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In this report we describe a chromatin immunoprecipitation (ChIP) protocol for two fully sequenced model diatom species Phaeodactylum tricornutum and Thalassiosira pseudonana. This protocol allows the extraction of satisfactory amounts of chromatin and gives reproducible results. We coupled the ChIP assay with real time quantitative PCR. Our results reveal that the two major histone marks H3K4me2 and H3K9me2 exist in P. tricornutum and T. pseudonana. As in other eukaryotes, H3K4me2 marks active genes whereas H3K9me2 marks transcriptionally inactive transposable elements. Unexpectedly however, T. pseudonana housekeeping genes also show a relative enrichment of H3K9me2. We also discuss optimization of the procedure, including growth conditions, cross linking and sonication. Validation of the protocol provides a set of genes and transposable elements that can be used as controls for studies using ChIP in each diatom species. This protocol can be easily adapted to other diatoms and eukaryotic phytoplankton species for genetic and biochemical studies.
Keywords:Phaeodactylum tricornutum; Thalassiosira pseudonana; Histone modifications; Chromatin immunoprecipitation; Epigenomics
Diatoms are a group of eukaryotic phytoplankton with a wide distribution and a large diversity in marine and fresh water ecosystems. It is estimated that there are between 10,000 and 100,000 extant diatom species [1]. Diatoms play essential roles in global biogeochemical cycles because they are believed to be responsible for 20% of global carbon fixation and 40% of marine primary productivity [2]. Diatoms capture CO2 through photosynthesis and act as a critical buffer against global warming by sequestering organic carbon in the ocean interior. They can live under different conditions in all oceans from the poles to the tropics, and are often the first group of phytoplankton to benefit from sporadic nutrient upwelling events, indicating their intrinsic adaptability to changing environments.
Completed whole genome sequences from three species, the centric diatom Thalassiosira pseudonana, and the pennate diatoms Phaeodactylum tricornutum and Fragilariopsis cylindrus are now available [3,4], ( http://genome.jgi-psf.org/Fracy1/Fracy1.home.html webcite). Additional pennate species, the toxic Pseudo-nitzschia multiseries, Fistulifera sp. and Seminavis variabilis are also being sequenced and will provide an additional source for comparative genomics. T. pseudonana is widely distributed in marine environments and is of significant ecological importance. P. tricornutum on the other hand is considered to be of little ecological relevance but is the model system for pennate diatoms because of a long history of physiological experiments and the availability of a wide range of tools for reverse genetics [5,6]. Furthermore, a digital gene expression database ( http://www.diatomics.biologie.ens.fr/EST3/index.php webcite) is available for both species.
The whole genome sequences have revealed a wealth of information about diatom genes. It was shown for example that diatoms have acquired genes both from their endosymbiotic ancestors and by horizontal gene transfer from prokaryotes. But while DNA is the substrate for mutations upon which natural selection can act, DNA sequence in itself may not explain adequately their ability to adapt to changing environments, and more flexible mechanisms based on epigenetic processes could provide additional control. These changes include DNA methylation and histone tail post-translational modifications that alter chromatin structure. Study of diatom epigenomes can therefore provide a more in depth look at the regulatory mechanisms underlying their natural phenotypic adaptability to environmental changes.
Several tools for studying chromatin have been developed. Among them, chromatin immuno-precipitation (ChIP) has become a powerful tool to detect in vivo interactions between a DNA-associated protein and genomic DNA. ChIP combined with microarray or massively parallel sequencing is used to study gene regulatory networks active during development and/or in response to the environment. ChIP is also a valuable tool for mapping genome wide epigenetic modifications such as histone marks, and has been used to characterize several eukaryotic genomes [7,8]. However, no ChIP protocol has been reported for marine phytoplankton. T. pseudonana and P. tricornutum were therefore chosen to set up a ChIP protocol in diatoms using two histone marks known to characterize active and repressive chromatin states in other organisms.
The principle of a ChIP procedure includes: (1) Cross linking of DNA and protein with formaldehyde to covalently combine DNA and attached proteins in vivo, (2) Fragmentation of the fixed chromatin to an average size of 500 bp ranging from 200 to 1000 bp, (3) Chromatin extraction by a succession of extraction buffers, (4) Immunoprecipitation with specific antibodies, (5) Purification of immune complexes after immunoprecipitation and reverse crosslinking, and (6) Analysis of bound DNA by PCR which involves comparison of the intensity of PCR signals from the precipitated template with positive and negative controls. Standard PCR on immunoprecipitated DNA from a specific genomic region provides a direct assessment of protein association with that region, whereas quantitative PCR can assess not only whether a protein binds to that region, but also further compare the relative abundance at different genomic regions.
The protocol described in this work was optimized for each of the steps described above. It is an adaptation of ChIP protocols used for yeast and Arabidopsis[9,10]. It is therefore not new in its principles but takes into consideration features inherent to diatoms such as their siliceous frustule, the cell wall and the chloroplast.
Materials and methods
Formaldehyde 36.5% (Fluka cat. No. 200018)
Glycine (Sigma cat. No. 2412)
Sucrose (Sigma cat. No. S-0389)
EDTA (Sigma cat. No. E-4884)
2-MERCAPTOETHANOL (Sigma cat. No. 125 k0165)
SDS (EUROMEDEX cat. No. EU660)
Triton X 100 (Amnersham bioscience)
Magnesium chloride (Sigma cat. No. 7786-30-3)
Sodium chloride (Sigma cat. No. 7647-14-5)
Sodium Acetate (Sigma cat. No. 127-09-3)
Lithium chloride (Sigma cat. No. 7447-41-8)
Sodium deoxycholate (Sigma cat. No. D-6750)
IGEPAL CA-630 (Sigma cat. No. 043 K0654)
NaHCO3 (Prolabo cat. No. 2 778.293)
Complete protease cocktail Inhibitor (Roche cat. No. 11873 580 001)
Proteinase K (Ambion cat. No. AM2546)
RNase A (Fermantas cat. No. EN0531)
Dynabeads protein G (Invitrogen cat. No. 100.04D)
Dynabeads protein A (Invitrogen cat. No. 100.02D)
Phenol (CAROLO ERBA Cat. no. 108-95-2)
Chloroforms (Prolab cat. No. 22 716.296)
Glycogen (Fermentas cat. No. R0561)
SYBER® Green master mix (Life Technologies)
Sonicator (Bioruptor® UCD-200 from Diagenode)
Rotating wheel
Freezers (−80 and −20°C)
Falcon tubes (50 ml)
Eppendorf tubes (1.5 and 2 ml)
Siliconized eppendorf
Magnet Dynal
Extraction buffer I: 0.4 M sucrose, 1 mini tablet Roche per 50 ml, 10 mM MgCl2, 5 mM 2-MERCAPTOETHANOL, 10 mM Tris–HCl pH 8
Extraction buffer II: 0.25 M sucrose, 10 mM Tris–HCl pH8, 10 mM MgCl2, 1% triton, 1 mini tablet Roche diluted in 1 ml (for 10 ml), 5 mM 2-MERCAPTOETHANOL
Extraction buffer III: 1.7 M sucrose, 1 minitablet diluted in 1 ml (for 10 ml), 0.15% Triton X-100, 2 mM MgCl2, 5 mM 2-MERCAPTOETHANOL, 10 mM Tris HCl pH8
Nuclei lysis buffer: 50 mM Tris HCl pH 8, 10 mM EDTA, 1 mini tablet of protease inhibitors diluted in 1 ml (for 10 ml), 1% SDS
ChIP dilution buffer: 1% triton, 1.2 mM EDTA, 167 mM NaCl, 16.7 mM Tris HCl pH8 CRITICAL keep at 4°C.
Low salt wash buffer: 150 mM NaCl, 0.1% SDS, 20 mM Tris–HCl pH8, 2 mM EDTA, 1% Triton X-100 CRITICAL keep at 4°C.
High salt wash buffer: 500 mM NaCl, 0.1% SDS, 1% Triton X-100, 20 mM Tris–HCl pH8, 2 mM EDTA CRITICAL keep at 4°C.
LiCl wash buffer: 0.25 M LiCL, 1% IGEPAL CA-630, 10 Mm Tris–HCl pH8, 1 mM EDTA, 1% sodium deoxycholate CRITICAL keep at 4°C.
TE buffer: 10 Mm Tris–HCl pH8, 1 Mm EDTA CRITICAL keep at 4°C.
Elution buffer: 1% SDS, 0.1 M NaHCO3 CRITICAL keep at 4°C.
CRITICAL: All the buffers should be prepared fresh and when required, the addition of the protease inhibitors, should be done directly before using the buffer.
Harvesting cells and cross-linking
1. Grow P. tricornutum culture in 400 ml artificial sea water under the standard conditions until cell density reaches around 1 million cells/ml.
2. Add 11.27 ml of 36.5% of formaldehyde to the culture to get final 1% concentration in the whole medium.
3. Stop fixation by adding 2 M glycine (final concentration is 0.125 M) for 5 min at room temperature.
4. Wash the cells with PBS solution twice by centrifugation at 4000 rpm for 5 min at 4°C.
Comment: Fixed pellet can be stored at −80°C for several months.
Chromatin extraction and sonication
5. Add approximately 5 ml of Extraction buffer I to 50 ml culture pellet.
6. Leave the falcon tubes on ice for 5 min.
7. Spin the solution at 4000 rpm for 20 min at 4°C.
8. Gently remove supernatant and resuspend the pellet in 1 ml of Extraction Buffer II.
9. Spin at 10,000 rpm for 10 min at 4°C.
10. Remove supernatant and resuspend pellet in 300 μl of Extraction Buffer III.
11. In a clean eppendorf, add 300 μl of Extraction Buffer III. Take the 300 μl solution (resuspended pellet) from last step and carefully layer it on top of the clean 300 μl of Extraction Buffer III.
12. Spin at 13,000 rpm for 1 hour at 4°C.
13. Remove the supernatant and resuspend the chromatin pellet in 300 μl (or 200 μl if small pellet) of Nuclei Lysis Buffer. Resuspend the pellet by pipetting up and down and vortexing (keep solution cold between vortexing).
14. Sonicate the chromatin solution for 9 cycles 30 seconds ON and 1 minute OFF for each cycle on full power. Keep 5 μl for DNA extraction to check sonication efficiency.
CRITICAL Supernatant is usually less than 300 μl following sonication due to liquid loss.
15. Check the sonicated chromatin after reverse cross-linking and recovering the DNA on 1% agarose gel. The DNA fragment should be around 200 bp-1000 bp.
Comment: Sonicated chromatin can be frozen at −80°C for 3 months or can be used directly for immunoprecipitation.
Immunoprecipitation and reverse crosslinking
16. Spin at full speed the chromatin solution for 5 minutes at 4 degrees to pellet debris. Remove supernatant to a new tube.
CRITICAL It is important to remove 20 μl for Total DNA Control for INPUT.
17. Measure out the remaining volume of sonicated chromatin and bring volume up to 3 ml with ChIP Dilution Buffer. The point here is to dilute the 1% SDS to 0.1% SDS with ChIP dilution buffer.
18. Split the chromatin solution into 3 tubes (1 ml each).
Tube 1. With DNA A beads labeled H3K4me2
Tube 2. With DNA A beads labeled H3K9me2
Tube 3. With DNA A beads labeled No Antibody (mock)
19. For each IP, mix 45 μl beads A and 45 μl DNA beads G in siliconized tubes. Wash DNA beads twice with ChIP Dilution buffer and resuspend beads in 90 μl Chip dilution buffer. Split the 90 μl resuspended beads into 30 and 60 μl volumes.
20. Add 5 μl of your antibody to the siliconized eppendorf tube that contains mixed 60 μl beads for Ig capturing. Add the diluted chromatin solutions into the tubes that contain 30 μl mixed beads for preclearing. Leave tubes with gentle rotation at 4°C for 2 hours.
Comment: Using precleared chromatin and siliconized eppendorf tubes can significantly decrease negative control (mock) noise background.
21. Wash the beads-Ig complexes once by 1 ml ChIP dilution buffer and resuspend in 60 μl ChIP dilution buffer. Transfer the precleared chromatin into the beads-Ig complexes. Leave tubes with gentle rotation at 4°C overnight.
22. Wash the DNA beads-Ig-antigen complexes 4 times using Low Salt Wash Buffer, High Salt Wash Buffer, LiCl Wash Buffer and TE Buffer in sequence. Use 1 ml of each buffer per wash and wash twice per sample. One wash is quick without agitation and the other one is at 4°C with gentle rotation for 5 min.
Comment: Put all the wash solutions on ice prior to immune complex collection.
23. Elute immune complexes by adding 250 μl of Elution Buffer to the washed beads. Vortex briefly for mixing and incubate at 65°C for 15 min (mix tubes during incubation). Put tubes on the magnet Dynal and carefully transfer the eluate to another eppendorf tube (not siliconized tube) and repeat elution once again. Combine them to obtain final 500 μl of elute.
25. Add 20 μl 5 M NaCl to the eluate and reverse crosslink at 65°C overnight.
Comment: Do not forget to get your total DNA out of the freezer and reverse cross-link with other samples (Add 500 μl of Elution buffer + 20 μl 5 M NaCl).
DNA recovery
26. Add 10 μl of 0.5 M EDTA, 20 μl Tris–HCl 1 M (pH 6.5), 2 μl of 10 mg/ml proteinase K, and 1 μl of 10 mg/ml RNase to the eluate and incubate for one hour at 45°C.
27. Recover DNA by phenol and chloroform (phenol; phenol/chloroform 1:1; chloroform) extraction (equal volume) and precipitate DNA with EtOH (2 vol 1/2) and 1/10 vol NaAc (3 M pH 5.3). Add 2 μl glycogen (20 mg/ml) to ethanol precipitation step. Incubate at - 80°C for 15 min or 2 hours at −20°C.
28. Centrifuge at 4°C at 13,000 rpm for 30 min and wash pellets with 400 μl 70% ethanol. Dry the pellet at room temperature.
29. Resuspend the pellet in 50 μl of distilled water or 30 μl to concentrate DNA.
Quantitative PCR
For both diatoms, specific primers were designed for two genes and a set of TEs (Table 1). The input DNA pulled out from ChIP was diluted 10 times before q-PCR. Quantitative PCR was performed using a Roche LightCycler® 480 machine on 1 μl of IP, input and mock DNA which were mixed to 5 μl LightCycler® DNA Master SYBR Green I 2X, 3 μl forward/reverse primers 1 μM, and 1 μL H2O. The PCR program was performed as follows: 10 min at 95°C; 45 cycles of 95°C for 15 seconds and 60°C for 1 min.
Table 1. Primer sequences used for QPCR analysis
Data analysis
The Ct value (number of cycles required for the fluorescent signal to cross the threshold) is recorded in the experimental report after analysis by Roche LightCycler® 480 software. The Ct values of the duplicates should show minimal variability, indicating that samples were properly handled (ideally, it should be below 0.2). Ct values were used for performing the calculation which consists on evaluating the fold difference between experimental sample and normalized input.
ΔCt (normalized to the input samples) value for each sample. ΔCt [normalized ChIP] = (Ct [ChIP] - (Ct [Input] - Log2 (Input Dilution Factor).
Where Input Dilution Factor = (fraction of the input chromatin saved)-1 × Input dilution factor before q PCR. Here the fraction of Input chromatin saved is 20 μl and the fraction for each IP is 90 μl. The IP fraction is 4.5 times the input fraction. For QPCR runs Input was diluted 10 times which makes the final dilution factor of the Input fraction (Input Dilution Factor) = 4.5 × 10 = 45. Then the equation above is as follows: ΔCt [normalized ChIP] = (Ct [ChIP] - (Ct [Input] - Log2 (45). Finally, the percentage (Input %) value for each sample is calculated as follows: Input % = 100/2 ΔCt [normalized ChIP]. The "Input %" value represents the enrichment of certain histone modification on specific region.
Peptide competition assay and western blot
