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Gov. Dannel P. Malloy's plan to move about 38,000 parents now covered by the state's HUSKY health insurance program to private insurance could result in more uninsured adults, a nonprofit health care advocacy group is warning.
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It has been long drilled into our minds to focus on form to improve results and prevent injury. But is the so-called "correct" way to train the best way?


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Please report any broken links or trouble you might come across to the Webmaster. Please take a moment to let us know so that we can correct any problems and make your visit as enjoyable and as informative as possible.
Korean War Campaigns
|Campaign and Dates
|Campaign and Dates
|North Korean Aggression, 4 August 1950
|Communist China Aggression, 10 to 20 December 1950
|Click On Image
For Full Size Image
|Image Description
|USNS Sgt. Andrew Miller (T-AK-242)) moored at the Port of Los Angeles, date unknown.
Photo by Ships Photographers, San Pedro, CA.
|Dick and Pat Morales for their uncle Jimmie Gilbert, USNS Sgt. Andrew Miller
|Vietnamese barges and sampans approach USNS Sgt. Andrew Miller (T-AK-242) off Vung Tau, Republic of Vietnam, during
Operation Eagle Pull and Operation Eagle Wind (evacuation of Saigon) 1975. Marines from AESF, Detachment Sierra, right foreground guard the boarding platform.
Department of Defense Photo (USMC) 7711275 from "U. S. MARINES IN VIETNAM, The Bitter End, 1973-1975".
|Robert Hurst
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This page is created and maintained by Gary P. Priolo


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This is your resource to discuss support topics with your peers, and learn from each other.
I just received the denial for my app written using WebWorks/Eclipse for the playbook. Please see the text below.
Dear App World Vendor, We are sorry but your request for 'No Decompression Dive Planner' to be a product in the BlackBerry App World has been denied for the following reasons : Items that need attention: - Please upload a signed copy of your application, appears that it is not signed. - no icon after installing application, please make sure to specify an icon no larger than 86x86 in the blackberry-tablet.xml file. Signing information: http://docs.blackberry.com/en/developers/deliverab
les/23959/Signing_your_application_1422721_11.jsp Commands RDK: blackberry-signer -verbose -cskpass -keystore -storepass RDK Author: blackberry-signer -keystore -storepass author You should also have the following files as a general rule as an end result when signing your application: - RDK.SF - RDK.EC - MANIFEST.MF - AURTHOR.SF - AUTHOR.EC These should all be contained within your .BAR file Regards; The BlackBerry App World™ Team
I am very confused because:
Lastly, I did submit this application prior to the free PlayBook deadline. Does this denial exclude me from gettting the playbook.
Please help.
Solved! Go to Solution.
05-04-2011 10:25 AM - edited 05-04-2011 10:26 AM
2) Those files will be added automatically to the .bar file once you properly sign your app
3) .bar file is actually an archive, you can open it with WinRar so you can check which files are inside
As for the PlayBook offer - you should receive one, my app was denied the same way (although it is an air app), and after signing the app and submitting once again, I got the free-playbook-mail
Awesome. Any clue on:
Is the config.xml file the WebWorks equivalent to AIR / blackberry-tablet.xml?
Yes, it seems so
You can check this documents
And this one
And of course the image size


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Indian Rock House at Fairfield Bay
Winter is the perfect time to explore the natural stone shelters where native Arkansans once lived
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This blog post is written in response to the post showing some of the worst practices of past. Well, just like last month's theme, everybody learns by doing it one step at a time. In my case, I started my career as a network engineer and had no database knowledge during that time. I can still remember my old code which became quite a laughingstock when it was sent for a code review. This story is indeed interesting, so instead of writing shortly, I am going to write today in detail.
It happened about 8 years ago when I was working as Network Engineer in United States. I was responsible for a large data center and we had more than 40 servers under my watch. In our SQL Server database we had one table for the inventory of all the servers. On that table we used to keep a lot of details of the server. To keep it simple, I will say that we have only two columns in that table: ServerID and ServerName. I was asked to write a script which was intended to retrieve all the data from this table and insert them into the new table that was created by the database administrator. I thought I could handle SQL Server script well enough because I learned SQL a bit when I was younger. I ended up writing a script where I wrote cursor to SELECT single row from the table and insert into new table. Well, this went very fine. The script was taking a while to perform its tasks; nevertheless, it worked. I used this script for a short while.
Here is the script that I wrote. I am using sample and simple database for example. Please note that this is not a good practice.
-- Create Table and Populate with Sample Data
CREATE TABLE ServerTable (ServerID INT, ServerName VARCHAR(100))
INSERT INTO ServerTable (ServerID, ServerName)
SELECT 1, 'First Server'
SELECT 2, 'Second Server'
SELECT 3, 'Third Server'
-- Creating New Table
CREATE TABLE NewServerTable (ServerID INT, ServerName VARCHAR(100))
-- Insert Logic
DECLARE @Flag INT
SELECT @Flag = COUNT(*) FROM ServerTable
WHILE(@Flag > 0)
INSERT INTO NewServerTable (ServerID, ServerName)
SELECT ServerID, ServerName
FROM ServerTable
WHERE ServerID = @Flag
SET @Flag = @Flag - 1
SELECT ServerID, ServerName
FROM NewServerTable
-- Clean up
DROP TABLE ServerTable
DROP TABLE NewServerTable
This script worked very well till we had changed in company policy. We then had a newly- appointed CTO (which I was about to become 3 years after this incidence), who introduced the requirement of Code Reviewing of all both old codes and new ongoing codes. When my code went to a code review, my Team Leader really laughed at my code. So he sent me new piece of the code which was much shorter and more efficient. The code which he proposed was as follows. I honestly say this was much better than my code.
