text:
Expanding the diagnostic use of PCR in leptospirosis: improved method for DNA extraction from blood cultures.
ABSTRACT Leptospirosis is a neglected zoonosis of ubiquitous distribution. Symptoms are often non-specific and may range from flu-like symptoms to multi-organ failure. Diagnosis can only be made by specific diagnostic tests like serology and PCR. In non-endemic countries, leptospirosis is often not suspected before antibiotic treatment has been initiated and consequently, relevant samples for diagnostic PCR are difficult to obtain. Blood cultures are obtained from most hospitalized patients before antibiotic therapy and incubated for at least five days, thus providing an important source of blood for PCR diagnosis. However, blood cultures contain inhibitors of PCR that are not readily removed by most DNA-extraction methods, primarily sodium polyanetholesulfonate (SPS).
In this study, two improved DNA extraction methods for use with blood cultures are presented and found to be superior in recovering DNA of Leptospira interrogans when compared with three previously described methods. The improved methods were easy and robust in use with all tested brands of blood culture media. Applied to 96 blood cultures obtained from 36 patients suspected of leptospirosis, all seven patients with positive convalescence serology were found positive by PCR if at least one anaerobic and one aerobic blood culture, sampled before antibiotic therapy were tested.
This study suggests that a specific and early diagnosis can be obtained in most cases of severe leptospirosis for up to five days after initiation of antimicrobial therapy, if PCR is applied to blood cultures already sampled as a routine procedure in most septic patients.
- SourceAvailable from: Jørgen Skov Jensen
Article: Novel TaqMan® PCR for detection of Leptospira species in urine and blood: Pit-falls of in silico validation.[Show abstract] [Hide abstract]
ABSTRACT: In the acute phase of leptospirosis, the diagnosis can be established with high sensitivity by testing blood and urine samples with polymerase chain reaction (PCR). However, only few real-time PCR assays have been validated for diagnostic use. The diagnostic accuracy of a novel TaqMan® PCR (LipL32 real-time PCR) targeting the lipl32 gene (or hap-1) and a previously described TaqMan® PCR (16S real-time PCR) targeting the rrs gene coding for 16S rRNA was evaluated when applied to both urine and blood specimens from humans suspected of leptospirosis. Applied to at least two blood cultures LipL32 real-time PCR had a sensitivity of 86%, and a specificity of 100%; and 16S real-time PCR had a sensitivity of 100%, and a specificity of 97%. Applied to urine samples, patients that were positive by the reference methods were also positive by both real-time PCR assays (n=4). For LipL32 real-time PCR the specificity was 100%, while for 16S real-time PCR it was only 91.5% due to unexpected cross-reactions with other bacteria. The analytical sensitivity was close to the theoretical limit-of-detection for both assays detecting all described human pathogenic species. We report a specific real-time PCR assay for detection of Leptospira, i.e., LipL32 real-time PCR that has been validated for diagnostic application in both urine and blood specimens from humans. We further show that a previously described 16S real-time PCR no longer can be recommended for diagnostic use due to a low specificity.Journal of microbiological methods 06/2012; 91(1):184-90. · 2.43 Impact Factor
Article: Ribosomal PCR and DNA sequencing for detection and identification of bacteria: experience from 6 years of routine analyses of patient samples.[Show abstract] [Hide abstract]
ABSTRACT: The use of broad range PCR and DNA sequencing of bacterial 16S ribosomal RNA genes for routine diagnostics of bacterial infections was evaluated. Here, the results from more than 2600 analyses during a 6-year period (2003-2009) are presented. Almost half of the samples were from joints and bones, and the second most frequent origin of samples was from the central nervous system. Overall, 26% of all samples were positive for bacterial DNA and bacterial identification was obtained in 80% of the PCR-positive samples by subsequent DNA sequencing. Ambiguous species identification was noticed among non-haemolytic streptococci, especially within the mitis group. The data show that ribosomal PCR with subsequent DNA sequencing of the PCR product is a most valuable supplement to culture for identifying bacterial agents of both acute and prolonged infections. However, some bacteria, including non-haemolytic streptococci, may not be precisely identified.Apmis 07/2013; · 2.07 Impact Factor
Page 1
Expanding the Diagnostic Use of PCR in Leptospirosis:
Improved Method for DNA Extraction from Blood
Steen Villumsen, Rebecca Pedersen¤, Karen Angeliki Krogfelt, Jørgen Skov Jensen*
Department of Microbiological Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark
Background: Leptospirosis is a neglected zoonosis of ubiquitous distribution. Symptoms are often non-specific and may
range from flu-like symptoms to multi-organ failure. Diagnosis can only be made by specific diagnostic tests like serology
and PCR. In non-endemic countries, leptospirosis is often not suspected before antibiotic treatment has been initiated and
consequently, relevant samples for diagnostic PCR are difficult to obtain. Blood cultures are obtained from most
hospitalized patients before antibiotic therapy and incubated for at least five days, thus providing an important source of
blood for PCR diagnosis. However, blood cultures contain inhibitors of PCR that are not readily removed by most DNA-
extraction methods, primarily sodium polyanetholesulfonate (SPS).
Methodology/Principal Findings: In this study, two improved DNA extraction methods for use with blood cultures are
presented and found to be superior in recovering DNA of Leptospira interrogans when compared with three previously
described methods. The improved methods were easy and robust in use with all tested brands of blood culture media.
Applied to 96 blood cultures obtained from 36 patients suspected of leptospirosis, all seven patients with positive
convalescence serology were found positive by PCR if at least one anaerobic and one aerobic blood culture, sampled before
antibiotic therapy were tested.
Conclusions/Significance: This study suggests that a specific and early diagnosis can be obtained in most cases of severe
leptospirosis for up to five days after initiation of antimicrobial therapy, if PCR is applied to blood cultures already sampled
as a routine procedure in most septic patients.
Citation: Villumsen S, Pedersen R, Krogfelt KA, Jensen JS (2010) Expanding the Diagnostic Use of PCR in Leptospirosis: Improved Method for DNA Extraction from
Blood Cultures. PLoS ONE 5(8): e12095. doi:10.1371/journal.pone.0012095
Editor: Stefan Bereswill, Charite ´-Universita ¨tsmedizin Berlin, Germany
Received June 28, 2010; Accepted July 14, 2010; Published August 11, 2010
Copyright: ? 2010 Villumsen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This study was financed by Statens Serum Institut, Copenhagen, Denmark, only. No external funders supported this work. The funders had no role in
study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: The authors have declared that no competing interests exist.
* E-mail: firstname.lastname@example.com
¤ Current address: Department of Microbial Ecology and Animal Health, National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark
Leptospira species is the causative agent of the leptospirosis, one
of the world's most wide-spread zoonosis [1]. Signs and symptoms
of the disease are often non-specific and range from flu-like
symptoms to multi-organ failure [1]. Carrier animals excrete the
bacteria in large numbers with the urine and transmission to
humans occurs mainly through contact with water or crops
contaminated with infected urine [1]. The disease is endemic in
developing countries mainly in the tropics where outbreaks occur
frequently after heavy rainfalls [2]. Travellers may be exposed
during activities in fresh water, and leptospirosis has recently been
shown to be a relatively common cause of fever in Swedish
travellers [3]. However, only few cases of leptospirosis are
diagnosed every year in the developed countries. These cases are
likely to represent an underestimate, since the diagnosis can only
be established by leptospira specific tests. The gold-standard in the
diagnosis of Leptospira spp. is detection of specific antibodies by
the microscopic agglutination test (MAT) [1]. In most cases, a
diagnostic serum sample can not be obtained before the 7thday of
disease and the diagnosis is thereby delayed for the same period. In
the early phase of the disease, a rapid diagnosis can be obtained by
PCR of Leptospira spp. This method has a sensitivity of 28–96% in
severe leptospirosis when applied to whole blood samples [4,5].
However, to ensure a high sensitivity, samples have to be obtained
before or shortly after the start of antibiotic therapy, since
antimicrobials quickly remove Leptospira spp. from the blood. In a
non-endemic area, leptospirosis is rarely a first line diagnosis, and
as symptoms can be severe, antibiotic treatment is often initiated
before leptospirosis is suspected. It is, therefore, often impossible to
obtain a relevant sample for diagnostic PCR.
In a hospital setting, blood cultures (BCs) are sampled from
most septic patients before antimicrobial therapy is initiated and
incubated for at least five days. BCs are closed container systems
and consequently not prone to DNA-contamination but they
contain inhibitors of the PCR that requires special procedures to
remove [6]. In previous studies where PCR has been applied to
BCs, only microorganisms that actually multiply in the BCs have
PLoS ONE | www.plosone.org1August 2010 | Volume 5 | Issue 8 | e12095
Page 2
been targeted. Even though, the sensitivity of the assays is highly
dependent on the DNA-recovery, only little has been done to
optimize these procedures. In whole blood samples, a high
recovery is especially important, since the density of bacteria is
very limited. Further, only a small fraction equivalent to 5–10 ml of
the original sample is included in the final PCR assay.
The aim of this study was to evaluate five DNA extraction
methods for their effectiveness in recovering Leptospira DNA and in
removing inhibitors from spiked BCs. Moreover, we aimed to
evaluate if BCs sampled before antimicrobial therapy could be
used in the diagnosis of leptospirosis.
Materials and Methods
DNA extraction methods
The following five DNA extraction methods were used in the
Method 1 (M1).
DNA was extracted from 200 ml of the tested
specimens using the DNeasyH Blood & Tissue Kit (Qiagen,
Hilden, Germany) according to the manufacturer's instructions
using the protocol for animal blood or cells. The final elution of
DNA was done in 200 ml buffer AE.
Method 2 (M2).
DNA was extracted from 200 ml of the tested
specimens using the MolYsis Plus Kit (Molzym GmbH & Co. KG,
Bremen, Germany) according to the manufacturer's instructions
for direct bacterial DNA isolation from blood culture. The final
elution of DNA was done in 100 ml buffer EB.