The antibody was pre-incubated with the peptide prior to use in immunoblotting assays. Different amounts (depends on different peptides and antibody) of peptides were added to a 10 ml BSA solution containing antibody and incubated under gentle agitation for 4 h at room temperature and an additional 1 hour at 4°C before the immunoblotting assays. Calibration curve were performed for each antibody to determin the amount of antibody to use (data not shown). The antibodies and peptides used in this work include H3K4me2 (Millipore Ref: 07–030), H3K9me2 (Millipore, Ref: 17–681), H3K4me1 peptide (Abcam Ref: ab1340), H3K4me2 peptide (Abcam Ref: ab7768), H3K4me3 peptide (Abcam Ref: ab1342), H3K9me1 peptide (Millipore, Ref: 12–569), H3K9me2 peptide (Millipore, Ref: 12–430), H3K9me3 peptide (Millipore, Ref: 12–568). The references of H3K9me2 antibodies that were discarded because of the lack of specificity are Millipore Ref: 17–648 and Millipore ref: 05–1249. Different concentrations of antibody and peptide concentration were compared. The nuclear enriched protein used for immunoblotting was extracted following the chromatin extraction protocol with minor modifications: the culture was not fixed by formaldehyde and sonication was not needed.
Results and discussion
Chromatin extraction and processing
ChIP is a very powerful technique for revealing association of specific DNA regions with proteins of interest. However, it is not a trivial technique, and highly specific antibodies against the protein of interest or a particular histone modification are required. Furthermore, false-negative signals may originate from inefficient antibody binding, and the beads used in ChIP can bind non-specific sites and cause background noise in negative controls. The starting materials for immunoprecipitation are also critical in terms of number of cells in a given volume, and sample dilution can effectively decrease background noise. To avoid saturation of antibody in ChIP assays, calibration curves should be built before precipitation with the antibody to determine the optimal amount of antibody to use.
The protocol described herein (Figure 1) has taken into consideration these issues as well as the particularities of diatoms. Diatoms have the unique ability to precipitate soluble silicic acid into a finely patterned cell wall built from amorphous silica, and they also possess photosynthetic chloroplasts. Chromatin extraction buffers modified from Arabidopsis ChIP protocols were used for P. tricornutum and T. pseudonana which have plant features, such as chloroplasts and a cell wall. It is important in this protocol to use artificial sea water instead of natural sea water in order to control the different components of the medium such as silica. Extraction of chromatin from diatom species can be difficult because of the silica based rigid cell wall [11], which can interfere with chromatin extraction because it binds DNA. In this study, we have chosen species that have different requirements for silicon. P. tricornutum has a facultative requirement for silicic acid, whereas T. pseudonana needs a silicon source to grow. A growth medium without silicic acid was used for P. tricornutum while T. pseudonana grew in medium containing a relatively high concentration of silica (~ 88 μM) reflecting the amount found in natural environment.
Figure 1. Outline of the ChIP QPCR protocol. Timing for each step is indicated between parentheses.
To preserve cell integrity, diatom cells were fixed in the growth medium prior to any handling. Formaldehyde was used for cross linking. Cross linking is a time dependent procedure and our trials have established 10 minutes as an optimal time for both P. tricornutum and T. pseudonana. Excessive cross linking might reduce antigen accessibility and sonication efficiency. For a different diatom species, we recommend 5 minutes for a start, proceed with sonication, reverse cross link and run a gel to see how much DNA is recovered and whether the size is optimal. If not, we recommend trying longer or shorter times for cross linking.
Sonication time was determined after trying three different times, 7 cycles of 25 seconds ON and 1 minute OFF, 9 and 12 cycles with 30 seconds ON and 1 minute OFF. Nine cycles gave the best range of DNA sizes which is between 220 and 1000 bp with a maximum of DNA fragments at 500 bp (Figure 2). Different dilutions of 200 ml sample containing 5 million cells per ml were used for sonication as high cell density can interfere with the efficiency of sonication. Four times dilution gave the best range of fragment sizes (Figure 2).
Figure 2. Sonication efficiency. Different times and number of cycles were used for sonication. After crosslinking, eluted DNA was loaded in a 2% gel to monitor the efficiency of sonication. From left to right, ladder, 10 times diluted sample containing 500000 cells/ml with 7 cycles 25 seconds ON and 1 minute OFF, original sample from 200 ml culture containing 5 million cells/ml, 4x and 2x diluted samples with 7 cycles, 25 seconds and 1 minute OFF and 4x diluted sample with 9 cycles, 30 seconds ON and 1 minute OFF.
Peptide competition assay
The nuclear-enriched protein used for immunoblotting was extracted following a chromatin extraction pro-tocol with minor modifications (see Methods section). Antibodies for two histone marks H3K4me2 and H3K9me2 were used for validation of the ChIP protocol. A peptide competition assay was performed to confirm the specific band reactivity of the antibodies. This is an important issue especially for genome wide studies as unspecific antibody binding will lead to non-specific signals increasing background noise and the occurrence of false positives. Both antibodies were pre-incubated with three different concentrations of the corresponding peptides (see Methods section) prior to immunobloting. For the H3K4me2 histone mark, the peptide competition assay did not detect the presence of a band for peptide H3K4me2 while a band was seen for the two other peptides, indicating the absence of competition with the other modifications of lysine 4 (Figure 3A). Three different H3K9me2 antibodies from Millipore were tested in this assay (see methods section for references). Two of them were discarded because of a lack of specificity while the third one gave better results in terms of specificity and noise (Figure 3B).
Figure 3. Analysis of antibody specificity using peptide competition assays on western blots of P. tricornutum nuclear extracts. (A) Upper lane contains from left to right 1 μg of H3K4me2 antibody alone or with 0.25 μg of one of the different modified peptides, H3K4me1, H3K4me2 and H3K4me3. Lower lane contains H4 antibody as internal loading control. (B) Upper lane contains 1 μg of H3K9me2 antibody alone or with 0.25 μg of one of the different modified peptides, H3K9me1, H3K9me2 and H3K9me3. Lower lane contains H4 antibody as internal loading control.
H3K4me2 is enriched in genes in P. tricornutum and T. pseudonana
Quantitative real time transcriptase polymerase chain reaction (Q-PCR) coupled with the ChIP protocol described herein was used to investigate the enrichment of H3K4 and H3K9 dimethylation on two genes, histone H4 and diatom phytochrome (Dph), and four transposable elements (TEs) in P. tricornutum and T. pseudonana. A clear enrichment of genes in H3K4me2 shown by a set of primers spanning promoter and gene body was demonstrated by Q-PCR (Figure 4A). Both histone H4 and Dph show significant differences in the enrichment in H3K4me2 between immunoprecipitated sample and mock which is the no antibody control. However, no significant differences were observed for the five TEs chosen in this study. Likewise, in T. pseudonana, the Dph and histone H4 show enrichment in H3K4me2 in both promoter and gene body, whereas the four chosen TEs (C12, C4, C19 and G1) show no enrichment in H3K4me2 (Figure 4B). Additional genes and TEs were tested and showed similar results (data not shown). Altogether, our data show a conservation of H3K4me2 location between the two diatom species and multicellular organisms, because genome wide studies in plants and mammals have indeed shown the presence of H3K4me2 on genes [12-14].
Figure 4. H3K4me2 histone modification on different regions of genes and TEs in P. tricornutum (A) and T. pseudonana (B). % IP indicates the enrichment. H4 P: promoter region of H4 histone gene. H4 B: body region of H4 histone gene. PHY P: promoter region of Dph gene. PHY B: body region of Dph.
H3K9me2 shows a different enrichment profile in P. tricornutum and T. pseudonana
ChIP analysis of H3K9me2 in P. tricornutum revealed that TEs contain a significant enrichment for this mark. We particularly focused on two previously characterized diatom-specific copia-like retrotransposable elements known as Blackbeard (Bkb) and Surcouf (Scf) [15]Bkb was particularly enriched in H3K9me2 while Scf was the least enriched. Intermediate levels were observed among the remaining TEs (Figure 5A). On the other hand, the two protein-coding genes histone H4 and Dph were clearly depleted of H3K9me2 in both promoter and gene body regions (Figure 5A). Both genes were shown by us and others to be transcriptionally active ( [5], unpublished results). It was previously shown that Bkb and Scf are also transcriptionally inactive in normal growth conditions [15]. This is consistent with the primary function of H3K9me2 in repressing TEs in order to maintain genome stability. In P. tricornutum, the ChIP assay shows that repressed TEs are marked by H3K9me2 while active genes are depleted in this mark which is similar to what has been observed in plants and mammals [16,17].
Figure 5. H3K9me2 histone modification on different regions of genes and TEs in P. tricornutum (A) and T. pseudonana (B). % IP indicates the enrichment. H4 P: promoter region of H4 histone gene. H4 B: body region of H4 histone gene. PHY P: promoter region of Dph gene. PHY B: body region of Dph. Mock values are indistinguishable from 0% in T. pseudonana.
In T. pseudonana, a similar pattern of enrichment of TEs by H3K9me2 was observed (Figure 5B). The four tested TEs were highly enriched for H3K9me2, indicating a conserved profile for this mark among both diatoms. Surprisingly, some genes also showed a significant enrichment for H3K9me2, particularly at promoter regions (Figure 5B). This unusual association of H3K9me2 with active genes can be due to an intrinsic feature of the centric diatom T. pseudonana which is believed to have diverged from its distantly related pennate diatom P. tricornutum 90 million years ago. Comparative genomics and analysis of molecular divergence has shown indeed that both genomes are as different as those of mammals and fish [4]. Furthermore, combinatorial patterns of antagonistic chromatin marks are known to occur [18-21]. In Drosophila S2 cells, clusters of transcriptionally active genes were reported to be enriched in H3K9me2 [22]. Similarly, differentiated mouse ES cells were reported to contain large domains of H3K9me2 [23]. A hypothesis is that combinatorial chromatin marks can poise genes for transcription, creating more flexible chromatin states ready to adjust for subtle changes in the microenvironment of regulated genes [22,24]. The presence of chromatin marks known to be silent on euchromatin regions or active genes is intriguing and future analysis will be particularly important for elucidating this question.
The ChIP protocol described herein provides reproducible results with two different diatom species grown in different media. The quality of the procedure monitored by the no antibody control is high as no or insignificant background noise was observed. This protocol is also rapid, and can be completed within 3 days. The quantity and quality of eluted DNA from immunoprecipitation are also satisfactory. Using the described ChIP method combined with real time quantitative PCR, we have demonstrated the existence in diatoms of two types of histone modifications, H3K4me2 and H3K9me2. The H3K4me2 mark is associated with transcriptionally active genes in P. tricornutum and T. pseudonana. This result is consistent with the distribution of H3K4me2 in plants and mammals. H3K9me2 binds TEs whereas no association with genes was detected in P. tricornutum. In T. pseudonana, H3K9me2 also correlated significantly with TEs, although it also appears to bind protein-coding genes. This is different from the distribution pattern of H3K9me2 in P. tricornutum. The differences of H3K9me2 distribution pattern between P. tricornutum and T. pseudonana may be inherent to the genetic and/or epigenetic background of the two species which belong to pennate and centric diatoms, respectively. Gene enrichment with H3K9me2 could be further confirmed by a genome wide study of H3K9me2 distribution. Our results show that our experimental and data analysis approach are indeed highly sensitive to detect differences between the two species if they do occur. The ChIP protocol we describe can be combined with microarray or massively parallel sequencing for further genome-wide studies. This protocol has been indeed successfully combined with Illumina sequencing for studies of global histone modifications in P. tricornutum (unpublished results). Furthermore, our ChIP assay can likely be easily adapted to other eukaryotic phytoplankton species for in vivo protein DNA interaction studies.
Funding for this work was provided by the Agence Nationale de Recherche (France) and a European Research Council Advanced Grant award to CB.
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Tachibana M, Sugimoto K, Nozaki M, Ueda J, Ohta T, Ohki M, Fukuda M, Takeda N, Niida H, Kato H, Shinkai Y: G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis.
Genes Dev 2002, 16:1779-1791. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Bapat SA, Jin V, Berry N, Balch C, Sharma N, Kurrey N, Zhang S, Fang F, Lan X, Li M, et al.: Multivalent epigenetic marks confer microenvironment-responsive epigenetic plasticity to ovarian cancer cells.
Epigenetics: off j DNA Methylation Soc 2010, 5:716-729. Publisher Full Text
Bernstein BE, Mikkelsen TS, Xie X, Kamal M, Huebert DJ, Cuff J, Fry B, Meissner A, Wernig M, Plath K, et al.: A bivalent chromatin structure marks key developmental genes in embryonic stem cells.
Cell 2006, 125:315-326. PubMed Abstract | Publisher Full Text
Roudier F, Ahmed I, Berard C, Sarazin A, Mary-Huard T, Cortijo S, Bouyer D, Caillieux E, Duvernois-Berthet E, Al-Shikhley L, et al.: Integrative epigenomic mapping defines four main chromatin states in Arabidopsis.
EMBO J 2011, 30:1928-1938. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Weishaupt H, Sigvardsson M, Attema JL: Epigenetic chromatin states uniquely define the developmental plasticity of murine hematopoietic stem cells.
Blood 2010, 115:247-256. PubMed Abstract | Publisher Full Text
Riddle NC, Minoda A, Kharchenko PV, Alekseyenko AA, Schwartz YB, Tolstorukov MY, Gorchakov AA, Jaffe JD, Kennedy C, Linder-Basso D, et al.: Plasticity in patterns of histone modifications and chromosomal proteins in Drosophila heterochromatin.
Genome Res 2011, 21:147-163. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Wen B, Wu H, Shinkai Y, Irizarry RA, Feinberg AP: Large histone H3 lysine 9 dimethylated chromatin blocks distinguish differentiated from embryonic stem cells.
Nat Genet 2009, 41:246-250. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Lanzuolo C, Lo Sardo F, Orlando V: Concerted epigenetic signatures inheritance at PcG targets through replication.