-- Create Table and Populate with Sample Data
CREATE TABLE ServerTable (ServerID INT, ServerName VARCHAR(100))
INSERT INTO ServerTable (ServerID, ServerName)
SELECT 1, 'First Server'
SELECT 2, 'Second Server'
SELECT 3, 'Third Server'
-- Creating New Table
CREATE TABLE NewServerTable (ServerID INT, ServerName VARCHAR(100))
-- Insert Logic
INSERT INTO NewServerTable (ServerID, ServerName)
SELECT ServerID, ServerName
FROM ServerTable
SELECT ServerID, ServerName
FROM NewServerTable
-- Clean up
DROP TABLE ServerTable
DROP TABLE NewServerTable
If you've noticed, the new insert code is just reduced to two lines only. Today you might think it is a very simple code, but for me it was a very new thing during those times. Now comes the best part of this whole story.
My team leader, who was very supportive and extremely positive person, asked me to demonstrate this whole incidence on our next team meeting. Honestly, I did not feel bad about accepting my mistake and learning new things. During the team meeting, I first accepted the fact that I was wrong with my code and thanked the Team Leader for giving me the opportunity to improve and develop myself. Because of his encouragement to accept my own mistake and improve from that incident, today I have reached this place.
Please get rid of cursors and use INSERT INTO…SELECT or SELECT INTO logic.
Reference: Pinal Dave (http://blog.SQLAuthority.com)
37 comments. Leave new
Hi Pinal, both the codes are same .
We can make this even simple by using Select Into…
SELECT ServerID, ServerName into NewServerTable
FROM ServerTable
This is nice until the schema of NewServerTable changes. I like named inserts:
insert into [NewServerTable]
select ServerID, ServerName
from [ServerTable]
This way, the insert still works should the DB team reorder the columns in NewServerTable or add nullable columns between ServerID and ServerName.
Great post, Pinal, however there are times where cursors are a better fit than selection inserts.
I read this topic and I think I lost my time.
Conclusion, you don't be a genius to be a sql guru :D …
Very nice that you show your own example.
Great job Pinal!
hi pinal,
i am java core developer,i like very much your blog
Hi Pinal,
I read this blog, to know ur improvements in ur knowledge and career.
No one share their Negative points. But you only shared to show your simplicity.
Really i proud of you.
and now You are the Team Leader!
Thanks a lot sir! You have saved my blushes countless times. I'm humbled that you'd dare to share such a simple lesson you learnt years ago. Once again, thank you!
You can refer the this also
Yes Pinal sir, very true, we learn from our mistakes.
One more thing i whould like to add here
we can also use 'Select Into' statement for the same purpose and it is much simpler
You are right ! After googling I found out – there are some advantages and disadvantages as well, using 'SELECT . INTO' ahead of 'INSERT INTO .. SELECT' like it creates a new table if not already exists but with no constraints. Again for the former one you need to change your recovery model.
But the former one is faster and can produce excellent result if you really need it for some specific situation.
Very useful to freshmen like me. Thanks for sharing.
In my three years of experience, I never used cursor. I used table variables, #tables to store data temprary using insert into statement. And after use of them, droped them(#tables)
Nice work.
Hi Sir, this is karthik… I need to know , to update the row using by CURSOR… I have written some query but it doesnt work properly.. you just rectify the mistake and then send me the correct query…
Thank you…
create proc stt
declare @name varchar(50)
declare @roll_no int
set @roll_no=100
declare stu_cursor cursor for
select name,roll_no from xyz order by name
open stu_cursor
fetch next from stu_cursor into @name,@roll_no
while @@FETCH_STATUS=0
print @name
print @counter
update xyz set roll_no=@roll_no where name=@name
set @roll_no=@roll_no+1
fetch next from stu_cursor into @name,@roll_no
close stu_cursor
deallocate stu_cursor
hi karthik
this is your solution for sql query and execute it…
frm jagan
alter procedure hi
declare @emp_name varchar(50)
declare @date date
declare @sal int
declare @emp_ad numeric
declare emp1 cursor for
select emp_name,date_of_joining,salary from emp order by emp_id
open emp1
fetch next from emp1 into @emp_name,@date,@sal
set @emp_ad=100
while @@FETCH_STATUS = 0
print @emp_name
update emp set emp_ad_id=@emp_ad from emp where emp_name=@emp_name
select @emp_ad=@emp_ad+1
print @emp_ad
fetch next from emp1 into @emp_name,@date,@sal
close emp1
deallocate emp1
hi karthik
this is your solution for sql query and execute it…
frm jagan
alter procedure hi
declare @emp_name varchar(50)
declare @date date
declare @sal int
declare @emp_ad numeric
declare emp1 cursor for
select emp_name,date_of_joining,salary from emp order by emp_id
open emp1
fetch next from emp1 into @emp_name,@date,@sal
set @emp_ad=100
while @@FETCH_STATUS = 0
print @emp_name
update emp set emp_ad_id=@emp_ad from emp where emp_name=@emp_name
select @emp_ad=@emp_ad+1
print @emp_ad
fetch next from emp1 into @emp_name,@date,@sal
close emp1
deallocate emp1
I want to insert data in new table and data are coming from two different tables which i already created so can you suggest me how can i copy data of two table in new table???