Method 3 (M3).
DNA was extracted from 100 ml of the tested
specimens using the method described by Fredricks and Relman
[6]. Briefly, cell lysis was obtained by a guanidine hydrochloride
based buffer. SPS (sodium polyanetholesulfonate) was removed by
adding benzyl alcohol to the solution and subsequently separated
from the aqueous phase containing the DNA by centrifugation.
DNA was precipitated from the supernatant by sodium acetate in
isopropanol. The pellet was ''washed'' in 70% ethanol and again
resuspended in 100 ml Tris-EDTA buffer.
Method 4 (M4).
Benzyl alcohol based removal of SPS and
other inhibitors in M3 was combined with the column based DNA
extraction of M1 in a new protocol, M4. A 100 ml aliquot of the
specimen was mixed with 100 ml lysis buffer (5 M UltraPureTM
guanidine hydrochloride (Invitrogen, CA) in 100mM UltraPureTM
Tris-HCl (pH 8.0; Invitrogen, Paisley, UK) and 10 ml proteinase
K (20 mg/ml; Qiagen) and incubated for 10 minutes at room
temperature. A total of 400 ml ultrapure water (Invitrogen) was
added followed by 800 ml .99% benzyl alcohol (ReagentPlusH,
Sigma-Aldrich, Brøndby, Denmark) and mixed. In order to
separate the phases, the sample was then centrifuged at 20.0006g
for 5 minutes at room temperature and 200 ml of supernatant (the
aqueous phase) was transferred to a new tube as described in
method 1, except that no proteinase K was added and that the
sample was not incubated during the lysis step. Only 100 ml of
buffer AE was added in the final elution step. This method was
developed to be used with anaerobic and paediatric blood culture
Method 5 (M5).
This method was equal to M4 except that
600 ml of ultrapure water was added before the phase separation.
This method was developed for use with all blood culture media.
Experimental set-up
The study consists of three experiments outlined in figure 1.
Experiment 1; evaluation of M1–M5 applied to one
One sample of BACTECTManaerobic Plus (BD
diagnostics, NJ) collected from an anonymous patient for
diagnostic purposes where no bacterial growth was detected
after at least 5 days of incubation at 37uC was spiked with Leptospira
interrogans (senso stricto) serovar Icterohaemorrhagiae strain M695
(availablefromKIT Biomedical
Collaboratingcentre forReference
leptospirosis, the Netherlands). The bacterium was grown in
DifcoTMLeptospira medium - EMJH (Becton-Dickinson, Le Pont
de Claix, France) until late log phase with a final concentration of
1.46106Leptospira/ml. Bacterial cells were counted by dark field
microscopy using a Helber counting chamber (Hawksley, Lansing,
UK) as described by the manufacturer. The quantified culture was
then frozen at 280uC in smaller aliquots. Just before use, one
aliquot was thawed and added to blood culture material to yield a
final concentration of 10,000 Leptospira/ml. The concentration of
bacteria in the spiked samples was chosen to be at a level that
would allow quantification of the recovery in methods with high
and low recovery. The spiked sample was simultaneously extracted
10 times by each of the evaluated methods, M1–M5. The recovery
of Leptospira DNA was assessed in triplicates by the quantitative
PCR (qPCR) assay, described below. A total of 30 qPCR assays
was made for each evaluated method. All DNA extractions were
performed simultaneously.
Experiment 2; evaluation of M5 applied to six different
blood cultures media.
In pre-study tests of M4, problems of
carry-over of inhibitors to the final eluate were observed when M4
was applied to BACTECTMaerobic Plus and BacT/ALERTH SA
(data not shown). The problem was identified as a very small water
phase in the phase separation step leading to carry-over of the
benzyl-alcohol phase on to the subsequent DNA extraction. The
changes made in M5 completely solved all problems of carry-over
of inhibitors and for that reason M5 is used in this experiment.
BACTECTManaerobic Plus, BACTECTMPaed Plus, BAC-
TECTMaerobic Plus (BD), BacT/ALERTH SA, BacT/ALERTH
SN and BacT/ALERTH PF (BioMerieux Inc., NC) collected for
diagnostic purposes as described under experiment 1 were
obtained from two Danish departments of clinical microbiology
(Hillerød Hospital, Hillerød, Denmark and Statens Serum Institut,
Copenhagen Denmark). For each of the six BC media tested,
material from five different anonymous patients was obtained and
spiked with Leptospira as described under experiment 1. DNA was
extracted once by M5 from each of the 30 spiked samples. All
DNA extractions were performed simultaneously. The recovery of
Leptospira DNA was assessed as triplicates by qPCR, as described
Experiment 3; clinical evaluation of M4 and M5.
material: Statens Serum Institut (SSI) is the only laboratory
performing routine diagnostic testing for Leptospira in Denmark. In
the period from January 2005 to June 2008, all BCs submitted for
culture of Leptospira for diagnostic purposes were inoculated in
DifcoTMLeptospira medium - EMJH in order to isolate Leptospira,
and the remainder of the BCs was stored at 220uC. When
diagnosticfollow-up had been
identifications were removed from the blood cultures and the
samples linked to the results of corresponding MAT and culture by
a unique, anonymous code and included in the ''Leptospira: PCR
diagnostics'' material collection (Danish Data Protection Agency:
journal number 2010-41-4862).
Some of the BCs had been incubated for at least 5 days at 37uC
at the local clinical microbiology laboratory before they were sent
to SSI for isolation of Leptospira spp. In all of these cases, contact
with SSI and the local clinical microbiology laboratory had been
established for diagnostic reasons, and samples were only
submitted if sampled before initiation of antimicrobial therapy.
Ninety-six blood cultures (BACTECTMaerobic Plus n=20;
BACTECTManaerobic Plus n=21; BACTECTMPaed Plus
Research on
completed, thepatient
Expanded Use of Leptospira PCR
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Page 3
n=2; BacT/ALERTH SA n=23, BacT/ALERTH SN n=21;
BacT/ALERTH FA n=8; BacT/ALERTH FN n=1) were
obtained from 36 patients (4 (11%) females and 32 (89%) males;
median age 38 years; range 2 years to 78 years). Twenty-nine of
these patients had a serum sample tested for antibodies against
Leptospira spp. at some point during the acute phase of the disease.
Ten patients had a repeated test for antibodies taken during
Reference method: The test for antibodies against Leptospira spp.
in a convalescence serum sample by a microscopic agglutination
test (MAT) is a recognised reference standard in the diagnosis of
leptospirosis [1]. Where requested by the clinicians, serum samples
were tested for antibodies against Leptospira spp. by a ISO 17025
certified microscopic agglutination test (MAT) conducted as
previously described by experienced laboratory technicians [7].
The MAT included Leptospira spp. of the following 13 serovars:
Patoc, Icterohaemorrhagiae (3 strains), Sejroe (2 strains), Saxkoeb-
ing, Javanica, Canicola, Ballum, Bratislava, Grippotyphosa,
Pomona, Bataviae, Hardjo and Hurstbridge. The following
dilutions of sera (in titer): 30, 100, 300, 1,000, 3,000, 10,000,
30,000 and 100,000 were tested by MAT. When only a single
serum sample was available for MAT, the test was considered
positive if the highest observed titer was above 1,000 and possibly
positive if the highest titer was with-in the range of 100–300.
When two serum samples were available and these were obtained
up to 4 weeks apart in the acute phase of the disease, then the test
was considered as positive when a seroconversion from no reaction
to a titer of at least 100 or a rise in the highest titer of two titer-
Figure 1. Flowchart for experiment 1–3. Figure shows flow over three independent experiments performed in the evaluation of five methods for
DNA extraction from blood cultures media. BD-AE: BACTECTMaerobic Plus, BD-AN: BACTECTManaerobic Plus, BD-Ped: BACTECTMPaed Plus, BacT-SA:
BacT/ALERTH SA, BacT-SN: BacT/ALERTH SN,BacT-PF: BacT/ALERTH PF, BacT-FA: BacT/ALERTH FA, BacT-FN: BacT/ALERTH FN. M1: Qiagen, DNeasyH
Blood & Tissue Kit; M2: Molzym, MolYsis Plus Kit; M3: Protocol described by Fredricks and Relman; M4: Improved method for anaerobic and paediatric
blood culture media; M5: Improved method adapted for all blood culture media.
Expanded Use of Leptospira PCR
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Page 4
steps, was observed. If no change or a change of only one titer-step
were observed, the results were interpreted according to the results
of the last sample as described above. Finally, the test was classified
as false positive, if a decrease in titer of two titer steps or more were
observed during this period, a finding sometimes observed with
this test. All serological tests and culture were conducted before the
results of the PCR assays were known.
Procedure for DNA extraction and qPCR: DNA extraction
was performed by M4 for anaerobic and paediatric BCs while M5
was applied to the aerobic BCs. Recovery of Leptospira DNA was
assessed as duplicates by qPCR as described below. M4 and M5
were evaluated for their ability to remove inhibitors of the qPCR
by comparing the Ct value of the no-template controls (NTC) with
the Ct value of samples where no amplification of the target DNA
Ethical considerations
Exemption for review by the ethical committee system and for
obtaining informed consent was obtained from the Committee on
Biomedical Research Ethics for Capital Region (protocol number
H-1-2010-FSP-20) in accordance with Danish law on quality
development projects.
Quantitative PCR assay
The Leptospira sp. DNA recovery of each extraction was assessed
by a quantitative PCR (qPCR) assay with previously described
primers and probe [8]. An in-house master mix was used [9] and
included an internal amplification control (IAC) designed
according to principles previously described [9]. All assays were
performed on an Applied Biosystems 7500 quantitative PCR
System platform using a previously described protocol [9]. Strict
precautions to avoid PCR product carry-over was observed
according to principles previously described [10]. A standard
curve was constructed from 10-fold dilutions of purified genomic
DNA of L. interrogans serovar Icterohaemorrhagiae strain M695
ranging in concentrations from approximately 100,000 leptospira
DNA-copies to 1 leptospira DNA-copy/assay by principles
previously described [9]. The limit of quantification (LOQ) of
the assay was 100 Leptospira sp. DNA copies/assay and the limit of
detection was ,10 Leptospira sp. DNA copies/assay. Ultrapure
water (Invitrogen) was used as NTC. Seven to eight NTC were
included on each of the three qPCR plates used in the experiment.