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Building on a Strong Foundation
Join The River Community Church as they celebrate moving into their "New" building and honoring the people that have gone before them who assisted in building the strong foundation that currently exists at TRC.
The Grand Opening for TRC will be on Sunday, March 10, 2013. The River Cafe will offer complimentary fresh gourmet coffee and pastries from Panera Bread at 9:15 a.m.
The program will begin 10 a.m. with an address from Ventura Mayor Mike Tracy. This will be followed by live music from TRC Worship Team, exciting videos and keynote speaker Pastor Dennis Easter. Pastors Dennis & Patsy Easter helped build the foundation of this church as Senior Pastors for over 15 years.
A very tasty lunch will be served after the program hosted and sponsored by RedBrick Pizza of Ventura.
"This is a very exciting season for TRC since we now have a permanent location! There is so much history on this property that people need to know about. 80 years ago, this church was the hub of Ventura and our goal is to be that resource again, to partner with the city in serving our community," says Jim Duran, Lead Pastor of TRC.
On March 7, 2013, the Ventura Chamber of Commerce will hold a Ribbon Cutting at TRC at 9:00 AM. All are invited and The Greek at the Harbor, The Pierpont Inn and Domino's will provide lunch. Historical tours will be given throughout the day.
Come join the celebration at 889 East Santa Clara Street, Ventura. Classes are available for children of all ages, as well as a nursery for infants and toddlers. To RSVP for this event please call (805) 648-7944 or visit the website at www.TheRiverCommunity.org .
This story is contributed by a member of the community and is neither endorsed nor affiliated with Ventura County Star.
Subscribe to the Ventura County Star
Need Help? Call us at 1-877-710-6182.
Monday-Friday: 6am-3pm / Saturday: 6:30am-10:30am / Sunday: 7am-11am