@Hardik, Refer link
Read this wonderful article and I'm sure you will get your answer!!
It's an awesome paragraph in favor of all the web users; they will get advantage from it I am sure.
Very good written article. It will be beneficial to everyone who employess it,
including myself. Keep up the good work – looking forward to more posts.
Pinal Dave, I appreciate your straightforward and modest approach to everything. You are a good man.
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be the first step so that you can lose weight. They will often taste beneficial, but
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enough strength to get through the day. In case
you are constantly taking in these foods, transferring to whole grains and other complex carbohydrates will aid you
to have more energy while eating less. Thanks alot : ) for your
blog post.
Its like you learn my thoughts! You seem to grasp a lot about this,
like you wrote the book in it or something. I think that
you could do with a few p.c. to force the message house a little bit, but other
than that, this is wonderful blog. A great read.
I will definitely be back.
hello!,I really like your writing so much! share we keep in touch more about your post on AOL?
I require an expert in this space to resolve my problem.
Maybe that's you! Taking a look ahead to see you.
thank you for your small but important lessons
Thanks a lot for sharing this with all of us you really know what you are talking about!
Bookmarked. Please also visit my website =).
We could have a link exchange arrangement between us!
SELECT .. INTO and INSERT INTO … SELECT * are both bulk logged operations and can benefit from it if the recovery model is SIMPLE or BULK LOGGED
BTW: Pinal, great blog.
I've learned vey much from it within the last 5 years :)
Though these are fast but create table lock. So in highly concurrent scenarios these does not scale. In scenario where high concurrent insert with bigger row set create problem if use build insert. In these scenarios cursor bases operation is better. So build operation is not always good. Any inputs?
Hi Pinal,
Need your guidance in solving below query
-1. Current process has ONE MS Access Database table with all fields of information in it. Lets say STUDENTS ( has all info about students bunched togather)
-2. Designed a new SQL server 2008 DB with all info grouped togather in resp tables- so there are more than one table now – Student, Personal, Courses, Teachers, Grades, Research
-3. Need to migrate data from ONE old table and distribute it over into these multiple tables with 'Student_id' as primary key of Students table, and 'Student_id' – ref as Foreign Key in all other tables
– But some columns from records of old table has data which needs to be checked for before inserting into new tables ( some data has been added to Static data – Such as Course list, so sourse name has to be one othe entires from COURSE_NAME table, while old data has some free text data inti so insert would fial on FK contraint)
What is the best way to achieve this data migration into new table stucture?
Thanks for (any) pointers
Say i am inserting 50 rows using insert… select , if any error occurs will it roll back automatically?
Hi Pinal, first I would like to thank you for such an excellent site. I have learned many things from you concerning SQL server. Right now I am honestly getting confused by cursor performance.
Everywhere I look I see that Cursors are bad… I am trying to reconcile these statements with my testing results against a foreach loop in SSIS 2012. Any clarity or if I am doing something incorrectly in SSIS would be appreciated.
This was my test cursor with 1000 records.
/************ Test Cursor Performance ********************/
DELETE testImportDatabaseMaintenance
DECLARE @DBMRecordid AS INT , @Endpointid AS VARCHAR(50)
DECLARE test_cur CURSOR FAST_FORWARD READ_ONLY FOR
SELECT idm.DBMRecId ,idm.EndpointId
from ImportDatabaseMaintenance idm
OPEN test_cur
FETCH NEXT FROM test_cur INTO @DBMRecordid , @Endpointid
INSERT Testimportdatabasemaintenance
(testImportDatabaseMaintenance.DBMRecId , testImportDatabaseMaintenance.EndpointId )
VALUES (@DBMRecordid , @Endpointid )
FETCH NEXT FROM test_cur INTO @DBMRecordid , @Endpointid
CLOSE test_cur
DEALLOCATE test_cur
this cursor completed in less than 1 second from SSMS.
I used this sp with a foreach loop in SSIS.
ALTER PROCEDURE A_Test_DBM_SSIS_3 @DBMRecId AS INT ,
@EndPointId as varchar(50)
INSERT testImportDatabaseMaintenance
VALUES ( @DBMRecId , @Endpointid)
The package does the same functionality as the cursor.
in a container I load the variable objIdbm with the recordset and then execute a foreach ADO Enumerator against the Stored Procedure referenced above. time to complete 388 seconds.
I started to search the web and found many other blogs and articles that are basically having the same results.
Am I missing something?