The qPCR targets Leptospira spp. specific sequences of the 16S
rRNA gene present in two copies in the genome [1,8]. The IAC
included in the assay made it possible to detect even partial
inhibition in assays, where no amplification of the target DNA
Statistical analysis
The effect on the DNA-recovery by five different extraction
methods (n=50) and the six different blood culture systems
(n=30) was estimated by the copy-number and was assessed by
multiple t-tests on log-transformed data (Logn+1) under Bonferroni
multiple-comparison-test (LSMEANS analysis associated with
PROC GLM, SAS 9.1; SAS Institute, US). The effect of blood
culture media on the Ct value of the IAC was compared by a two-
sample t-test of the mean for each PCR plate.
Experiment 1.
figure 2 and table 1. M2–M5 removed inhibitors of the PCR to an
extent that allowed amplification of Leptospira DNA in all of the
corresponding qPCR assays. When M1 was applied to the samples,
no amplification of Leptospira DNA was possible and the IAC was
only amplified in two of the 30 corresponding qPCR-assays.
The results of experiment 1 are shown in
Figure 2. Leptospira sp. DNA recovery and inhibition of PCR.
Estimated total copy-number of Leptospira interrogans DNA recovered
from a spiked sample of BACTECTManaerobic Plus containing
approximately 50.000 Leptospira/5 ml by five DNA extraction methods.
The mean copy-numbers are given above each column. Error bars
indicate the 95% confidence interval. M1: Qiagen, DNeasyH Blood &
Tissue Kit; M2: Molzym, MolYsis Plus Kit; M3: Protocol described by
Fredricks and Relman; M4: Improved protocol for anaerobic and
paediatric blood culture media; M5: Improved protocol adapted for
all blood culture media.
Table 1. Effectiveness of DNA extraction method, M1–M5 in recovering Leptospira DNA from BACTECTManaerobic Plus.
quantitative PCR
Positive internal
amplification control
Leptospira DNA recovered in percent
of M4 (95% confidence interval) P-value
M1 10020% (0 to 0.02%)P,0.001
M2 1030300.22% (0.16% to 0.29%)P,0.001
M310300 11% (9% to 14%)P,0.001
M5 1030058% (44% to 74%)P,0.031
M1: Qiagen, DNeasyH Blood & Tissue Kit; M2: Molzym, MolYsis Plus Kit; M3: Protocol described by Fredricks and Relman; M4: Improved method for anaerobic and
paediatric blood culture media; M5: Improved method adapted for all blood culture media.
Expanded Use of Leptospira PCR
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The highest effectiveness in recovering Leptospira DNA was
obtained by M4, while M5 resulted in a slightly lower recovery,
58% (95% CI, 44% to 74%; p,0.031) of M4. Due to the
increased dilution of the sample in M5 compared to M4, a
recovery of 76% was expected. Both methods were equally simple
to use and the results were highly reproducible.
The three established methods (M1–M3) all had a significantly
lower effectiveness in recovering Leptospira DNA than both M4 and
M5. For M3, this was reduced to 11% (9%–14%; p,0.001) of M4
and for M2 only 0.22% (95% CI, 0.16% to 0.29%; p,0.001) of
Experiment 2.
Results of experiment 2 are shown in figure 3.
There was no sign of inhibition of the qPCR in any of the tested
specimens, when M5 was applied to all of the tested blood culture
media. There was no significant difference (n=30; p=0.54) in the
Leptospira DNA-recovery obtained between the six different blood
culture media tested.
Experiment 3.
Seven of the 36 patients were diagnosed with
leptospirosis by serological findings of either a single serum MAT
titer $1,000 (n=6) or by seroconversion to a MAT titer of 100 in
several serovars in an early convalescence serum sample (n=1). In
one of the seven patients, EMJH-subculture of one of the BCs
(BACTECTMaerobic Plus) was found positive for Leptospira
borgpetersenii senso stricto. From the remaining 29 patients, a
serum sample was available from 21.In five of the 21 patients, a
repeated serum sample taken and tested by MAT during
convalescence and in one of the 21 patients the primary sample
was taken 12 days after the blood cultures. Of the 21 patients, two
had a MAT titer in the acute phase of the disease of 100 and 300
respectively, but on repeated testing during convalescence these
were normalized. These patients were classified as false positive by
All of the seven patients diagnosed with leptospirosis by serology
were positive by qPCR. One additional patient was positive by
qPCR, but negative by MAT. He had only one serum sample
taken at the same time as the blood cultures, and thus, it is likely
that antibodies were not present at the time of sampling. All the
remaining patients were negative by qPCR.
In 7 of the 8 positive patients the DNA-recovery was below
LOQ i.e. they contained less than 100 DNA copies pr. asssay. A
total of 17 BCs were positive by qPCR. The approximate median
recovery in all positive samples was 11 Leptospira DNA-copies pr.
assay (range 1 to 262). In four patients, all of the 2–5 analysed BCs
were positive, while in four patients, where the DNA-recovery was
close to LOD, only half of the 2 or 4 tested BCs were positive. In
one of these last four patients, only one of the duplicate qPCR
assays was positive in one BC.
The Ct value for the IAC was recorded for all 158 qPCR assays
performed on the 79 blood cultures (nBC) where no amplification
of leptospira DNA occurred in any of the two corresponding PCR
assays. The 79 BCs included BACTECTMaerobic Plus, n=17;
BACTECTManaerobic Plus, n=16; BACTECTMPaed Plus,
n=2; BacT/ALERTH SA, n=19, BacT/ALERTH SN, n=19;
BacT/ALERTH FA, n=5; BacT/ALERTH FN, n=1). The
results were compared to the Ct value recorded from the NTC
and the results shown in table 2. A total of 3 plates were used for
testing all clinical samples by qPCR.
In this study we present two improved DNA extraction methods
designed for extraction of DNA from blood culture media, M4 and
M5. The amount of Leptospira sp. DNA recovered by these
methods were 9 and 5 times higher, respectively than what was
obtained by M3, the best performing of the three previously
described methods. This difference can largely be explained by the
use of a column based DNA extraction in M4 and M5 in contrast
to the DNA precipitation method used in M3. The sensitivity of
the diagnostic set-up was remarkable when M4 and M5 were
validated on samples from patients suspected of leptospirosis. At
least one BC, from all of the seven patients that were tested
positive by convalescence serology was also positive by qPCR in at
least one of the duplicate qPCR assay wells. In addition, one
patient was positive by qPCR but never had convalescence
serology performed. These findings indicate that a very high
sensitivity close to 100% can be obtained by this diagnostic set-up.
The results are comparable to what have been found in a recent
study where qPCR was compared to culture of Leptospira spp. [5],
but remarkably higher than the 28–50% sensitivity that has
previously been reported when PCR has been applied to whole
blood samples in severe leptospirosis and compared with MAT
[4,11]. Four factors are likely to be important for this difference. In
Figure 3. DNA recovery from different blood culture media.
Estimated total copy-number of Leptospira interrogans DNA recovered
from spiked samples of six different blood culture media containing
approximately 50.000 Leptospira/5ml. All DNA extractions were per-
formed with M5, the improved protocol adapted for all blood culture
media. The mean copy-numbers are given above each column. Error
bars indicate the 95% confidence interval. BD-AE: BACTECTMaerobic
Plus, BD-AN: BACTECTManaerobic Plus, BD-Ped: BACTECTMPaed Plus,
BacT-SA: BacT/ALERTH SA, BacT-SN: BacT/ALERTH SN and BacT-PF: BacT/
Table 2. Comparison of the mean Ct value of the internal
amplification control in no-template controls and blood
cultures tested negative for Leptospira sp. DNA.
qPCR run No-template controlNegative blood culture P-value
n CtSDn Ct SD
1733.8 0.4 52 33.5 0.3p.0.05
2833.70.4 3433.50.2p.0.05
37 34.10.37234.0 0.2p.0.05
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this study, at least two blood samples from each of the qPCR
positive patients were tested. Only in 4 of the 8 positive patients,
all BCs were positive by qPCR and this indicates that it is
important to test at least two samples. Further, in one patient only
one of duplicate qPCR assay wells was positive close to LOD.
Even though such a result in a clinical situation needs further
confirmation, the diagnosis might have been missed if the qPCR
assay had not been tested in duplicate. Previous studies do not
report having tested more than one sample from each patient nor
do they report having performed more than one PCR assay for
each sample [4,11]. Furthermore, in this study BCs were tested
and not serum or whole blood samples and this might affect the
number of Leptospira spp. present in the samples. Five of the eight
PCR positive patients were positive in more than one BC. The
recovery of Leptospira DNA from the aerobic and the anaerobic BC
were comparable in four of these patients but in one patient, the
recovery of Leptospira DNA in BacT/ALERTH SA and BacT/
ALERTH FA was .10 times higher than what was obtained from
BacT/ALERTH SN, sampled at the same time. This suggests that
some multiplication of the bacteria had occurred. The use of BCs
and time of incubation might, therefore, at least in some cases play
a role in the sensitivity of the assay, but we did not record the exact
incubation period. Lastly, DNA precipitation technique has been
used in previous studies for DNA-extraction and as we have shown
in experiment 1, this is likely to result in a lower recovery of DNA
compared with the column based method used in M4 and M5.
