HDTenEightyP/NetNews huggingface dataset:
From: brian@bio.aau.dk
Newsgroups: bionet.immunology
Subject: Hybridoma problems more info.
Date: 25 Jul 92 02:16:07 GMT
Organization: BIOBASE, Denmark

Hello again.

In continuation of my problems concerning growth of hybridoma cells,
I will try to give a more full story of what we actually did. This
is due to the responses that I've got (and I would like to thank the
people who showed interest).

1) 10 e8 spleen cells and 5x10 e7 myeloma cells were mixed in 10 ml of
   RPMI 1640 + L-Glutamin, and the cells were spun down.
2) 50% PEG 4000 (Gibco) were added to the pellet over 1 min. and the
   cells were allowed to rest for 1 min.
3) Then 10 ml of RPMI 1640 + L-Glutamin were added and the cells were spun 
   down again.
4) Cells were resuspended in RPMI 1640 + L-Glutamin + 20% FCS + 10 % HECS
   (Human endothelia cell supernatant). The cells were plated out in 96 wells
   plates coated with collagen, 100 ul in each. and allowed to rest for 1 day.
5) Then 100 ul of The same medium as in 4) supplemented with 1xHAT and 
   Pen/strept. were added on top of the old medium. then rest for 1 day.
6) Half of the medium were then taken off and replaced with fresh medium
   as in 5). This is continued until day 8. 
7) on day 11 the cells are tested for antibody production and the positive
   are transfered to 6 wells plates which is still coated with collagen. and
   the medium is still RPMI 1640 + HAT. (The passage is with 0,1% trypsin)
8) 3 to 4 days later the cells are passaged. Again by 0,1% trypsin after which
   the cells are collected in RPMI 1640 + 20% FCS and at this stage we change
   from HAT to HT as supplement, and spun down. They are then resuspended in
   in the same medium RPMI 1640 + L-Glutamin + 20 % FCS + 10% HECS + 1xHT.

It is then our problems arise. The cells attach to the plates but they will
no longer proliferate.

I hope this helps some of you more experienced people out there, in relating
to our problem and maybe give some suggestion how to overcome the problems.

In advance I thank for the interest.

Peter Kristensen
Aarhus University
Department of Chemistry
Division of Biostructural Chemistry
Denmark

E-mail : Brian@bio.aau.dk
From: kristoff@genbank.bio.net (David Kristofferson)
Newsgroups: bionet.announce
Subject: CALL FOR VOTES: bionet.bionette
Date: 24 Jul 92 17:18:51 GMT
Reply-To: biovote@genbank.bio.net

Voting is now open on bionet.bionette.  Here is the charter as
presented previously.

----------------------------------------------------------------------

Proposal to establish bionet.bionette

Discussion leader:   Cassandra Smith
                     Lawrence Berkeley Laboratory

bionet.bionette charter:

The purpose of this discussion group will be to promote networking
among professional women in the biological sciences. In essence what
is proposed is a bioscience new girl network.  A major problem that
women in science face is isolation. Thus, the goal of this newsgroup
is to provide a forum where women in the biological sciences can (1)
share experiences and interests and (2) provide advice and support to
encourage women, despite obvious and not so obvious obstacles, to
enter and continue scientific careers.
------------------------------------------------------------------

Voting is now open on bionet.bionette and will run through 23 August
1992.  Please send your vote to any of the following addresses:

Address                               Location        Network
-------                               --------        -------
biovote@daresbury.ac.uk               U.K.            JANET
biovote@genbank.bio.net               U.S.A.          Internet/BITNET
biovote@irlearn.ucd.ie                Ireland         EARN/BITNET

Your vote should contain a single line:

YES on bionet.bionette

if you favor creation of the newsgroup or

NO on bionet.bionette

if you think that the creation of this newsgroup will adversely affect
the BIOSCI system.  If you are simply not interested in participating
in this newsgroup, please don't cast a NO vote, but instead just don't
vote at all.

The newsgroup proposal must receive at least 80 YES votes to pass and
the number of YES votes must be greater than the number of NO votes by
at least 40.  Discussion of the newsgroup proposal is now closed and
we strongly discourage posting any messages in other forums about the
fact that a CALL FOR VOTES has been issued.

				Sincerely,

				Dave Kristofferson
				GenBank Manager

				kristoff@genbank.bio.net
Xref: sparky bionet.announce:142 bionet.molbio.genbank:431
From: kristoff@presto.ig.com (David Kristofferson)
Newsgroups: bionet.announce,bionet.molbio.genbank
Subject: GOS off-line due to genbank.bio.net disk crash
Date: 22 Jul 92 17:32:26 GMT
Followup-To: poster
Organization: IntelliGenetics Inc., Mtn. View, Ca.

We had a power problem during the night that led to a disk crash, so
genbank.bio.net is off-line until probably mid afternoon today, 22
July.  BIOSCI mail distribution and the GenBank e-mail servers will
all be affected.  Mail to the various genbank.bio.net addresses will
be queued, but we will not be able to answer your queries until after
the system comes back up.  I will also send a copy of this posting to
the Daresbury addresses to increase its chance of getting distributed
widely.

Dave Kristofferson
From: santulli@europa.informatik.uni-dortmund.de (Marco Santulli)
Newsgroups: bionet.neuroscience
Subject: activation dinamics of neurons
Date: 24 Jul 1992 17:03:16 GMT
Organization: University of Dortmund, Germany
Distribution: world
NNTP-Posting-Host: europa

I'm looking for a detailed description of the activation
dynamics of the real neuron.