Thank you in advance for your time.
thanks for sharing, it is a good article
You are welcome.


text:
Helen "JoAnne" Berg, 90
Helen "JoAnne" Berg, 90, of Marshalltown, IA passed away Saturday, November, 23, 2013 at Fountain West Nursing Home in West Des Moines, IA. Private graveside services will be held at Riverside cemetery, in Marshalltown. The Main St. Chapel of the Anderson Funeral Homes in Marshalltown has been entrusted with the arrangements. Online condolences may be sent www.andersonfhs.com.
JoAnne was born April 29, 1923, in Sigourney (Keokuk County) Iowa to Raymond J. and Addie C. (Foster) Hawk. At an early age, she moved to Hedrick, Iowa. JoAnne graduated from Hedrick Consolidated School in 1940. She attended William Penn University in Oskaloosa. JoAnne was employed with Burlington Hawk-Eye Gazette in editorial department 1942-1946 and had a similar position in the editorial department at the Marshalltown Times Republican in 1947 until 1949. JoAnne was united in marriage to Cyril N. Berg in Marshalltown December 24, 1948. In the later years she worked in the business office at Marshalltown Medical Clinic and in medical records department at Marshalltown Medical and Surgical Center, retiring from that employment December 31, 1984. More recently worked part-time as a homemaker home health aide for the Community Nursing Service, retiring in August of 1999. Since July of 2003 she has lived in West Des Moines. Jo Anne was a member of the First Congregational Church, having served as a deaconess and teacher in the Sunday School, active in the Women's Fellowship of the church, member of the American Legion Auxiliary and MMSC Auxiliary.
JoAnne will be missed by a son, Don K. (Mary) Berg of Chicago; two daughters, Kristine Berg of West Des Moines; and Carol (Arnold) Lemker of West Des Moines; four grandchildren, Alissa (Steve) Lilla , Arnie Lemker, Mary Alice Berg, and Timothy Berg and three great grandchildren, Jack and Anne Marie Lilla, and Ellie Berg. Preceded in death by her parents; and husband on June 8, 1961.


text:
Contracts for Creative Professionals
Saturday, June 15, 2013 from 9:00 AM to 1:00 PM (PDT)
Event Details
Contracts mystify many creative professionals, yet they are the lifeblood of the trade. In this workshop, geared towards freelance creatives and small agencies, we will work together to understand contracts and see how they can be used as a tool to do better work, make more money and keep clients happy.
This workshop will teach the basics of how contracts work, how to approach contract negotiation as a collaborative process, explain important and common contract clauses and offer guidance on when and how to seek help drafting or enforcing contracts.
Here is what you will learn, at the very least:
- What a contract is and how it is formed.
- How contracts are structured.
- Common contract clauses.
- Typical issues in creative contracts.
- How to approach contracts with your clients in a non-adverserial way.
- Using online signing services to make the process painless.
- How to find and adapt basic contracts.
- How to enforce contracts.
- How to work with an attorney to draft, modify and enforce contracts.
Since this is an early version of what we hope will become a valuable, repeatable event for the creative community, participation in this first workshop will be limited to just 10 people and cost only $89.
When & Where
The Hub Los Angeles
830 Traction Ave. #3A
Los Angeles,
Saturday, June 15, 2013 from 9:00 AM to 1:00 PM (PDT)
Add to my calendar
Semi-Aware is a forward-thinking service that aims to connect people in creative industries with legal knowledge tailored to the needs of the independent, professional creator. It was started in 2013 by Jonathan Tobin and Spencer Cross.
Jonathan Tobin is a California attorney with deep roots in the creative communities of Los Angeles. Working as a web developer and graphic designer since the late 1990s, he has practical insight into the issues facing creative professionals.
Spencer Cross is a Los Angeles-based designer and community organizer. He works to foster community and camaraderie among LA-area creative professionals by coordinating a variety of social and educational events and discussion forums.


text:
No Evidence for Interference of H&E Staining in DNA TestingUsefulness of DNA Extraction From H&E-Stained Archival Tissue Sections
Although histochemical staining has been believed to inhibit the DNA amplification reaction, no previous study has systematically evaluated the influence of histochemical staining on downstream molecular assays. To evaluate an influence of H&E staining on DNA testing, we isolated DNA from 10 unstained, 10 hematoxylin-stained, 10 eosin-stained, and 10 H&E-stained tissue sections (ie, 4 groups), from each of 5 colon cancers. Among the 4 groups, we did not observe any significant or appreciable difference in DNA fragmentation by agarose gel electrophoresis, in DNA amplification by real-time polymerase chain reaction (PCR), in microsatellite PCR fragment analyses, or in a PCR-pyrosequencing assay. As a proof-of-principle study, we successfully performed microsatellite instability analysis and sequencing of KRAS and BRAF on more than 1,300 colorectal cancers using DNA extracted from H&E-stained tissue sections. Our data provide no evidence for an interfering effect of H&E staining on DNA testing, suggesting that DNA from H&E-stained sections can be effectively used for routine DNA testing.