The low DNA-copy-number present in the clinical samples stresses
the importance of optimal microbial DNA recovery. It is therefore
likely that neither M2 nor M1 can be used for diagnostic PCR in
Fredricks and Relman were the first to effectively solve the
problems of removing all inhibitors of the PCR from BCs. They
indentified the anti-coagulant sodium polyanetholesulfonate (SPS)
present in a anaerobic media of BacT/ALERTH as a potent
inhibitor of PCR not readily removed by most extraction methods
[6] and came up with a benzyl alcohol based extraction method,
M3 to solve this problem [6]. However, M3 is, in our experience,
not very robust in a clinical setting, especially when applied to
aerobic BCs and for that reason the method has previously been
rejected [12]. M4 and M5 have been developed on the basis of
M3. By incorporating Proteinase K treatment in the lysis step,
increasing the speed of centrifugation to 20.0006g in the phase
separation step and for M5 by further adding extra water in the
phase separation step all problems of carry-over of inhibitors to the
final DNA preparation were completely solved. Throughout all
the experiments, M4 and M5 were very robust and easy to use and
as they are based on column-based DNA extraction, both methods
are amenable to automation.
The IAC is a very sensitive way to detect inhibitors of the PCR,
but in none of the experiments were there any signs of inhibition of
the qPCR assay when M4 and M5 were used. A more exact
estimate of this was given in experiment 3, by comparing for each
qPCR plate the Ct value of the IAC of the NTC with the Ct value
of the IAC in the BC samples where no amplification of Leptospira
spp. DNA occurred. These 79 BCs included 5 BCs containing
charcoal. No significant difference was observed in any of the
assays and, thus, no sign of inhibition of the qPCR assays (table 2).
Furthermore, no significant difference between in the recovery of
Leptospira sp. DNA was observed when M5 in experiment 2 was
applied to 6 different blood culture media from a total of 30
different patients.
To our knowledge, only one study by Gebert et al. has
previously addressed the issue of recovering low amounts of
bacterial DNA from blood cultures [13]. In that study, a near full
recovery of Staphylococcus aureus DNA was obtained by M2, but
when the method was applied to Escherichia coli the recovery was
somewhat lower. In general, Gram-negative bacteria like E. coli
and especially Leptospira spp. have a more fragile cell wall than
Gram-positive bacteria. In M2, human cells are lysed and the
DNA removed in the initial steps. Preliminary results suggest that
the low recovery obtained by M2 in our study partly can be
explained by a huge loss of bacterial DNA during these
If used with the described or slightly modified protocols, it is
likely that BCs can be used in the diagnosis of other fastidious
microorganisms like Coxiella burnetii [13] and dengue virus [14] in
the future. It is also likely that the protocols will find use in severe
infections like meningitis, where antibiotic treatment sometimes
foregoes sampling of BCs and there by prevents the causative
agent from growing in the media. Preliminary results suggest that
M4 and M5 can be applied to Neisseria meningitidis and Streptococcus
pneumoniae with a similarly high DNA recovery (data not shown).
The diagnosis of leptospirosis in non-endemic countries will
often rely on convalescence serology since relevant samples for
diagnostic PCR are difficult to obtain. In this study, two improved,
easy and robust DNA extraction methods are presented. Both
methods effectively removed all inhibitors of the PCR and they
were found superior to three previously described methods in
recovering Leptospira sp. DNA from blood cultures. This study,
further, suggests that most cases of severe leptospirosis can be
diagnosed by specific PCR if the described DNA extraction
methods are applied to blood cultures routinely obtained from
most septic patients and incubated for at least five days.
Consequently, in the future, most cases of severe leptospirosis
can be diagnosed by PCR in the early phase of the disease.
Laboratory technician Birthe Dohn is thanked for assisting during the
laboratory experiments. Per Moestrup Jensen is thanked for critic statistical
review. Klaus Leth Mortensen is thanked for critical review of the
Author Contributions
Conceived and designed the experiments: SV KAK JSJ. Performed the
experiments: SV RP. Analyzed the data: SV RP JSJ. Contributed
reagents/materials/analysis tools: SV KAK JSJ. Wrote the paper: SV
1. Levett PN (2001) Leptospirosis. Clin Microbiol Rev 14: 296–326.
2. Bharti AR, Nally JE, Ricaldi JN, Matthias MA, Diaz MM, et al. (2003)
Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3:
3. Askling HH, Lesko B, Vene S, Berndtson A, Bjorkman P, et al. (2009) Serologic
analysis of returned travelers with fever, Sweden. Emerg Infect Dis 15:
4. Fonseca CA, Teixeira MM, Romero EC, Tengan FM, Silva MV, et al. (2006)
Leptospira DNA detection for the diagnosis of human leptospirosis. J Infect 52:
5. Slack A, Symonds M, Dohnt M, Harris C, Brookes D, et al. (2007) Evaluation of
a modified Taqman assay detecting pathogenic Leptospira spp. against culture
and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical
environment. Diagnostic Microbiology and Infectious Disease 57: 361–366.
6. Fredricks DN, Relman DA (1998) Improved amplification of microbial DNA
from blood cultures by removal of the PCR inhibitor sodium polyanethole-
sulfonate. J Clin Microbiol 36: 2810–2816.
7. Hartskeerl RA, Smits HL, Korver H, Goris M, Terpstra WJ (2006) International
course on laboratory methods for the diagnosis of leptospirosis. Amsterdam, the
Nederlands: KIT Biomedical Research. 109 p.
Expanded Use of Leptospira PCR
PLoS ONE | www.plosone.org6 August 2010 | Volume 5 | Issue 8 | e12095
Page 7
8. Smythe LD, Smith IL, Smith GA, Dohnt MF, Symonds ML, et al. (2002) A
quantitative PCR (TaqMan) assay for pathogenic Leptospira spp. BMC Infect
Dis 2: 13.
9. Jensen JS, Bjornelius E, Dohn B, Lidbrink P (2004) Use of TaqMan 59 nuclease
real-time PCR for quantitative detection of Mycoplasma genitalium DNA in
males with and without urethritis who were attendees at a sexually transmitted
disease clinic. J Clin Microbiol 42: 683–692.
10. Krojgaard LH, Villumsen S, Markussen MD, Jensen JS, Leirs H, et al. (2009)
High prevalence of Leptospira spp. in sewer rats (Rattus norvegicus). Epidemiol
Infect 137: 1586–1592.
11. Gravekamp C, van de Kemp H, Franzen M, Carrington D, Schoone GJ, et al.
(1993) Detection of seven species of pathogenic leptospires by PCR using two
sets of primers. J Gen Microbiol 139: 1691–1700.
12. Millar BC, Jiru X, Moore JE, Earle JAP (2000) A simple and sensitive method to
extract bacterial, yeast and fungal DNA from blood culture material. Journal of
Microbiological Methods 42: 139–147.
13. Gebert S, Siegel D, Wellinghausen N (2008) Rapid detection of pathogens in
blood culture bottles by real-time PCR in conjunction with the pre-analytic tool
MolYsis. Journal of Infection 57: 307–316.
14. Halstead SB (2007) Dengue. The Lancet 370: 1644–1652.
Expanded Use of Leptospira PCR
PLoS ONE | www.plosone.org7 August 2010 | Volume 5 | Issue 8 | e12095


text:
Michael Clarke Duncan: 'The Green Mile' and 11 Other Memorable Roles
5:58 PM PDT 9/3/2012 by THR Staff
The Oscar-nominated actor died Monday, Sept. 3, nearly two months after being hospitalized for a myocardial infarction, from which he never fully recovered. THR takes a look back at some of his most well-known films and TV shows, from "Armageddon" to "The Finder."
More Galleries
Joan Rivers' Best Quotes About Plastic Surgery
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PHOTOS: Maya Angelou and Her Hollywood Friends
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text:
Clown Had iPad Stolen From Steve Jobs' House
Don't Miss This
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- Jackson Teen Leading Rally Against Washington Redskins' Name At San Francisco 49ers Game
PALO ALTO, Calif. (AP) — An iPad stolen from the home of the late Steve Jobs ended up in the hands of a professional clown who had no idea it was pilfered from the Apple co-founder's house.
Kenneth Kahn, also known as Kenny the Clown, says he unwittingly received the stolen tablet from a friend who was later arrested for breaking into the Jobs residence in Palo Alto.
The San Jose Mercury News reports that Kahn says he never examined the device's contents. Instead he downloaded songs to play while entertaining kids and tourists.
Kahn says he had no idea where the iPad came from until his friend, 35-year-old Kariem McFarlin of Alameda, was arrested Aug. 2.
Kahn says he had it for a few days before police came asking for the purloined tablet.
Copyright 2012 The Associated Press.


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Average Customer Reviews
Energy Star Rated - No
Most Usable Capacity¹
Most usable space inside, same space on the outside, so you'll have space for everything from party platters and beverages to kids' snacks and juice boxes.
PureAdvantage® Filtration Systems
Our filtration system keeps clean, filtered air circulated throughout to prevent odor transfer and keeps fresh, clean water and ice at your fingertips.
Luxury-Glide® Cool Zone™ Drawer
Keeps contents chilled, so it's perfect for wine and beer or even snacks and juice boxes.
IQ-Touch™ Controls
For those who like to have their options always visible, the IQ-Touch™ control panel displays your options at all times.
Luxury-Design® Lighting
Our ramp-up designer lighting is as beautiful as it is functional.
Maximum Efficiency ENERGY STAR®²
Luxury-Glide® Humidity-Controlled Crisper Drawers
Drawers smoothly glide out and keep produce fresh with adjustable humidity control and minimal temperature variation.
Sliding SpillSafe® Glass Shelves
Designed to keep spills contained, making cleanup easy.³
SpillSafe® Glass Shelves4
Designed to keep spills contained, making cleanup easy.
Luxury-Glide® Freezer Drawers
Glide out smoothly and fully extend, giving you easy access to what's inside.
Alarm System
Notifies you if there's been a power failure, if a door stays open longer than five minutes and if the interior temperature of the refrigerator ever rises above 56 degrees for more than 20 minutes.