subset: commoncrawl


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Iver Bogen, Ph. D.
- Longtime educator, community activist, and humanitarian, Iver Bogen, died peacefully on Tuesday, April 24, 2012, surrounded by his family at Essentia Health Center.
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Alumni/ae Notes
Please send your news to email@example.com.
Ann Drake (MDiv 1964) had a monthlong art exhibit of her paintings at the Faulkner County Library in Conway, AR, where she has retired. "To express oneself in visual form you must do it over and over again until you develop your way. You are always developing and learning each time you do something, even a doodle," she said in a press release at the time of the show. In 2004 her home burned to the ground, and she lost everything, including her paintings. The paintings on exhibit are ones that were in the possession of others at the time.
Charlie Ensley (MDiv 1973) has begun his 35th year in parish ministry. He currently serves the Bay Shore United Church of Christ in Long Beach, CA.
Rev. Dr. Charles Hambrick-Stowe (MDiv, MA 1973) has been called as the new senior minister at the First Congregational Church of Ridgefi eld, CT. The church was established in 1713, and in 295 years has had just 20 ministers. Dr. Hambrick-Stowe is the author or editor of 8 books and has published numerous articles in the fi elds of church history, Christian faith, and American religious experience.
Maria Hoaglund, who attended PSR in the early 1980s, has published a book based on her 12 years in hospice ministry. The Last Adventure of Life was published under the pen name Maria Dancing Heart.
Roberta Sears Romiger (MDiv 1982) was named general secretary of the United Reformed Church of England in July. She is the fi rst woman ever appointed to this position.
Anne Dilenschneider (MDiv 1987) is director of Caminar-Eucalyptus House, a residential program in San Mateo County that helps people with psychiatric disabilities live independently.
Toshi Yamamoto (MDiv 1987) has taken a position as a professor and a chaplain at Kwansei Gakuin University, a Methodist-related university in Kobe. He spent the last 15 years working for NCC Japan, the fi rst 10 of which he was in charge of international affairs and the last 5 as general secretary.
Jurgen Schwing (MA1993), an ordained UCC minister, recently published a chapter on "Spiritual Guidance at the End of Life" in Sharing Sacred Stories: Current Approaches in Spiritual Direction and Guidance ,edited by Robert Frager (Crossroad, 2007). Schwing presented the chapter at the World Congress on Psychology and Spirituality in New Delhi, India.
Helen Masek (MA 1995) drove across the country for eight weeks this summer with her husband. She attended the "Collection 2008" of the Guild of American Paper Cutters in Lancaster, PA and presented two workshops for people from 18 churches in southwest and central Pennsylvania.
Floyd Bland (DMin 1999) has written a book, The Christian Heritage. Through the years, he has served as pastor, teacher, evangelist, and administrator, helping to equip others for ministry. He currently serves as a prison chaplain in Crestview, FL.
Kimberly Brown Montenegro (MDiv 2008) has been called to be the pastor at the North Sacramento United Methodist Church in Sacramento, CA.
James Latimer (MDiv 2002) has accepted a call to be the transition minister for a three-year period at the Community United Church of Christ in Reading, PA.
Wakoh Shannon Hickey (MDiv 2003) successfully defended her dissertation in May and is starting a teaching position at Alfred University in Alfred, NY. There was a celebration of her accomplishments on the PSR campus August 3.
Tevita "Fili" Vaikona (MDiv 2003) has accepted a call to be pastor at First United Methodist Church in Red Bluff, CA.
Jen Rude (MDiv 2005) has received the "30 Under 30" award from the Windy City Times in Chicago, acknowledging her as a mover and a shaker in the LGBT community. She serves at associate pastor at Resurrection Lutheran Church in Lakeview and on the youth outreach team at the Night Ministry, working with queer youth and those who are experiencing homelessness.
Matthew Seeds (MDiv 2008) has accepted a call to be the pastor of Zion United Church of Christ in York, PA. He was ordained at Dubbs Memorial United Church of Christ in Allentown, PA (his home church).
Sara Steen (MDiv 2006) has been appointed director of outdoor ministries for the Northern California Nevada Conference of the United Church of Christ. She said of her new task, "I am committed to fi nding creative solutions that continue to allow youth and adults to encounter God out-of-doors in safe and nurturing environments."
Kacey Alexander (MDiv 2007) was ordained in the Disciples of Christ Church June 7 at the PSR campus. There also was an ordination dedication and reception at First Christian Church in Albany, OR on June 22.
Rachel Ciupek-Reed (MDiv 2007) has accepted a call to be senior pastor at the Mount Hollywood Congregational Church (founded in 1905) in Los Angeles.
Niccole Coggins (MA 2007) began her PhD studies in history at UC Santa Barbara this fall. She will be building on the work of her master's thesis at PSR, which examined the historic and current relationship between the United Church of Christ and native Hawaiians.
Andrew Metcalfe (MDiv 2008) was ordained May 4 at First Congregational Church of Berkeley. He received a call to Kenwood Community Church.


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The EFIX box contains:
• EFI-X module (EFIX USB V1.01, V1.1, & V3)
• USB header extension cable
• 3M double sided tape.
NOTE: You only have to use the extension cable if EFI-X module doesn't fit. If you don't need the extension cable, just connect the EFI-X module directly to USB1 header on your motherboard.
Always disconnect the power cable from your PC when installing the EFI-X module.
EFI-X Installation with the USB extension cord:
When using the extension cord, please connect it to the EFI-X module as shown above before continuing. Note that one pin is missing on the USB extension cord and there is one blocked pin on the EFI-X module which corresponds to the missing pin on the USB extension cord. Please align accordingly and connect the extension chord to the EFI-X module.
Plug the other end of the EFI-X module extension cord into an available USB1 header on your motherboard. Note that one hole on the USB extension cord is blocked. This blocked hole corresponds to the missing pin in the motherboard USB1 header socket. Please align them accordingly and connect.
The EFi-X module installation is now complete.
Now it's time for the next step: EFIX MOTHERBOARD'S BIOS CONFIGURATION


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Proposed 2014-04-26: Investment Policy - ADOPTED 2014-05-21
A draft investment policy.
A draft investment policy.
Donate your used car through the Car Talk vehicle donation program. They will arrange for removal of your vehicle which will be sold and the proceeds given to WJFF. Click here for more information!
|Local News - New York Focus
|Local News - Pennsylvania Focus