- Histochemical staining
- Polymerase chain reaction
- Molecular diagnostics
- Mutation detection
- Microdissection
- Solid tumor
Screening and identification of genetic alterations in formalin-fixed, paraffin-embedded (FFPE) tumor materials have been important in molecular diagnosis and clinical medicine.1–6 Solid tumors are inherently heterogeneous tissues that contain neoplastic cells admixed with non-neoplastic cells, including inflammatory cells, vascular and lymphatic endothelial cells, smooth muscle cells, and stromal fibroblasts. Therefore, an enrichment of neoplastic cells is commonly performed before DNA extraction in molecular testing to avoid false-negative results due to low neoplastic cellularity.7
Macrodissection from tissue sections on glass slides has been a widely used method to exclude pure nonneoplastic areas and enrich neoplastic cellularity for molecular testing. For macrodissection of tumor areas, an accurate morphologic identification of tumor areas is essential.8,9 Thus, macrodissection should be guided by an H&E-stained tissue section with tumor areas marked by a pathologist under a microscope Image 1A. It would be ideal to dissect tumor tissue from an H&E-stained section (without coverslip) because the H&E stain enables us to easily identify tumor areas Image 1B, particularly next to the H&E slide with the marked tumor areas (Image 1A). In contrast, it is difficult to accurately identify tumor areas of an unstained tissue section Image 1C, and tumor areas can be missed.
With regard to the influence of histochemical staining on downstream molecular assays, some previous studies have shown interfering effects of histochemical staining, especially hematoxylin stain,10–12 while another study failed to show a significant difference in polymerase chain reaction (PCR) amplification between DNA specimens from H&E-stained and unstained tissue sections.13 To avoid possible interfering effects of histochemical staining on molecular assays, macrodissection of tumor from unstained tissue sections is commonly performed, despite the difficulty in identifying tumor areas (Image 1C) and the risk of missing tumor areas.1,14–17 To our knowledge, no previous study has systematically evaluated the influence of histochemical staining on downstream molecular assays.
We therefore conducted this study to evaluate various molecular assay results on DNA extracted from H&E-stained, hematoxylin-stained, and eosin-stained FFPE tissue sections, in comparison with DNA extracted from unstained tissue sections. We compared results of agarose gel genomic DNA fragment analysis (without PCR), quantitative real-time PCR assay, PCR-pyrosequencing assay, and PCR–capillary electrophoresis fragment analyses. We did not observe an appreciable difference in any of the results on DNA specimens from stained tissue. Furthermore, as a proof-of-principle study, we successfully used DNA from H&E-stained tissue sections and obtained DNA analysis data from more than 1,300 colorectal cancers. Our data support the feasibility of DNA extraction from H&E-stained tissue sections for routine clinical laboratory workflow.
Materials and Methods
FFPE Tissue Specimens
Specimen collection and analysis in this study were approved by the Harvard School of Public Health, Brigham and Women's Hospital, and Dana-Farber Cancer Institute institutional review boards (Boston, MA). FFPE colorectal cancer tissue specimens (5 cases) were anonymized after collection from the archival file of the Department of Pathology, Brigham and Women's Hospital, to assess the effects of histochemical staining on downstream molecular testing Figure 1.
As a proof-of-principle feasibility study, we tested DNA extraction from H&E-stained tissue sections in a large cohort of colorectal cancer cases. We used 2 US nationwide prospective cohort studies, the Nurses' Health Study (N = 121,701 women followed up since 1976) and the Health Professionals Follow-up Study (N = 51,529 men followed up since 1986).18,19 We collected FFPE tissue blocks from hospitals and pathology laboratories throughout the United States where cohort participants with colorectal cancer underwent cancer resections.18,19 H&E-stained tissue sections were reviewed by a pathologist (S.O.). Genomic DNA was extracted from H&E-stained tissue sections, using another H&E-stained section with marked tumor areas as a guide (as in Images 1A and 1B), for downstream KRAS, BRAF, and MSI testing, and results were available for 1,314, 1,313, and 1,293 cases, respectively.
Overall Study Strategy, DNA Extraction, and Agarose Gel Electrophoresis
Deparaffinized and hydrated 10-μm sections from the same tissue block were stained by hematoxylin (Harris hematoxylin, Surgipath, Richmond, IL) alone for 3 minutes (group 2), by eosin (Surgipath) alone for 15 seconds (group 3), or by H&E (group 4; Figure 1). Unstained slides were prepared as control slides (group 1). Figure 1 illustrates our overall strategy of experiments to assess the effects of hematoxylin and/or eosin staining on DNA integrity and downstream molecular assays.
To avoid bias that could be introduced by tumor-directed macrodissection, a whole tissue section (including tumor and adjacent normal tissue) was entirely scraped off from a glass slide by using a sterile needle. Thus, DNA yields were very similar between the groups. The scraped tissue was collected into a microtube, and genomic DNA was extracted using QIAamp DNA Mini Kit (Qiagen, Valencia, CA). As a result, each of 10 tissue sections was "aliquoted" to 1 tube, to yield 10 aliquoted DNA specimens for each group (Figure 1). The 260/280 nm absorbance ratio of extracted DNA was approximately 1.8 (NanoDrop, Thermo Scientific, Waltham, MA). Extracted DNA from unstained tissue and H&E-stained tissue from all 5 cases was analyzed by electrophoresis in 0.8% agarose gel Image 2. Three cases were used for real-time PCR and PCR-pyrosequencing assay, and the other 2 cases were used for microsatellite PCR fragment analysis.