Signature Soft-Arc Door And Handle Design
Full-Extension Freezer Baskets
Sabbath Mode (Star-K® Certified)
Platinum® Star Warranty
Guaranteed performance provides peace of mind and service from our national network of authorized independent service professionals.
Certifications & Approvals
Electrical Specifications
Exterior Specifications
Freezer Specifications
General Specifications
Ice Maker
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Approximate Dimensions
Approximate Weight
Warranty Information
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(based on 25 reviews)
Ratings Distribution
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of respondents would recommend this to a friend.
Most Liked Positive Review
Our refrigerator is good except for the fact that the cold water tank must be small and we only get about 1 cup of cold drinking water at a time, and need to wait...Read complete review
Our refrigerator is good except for the fact that the cold water tank must be small and we only get about 1 cup of cold drinking water at a time, and need to wait for tank to replenish, also wish there was a temp gauge on the center drawer inside.
Most Liked Negative Review
somewhat pleased
I have had a lot of problems with my refrigerator. I thought that purchasing a higher priced would give me less problems. You cannot buy filters in the store and a lot of times...Read complete review
I have had a lot of problems with my refrigerator. I thought that purchasing a higher priced would give me less problems. You cannot buy filters in the store and a lot of times the ice maker shuts down and I have to contact company to help re=set as it is to difficult to do. It makes me wish I BOUGHT SOMETHING ELSE. I do love my electrolux dishwasher.
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Shake my head
By nadremmak
from Undisclosed
Comments about Electrolux Standard Depth 27 Cu.Ft. Black French Door Bottom Freezer Refrigerator:
I bought my fridge 5 years ago and in all honesty it stinks. I have had many service calls to repair the ice making machinery they have replaced my complete refridgerator and many ice making parts. Now my display is gone and it just crushes ice no cubes any more. This is the worst refiridgerator I have ever owned. Painful although it keeps food the ice situation is stupid.
Bottom Line No, I would not recommend this to a friend
Was this review helpful? Yes / No
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Comment on this review
Good fridge
By Susan
Everything works as it should. Love the water filter and ice machine.
Bottom Line Yes, I would recommend this to a friend
our Electrolux experience
By candy
The design of the refrigerator is excellent for storage love it. The ice maker design takes this product from excellent to good. We have had issues with our ice maker that resulted in electrolux replacing our unit.
By Whitey
I think Electrolux has the "Optimum Refrigerator" with the IQ Touch French Door Model. It has TWO Ice-makers which is extremely handy for us. We have a swimming pool and are constantly entertaining. The second ice-maker is in the bottom freezer and easy to access to fill coolers to take outside!
Very pleased with appliance package
By J-Dub99
Very pleased with the overall performance of the Electrolux appliances. The lighting in the refrigerator is superb!! Do not like the alarm that sounds when you have a power failure. Especially when sleeping at 2 am. The French door style is very sleek and stylish! Ice maker produces plenty of ice to supply the family. Stainless appliances is a very nice upgrade to any kitchen!
We have enjoyed our appliance
By mike
This is one of the best appliances that we have ever had.
I made a great choice
By Dennis
I'm building a new house and we are buying all new appliances for it. I got a email from a club I belong to and it had a great buy on this refrigerator. I told my wife I should buy this now as we can take it with us and I'm really glad we did. It works great and we love it. I read reviews that said it was loud, the drinking water didn't work right and the ice maker didn't work right and I found these reviews to all be false.
By Lisa
My first refrigerator was wonderful but would not produce any ice even though there were 3 repairs/replacements. They sent me a whole new fridge but the doors do not work. The plastic part that flips into the groove has to be manually moved before you can shut the door frequently. It's going to break eventually. Too bad. This is a beautiful fridge.
Works Great
I recently bought this refrigerator and got a really good price on it. At first I couldn't get the ice or water to work and I found out I needed to return it to the factory settings and everything worked fine. I like the way its layed out, its not loud and everything is working great. I do like the styling and would reccommend this to a friend.
very unhappy
By unhappy
for a quality brand name like Electrolux to put out a refrigerator with an ice maker doesn't work is a major disappointment.
Delivery is subject to reasonable access to the area without damaging the purchaser's property or the product, or jeopardizing the safety of the delivery personnel.
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Bad carrots?
- goodhealthgourmet Aug 27, 2008 05:58 PM
i just opened a bag of organic yellow peeled & cut carrots that i bought yesterday, and immediately knew that they were bad - slimy, with a funky odor. checked the date - Sept 8, 2008.
this is the FOURTH time this has happened to me in the past few weeks. i've tried different brands, different varieties [orange & yellow], organic, conventional, different stores...
the one large bag of full-size unpeeled carrots i bought was fine, but all the baby or cut & peeled carrots are bad! what's going on??? has anyone else had a problem...?
The last time I bought yellow baby carrots, my SO happily munched through them until he picked up one wiggly one . . . needless to say the rest of the bag got thrown out.
Ah well, at least we can take comfort in the fact that whole carrots, cut into sticks, generally taste much better, and much more like carrots! I think it's the end of baby carrots around here.
I had to give up buying baby carrots because of the same problem. Stuck my hand into slime too many times. You can see all the moisture in the bag when you buy them. Too yucky. The whole ones you peel yourself taste better anyway.
i'm glad it's not just me. and you're both right - the big ones really do taste better anyway.
those prepared carrots have always been one of my few go-to "convenience" purchases...but it's hardly convenient if they're inedible :)
That's happened to me twice now in the past to weeks - it first happened with a 3lb bag of baby carrots, so I figured that I would try a smaller bag next time. Yesterday I picked up a 1lb bag from Whole Foods and had the same issue!
I might also revert back to peeling and cutting my own regular carrots.
re: TripChick
the first one was from WFM - i picked up my usual 1-lb bag from them - i think they're called "Whole Fields" [probably the ones you bought]. i returned it, and when i went to pick up another one i noticed that there were no longer any bags of those on the shelf. instead, they had only larger 3-lb resealable bags from a different company ["Bio" something or other] that i've never seen before. anyway, i bought one of those instead, got them home...same problem. also happened with a 1-lb organic bag from TJ's, as well as yesterday's purchase from a Kings supermarket in NJ.
i just think it's odd that it happened so many times in such a short span...i think in all the years i've been buying them i've only ever gotten one bad bag before this.
oh well, no more baby carrots for me!
re: lgss
it goes beyond that. i'm sick of taking them back - this is the fourth time - and the third store - in a month. and i feel like there may be something going on with the crops - it's not likely to be a quality control or freshness issue at the processing or packaging level because it has happened with several different brands.
re: swsidejim
yes, I just had this happen to me last week.
re: The Old Gal
i've already said this a couple of times further up-thread...i DID return the carrots...and it happened at THREE different, completely unrelated stores.
if it was only happening at one place or with one brand, i wouldn't think it was such an issue.
I have recently started buying baby carrots and I was having the same slime problem. I did some research and found this statement from Devon Zagory, Ph.D. who is the Vice President of Davis Fresh Technologies. I felt a lot better once I read this, so hopefully this will help you guys out!
"In my experience the sliminess on peeled carrots is due to the
proliferation of lactic acid bacteria. Many processors of carrots prefer
that the carrots be very moist in the bag to prevent development of dry,
white cells on the surface. These bacteria are normal components of the
carrot microflora and come from the soil that the carrots were grown in.
They are generally not harmful and produce little odor. They tend to thrive
in low oxygen, high moisture environments which often develop in bags of
peeled carrots. You can probably wash the carrots and eat them without ill
effect, but most people find them unsavory once they become slimy. To
prevent the development of the sliminess I would recommend keeping the
carrots colder (measure the temperature in your refrigerator and if it is
above about 40F, turn it down) and eating them sooner."
Devon Zagory, Ph.D.
Senior Vice President
Food Safety & Quality Programs
Davis Fresh Technologies, LLC
re: Laura1
I used to look for the dry, whitish looking ones. They were great. Lately I have only seen the wet looking carrot, and they are yuck.


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Alsarah & the Nubatones
Sat, Nov 10, 2012
Times: 8pm
Venue: MASS MoCA
1040 Mass Moca Way, North Adams, MA 01247Contact:
Jodi Joseph
0 people saved this
Powerful Sudanese-American singer Alsarah fronts this extraordinary band bound together by a collective love for Nubian music and the genuine belief that soul transcends all cultural and linguistic barriers. They blend a selection of Nubian 'songs of return' from the 1970s to today with original, Western soul-inspired material and vintage Sudanese pop. Their set is a musical journey through diaspora and migration viewed through an urban lens.
"A blend of original compositions and traditional pieces, grounded in the Sudanese and Nubian musical traditions but sails in the pocket of funk and soul music." -Afropop Worldwide
7 Reasons to Visit the Berkshires This Winter
As evidenced by the snowflakes, shopping sales and Christmas lights, winter has pretty much arrived....
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BPL Community Profile
todd harper (funnymoney) - BPL Lifetime Member
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Showing 231 - 240 of 1790 | Page: 1 ...15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 ...179
Re: resurrection - shifting the load South
Doug I's "Half Pack" is a cool design. Can find it under "half pack" I believe. It is built by Chris Zimmer and made of cuben & dyneema. Very cool. Check it out. ...
Re: the opposite of ultralight
somewhere on this site there's a similar thread - complete w/pics of a few "non-UL" people. Great stuff just don't have time to find it.
Re: Epic trips in states that don't get the glory
Head a little further East from Louisiana and hike the Florida Trail. It oughta be long enough for ya! Well over 1,000 miles through such a vast array of wildlife and (really!) some very challeng...
Re: FS Brawny Silnylon Ultralight Tarp Tent
Can you post pics of it set up?
Re: Attention Flat Tarp and Net Tent Users
The waterproof material on the ends does its job. The only time you'll have an issue is when you pitch the tarp way high.
Re: Gear Shelf
Nice job Maggie!
Re: Zlite Comfort?
Zlite Comfort? No such thing! :)
Re: Good Lid Designs?