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We explore heli-boarding in Europe on the cheap
Homeopathic predictions for winter!
New and crude limited graphics for 2016!
Let's turn this into a competition - see within for details
For people who want to listen to music and use a walkie at the same time
A proper DIY powder board from one of our readers
UK based snowboard pioneers DOUK are offering the ultimate in snowboard gluttony; building your own snowboard.
See ya Tignes! The British Championships head home to Switzerland.
Burton Riglet park is returning to Tamworth Snowdome this October with another great chance to give your kids an introduction to snowboarding.
Download this dramatic black-and-white snowboard photo for your desktop here.
Shot by Ed Blomfield
Shot by Remi Petit
Photo by Dominic 'Howzee' Zimmerman


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Hoo-Loo-Choo. Alias John Bull and the Doctors
1 portrait of John Charles Spencer, 3rd Earl Spencer
© National Portrait Gallery, London
Hoo-Loo-Choo. Alias John Bull and the Doctors
by John ('HB') Doyle, printed by Charles Etienne Pierre Motte, published by Thomas McLean
lithograph, published 2 May 1831
11 1/4 in. x 16 3/8 in. (286 mm x 415 mm) overall
acquired Unknown source, 1900
Reference Collection
Click on the links below to find out more:
Artistsback to top
- John ('HB') Doyle (1797-1868), 'HB'; caricaturist. Artist associated with 702 portraits, Sitter in 1 portrait.
- Thomas McLean (1788-1875), Publisher and dealer. Artist associated with 897 portraits.
- Charles Etienne Pierre Motte (1785-1836). Artist associated with 83 portraits.
Sittersback to top
- John Bull (active 1712-present), Fictitious epitomist of Englishness and British imperialism. Sitter associated with 104 portraits.
- Charles Grey, 2nd Earl Grey (1764-1845), Prime Minister. Sitter associated with 173 portraits.
- Sir Robert Peel, 2nd Bt (1788-1850), Prime Minister. Sitter associated with 195 portraits.
- John Russell, 1st Earl Russell (1792-1878), Prime Minister and writer. Sitter associated with 229 portraits.
- John Charles Spencer, 3rd Earl Spencer (1782-1845), Whig statesman. Sitter in 62 portraits.
- Arthur Wellesley, 1st Duke of Wellington (1769-1852), Field Marshal and Prime Minister. Sitter associated with 391 portraits.
Portrait setback to top
Subject/Themeback to top
Tell us moreback to top
Can you tell us more about this portrait? Spotted an error, something missing, or do you know anything that we don't know? If you have any information to share please complete the form below
Related pages


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Your cart is empty!
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When using this product
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Colgate Baking Soda And Peroxide Whitening Oxygen Bubbles Fluoride Toothpaste, 6.4 Oz
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Grandma's Tattoo from Miranda Harple on Vimeo.
Role: Videographer, Producer
From AARP -
In the Harple family, coordinating a family gathering is akin to planning a military operation. Mom, dad and their five children are scattered from Amsterdam to London to Washington to New England. But an impromptu e-mail request from daughter Laurel to her two sisters sparked an unusual family outing: What if the women got a small tattoo of an anchor, each on different places of their bodies, to symbolize the family's deep connection to sailing?
After all, the young women had grown up surrounded by avid sailors. Their parents, Caren and Dan, had met as teenagers through sailing, and maternal grandparents Phyllis and Bob enjoyed sailing throughout their marriage. Also, two of their uncles are world-class sailors.
Earlier this year, when grandma Phyllis noticed the tattoo of an anchor on her daughter Caren's foot, her response was exactly opposite of what Caren had expected. "I really like that. I'm going to get it," she simply said. Caren knew her mother was sentimental about the sea, but certainly never thought she'd approve of body art, much less be inspired to get her own tattoo. She's a proper New England woman, after all.
Katrina, the youngest sister, plans to get her tattoo once she turns 18. "I'm going to get it with all of my sisters, but I think it really doesn't matter when I'm going to get it … it's the significance."


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Curbsyde Productions and OneWheelDrive.Net are proud to present our newest project: "Reconnecting with Canada" (RWC). RWC features travelers Joe & Flora Lloyd on a three month 30,000 kilometer journey reconnecting with their homeland after a 8 year absence.
Reconnecting with Canada is destined as a major multi episode series on a "pay-per-download" model similar to the newly released Australia Inside Out.
On top of filming the journey across Canada for commercial distribution we will also be providing a weekly web-series "on-the-go". The web series will be in ride-reports, on OneWheelDrive.Net, and our Openfilm channel.
As with all of our projects, we self-fund the film production so we're always on a tight budget. The route, and our schedule are pretty much set in stone once we are underway
We'd love to have your input! Feel like meeting up for a short ride along the way? Know of something or someone film worth on our route? Do you have space for our puny tent on the lawn?
Reconnecting with Canada is a story of Canadians Reconnecting with fellow Canadians, as such the social aspect is very important to us.
Thanks for all your support in the past with Australia Inside Out
For media kit, press, or sponsorship enquiries email Joe Lloyd.


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Game Developer, Student and Philanthropist. My primary job here is to help maintain the page for 001 Game Creator.
You are not logged in, your comment will be anonymous unless you join the community today (totally free - or sign in with your social account on the right) which we encourage all contributors to do.
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FaceLock ProAdd to My Apps
|SmartApps Mobile
|App Views
|1 views
|0 votes
|Google Rating
With FaceLock Pro, get a better face unlock even on Android 2.3! FaceLock also lets you choose individual apps you want to protect. Your face is the key to unlock them. See our tips and FAQ for help on how to do a proper face training: http://www.facelock.mobi/tips-and-faq A bad training can impair recognition or recognize other people.There are *NO ADVERTISEMENTS* in ANY version of FaceLock for apps.Pro version comes with all these features you'll love: - Lock as many apps as you want with your face - Lock screen replacement (like FACE UNLOCK) (experimental, lock screen might not work perfectly on all devices). - PATTERN lock is now available! - Set PIN, Pattern or password as alternative access method - More locking options, delayed lock - Hide notification, show icons in lock screen and lots of other visual customization options. - Tasker plug-in - And we are working on even more features...About security: task manager, settings and market are protected so no one can kill or uninstall FaceLock out of the box. Face recognition is reliable and safe with the default settings. For recognition in difficult lighting you can train more images whenever you are not recognized. For increased security you can raise the security level. Tasker, Locale, EasyProfiles and Llama integration has been tested with Tasker so far.By downloading this software you agree to the terms of the end-user license agreement: http://www.facelock.mobi/facelock-for-apps/eulaFaceLock Keywords: face unlock, gallery lock, face recognition, computer vision, visidon, face lock, gotya, applock, smart lock, msecure, cerberus, anti theft, find my phone, phone protection, phone locator, where is my droid, phone tracker, security


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South Point School marks Roald Dahl Day
Bookstore chain Starmark in association with South Point School on Friday celebrated Roald Dahl Day in order to mark the British novelist, short story writer, poet, fighter pilot and screenwriter - Roald Dahl's 97th birth anniversary here.
The celebration featured enactment of selected scenes from Roald Dahl's children's stories by the students of South Point.
"Actually I was invited to do this event by Starmark. Roald Dahl is one of my favourite authors till date. He was my favourite when I was a child. I love reading his stories to my sons, I have two boys and love reading them for my own pleasure" said the trainer, Baishali Chatterjee Dutt.
Commenting on her favourite set of books, she said, "The energy was amazing among the kids. All three sessions that I have had were wonderful. I had one in Modern High School for Girls and two sessions in South High Point School. If the children don't react it doesn't work. But the reacted beautifully."
"It was an excellent program. These days' children do not read much. Program like this initiate them in reading more. When Starmark approached us, we thought this will do the children very good" said Dalbir Kaur Chadda, the Principal of the school.
"Being a principal of this school I obviously want my students to learn discipline and to be a good human being. And I think apart from discipline, kindness is important. Today the world is not kind, so I think I want my students to be a good human being, well mannered and kind" Chadda added.
Some of the books that Dutt read out are 'Charlie and the Chocolate Factory', 'The Magic Finger' etc.
(Posted on 14-09-2013)


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Whether it's a fully lined coat or just a touch of trim on the collar, shearling adds extra warmth and a bit of rugged luxury to your outerwear choices for winter.
Goodman's Guide: Chunky Knits
Whether it's a classic ivory fisherman sweater, a cabled knit, or a more colorful Icelandic or Fair Isle style, one of these cozy sweaters is sure to become one of your favorites.
The Made To Measure Event: Thom Sweeney
Mark your calendar for the Made-To-Measure event beginning this Friday, September 25.
#GoodmansGuide: Tips on being a Goodman.
The debut issue of our new men's magazine has just launched and some of our favorite designers and friends have taken to Instagram to share their excitement.
5 Questions: Public School
Meet designers Dao-Yi Chow and Maxwell Osborne during #AGoodtimeAtGoodmans this Saturday, September 19 from 1-6pm at the Goodman's Men's Store.
5 Questions: Buscemi
Though Jon Buscemi says he isn't really in the "fashion business" the fashion world has certainly taken to his highly coveted sneaker line with its signature hardware.
For The Modern Man: Black Tie Dressing
From a classic peak lapel tux, to a more festive velvet or silk evening jacket we are here to help you with all of your special occasion wardrobe needs. Whether it's a special date night or a bl
In the Spotlight: Thom Browne
With the utmost attention to detail and craftsmanship, Thom Browne's most recent show in Paris left everyone in awe. Inspired by Japanese culture, his pieces incorporated the traditional symbols
7 Easy Father's Day Gifts
What to get the guy who has everything? Here are 7 Easy Father's Day Gift ideas he'll be sure to love.
5 Questions: AMI
An alum of Dior, Givenchy and Marc Jacobs, Alexandre Mattiussi launched his line AMI in 2011, inspired by his friends, his travels, and what he saw cool young guys wearing on the street.
Instagram for women's fashion & lifestyle
Instagram for men's fashion & lifestyle


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SUNY Kicks Off Smart Track Campaign to Ease Loan Defaults
State University of New York chancellor Nancy Zimpher and the SUNY board of trustees announced on Tuesday the SUNY Smart Track campaign to combat student debt throughout New York and set a national model as college students across the country are taking on more loan debt than ever before.
Schneiderman Issues Tips to Avoid Texting Scams Following Game Theory Settlement
Attorney General Eric Schneiderman on Saturday announced a settlement with Game Theory, a mobile-content corporation based in San Jose, Calif., following an investigation that revealed the company was sending spam text messages to New Yorkers and then tricking them into signing up for texting services.
New Yorkers Put Trust in Governor Cuomo
Schumer Cracks Down on Youth Football Concussions with New Legislation Efforts
U.S. senator Charles Schumer was joined by local high school football players, coaches and medical professionals in Rochester earlier this week as he launched his push for legislation to improve high school and youth football helmet safety standards, to help cut down on concussions.


