when braking, so it seems it would be awkward to have a pole in between there. A steel pole, I mean. And is it just me, or is not providing long enough screws in a product that normally sells for $550 really messed up? How hard is it? I mean, you can get a new 64GB iPad 3 for that. This is metal, screws and a seat. Last edited by dealstorm; 12-13-2012 at 11:56 PM.. |
Why did this get moved to dealtalk? The deal is all the way through December ........................
Anyway, the pole isn't too bad in this one. it was a bigger issue in the older playseat model and especially bad in wheelstands. The screws issue is pretty annoying if you ask me. There aren't that many wheels for 360 and you would expect them to be a bit more diligent if they're going to put the forza branding on there that they make it compatible with forza branded wheels. But fixed with $1 so I don't mind in the long run given the price with this deal |
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High Rise Cat
I'm 3 today!October 12th 2013 8:34 am[ View A Comments (1) ] Today I'm 3 years old! And mom's birthday is in 4 more days, what fun! Next year we should be equal in age according to the age chart. Happy Birthday Dusty! xoxo Kaci |
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The Shadow Collectors Edition Blu-ray Review (Mania.com)
Tim Janson
Review Date:
Monday, March 10, 2014
When I look back at the 1994 film, The Shadow I do so with one part fondness and three parts disappointment over what could have been. As a fan of The Shadow for many years I was especially enthusiastic about the production since it had been nearly 50 years since the character last appeared on screen in a "B" movie The Shadow Returns. On paper the film had a lot going for it…Alec Baldwin in the title role was one of Hollywood's biggest starts of 1990s and was surrounded by a cast featuring John Lone, Penelope Ann Miller, Tim Curry, Peter Boyle, and a pre X-Men and Lord of the Rings Ian McKellan. It also had a screenplay written by David Koepp who had previously written successful films such as "Jurassic Park", "Carlito's Way", and "The Paper". Yet despite all that it had going for it The Shadow failed to connect with fans or critics. While the film made back (barely) its $40 million dollar budget, Universal films considered it a disappointment and a potential franchise never materialized.
Now, nearly 20 years after its theatrical release Shout Factory delivers a new Collector's Edition Blu-Ray featuring a 1080p High-Definition Widescreen transfer and new bonus material. Well, calling this a Collector's Edition might be somewhat of an ambitious title. The only bonus material included other than the original trailer and still gallery is a new interview with the director and cast which we will talk about later. The HD transfer looks gorgeous but the film always did look great and it is only 20 years old.
Still the cinematography and set design are the film's strongest points. The look and feel of the glamorous 1930's is perfectly captured with his glitzy nightclubs, women dressed in flowing gowns and men in tuxedos. The streets, the set pieces all help bring the pulp hero's era to life. Koepp's script build's a backstory to Lamont Cranston, aka The Shadow showing us his dark background and how he is able to know the "evil that lurks in the heart of men". The Shadow's first appearance, rescuing a man from being murdered by a gang of mobsters and using his signature twin .45s, was one of the highlights of the film.
Unfortunately, things would derail from there. The Shadow's rival is Shiwan Khan (Lone) a descendent of Genghis Khan who has same mental/supernatural abilities as the Shadow does. Mulcachy tries to hit a grand slam with no runners on base however. Rather than keeping it simple he goes overboard with a plot involving Khan's plan for world domination and kidnapping an atomic scientist (Mckellan) to build a nuclear weapon to destroy New York. The plot is further muddled by giving Margo Lane (Miller) telepathic abilities to read Cranston's mind yet he cannot use his powers on her. The over complication of plot elements prevents the film from truly embracing its gritty pulp magazine roots.
While there has been talk of producing a new Shadow film, including by Sam Raimi, at this time nothing is in the works. The 1994 had its share of faults but it remains a gorgeous looking, and generally well-acted film that could have and should have been much better.
Looking Back at the Shadow (23:44) –
This new documentary was produced for the collector's edition blu-ray and features interviews with Alec Baldwin, Penelope Ann Miller, Director Russell Mulcahy, and screenwriter David Koepp. All share their memories of working on the film.
Mania Grade:
Alec Baldwin, John Lone, Penelope Ann Miller, Ian McKellan, Tim Curry, Peter Boyle
Written By:
David Koepp
Directed By:
Russell Mulcahy
Original Year of Release:
Universal Studios
Special Features:
See Below


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- Posts: 1378
Right. But you do need secondary stats.
If you ever have an example of something that you don't think is right, let me know, and I'll explain what's going on. I need the two items and which scale you're using. (A screenshot can work for all three of those.)


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Haskins, Heather (1995) Bending the rules : the Montreal Branch of the Woman's Art Association of Canada, 1894-1900. Masters thesis, Concordia University.
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The first six years of the Montreal Branch of the Woman's Art Association of Canada are examined in light of women's expected class and gender roles in late nineteenth century Montreal society. The Branch's activities are described, and compared to their aims. The many reasons why handicrafts were supported and promoted are explored. A case study or the Exhibition of Arts and Handicrafts (October 22-November 3, 1900) is used to reveal this support and promotion in action. Attention is drawn to the organization and display of the exhibition, and the role of the lender, as they reflect contemporary social concerns. The Basketwork Section is discussed as it relates to contemporary political and ethnographic concerns for Native Canadians. Despite the appearance of conforming lo the status quo, the Montreal Branch's actions bely an ambivalence toward accepted standards of behaviour women. In effect, they were able to achieve many of their goals by bending the rules.
Divisions: | Concordia University > Faculty of Fine Arts > Art History |
Item Type: | Thesis (Masters) |
Authors: | Haskins, Heather |
Pagination: | vi, 106 leaves : ill., ports. ; 29 cm. |
Institution: | Concordia University |
Degree Name: | Theses (M.A.) |
Program: | Art History |
Date: | 1995 |
Thesis Supervisor(s): | Helland, Janice |
ID Code: | 6199 |
Deposited By: | Concordia University Libraries |
Deposited On: | 03 Sep 2009 09:49 |
Last Modified: | 08 Dec 2010 10:49 |
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