Joel, Cut the wall out of an aluminum can and use Intense Angler's instructions to make a lid. You can find him on Youtube. Light, free, fun, & cool. Please show us what you've ...
Re: Emberlit Fire Ant Stove - lightest stove in the world?
Somehow I missed it - thanks for posting! His Emberlit stove has a big following. Very nice!
SOLD: Caldera Cones and BPL 550 pot w/handles, etc
I have two Calderas plus a ti pot for sale: ** Both are for the old style BPL 550 / Tibetan / Brasslite / current LiteTrail & probably Toaks pots *** First $100 takes ALL!!!! I would lik...
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Humorist Loretta LaRoche in Let's Stop Global Whining
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The last date listed for Loretta LaRoche: Let's Stop Global Whining was Saturday March 31, 2012 / 8:00pm.
Currently at The Hanover Theatre
Country Music Legend Clint Black Performs
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With more than 20 million sold records under his hat, Clint Black is one of country music's brigh...Learn More
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There's a parking garage right across the street, but you have to do a u-turn to get there.Cirque Éloize: Cirkopolis info • Mar 06 2014 star this tip starred
Park at the outdoor lot behind the Hanover Theater, easy in, easy out, $10.Jersey Boys info • Jan 28 2014 star this tip starred
Every day we are assaulted with messages of fear. Our lives are permeated with doing to the point where we are no longer being. Busyness and stress are part of our daily conversations. We need to stop whining, stop talking about what we can't control, and start having some fun! No one whose dead ever got a reprieve because they were too busy! Come and laugh at the absurdity that has become the "American Way" and discover that laughter is truly the best medicine.


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The Bulletin by Firdose Moonda
May 5, 2011
Kochi Tuskers Kerala 156 for 5 (Jayawardene 55, Unadkat 2-25) beat Kolkata Knight Riders 139 for 7 (Morgan 66, Gomez 2-14 by 17 runs
Scorecard and ball-by-ball details
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Series/Tournaments: Indian Premier League
Teams: India | Kochi Tuskers Kerala | Kolkata Knight Riders
A disciplined performance in the field and an explosive 35 off 19 balls from Brad Hodge helped Kochi Tuskers overcome Kolkata Knight Riders in the last match at the Nehru Stadium this IPL season.
Hodge's last over blitz, in which he took 21 runs off countryman Brett Lee, proved to be the difference between the two sides, as Kochi defended 156 by 17 runs. In the chase, RP Singh and Sreesanth failed to get the same kind of movement that Brett Lee extracted early in the Kochi innings. Jacques Kallis and Eoin Morgan didn't have to take many risks early on as there were many poor deliveries that were smacked to the boundary. In the first three overs, the bulk of the short and wide ones came from RP Singh.
R Vinay Kumar and Prasanth Parameswaran pulled back the chase before it raged out of control with a selection of back of a length deliveries that proved difficult to get away. Although they kept the boundaries down, they didn't trouble the batsmen much and failed to get a breakthrough until after the halfway stage, when Kolkata were well set. Kallis was the senior partner and easily outscored Morgan in that phase. Seven times in the first ten overs Kallis stole the strike at the end of the over.
Just as Kallis looked as though he had grown roots, Raiphi Gomez rattled Kolkata with a double strike in his second over. He bowled Kallis with a legcutter and had Gautam Gambhir caught in the covers off consecutive balls, which left Morgan to assume the senior role. Manoj Tiwary could not last long, and Yusuf Pathan was expected to counterattack, but he and Morgan were frustrated by Gomez's variations and Parameswaran's accurate fuller deliveries. Sreesanth let the noose loosen, giving Morgan back-to-back boundaries but Vinay Kumar was on hand to tighten it. Confusions between Morgan and Yusuf mounted in Vinay's last over, and Morgan was run out when both batsmen ended up at the wicketkeeper's end.
It brought Brett Lee to the crease, in poetic justice for the last over he bowled, which went for 21. There were 25 to get off the last over of Kolkata's innings. Lee was run-out and the task proved too steep.
Kochi's innings was anchored by a third-wicket partnership between Mahela Jayawardene and Michael Klinger before being given momentum at the death by Hodge. It didn't look as though Kochi would get over the 150-run mark, especially after the way things started. Lee's first over was a whole bag of peaches. He got impressive away movement and started the innings with a maiden.
Some success seemed inevitable after the start Kolkata got and it came from Jaidev Unadkat, although he hardly deserved it. He banged one in, too short and too wide outside off that Brendon McCullum chased and his fine edge nestled in Kallis' hands at slip.
Parthiv Patel came in at No. 3 and opened his account with two stunning boundaries. He looked energetic and confident in his strokeplay and dealt with Unadkat's bouncer and the introduction of spin, in the form of Iqbal Abdulla, with relative ease. Surprisingly, it was the short ball that undid him, when he charged down the track and miscued a pull shot to midwicket.
The stage was set for Jayawardene to play an innings of authority and, with Klinger at the other end, he did exactly that. They played creative cricket, managing a boundary off five of the seven overs they were together for and pushed each other to take singles before Klinger holed out. When Abdulla got the wicket of Ravindra Jadeja for eight, Kochi were being pegged back and some impetus was needed.
The floodgates were opened with Jayawardene's six over long leg at the start of the 17th over and Kochi put on 54 runs in the final four overs, with Hodge's fireworks yielding almost half of those.
Firdose Moonda is ESPNcricinfo's South Africa correspondentFeeds: Firdose Moonda
© ESPN Sports Media Ltd.
|Comments have now been closed for this article
Pakistan should not welcome Amir back
The serene team culture cultivated by Misbah and his men shouldn't be allowed to be disrupted by a player with a tainted past
November games need November prices
An early start to the international season, coupled with costly tickets, have kept the Australian public away from the cricket
The inherent dangers of batting
The sickening blow that struck Phillip Hughes is a reminder of the ever-present dangers associated with facing fast bowlers, even while wearing a helmet
Hope for Hughes, feel for Abbott
It is impossible to imagine how Sean Abbott must feel after sending down that bouncer to Phillip Hughes. While the cricket world hopes for Hughes' recovery, it should also ensure Abbott is supported
Why the Indian opener would be well advised to shelve the hook and pull in Australia
November games need November prices (85)
An early start to the international season, coupled with costly tickets, have kept the Australian public away from the cricket
The inherent dangers of batting (43)
The sickening blow that struck Phillip Hughes is a reminder of the ever-present dangers associated with facing fast bowlers, even while wearing a helmet
A crisis that defines the age (40)
Never mind cricket's absence from free-to-air TV - changes in social attitudes, the demands of work, and an individualistic age are all contributing to a decline in participation
Misbah's Pakistan or Imran's Pakistan? (38)
Pakistan have notched up some fine wins under Misbah-ul-Haq's leadership, but they haven't yet achieved consistent results outside the UAE
Going out to play cricket today would have been near enough to impossible. Even doing so next week in the nets and at the Gabba for the first Test will be difficult


text: ?Color is a strong weapon. With a gallon of paint you can change the atmosphere of a room. It's necessary to seek color consultation before starting even a simple faux painting in Los Angeles. Color consultation would end your indecisiveness about which color is most suitable for your home or room. It helps you figure which shade satisfies you most, relaxes your senses, and sets up the right mood. Murals, especially those made in LA, also uplift one's spirit. You may call a professional muralist in Los Angeles to work on an amazing mural collection in your house You can effortlessly impress visitors with murals. Why not have breathtaking trompe l'oeil murals at your front door so your visitors will feel that they are, indeed, welcome? Paint color serves as a powerful tool aside from other decorative elements. It brings a dramatic impact to every living space. It puts tranquility in every room or produces a profound feeling. Murals and painting can set a specific mood such as relaxation. Murals can inform or create an illusion, and uplift one's spirit. It can also let the mind travel to a certain perspective. It can bring you to places and make you feel closer to nature. Let these paintings and murals rule your world; travel while in your room, in your office, and even in your commercial space. Personalized paintings and murals are the best to have. If you are a nature lover, then why not have sceneries of the Niagara Falls or the Mount Fuji in your room? This will bring you closer to nature. Are you some kind of an entrepreneur? Then inspire your senses while in your restaurant, hotel, or retail establishment. This is an important element which could lead to the success of your business. Painting your bathroom with images you love creates a window or doorways to a world that adds depth to your life, which was nonexistent before. Bathroom murals are ideal for rooms that have no windows to bring memories of favorite places into one's home. Let your children appreciate artistic paintings and murals. Start stimulating your kids' imagination with a wall mural art. Brighten up your home or special nook with a wall art. Kids love wall murals for their rooms. There are themes and designs to choose from, or you may even personalize your own! Trompe L'Oeil murals enhance the illusion of depth and erase transition between real and painted worlds. As a stand along element, Trompe L'Oeil alters the appearance of architectural features and real objects adding depth, relief, ornamentation to an otherwise flat surface. Want to bring those fantastic blue skies inside your room? Authentic illusion of skies can transform confined interiors into spaces of natural beauty and freshness. Transform enclosed spaces into more spacious, inviting and memorable environments. The author is an expert writer associated with a company offering color consultation in Los Angeles. Visit the site for more information.


text: 500 pts Greg's Flashback Trivia 5/6 <Classic Rewards Club Frederick M. Brown/Getty Images In 1972, "Song Sung Blue" entered the U.S. Billboard charts. Who is responsible for this hit? Sign in to Participate Category: Trivia Share on Facebook Share on Twitter More Leave a comment Send to a friend! Print this page Pin it! Reddit This! Share on Tumblr! More Ways To Get and Use Points Greg's Flashback Trivia 11/25/2014Greg's Flashback Trivia 11/24/2014Oldiez Person Of Interest – Week of November 24thBeer Break of the Day – Week of November 24thWODZ Listen Live Word Of The Day – Week Of November 24thWeekly Bonus Code Week of November 24th


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We are a College and Career Readiness APEX* magnet school.See it in action here.*- APEX: Academic Programs for Equity and eXcellence
No Place For Hate Communities In Schools Boys' Athletics
Girls' Athletics
Yearbook National Junior Honor Society Student Council
Spirit/Dance Team
Robotics Club
Skyward Gradebook Parent Access
2013 Accountability Summary
TEA Released Test
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Scott AEIS Report 2011-2012
Distinctions 2012-2013
Campus Comparison Groups 2012 - 2013
Index Calculation & Data Table 2012-2013
Parents: please call the office to schedule a conference with teachers.