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Quality CareFind out why Mayo Clinic is the right place for your health care. Make an appointment.
Meet the StaffFind a directory of doctors and departments at all Mayo Clinic campuses. Visit now.
Research and Clinical TrialsSee how Mayo Clinic research and clinical trials advance the science of medicine and improve patient care. Explore now.
Visit Our SchoolsEducators at Mayo Clinic train tomorrow's leaders to deliver compassionate, high-value, safe patient care. Choose a degree.
Professional ServicesExplore Mayo Clinic's many resources and see jobs available for medical professionals. Get updates.
Give to Mayo ClinicHelp set a new world standard in care for people everywhere. Give now.
Mayo Clinic offers appointments in Arizona, Florida and Minnesota and at Mayo Clinic Health System locations.
See listing in Cancer Education.
Below are support groups facilitated by Mayo Clinic social workers and open to the public.
Mayo Clinic Hospice offers adult grief support groups two times a year. The groups are free and open to any adult who has experienced:
Contact Mayo Clinic Hospice at 507-284-4002 or 800-679-9084 (toll-free).
This support group is for people who have a left ventricular assist device (LVAD) and their family members. The groups meets from 1:30 to 2:30 p.m. on Tuesdays in Room 5-502, Mary Brigh Building at Mayo Clinic Hospital, Saint Marys Campus. Contact Tami J. Halverson, L.I.C.S.W., at 507-284-2131.
For siblings of children with chronic medical conditions, ages 5 to 18. The younger children's program meets twice a year, and the teens meet twice a year. Contact the Child Life Program at 507-255-4091.
The transplant solid organ support group meets every Monday from 4 to 5 p.m. in Room 4-510, Mary Brigh Building at Mayo Clinic Hospital, Saint Marys Campus. It is an open group for pre- and post-transplant patients and their caregivers. Contact Claire M. Dudzinski, L.G.S.W., with questions at 507-284-2131.
This group is for patients and families who have been seen or are being seen in the Mayo Clinic outpatient brain rehabilitation program. The group meets the second Wednesday of every month from 7 to 8 p.m. Contact Anita M. Milburn, L.I.C.S.W., at 507-284-7241.
Any use of this site constitutes your agreement to the Terms and Conditions and Privacy Policy linked below.
Mayo Clinic is a not-for-profit organization and proceeds from Web advertising help support our mission. Mayo Clinic does not endorse any of the third party products and services advertised.
A single copy of these materials may be reprinted for noncommercial personal use only. "Mayo," "Mayo Clinic," "MayoClinic.org," "Mayo Clinic Healthy Living," and the triple-shield Mayo Clinic logo are trademarks of Mayo Foundation for Medical Education and Research.


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*Editors' Choice* The Gun Metal red is a Bordeaux-style blend-a smooth, seamless mix of spicy red fruits with new barrel highlights of coffee grounds, toasted walnuts and baking spices. There's a lick of citrus running through it also, a testament to the natural acidity that adds life and lift to Washington state reds.
The 2010 Gun Metal (Cabernet Sauvignon, Merlot, Cabernet Franc and Petit Verdot) is a classic Bordeaux blend that gives up knockout aromas and flavors of leaf tobacco, black currants, pencil shavings and assorted spice as well as a full-bodied, balanced palate that has outstanding mid-palate depth, excellent overall purity and freshness, and ripe tannin. Seeing the most oak of the lineup, with 20 months in 80% new French oak, you wouldn't know it and it has ample fruit. It should be even better in another 2-3 years and have 15 years or more of total longevity. Drink 2015-2025.
Finely wrought, with a supple feel to the black cherry, nutmeg and black tea flavors as they course through the expressive finish. Cabernet Sauvignon, Merlot, Cabernet Franc and Petit Verdot. Drink now through 2018. 450 cases made.
The item you have chosen is not in stock in our retail stores or within our main warehouse.
2012 Novelty Hill Columbia Valley Cabernet Sauvignon
2013 Caymus Napa Valley Cabernet Sauvignon
2013 Wines Of Substance Columbia Valley Cabernet Sauvignon
2012 Ancient Peaks Paso Robles Cabernet Sauvignon
2012 Beaulieu Vineyard "Tapestry Reserve" Napa Valley Bordeaux Blend


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Canals of Birmingham - worth capturing on film or an eyesore?
It is said that Birmingham has more canals than Venice. However, think of Venice and you think of romance, gondolas and Venetian balls. Think of Birmingham and you think of… well, very little. So is this because the canals are an absolute eyesore, or are they actually worth photographing? Should Birmingham do more to become a little Venice, or should they just give up?
Whether or not Birmingham actually has more canals than Venice is something of a discussion point. What we do know is that there were 174 miles of canals in Birmingham by the middle of the 18th century. This would have been more than Venice at the time. Now, however, a large proportion has actually disappeared, leaving only 114 miles of navigable water. That's still a lot, but perhaps not as much as Venice.
Those who walk the towpath have the opportunity to see not just Birmingham, but also all the surrounding towns. That is certainly a perfect opportunity for some beautiful photography as it doesn't just show 15 entire miles of improved canals (for safety and nicer walks), but it also means you don't have to take a boat.
We know that the canals have been partially responsible for the development of Birmingham as a city, and for the Black Country as a whole. It helped to develop a range of different industries in the area and had it not been for these, Birmingham would probably have been nothing but a small village, if that.
Birmingham will never be Venice. It was never, nor will it ever be, a city of art and culture. It is not romantic, but rather industrial. You don't want to go through a canal in Birmingham on the front of a gondola with your loved one. For one, the weather in Birmingham doesn't really allow for that sort of behaviour.
However, perhaps we should also stop romanticising Venice. It is, all things put together, a very dirty city with a sordid history. The Venetian balls, now so popular and emulated across the world (think of the Rio carnival and Mardi Gras in New Orleans), was actually a punishment for those people who were masked every day of the week, getting involved in orgies and criminal activity. Birmingham isn't like that, so perhaps it is actually better than Venice. It certainly is worth going over and taking a few good shots, even if it is just for fun. If you are interested in architectural photography, talk to Paul Arthur, an architectural photographer in Birmingham. Paul is located at Paul Arthur Photography, The Fireworks, The Old Fire Station, 68 Albion Street, Birmingham, B1 3EA. Call: 0121 4 050505. Or maybe you're looking for an established corporate photographer like Seven Star Photography?


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– At MLB.com, Jon Mayo has a piece up on the final preparations for the draft. Nice line from a scouting director about team-specific workouts, "It's like the last piece of the interview."
– Kiley McDaniel, who was our guest on the Mostly Mets Podcast this week has a new Mock Draft up at Scout.com. The nice thing about this one is that it includes a range of selection for each pick.
His grid for the Mets:
The Pick 2nd Option 3rd Option 4th Option
D.J. Peterson Hunter Renfroe Dominic Smith Phil Ervin
In Kiley's world, OF Austin Meadows, OF Clint Frazier and C Reese McGuire are all taken a few spots in front of the Mets. Kiley spends most of his time going to games where he sees the players himself, and talks to folks in the industry. That, combined with his time in a few different front offices, give him solid network of sources to draw on.
We spent a lot of time on the podcast talking about Peterson and Renfroe. I'm still skeptical about Peterson, but Kiley has been getting excellent scouting reports on him. He's a good interview and opinionated; for example, he pretty strongly dismissed the Mets' interest in Arkansas RHP Ryne Stanek.