Real-Time PCR Assay for GAPDH
To assess PCR amplification efficiency, we performed real-time PCR using the primers for GAPDH and SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA) by the ABI 7300 Real-Time PCR system (Applied Biosystems). Primer sequences were 5′-GTCATGGGTGTGAACCATGAGAA-3′ and 5′-TGGT-CATGAGTCCTTCCACGAT-3′. The PCR reaction was repeated 4 times on each of the 10 DNA aliquots for groups 1 through 4, and cycle threshold (Ct) values were compared.
Microsatellite PCR Fragment Analysis
To assess the effects of hematoxylin and/or eosin staining on PCR fragment analysis, we performed PCR for dinucleotide markers, D2S123 and D5S346,20 after PCR-based whole genome amplification on genomic DNA as previously described.21 Forward primers were labeled with fluorescence, and PCR product sizes were 180 base pairs (bp; D2S123) and 129 bp (D5S346). PCR products were electrophoresed and analyzed by the ABI 3730 DNA Analyzer (Applied Biosystems). PCR fragment analysis was repeated twice on each of the 10 DNA aliquots.
For the large feasibility cohort, we performed microsatellite instability (MSI) analysis using a 10-marker panel (D2S123, D5S346, D17S250, BAT25, BAT26, BAT40, D18S55, D18S56, D18S67, and D18S487).20 MSI-high was defined as the presence of instability in 30% or more of the markers and MSI-low/microsatellite stability as instability in fewer than 30% of the markers.20
PCR-Pyrosequencing Assay for KRAS and BRAF
To assess the effect of hematoxylin and/or eosin staining on pyrosequencing, we performed a PCR-pyrosequencing assay for KRAS (codons 12 and 13), which was previously developed and validated.21 The size of PCR product was 82 bp, and 10 μL of each was sequenced by using the Pyrosequencing PSQ96 HS System (Qiagen). The PCR-pyrosequencing reaction was repeated 3 times on each of the 10 aliquots. In addition, in the feasibility cohort, the PCR-pyrosequencing assay for BRAF (codon 600) was performed as previously described.22
Statistical Analysis
For all statistical analyses, we used the SAS program (version 9.1, SAS Institute, Cary, NC). All P values were 2-sided. The Ct in real-time PCR, which reflected amplification efficiency of each PCR reaction given similar amounts of scraped tissue and extracted DNA, was compared by using the analysis of variance (ANOVA) test, adjusting for case, spot of each reaction, and plate. We performed the ANOVA test for comparing peak heights of the fluorescence signal on capillary electrophoretograms in microsatellite analysis. The distribution of the peak height values (median, 3,028.5; range, 0–10,618) was normalized by logarithmic transformation after adding 0.5 to each value (to log-transform the value of "0"). A deviation from the null hypothesis in any of the comparisons might imply an interfering effect of histochemical staining on the DNA testing process.
Integrity of DNA From Unstained and Stained Tissue
Our overall strategy to assess the influence of H&E staining on subsequent DNA testing for FFPE tissue is shown in Figure 1. First, we assessed the influence of H&E staining on DNA integrity. Extracted DNA specimens of group 1 (unstained) and group 4 (H&E-stained) were loaded onto 0.8% agarose gel (Image 2). There was no appreciable difference in DNA integrity between the DNA specimens from H&E-stained and unstained tissue sections from all 5 cases.
Comparison of Ct Values in Real-Time PCR
We compared Ct values in real-time PCR for GAPDH in the 4 groups using 3 cases (× 10 sections × 4 repeated runs) by the ANOVA test, which adjusted for case, spot of each reaction, and plate Figure 2A. There was no significant difference in Ct values between groups (P = .19).
Microsatellite PCR Fragment Analysis
We performed PCR fragment analysis for the 2 microsatellite markers, D2S123 and D5S346, and assessed the potential influence of tissue staining by using 2 cases (× 10 sections × 2 repeats). We measured the peak height in electrophoretograms, and compared the mean peak height value for the 4 groups. There was no significant difference in the mean peak height in the 4 groups (overall P = .35) Figure 2B.
PCR-Pyrosequencing Assay for KRAS
We performed the PCR-pyrosequencing assay for KRAS codons 12 and 13 on 3 cases (× 10 sections × 3 repeats) and compared results for the 4 groups. In 2 cases, there was a c.35G>A (p.G12D) mutation, and 1 case showed only a wild-type sequence Figure 3. All pyrograms for KRAS codons 12 and 13 showed clear peaks and little background, and there was no appreciable difference in the 4 groups.