Click on the STAAR icon below to go to TEA's Test Resources page:
Click on the links below to access the following documents:
A TEA Recognized School, 2011 A TEA Exemplary School, 2010
Debra Owens, Principal
Salvador Chapa, Assistant Principal Tracy McCarley, Counselor Mary Ann Badalamenti, Campus Facilitator Joanna Nallie, Secretary Roberta Gonzalez, AP Secretary Phone: (409)761-6100 Fax: (409)770-0339


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- WOW! Samir Handanovic saves 6th (sixth!) consecutive penalty for Inter Milan v Dnipro
- TrollStation take credit for 3 pitch invasions during Spurs-Partizan 1st half
Great move! Omar Gonzalez (LA Galaxy) goal v Juventus
- Updated: August 4, 2013
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The best jokes as 3 pitch invaders halt Spurs v Partizan


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Tel: (610)687-4100
The Armenian Sisters Academy is dedicated to providing a quality education according to the Montessori Philosophy in a safe, caring, Christian environment where each child is nurtured to reach his/her full potential and to preparing the child to face the complex challenges in our society and to become a positive force in the community. The Academy promotes a strong sense of family, with the faculty, parents and children working together toward a common goal; the self-realization and academic happiness of each child
School Overview
|School Membership(s)School Assoc.
|American Montessori Society (AMS)
Association of Delaware Valley Independent Schools (ADVIS)
Pennsylvania Association of Independent Schools (PAIS)
|Religious Affiliation
|Grades Offered
|Grades Prekindergarten - 8
|Year Founded
Student Body
|Total Students
|113 students
|Student Body Type
|Students by Grade
Academics and Faculty
|Total Classroom Teachers
|23 teachers
|Student : Teacher Ratio
National avg.: 13:1
|% Faculty w/Advanced Degree
|Classroom Dress Code
Finances and Admission
|Admission Deadline
|None / Rolling
|Yearly Tuition Cost
|Tuition Notes
|Tuition for first and second child is $4,750; for third child, tuition cost is $4,350
|% on Financial Aid
|Average Financial Aid Grant
|Admissions Director
|Susan Pogharian
|Total Sports Offered
|3 sports
|Basketball, Dance, Track and FieldBasketball, Dance,
Track and Field
|Total ExtracurricularsTotal Extra-curric.
|2 extracurriculars
|Girl Scounts, Club Time Girl Scounts, Club Time,
School Notes:
What makes the Armenian Sisters Academy special? Simply put, in addition to a strong academic program, students learn essential values. At the Academy, respect is fostered towards other students, teachers and the school itself. ASA children are taught life lessons in kindness and helping others. Small class sizes ensure they receive individualized attention - there is no getting lost in the shuffle at the Academy. Young children are mentored and as they mature, they guide other youngsters. Students learn and grow spiritually and culturally in a comfortable environment where they are part of a large, extended family. And in the process, they become fluent in a second language and have a solid sense of identity and belonging in their community.
Nearby Schools:
All Schools | High Schools | Elementary Schools | Preschools | Public Schools
Show me:
- School Location Miles Students Grades
WayneWoodlynde School
445 Upper Gulph Road
Wayne, PA, 19087
(610)687-9660Special Program Emphasis0 mi | 255 students | Gr. K-12
WayneGoddard School
95 Crestline Rd
Wayne, PA, 19087
(610)688-5229Daycare / Preschool0.4 mi | 10 students | Gr. NS-K
StraffordOur Lady Of The Assumption Preschool & Kdgn
135 Fairfield Ln
Strafford, PA, 19087
(610)688-5277Daycare / Preschool (Roman Catholic)0.6 mi | 20 students | Gr. PK-K
WayneHeritage School
675 N Wayne Avenue
Wayne, PA, 19087
(610)688-6342Daycare / Preschool (Christian)0.8 mi | 24 students | Gr. PK-K
DevonDevon Preparatory School
363 N. Valley Forge Road
Devon, PA, 19333
(610)688-73370.9 mi | 259 students | Gr. 6-12
All-boys (Roman Catholic)
School Reviews
Area Statistics
Recent Articles
Updated October 26, 2014
What do you do when you are dissatisfied with your local public schools? Homeschool your children? Send them to private school? Let's look at your options.
Your religion matters to you. You want your child to attend a school which will combine academics and religious education. Here is an overview of what is available together with some links to denominational web sites.
Add things up and you'll quickly find the answer- math is crucial both in academics and the real world. By following these strategies and following personal training programs, students can uncover their weaknesses and conquer math.


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You're leaving Wedding Paper Divas to see the Deal of the Day at our parent site, Tiny Prints.
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"I just got my save the dates in the mail, and they are so gorgeous! I am impressed with the quality of it all. I got them in the pink and black color, with the shimmer paper and shimmer white envelopes, and i love how they came out. I also asked if i could have my wedding website added to the back of the card, which was done and it looks lovely. I am very pleased, and look forward to ordering my invitations!"
"Loved how my Save the Dates came out! I did have to change the font because our names were hard to read, but it turned out great!"
"I ordered all of my wedding invitations, save-the dates, thank you cards, etc from wedding paper divas and EVERY SINGLE thing I ordered was perfect. My proofs were done quickly, I received the items early and was beyond excited to open every one. WPD is a wonderful company, great customer service and very professional. Highly, highly recommend them."
"This is a beautiful design - ordered the silver Save The Date cards with an upgrade to the pearlized white paper, and we are thrilled with the results! We used one full size image for the back and we love how it looks. Would highly recommend as a simple, classic, and sophisticated design that isn't overwhelming, but lets you focus on the information and the image."
"I searched many sites to find a design that was simple yet elegant and I finally found it with the Simplified Deco. I was very pleased with the design process which was very easy, and the complimentary design/etiquette review caught several errors which saved me a headache! I have received tons of compliments from friends and family who say they are beautiful! I plan to use this whole suite for my wedding and look forward to working with WPD again!"


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Kihei Condo Rental Photos and Description
2 Bed, 2 Full Bath With Easy Access To Swimming Or Surfing
Aloha to our future guests and returning friends! We are so excited to share our warm and wonderful Maui condo with you. It was renovated in August 2012 and needs to be enjoyed.
Pacific Shores is a small condo complex superbly located across from Kamaole I and Charley Young Beach, a long beautiful beach ideally suited for swimming, snorkeling, sunbathing, toasting the sunset, or building giant sandcastles. Just a micro-walk to the north is another beach tailor-made for learning and practicing the zen of surfing or the coolest new activity, paddle boarding.
Start your days with coffee or tea on the lanai, a surf lesson, breakfast at the Kihei Caffe and continue on with beach time, Tuesday tacos at Fred's, a little volleyball or whatever your heart desires. We've tried to provide everything you'll need - to know you're on vacation but have all the comforts of home - including hotel-sized toiletries, fresh fluffy linens, and a fully equipped kitchen, as well as beach toys, chairs, and umbrellas.
In addition to an ideal location, Pacific Shores sports a clean and beautiful pool with sea turtle mosaics in a wash of royal blue tile as well as a large, well heated, relaxing hot tub. Alongside the pool are two BBQ grills, ready for your use, and plenty of tables for dining, picnicking, or just keeping your water bottles upright while you swim.
For evening entertainment, we offer Netflix and HuluPlus subscriptions (always logged on), a DVD player, complementary very high speed Wireless Internet access, and a game table complete with a variety of games and puzzles for all ages.
Whether you are honeymooning, vacationing with the kids, or just getting away with friends for fun in the sun, our location is ideal and our condo is inviting you to stay.
- Suitability:
- Minimum Age Limit for Renters
- alternative lifestyle
- children welcome
- limited accessibility
- non smoking only
- pets not allowed
About Jan Hanson
We are a group of four friends who could never have afforded a vacation home on Maui on our own. We are so excited to have found a way to spend large portions of our retirement years in paradise.
Jan Hanson purchased this condo in 2012
Why Jan Hanson chose Kihei
The location and amenities of this complex were exactly what we wanted. Two bedrooms AND two bathrooms allows us to have visitors when we are using the unit, but also allows them privacy. Our main objective was to find a place where WE would want to spend a lot of time. We renovated it right away and couldn't be more thrilled with the results.
What makes this condo unique
Two beaches with distinctly different offerings. It is wonderful to be able to get up in the morning and walk out to either a swimming beach OR a surfing beach with easy board rental. The unit is also about evenly situated between walkable restaurants and shops to the south and to the north. We may never get a car!


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PodcastOne has issued a demand to Stitcher (recently acquired by Deezer) to cease distribution of programming that PodcastOne produces or has the exclusive sales rights to. Norm Pattiz (pictured), PodcastOne Chairman/CEO, commented on the dispute saying, "In some cases, Stitcher has absolutely no rights to carry our programs, and in others, no rights to sell or monetize our exclusively represented content in any way. This has been an ongoing problem, and enough is enough. You can't just steal content without permission." … TechCrunch East Coast Editor John Biggs and Jordan Crook, TechCrunch writer and reporter, will host TechCrunch Radio as an exclusive weekly series for SiriusXM subscribers. The weekly show will be available each Tuesday at 6pm (ET) starting today (11/25) … Also effective today, TuneIn launches as the newest app now supported for the Google Chromecast device. The integration will allow Chromecast users to easily stream TuneIn radio content on their television screen.