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Dr. Craig Watts's Ratings
The best doctor I've ever know dot. And I have seen a lot of them since over 50 years. Sure he's a top guy but he treat also the person not just the disease.
Super Doc, thanks you.
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Cela faisait six ans que je souffrais d'un ma de pied. C'est le seul qui m'a crue. Il a fait un examen complet pour se rendre compte que la source du mal était mon dos. Je me fous de la ponctualité. Cela faisait six ans que j'attendais un diagnostic. J'adore ce médecin et je ne l'oublierai jamais.
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Je suis une patiente du Dr Watts depuis plusieurs années et si je ne l'avais pas eu comme rhumatologue sur mon passage de vie je ne sais pas ce que ma vie serait devenue. Il est un expert dans son domaine et il sait quand il est temps d'agir pour vous aider. Chaque patient a le temps qu'il lui est nécessaire. Il lui arrive de perdre patience mais souvent c'est pour des raisons administratives tel les coimpagnies assures qui harcèlent les patients pour faire compléter des formulaires au détriment de la santé des patients. Ce médecin est dévoué à son métier et jamais avare de son temps. Il a fait d'excellentes références pour mon dossier et comme je l'ai mentionné, il fut une bénédiction pour moi. Son personnel administratif est d'un professionnel exceptionnel et fait preuve de beaucoup d'empathie. Merci Dr. Watts à vous et votre équipe de m'avoir accepté comme patiente car je commence à voir le bout du tunnel.
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I know he is an excellent rhumatologist. The clinic was nice and the secretaries too, which is very important. But, I wasn't told that there were more than 2 hours of waiting time in advance and that I could do a phone check-in.... In fact, he was 2h30 late! He answered to his cell phone during my exam, had a (personnal) conversation with his secretary and was kind of rude at first. But he knows his stuff and was very knowledgable and took his time with me (1hour). He sent me for other tests, told his secretary he would see me again soon but he didn't say goodbye or anything to me. I felt a little bit like I was not intelligent enough for him...
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Dr. Watts decides who is worthy of his time and efforts and if he does not like you he is outrageously rude bordering on cruel. It does not take much to offend him either. Doctors in Canada generally are arrogant and behave as though only they can read, everyone else is a moron and they are doing you a big favour affording you their time. The medical system is corrupt. We entrust resources to doctors and they have become like politicians; they somehow believe it all belongs to them now, it is primarily for them and who they deem worthy and for them to give anything back is doing the peons a favour. They forget that we pay for the all of it through our taxes and there is no favour involved in it at all. You know how corrupt our medical system has become when the receptionist talks to patients like they are morons. He has a receptionist who looks about 25 and is as rude and and condescending and treats sick people like cattle as Dr. Watts is when he decides you are not worth his time.
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G-d surely blessed this Dr with a huge amount knowledge, wisdom and personality. Very few drs go out of their way to think the way he does when he diagnoses a patient. He will think far further then the typical diseases and will be there to guide you through your hardships. Also Dr Watts no matter what will make sure to call you back to answer your questions and concerns,even on weekends or late at night. May g-d bless him with health longevity.
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I went ignored about joint issues from my pediatrician for years. Then when a medical professional finally listened I was referred to a different rheumatologist who was extremely rude. I found dr Watts through my aunt and he agreed to take me as a patient. He's phenomenal after years of pain I was diagnosed at 18 and he started exploring pain management options with me, explained me in detail about everything. Nicest doctor I've ever had, his staff is also top notch and super nice. You may wait in the waiting room for an hour or two but it's because he is super attentive with all his patients. He'll fit me in if I'm really in pain and I'm super thankful to have him as my doctor. Thank you so much dr Watts keep up the good work!
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Not only is he not punctual, but when you do finally get to see him, he is not attentive. I've seen him twice, and both times I felt like an afterthought. He has a phone on his desk and does not refrain from answering it, rather than allow the receptionist to take messages. The last time I saw him, I was in his office for an hour, he was on the phone for 45mins. It's very difficult to have a conversation and get answers to the questions you have when you're constantly interrupted. When I finally did get to see him after a second MRI and xray, he announced that my issue was untreatable and got snippy when I asked questions about it. I've been dealing with pain, and treating it for 2 years, I would appreciate some sensitivity when being told that I'd be stuck with it for the rest of my life. Needless to say, I'm looking for a second opinion.
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Doctor Watts might be late-why-he takes CARE of every patient-he is not only a medical card swipe kind of a doctor.I am so glad I can be in his care-I feel safe-yes I trust him with my life 110 %.I would wait not only hours but life time to see him. If you are lucky enough to be there - respect that in every way. The stuff is super friendly-follow up is more then above the duty.A very grateful patient -Brygida Melisa
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Outstanding doctor. Yes, you wait but he gives you his time once you're in his office. He will fit you in if you're in a lot of pain and takes the time to answer all your questions. I hope he never retires cause I don't know what I'd do without him.
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12 Things You Should Know When You Call Your Lender
Contact your servicer immediately if you are unable to make your mortgage payments. (Your servicer is the company that sends you statements and the people you make your payments to.) Even if you are already delinquent or in foreclosure, your servicer may be able to help you with a solution. Don't delay.
Ask your servicer about alternatives to foreclosure, including repayment plans, loan modification, postponements of regular payments (called forbearance) and other "workout" options. Visit our
Glossary of Foreclosure Terms
for more information on these terms.
Remember: Workout options are tailored to fit your individual situation, but you must qualify for the plan being considered.
Provide any information requested by your servicer quickly to avoid further foreclosure action – don't wait until the last minute. Servicers must evaluate each borrower's circumstances on a case-by-case basis and go through an approval process to arrange workout plans. This can take time, so it's important you act immediately and provide any information the servicer requests promptly and completely.
Be prepared to provide detailed financial information to assist your servicer in qualifying you for a potential workout option. Be honest about your circumstances and personal finances – it will help lead to the workout option that best fits your situation.
Be ready to change your spending habits. By creating a workable budget and understanding where your money is going, you are more likely to be successful in a workout plan.
If you need additional assistance or are uncomfortable calling your servicer, request financial counseling from a reputable third party first. Check to see if you qualify for the Obama Administration's modification program by visiting
. For the
U.S. Department of Housing and Urban Development's
(HUD) list of approved counselors call (800) 569-4287. For free counseling from
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Be sure you open all mail from your servicer or your servicer's law firm and return calls promptly. Failure to respond quickly may result in further foreclosure actions and additional costs.
Ask the critical questions:
What is the anticipated timeline to complete a workout?
Will the foreclosure sale be postponed while your servicer reviews the workout option?
What are your obligations under the work-out arrangement: due dates, amounts due, how long your servicer will postpone collection of payments, if applicable, and when such deferred payments must be paid back?
If you are not making payments into an escrow account for real estate taxes and insurance, your servicer will likely require that you begin doing so and will work with you to recover any payments already made on your behalf. You can usually save money and get better coverage if you obtain your own homeowner's hazard insurance policy instead of having your servicer arrange coverage for you.
Stay in contact with your servicer and/or counselor at all times. Notify your servicer and counselor of any change in your circumstances, including new employment or problems with making payments under a workout plan.
Be realistic about your own financial condition. If you cannot afford to keep your property, consider selling it to get your equity out, or talk to your servicer about other options to give up your property.
Understand that the servicer is trying to ensure a positive result for you. No one wins in a foreclosure. Be cooperative, honest and keep your promises on any workout arrangement.
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James Harden says he wants to stay in Oklahoma City and play with the Thunder.
He reiterated as much Monday at Thunder media day.
"Of course I want to be here," he said.
James Harden says he wants to stay in Oklahoma City and play with the Thunder.
He reiterated as much Monday at Thunder media day.
"Of course I want to be here," he said.
But the clock continues to tick. Harden and the Thunder must agree on terms to a contract extension by the end of this month. An offer has been made, the sides are talking, but if a deal is not done by Oct. 31, the versatile guard will be eligible to become a restricted free agent next summer.
If it gets to that point, there's no way the Thunder will be able to match the huge contract some team will surely offer him.
So, the question is this — is Harden ready to leave all this?
That is the question Harden must ask and answer in these contract negotiations. Is he ready to leave a still youthful but supremely talented team that went to the NBA Finals last season? Ready to leave a nucleus that only seems to get better? Ready to leave a franchise that has built a culture that players rave about?
Sure didn't sound like it Monday.
Harden repeated his willingness to sacrifice to stay with the Thunder, asserting that a max contract wasn't a necessity. He even went so far as to say he believed a deal would be done by the Oct. 31 deadline.
"That's why I'm not too worried about it," he said.
Maybe the Thunder can re-sign Harden after all.
Crunching the numbers and listening to the qualifiers thrown around by the Thunder brass, it hasn't seemed like keeping Harden would be possible. Surely, he would want more money than the team could handle.
Getting a deal done still seems like an extremely long shot — how much can the Thunder really offer without putting itself in luxury-tax jeopardy? — but listening to Harden on Monday, it didn't seem completely impossible.
"We've built a brotherhood here, a brotherhood that's hard to break," he said. "Other teams are just teammates, but we're really brothers."
More than anything, re-signing Harden is a test of the Thunder Way.
Sam Presti is the architect of that system. The Thunder general manager has always said it was his intention to build a successful franchise that would be sustainable over the long term, even in a small market like Oklahoma City. That meant drafting players who were not only great players but also great teammates, putting a higher value on sacrifice and team than on ego and self, then affording those players every advantage in training, fitness, nutrition and medical know-how.
The players, for example, are fed breakfast and lunch on practice days. That might seem small, but it's not something most teams do.
Continue reading this story on the...


text: Trial run I diluted as per N2 instructions and used certo. I took a home test 2 days prior to the real test that was at 50 nanograms. I BARELY passed with a VERY faint white line Test day I diluted as per N2 instructions and used certo. I smoked 10 days prior and am a regular smoker. I subbed clean urine as main but they wanted me to fill to a higher line than the 2 oz I had, so used about a 1/5th of my own dilurted urine. I just read somewhere that the military test for 15 nanograms. What is the cut off for pre-employement? The company I am working for is regulated by the FDA and the physical I got was REALLY hardcore. I am worried that if I was at 50 nanograms from the home test with the diluted if I mixed my sub it might mix together somewhere around 15-20. I might fail a test that goes that low. Thanks in advance! Getting real paranoid