MSI Testing and Pyrosequencing for KRAS and BRAF in the Large Feasibility Cohort
Finally, we performed a proof-of-principle feasibility study on FFPE tissue blocks that were fixed and processed in numerous pathology laboratories throughout the United States, using the 2 US nationwide prospective cohort studies.18 We dissected tumor tissue from H&E-stained sections (without coverslip) using a guide H&E slide with marked tumor areas (as in Images 1A and 1B), and extracted DNA for subsequent MSI analysis and the PCR-pyrosequencing assay for KRAS and BRAF. KRAS and BRAF mutations were detected in 470 (35.8%) of 1,314 cases and 186 (14.2%) of 1,313 cases, respectively Table 1. MSI-high was present in 197 (15.2%) of 1,293 cases. These frequencies are compatible with the published data on these molecular features in colorectal cancers,23–27 particularly in population-based studies.28–33 There was no evidence for an interfering effect of H&E staining on these molecular assays. These data suggest that DNA from H&E-stained sections can be used for routine tumor DNA testing on FFPE tissue.
We conducted this study to evaluate the potential influence of H&E staining on subsequent DNA testing. Solid tumor tissue is inherently heterogeneous tissue with an admixture of neoplastic cells and nonneoplastic cells, including inflammatory, stromal, endothelial, and smooth muscle cells, even within tumor tissue areas. Macrodissection from H&E-stained tissue sections enables accurate identification and dissection of tumor areas to obtain DNA specimens with a high content of neoplastic cell DNA. Especially for small tumor nests or metastatic tumor foci that must be carefully and precisely dissected, it is very useful to stain and clearly visualize the tissue section. H&E staining is a common and simple method to visualize the tumor area of interest. Although hematoxylin staining has been believed to inhibit DNA amplification reaction, only a small number of studies have been performed to assess the potential influence of tissue staining on amplification of DNA by PCR.10,11,13 Notably, no previous study has systematically evaluated the influence of H&E staining on downstream molecular assays. Our current study provided no evidence for interfering effects of H&E staining on subsequent DNA assays. Our data support routine clinical use of H&E staining of tissue sections for subsequent tumor tissue dissection, DNA extraction, and molecular analyses.
Previous studies evaluated potential influence of tissue staining on DNA assays.10,11,13 Burton et al10 and Chen et al34 showed that hematoxylin staining inhibited DNA amplification by PCR, and Diss et al11 reported that DNA from H&E-stained tissue was amplified less efficiently than DNA from unstained tissue. On the other hand, Murase et al13 reported no significant difference in PCR amplification between DNA specimens from unstained and H&E-stained tissue. However, none of the previous studies10,11,13 quantitatively evaluated the efficiencies of PCR amplification on DNA specimens from unstained and stained tissues. In the present study, there was no statistically significant difference in GAPDH amplification by quantitative real-time PCR between DNA specimens from unstained tissue, hematoxylin-stained tissue, eosin-stained tissue, and H&E-stained tissue.
Pyrosequencing is a nonelectrophoretic sequencing by nucleotide extension. Pyrosequencing is useful for sequencing of relatively short nucleotides (up to 40 to 50 bp). Therefore, pyrosequencing has been applied for single nucleotide polymorphism genotyping,35 bacterial strain typing,36 mutation detection in tumors,21,37,38 quantitative promoter CpG island methylation analysis,39,40 and measurement of LINE-1 methylation.41 We evaluated the influence of H&E staining for the PCR-pyrosequencing assay for KRAS and showed no appreciable difference in results between DNA specimens from unstained and H&E-stained tissues.
Capillary electrophoresis is commonly used for microsatellite analysis in FFPE clinical tissue specimens (eg, MSI and loss of heterozygosity analyses).42–45 We showed no significant difference in peak heights of the electrophoretograms of 2 microsatellite markers (D2S123 and D5S346) between DNA specimens from unstained and stained tissues.
Murphy et al46 reported an artifact (a peak at 71 bp) in capillary electrophoresis due to autofluorescence from eosin, which was probably contaminated by nonstringent DNA purification. A notable difference from our study was that Murphy et al46 experienced the artifact after biopsy tissue (not a tissue section on a slide) was stained with eosin "to enable identification of small tissue fragments during sectioning." We have not experienced this problem, despite the fact that we have performed microsatellite PCR fragment analysis on more than 1,300 cases using DNA from H&E-stained tissue. Nevertheless, we agree with Murphy et al46 that artifacts caused by eosin should be avoided by using stringent DNA purification steps, and that artifacts from histochemical staining should be considered when peaks of the same product size (eg, 71 bp) are present in multiple specimens or primary peaks contain additional underlying peaks of other colors.
As a potential limitation of our study, we used a relatively small number of tissue blocks (N = 5) to systematically compare the effects of histochemical staining in downstream molecular assays. Nevertheless, we used as many as 10 sections and 10 DNA aliquots for each condition to achieve a precise measurement estimate. Moreover, as the proof-of-principle feasibility study, we used DNA extracted from H&E-stained tissue of more than 1,300 colorectal cancers in 2 US nationwide prospective cohort studies (the Nurses' Health Study and the Health Professionals Follow-up Study) for MSI analysis and pyrosequencing of KRAS and BRAF (Table 1). The FFPE tissue specimens were retrieved from numerous pathology laboratories throughout the United States. Despite the diversity of laboratories that processed and stored tissues, we were successful in performing those molecular assays in more than 95% of cases tested, and the frequencies of MSI-high and mutations of KRAS and BRAF were compatible with published data on colorectal cancer,23–27 particularly in population-based studies.28–33 In addition, using DNA from H&E-stained tissue sections, it has been shown that BRAF mutation and MSI-high in colon cancer are associated with the CpG island methylator phenotype (CIMP20,28,47–52) and patient outcome,20,53 and KRAS mutation is associated with CIMP-low in our cohort studies.22 Our data indicate that H&E-staining is useful for DNA analysis on FFPE tissues from numerous pathology laboratories in multicenter clinical trials or population-based cohort studies,42,54 which can contribute to improvement of patient care and public heath.