Chicago Tribune reporter Robert Channick spoke with WGN Radio President and manager Jimmy de Castro about yesterday's announced decision to pull the plug on both it's FM Sports/Talk experiment at 87.7 The Game (WGWG-LP) and the recently launched streaming venture, WGN.fm. As you'd probably expect, de Castro (pictured) cited economics as the driving force behind the decision to shutter both operations that were launched with a fair degree of fanfare some nine months ago. Tribune has a deal to operate WGWG through next September via an LMA with Venture Technologies Group, which owns the frequency. Asked what, if anything, will be airing on the 87.7 frequency come January 2015, de Castro (pictured) told the newspaper he really didn't know yet but, "It's not going to be sports, that's for sure." Read more (subscription required) HERE.
Cumulus/Youngstown Market Manager Rick Parrish says he's headed for retirement after more than four decades in radio ownership and corporate management. Quarterback for the Youngstown cluster (including News/Talkers WPIC and WSOM, and Sports/Talkers WLLF-FM and WBBW-AM) since 2010, Parrish (pictured) says he believes the best is still to come for a guy who has "loved radio since the age of 15 when my mother drove me to KMAD in Madill, Oklahoma for my first day of on-air work!" … Former award winning ESPN podcast producer Jay Soderberg is named Head of Content for BlogTalkRadio, where he'll focus on cultivating and expanding the online radio platform's community of podcasters, and also take a leading role in shaping the company's future content portfolio … Jonathan Ghazal has been named Sales Manager for Greater Media-owned Detroit Sports 105.1 (WMGC). Ghazal joined the company's Motor City cluster as an Account Executive in October of 2012 … Alex Ferrer, best known for his nine year run on FOX TV's syndicated Judge Alex Show, will debut a new legal-focused radio show via Andy Dean's new ConnectPal digital venture. A frequent guest commentator on Fox News, CNN, and HLN, 'Judge Alex' is also highly active on social media, with a Facebook following closing in on a quarter-million fans.
Following his recent split with Cumulus Media-owned Westwood One, former Governor Mike Huckabee (left) has officially announced it's "on with the show" and confirms he will continue to host his daily commentary, The Huckabee Report, without interruption to affiliates. Huckabee spokesperson and McConnell Media namesake, John McConnell, reports that a new distribution and ad sales partner for the daily feature will be announced next week. "The support and interest from the Governor's current affiliates has been tremendous," McConnell told NTS MediaOnline Today. "He's beloved by his fans and his fan base is huge!" … Cumulus/Memphis has tapped popular local radio veteran Forrest Goodman as morning host at Newstalk 98.9. He'll be joined for wake-up duties by Steve Butler, weekdays from 6-9am … Westwood One ups veteran network sales exec Julie Boruchov (right) to Vice-President, Sports Sales and Marketing/East Coast, in its network sales division. She was formerly Director, Sports Sales, East Coast. In her expanded role, Boruchov will work more closely with the network's ports partners and agency clients to help drive revenue and maximize brand initiatives and activations, while also managing a team of east coast-based sports sales account executives.
After wrapping up weekend hosting duties for Premiere Networks syndicated Coast To Coast earlier this year, longtime Texas talker John B. Wells (pictured) debuted Caravan To Midnight, a daily three-hour subscriber-based cybercast. Produced Monday-Friday and available for broadcast in all day parts, Wells "explores and exploits critical news stories that the mainstream media often ignores or superficially covers." John B. is now taking the next step, announcing a national syndication rollout of 'Caravan' via his own C2M Network, with WRKO/Boston signing on as the show's the first Top 10 market affiliate (WRKO will air the show on Saturday nights). Heading up syndication efforts for Wells' new Dallas-based venture is former CBS Radio ad exec David Rubini who said, "John B. has a long track record for making money for his clients. Given the instant and overwhelming response and support for the Caravan to Midnight cybercast, I'm confident his track record is going to get longer." Wells also said his future plans also include developing new talent for syndication via his C2M Network.
Veteran Talk radio PD Paul Duckworth reports he's going to give retirement another try. "After two-and-a-half terrific years at KOMO and KVI in Seattle, I'm going to try my hand at that whole retirement thing again," said Duckworth (pictured), who previously tried to hang up his headphones a couple of years back, following a distinguished programming career that included prior stints at WMAL/Washington, KLIF/Dallas and KPAM/Portland, along with a previous run at KOMO from 1997-2005. You can wish Paul 'Happy Trails' HERE … Congrats to Salem Radio Network host Michael Medved on receiving the 2014 Quill & Badge Award from the International Association of Police Unions, AFL-CIO. The award was given to Medved to cite his ongoing on-air and print support of America's law enforcement officers … This Thursday (11/6), E! Television personality and best-selling author Ross Matthews will debut a new weekly podcast, Straight Talk with Ross Matthews, via PodcastOne … CONDOLENCES: To the family and friends of Tom Magliozzi, half of the wisecracking Car Talk duo known as Click and Clack, who died from complications of Alzheimer's disease. He was 77. Read more from NPR HERE.
In a recent post, New York magazine technology columnist Kevin Roose writes that after a few years of waning interest, today's world of podcasting is experiencing a "renaissance" in growth and argues that the medium is, "More alive than ever." In support of that claim, Roose cites the most recent Infinite Dial study from Edison Research and Triton Digital, which reported that some 39 million respondents had listened to a podcast in the 30 days prior to the study being taken — the highest number on record. So what's behind the resurging interest in a technology that was first introduced in the early 2000′s? Three key things: Improved quality/content, compelling economics and connected cars. "Connected cars are a boon for the entire streaming audio industry, but they're especially exciting for podcast makers, whose shows are perfectly suited to in-car listening," writes Roose. "Just as TV watchers can now choose Netflix or Amazon streams over surfing channels, radio listeners will soon have a bevy of on-demand options at their disposal." Read the full article HERE.
Paris, France-based worldwide digital streaming music service Deezer announced today that it will acquire Stitcher, the San Francisco-based personalized Internet Talk radio and mobile audio provider. Stitcher currently carries 8 of the top 10 broadcast radio talk shows, content from more than 12,000 providers and is enabled for the connected dashboard in more than 50 automobile models from BMW, Ford, GM, Jaguar and Mazda. It's also available via Apple CarPlay and Android Auto … Former WIBA/Madison host Mitch Henck (pictured) — who got 'RIF'd' at the iHeartMedia-owned News/Talker this past June after a 12-year run — will launch a new daily online talk show on Monday (10/27). The Mitch Henck Show will stream from 8-11am (CT) weekdays at MitchHenck.com … iHeartRadio joins the lineup of Apple's new CarPlay, placing the popular smartphone app directly into the automobile dashboard … The previously announced debut of Jenny McCarthy's new daily SiriusXM show – Dirty, Sexy, Funny with Jenny McCarthy — happens Monday on the satellite radio service's Stars channel, from 10am-noon (ET).
You've likely read about last weekend's Pumpkin Fest riots in Keene, NH, a town that also happens to be the home of Free Talk Live host Ian Freeman. Ian (pictured) was on the ground and witnessed crowds of drunken college students destroying property, as well as what he terms, "an unnecessarily aggressive police response" to the situation. Station newsrooms, hosts and producers who would like to get Freeman's up-close-and-personal views on the chaotic weekend scene in the small New Hampshire town can reach him HERE … WYD Media Ad Sales has inked a deal with Spreaker to rep ad sales for the digital talk media and audio content distribution platform. Additionally WYD will assist Spreaker with overall brand management and creation of alternative and customized opportunities for the advertising community … Carlos Lamadrid has been appointed Vice President of National Sales and Strategic Partnerships for Entercom. Lamadrid, most recently SVP at TC Media, will report to Entercom Station Group President Weezie Kramer.
Bridge Ratings, which has been trending News/Talk radio preferences since 1999, has released a new report that was recently posted at LinkedIn by company President/CEO Dave Van Dyke. So what is the study's top line finding? Information overload is causing fatigue for some News/Talk listeners. "Over-saturation, ever-shortening of news cycles and repetition of hard news with too-little updated information, is wearing thin on consumers who are seeking alternative sources for information that is fresh, entertaining and positive," writes Van Dyke (pictured). "Radio has become one of many pieces in the media landscape puzzle. This role has become uncomfortable for broadcasters but should be seen as an opportunity to reinvent, develop and serve their communities during a time of rich information flow and a vast funnel of information to consume." Bridge's latest study is based on interviews with some 3,200 News/Talk radio listeners who consume at least 3 hours of News/Talk radio weekly. Respondents were compiled through random digital dial, internet self-administered questionnaires and in-person interviews, with a margin of error of +/- 1.7%.


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You don't pay for tutoring
unless you find a good fit!
Jennifer C. has not yet had a background check. After sending a message to Jennifer, you will be able to order a background check for $7.99. The check will be processed by First Advantage. For more information, please review the background check information page.
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My name is Jennifer, and I am a fine artist who specializes in painting with oil.
I am always thrilled to have the opportunity to demonstrate to students the fascinating nuances of this very old craft.
The joy of painting comes from discovering the qualities of the paint and then learning to manipulate it, so that the paint transforms itself into the wonderful vehicle of your creative expression.
Painting opens and feeds the mind; it is beautiful, scientific, unassuming, forgiving, satisfying, meditative and challenging by it's nature.
It is also strenuous, because it requires the use of both sides of the brain simultaneously. Ideally, a painter becomes calm and introspective while painting, and learns to tap into the pliable, confident, inner-self to help make the many decisions which painting requires. Learning to confidently express and execute one's decisions is a valuable life-tool, indeed.
In addition to teaching the craft, I also consider it an honor to share with a student, the engaging historical, scientific and anthropological background associated with the art of painting.
Art is a source of lifelong enjoyment, and I sincerely hope that I may be the one to help foster that skill in you or your child.
Read all subject qualifications »
Louisiana Tech University
Louisiana Tech University (Other)
Standard Hourly Fee: $45.00
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Jennifer will travel within 25 miles of Derry, NH 03038.
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