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Greg Kroah-Hartman has announced the immediate availability for download of Linux kernel 3.4.73, which comes with a few changes and fixes.
The new maintenance version of the 3.4.x branch has been released and comes with fair number of changes and various improvements.
As usual, these point release versions of the Linux kernel don't bring anything major and are mostly comprised of driver updates and various fixes. This should not stop you from upgrading to the new build.
"I'm announcing the release of the 3.4.73 kernel. All users of the 3.4.x kernel series must upgrade."
"The updated 3.10.y git tree can be found at: git://git.kernel.org/pub/scm/linux/kernel/git/stable/linux-stable.git linux-3.4.y and can be browsed at the normal kernel.org git web browser: http://git.kernel.org/?p=linux/kernel/git/stable/linux-stable.git," Greg Kroah-Hartman said in the email announcement.
Highlights of Linux kernel 3.4.73:
• The "ip rule delete table 256" problem has been fixed (net);
• An unaligned access error fix has been implemented (connector);
• clamp ->msg_namelen instead of returning an error (net);
• strlcpy() is now used instead of strcpy() (isdnloop);
• The br's address entry has been flushed in fdb when removing the bridge dev;
• rt6_get_dflt_router is now used to get default router in rt6_route_rcv (ipv6);
• A possible seqlock deadlock has been fixed;
• The leakage of uninitialized memory to user in recv syscalls has been prevented;
• The recvmsg handler msg_name and msg_namelen logic have been reworked;
• addr_len/msg->msg_namelen assignment has been fixed in recv_error and rxpmtu functions;
• The uncompression of the traffic class field was incorrect (6lowpan);
• The interaction of VFIO_IOMMU_MAP_DMA with IOMMU address limits has been fixed;
• A bug of error handling in the MMC driver has been corrected.
Users of the Linux kernel 3.4.x branch are urged to update to the new version as soon as possible. More details about the changes in this branch of the kernel can be found in the official announcement. Download Linux Kernel 3.4.73 right now from Softpedia.


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Boise VA releases crisis line and medical center info
Image by MGN Online
After the deaths of Idaho National Guardsman Erik Jorgensen and Airman Sandy Bojo, the Boise VA Medical Center released the following medical and mental health resource information for veterans:
Boise VA Medical Center 208.422.1000
Veterans wishing to get help with PTSD, TBI or other health issues should call this number to ask questions or find out how to enroll for services. The Boise VA also has an emergency room that is open 24/7.
Veterans Crisis Line 800.273.8255
Veterans who are having thoughts of harming themselves and/or others, and want someone to talk with immediately can call this toll-free number 24/7 and speak with someone who can help. Representatives who speak with veterans at this Crisis Line are highly trained and able to escalate the response of resources in the veteran's local community if they are needed.
Boise VA Public Affairs Officer Josh L. Callihan stated, "Regardless of where a veteran goes or calls, if a veteran is having thoughts of harming themselves or others they should ask for help immediately."
- Latest


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I recently found this forum while looking for some resources to help me when i was making my first pack and i cant even tell you how helpful it has been. I have recently come down with the urge to have a nice day/overnight pack. This was a problem because i am all but broke and saving what i have for when i go back to school. So i decided o would make one. I am about 80% done at this point and plan to finish tomorrow and will post pics. I have really enjoyed this project so far and am hooked on MYOG now and will continue with it. (by the way my mom helped a lot with this so thats the reason why the stitching is so good. )
Ok so i just finished and here are some of the pictures. The pack is made from ripstop nylon and all the panels are two pieces quilted together. Im not sure the volume but i would guess around 25L.
Very cool design Andrew! Let us know how it performs.
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Princy Quadros-Mennella, PhD
Neuroscience Program Director
Bay Path University
588 Longmeadow Street
Longmeadow, MA 01106
Major in Neuroscience
The major in neuroscience consits of nine (9) courses in the sciences and mathematics and one course in ethics. Additionally, six (6) required specialized neuroscience courses and five (5) elective courses are required. The required courses will provide you with the foundation in essential neuroscience principles and theory as well as to develop skills associated with primary literature evaluation, experimental design, data collection, research, oral presentation and scientific writing. The elective courses will allow you to increase the breadth of your knowledge in specific areas of neuroscience and on topics that are of interest to you.
In total, if you are majoring in neuroscience you will be required to take sixty (60) credits of required and elective courses in addition to the forty-three (43) credits of general education core courses. As part of the curriculum, you will be required to participate in at least one summer research program (Neuroscience Internship; NEU 498) to provide you with research experience.
Minor in Neuroscience
The curriculum for the minor is designed for students from other majors whose career plans might be enhanced by an understanding of neuroscience. The minor in neuroscience will consist of six (6) courses (totaling 20 credits) which include the two semester introductory neuroscience course sequence, research methods in neuroscience, and three neuroscience electives. Following completion of these courses, students in the minor will have obtained basic knowledge in the neurosciences to apply in careers that might utilize neuroscience concepts.
Learn about our special WiSH Program which offers a four-year curriculum consisting of integrated and advanced study and research for dedicated future women scientists.


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Cringing in the rain-A A +A
Friday, September 5, 2014
WE ARE in the 'Ber months and we have started the countdown to Masskara, All Saints' Day, and of course Christmas. And lest we forget the season for splurging, even the malls have started playing Christmas songs.
And lest we forget, we are also entering the height of typhoon season.
The costliest Philippine typhoons were Yolanda and Pablo that struck in November, Pepeng in October, Ondoy in September. Much earlier, we had Ruping in November of 1990.
The Philippines sit astride the typhoon belt. Bagyó is the common name
in many of our local languages in referring to any tropical cyclone.
There are four recognized climate types in the country, and are based on the distribution of rainfall.
Negros experiences two types of typhoons. For those Type I, the southern parts of Negros Oriental and Negros Occidental have two pronounced seasons, dry from November to April, and wet during the rest of the year.
Maximum rain period is from June to September.
For those in Type III, Northern Negros Occidental has no very pronounced maximum rain period, with a short dry season lasting only from one to three months, either during the period from December to February or from March to May. This climate type resembles type I since it has a short dry season.
In the era of climate change, perhaps we to add Type V. The Asian Development Bank Independent Evaluation Department warned of more frequent and heavier rains in the coming years with dangers to population seen to rise, unless concerned authorities take appropriate steps to prevent disasters and address effects of climate change.
The study was prepared by Vinod Thomas of IE-ADB; José Ramón Albert of the Philippine Institute for Development Studies; and Cameron Hepburn of Oxford University and the London School of Economics.
The paper cited the greater vulnerability of people to growing dangers arising from harsh weather conditions. "This paper's empirical evidence, on an association between the rising frequency of intense natural disasters in Asia and the Pacific countries and climate change, stresses the urgency for reducing human-made greenhouse gases in the atmosphere," Thomas added.
"Prevention is critically important and, therefore, preparedness and investments in disaster-risk reduction must be prioritized," Thomas said. "(In the Philippines) disaster preparedness is underemphasized compared to disaster response."
It's like what the Americans would say, "Closing the barn doors after the cows have fled." Reactive, instead of being pro-active.
In another study, the Building Resilience to Natural Disasters and Major Economic Crises (Brace) recently released by the Economic and Social Commission for Asia and the Pacific (Escap), experts discussed strategies on how organizations and policymakers can address the twin threats of natural disasters and economic crises in an increasingly interconnected world.
Brace aims to strengthen community-based disaster-risk management and
support for vulnerable families living in hazard-prone areas. Yes, 'tis not the time for singing in the rain but the time for preparations. Are we prepared for the next big ones? If not, we can expect much cringing in the rain.
Published in the Sun.Star Bacolod newspaper on September 05, 2014.
Country News
Cagayan de Oro
Forum rules: Do not use obscenity. Some words have been banned. Stick to the topic. Do not veer away from the discussion. Be coherent and respectful. Do not shout or use CAPITAL LETTERS!


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By TERRY WILLISSun Advocate sports reporter
On Sept. 27 seven teams competed in an invitational cross country meet at the Carbon High outdoor sports complex.
It was a crowded field with 84 boys and 54 girls competing. Carbon placed fourth as a team in the meet.
With Paul Goodrich nursing an injured knee, the team wasn't as competitive as usual. Finishing ninth, two minutes off his record time, Goodrich was happy just to be able to compete.
Andrew Bates has been Carbon's most improved runner this so far this year. He finished 49th out of 84 with a very respectable time.
Finishing in the top 20 was Bea Morley 15th and Kent Olsen 18th. Elisabeth Smith and Samantha Smith were the only two girls running for Carbon and they finished 12th and 18th respectively.
The day before Carbon had participated in a meet in Duchesne. Kent Olsen placed fifth, while Travis Hepworth took home tenth. The team placed fourth out of seven teams in that meet as well.
Carbon will compete in Delta on Oct. 4.
The region meet will be held in Price on October 11.


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Yesterday, Leo Myers and I spent two hours (!) on a webinar going over what to expect at Jamboree. We had a wonderful audience that asked great questions.
The webinar will be online in a day or two, and I'll drop a note when it is available. In the meantime, and if you don't happen to have two hours to spend, you might want to download the PowerPoint presentation that we showed. If you are a first-timer at Jamboree, you will get some insights about our event that will help you make the most of the conference.
You might also want to look at the Jamboree website for some other hints on Getting the Most out of Jamboree.
NOTE: Most of you received this notice yesterday. We apologize for the broken link to the PowerPoint. Please feel free to download it now.
03 June 2013
Jamboree PowerPoint Available for Download (corrected link)
Posted by Paula Hinkel at 1:32 AM
Labels: #Genealogy, #SCGS13, #SCGS2013, Genealogy Jamboree, Getting the Most
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