Our present study represents the first systematic investigation on the effects of H&E staining in downstream molecular assays. Our results provide no evidence to support the common conception that H&E staining interferes with molecular assays on DNA from FFPE tissue. Our data support routine clinical use of H&E staining on tumor sections for subsequent tissue dissection and DNA extraction. H&E staining enables easy and accurate identification of tumor areas to obtain DNA with a high content of neoplastic cellular DNA. H&E staining can be easily incorporated into routine pathology workflow, so that laboratory personnel can easily identify and dissect tumor areas from H&E-stained tissue sections, guided by an H&E-stained slide with marked tumor areas. Thus, our data may have considerable influence on clinical molecular diagnostics and routine tumor molecular testing toward personalized medicine.
We deeply thank the Nurses' Health Study (NHS) and Health Professionals Follow-up Study (HPFS) cohort participants who have agreed to provide us with information through questionnaires and biological specimens; hospitals and pathology departments throughout the United States for generously providing us with tissue specimens; the staff of the NHS and the HPFS for their valuable contributions; and all of the US state cancer registries for their help.
↵* Drs Morikawa, Shima, and Kuchiba contributed equally to this work.
The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or NIH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Supported by US National Institute of Health (NIH) grants P01 CA87969 (S.E. Hankinson), P01 CA55075 (W.C. Willett), P50 CA127003 (Dr Fuchs), R01 CA118553 (Dr Fuchs), R01 CA124908 (Dr Fuchs), and R01 CA151993 (Dr Ogino).
- © American Society for Clinical Pathology


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The length of time for the expiration date or "best used before" date depends on the type of product, as well as the brand.
Perishable items (such as flax oils or certain probiotics) generally have shorter expiration dates. Although our warehouse is fully air-conditioned, these more fragile items are put in cold storage (freezer or refrigeration unit) for maximum freshness.
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- Product Code: HRP-82141
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By: Ovetta Sampson
| Posted: July 23, 2008
Brothers Jean Carlo and Jean Pier, so malnourished as toddlers they were the size of potato sacks, are alive today because of supporters of the Child Survival Program.
Like all of Compassion's programs, the primarily home-based Child Survival Program is holistic in nature. It addresses several areas of growth critical to a child's healthy development. Below are details on how the Celim Child Survival Program workers helped rescue Jean Carlo and Jean Pier Taco, brothers from Peru, from the brink of death using a holistic child development approach.
When Celim CSP workers found Jean Carlo and Jean Pier Taco, the toddlers were so malnourished each weighed less than 15 pounds. They could barely hold their heads up. They couldn't walk. And they rarely talked. CSP staff worked for months nurturing the boys' physical, developmental and intellectual development. Here's how the Child Survival Program helped the boys in each area:
Martho Taco says the Child Survival Program is a miracle, not only for rescuing her son but also for saving her soul. "I know God was the one who created the Child Survival Program," she says, "and I am very thankful."
What did you like about this story?
In fiscal year 2016, 82.4% of Compassion's expenses went directly to program activities helping children in our programs.
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Compassion International, Inc. is a registered 501 (c)(3) non profit organization.
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Compassion and Compassion International are Registered Trademarks of Compassion International, Inc.
We're here to serve you. See the many ways to reach us or choose from the three options below to contact us online:
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Your donation to the Where Most Needed fund will help us better serve children in extreme poverty.
"Whoever is kind to the poor lends to the LORD, and he will reward them for what they have done."
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Avery Chapel A.M.E. Church 3rd Bi-annual Christmas Concert
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London, United Kingdom
Florence, Italy
Event Details
Avery Chapel A.M.E. Church Choir in concert, featuring the Avery Chapel Male Chorus and Inspirational Choir. Guest include Evangelist Latonya Randle of Memphis Tn, and Psalmist Stacy Johnson of Coldwater, MS plus many many more!!!
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Saturday, December 14, 2013 from 6:00 PM to 9:00 PM (PST)
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Valid for Moodle 2.8. Updated on 7/14/2015.Blocks are the smaller boxes that appear on the sides of your Moodle course page. Some of these are specified by Moodle (such as Navigation and Settings) and some have been added here at Amherst (such as Need Help?). In addition you can add additional blocks to your course to show the information you want your students to see.
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This review is from: Avatar Desktop PC Gaming FX6365TI AMD FX-Series FX-6300 (3.50 GHz) 16 GB DDR3 1 TB HDD NVIDIA GeForce GTX 650Ti 1GB Windows 8 64-Bit
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