Are they equal for every neuron, or is there a differenciation
in the various functions of neurons with different acitvities??

Please post answers or mail them to:

-----------------------------------------------------------
Marco santulli                                   carpe diem
santulli@jupiter.informatik.uni-dortmund.de

Dept. of Computer Science I  University of Dortmund Germany
-----------------------------------------------------------
From: zador-anthony@CS.YALE.EDU (Tony Zador)
Newsgroups: bionet.neuroscience
Subject: NO and epilepsy
Date: 22 Jul 92 03:10:00 GMT
Distribution: bionet

Is anyone aware of any work on NO and epilepsy, e.g. in hippocampal
slices?

Tony Zador
zador@cs.yale.edu
From: pedro@mulab.physiol.upenn.edu (maldonado)
Newsgroups: bionet.neuroscience
Subject: Lucifer Yellow
Date: 30 Jul 92 21:43:54 GMT
Distribution: usa
Organization: University of Pennsylvania


	Lucifer Yellow releases free radicals inside the cell, so it is
not possible to keep the cell for long, physiologically intact. By long
I mean 30 minutes. Of course, if you are constantly using UV
illumination, the time is reduced an order of magnitude.
Newsgroups: bionet.neuroscience
From: andrew@uaneuro.uah.ualberta.ca (Andrew Penn)
Subject: Clinical Neuroscience
Reply-To: euro.uah.ualberta.ca (Andrew Penn)
Organization: University Of Alberta, Edmonton Canada
Date: Wed, 22 Jul 1992 00:54:42 GMT

	As a first step to establishing a repository for clinical  
guideline and consensus material I need an estimate of how many people  
following the Neuroscience News Group are Neurologists or Neurosurgeons.  
With some trepidation I would ask for short replies stating your  
specialty, though I suspect the number is small.

Andrew Penn
Assistant Professor of Neurology,
University of Alberta,
Edmonton, Alberta,
Canada
E-Mail:	andrew@uaneuro.uah.ualberta.ca
From: mark@uucp.nibr
Newsgroups: bionet.neuroscience
Subject: microtubule elongation
Date: 23 Jul 92 07:44:18 GMT
Distribution: bionet
Original-To: neur-sci@nl.nluug.sun4nl


I am working on a model concerning the elongation of microtubules
in neurons during outgrowth. My model so far shows that stopping
neuritic outgrowth by stopping microtubule outgrowth demands a stop
of the input of material in the outgrowing neuritic tips. I have two
questions about this, which I could not trace out in literature:

1 - Is elevated calcium able to (locally) block transport of soluble
    proteins at the site of the growth cone ?

2 - Is the production of tubulin (all isoforms) at the soma regulated
    by the concentration of tubulin ? More specifically: is there a
    negative feedback system ?

All responses would be welcome,

Mark van Veen
Neth. Inst. Brain Research
mark%nibr@nluug.nl
From: uqecd08@ucl.ac.uk (John Thomas)
Newsgroups: bionet.neuroscience
Subject: PhD seeks position near Brussels
Date: 30 Jul 92 16:50:04 GMT
Organization: Bloomsbury Computing Consortium

A friend of mine would like to do further work in neuroscience, or
a related field, within a few hours of Brussels. If the following is of
interest to you as an employer, reply by e-mail  with brief details of
the proposed job and institution and I will send a hard-copy full cv. 

Alternatively if you work in the field and know of useful contacts, a reply 
would be appreciated.


First degree: Upper Second Class Honours in pharmacology at the
University of Manchester, England.

Doctorate: Investigation of the involvement of eicosanoids in the
central inflammatory response with special reference to compounds
mediating the response to cytokines and bacterial endotoxins. Used in
vivo and in vitro techniques. Radioimmunoassay and Percoll density
gradients were among techniques used in studies on rats.

Studies were completed at the Department of Physiological Sciences,
University of Manchester, England.

Post-Doctoral experience: Brief continuation of PhD work at Manchester.
Followed by two years on the site and mode of action of cytokines in the
CNS using in vivo and in vitro systems at The Hospital for Sick
Children, Toronto, Canada.


--
|   v___v    | John Thomas, Programmer         e-mail: J.Thomas@ucl.ac.uk     |
|  / o-o \   | LEPRA Evaluation Project        Tel: 011 44 71 927 2217        |
|  \__Y__/   | London School of Hygiene & Trop. Med.                          |
| ( ~ ~ ~ )  | Keppel St, London WC1E 7HT, England                            |
From: BQWANG@MACC.WISC.EDU (Baoqing Wang)
Newsgroups: bionet.neuroscience
Subject: Lucifer Yellow filled electrode
Date: 27 Jul 92 22:51:00 GMT
Distribution: bionet

Dear friends:
I am trying to use Lucifer Yellow (LY) filled glasselectrode to do
intracellular recording, and then inject LY into the cell to label
it.  I find that it's difficult to keep the cell for a long time.
Would anyone please share your experience with me?  Any suggestion
would be appreciated.  Thank you in advance.
 
Sincerely,
Baoqing Wang
in University of Wisconsin-Madison
Email: BQWANG@macc.wisc.edu   or   BQWANG@wiscmacc.BitNet
From: HAINES@HELIX.MGH.HARVARD.EDU ("Jonathan L. Haines")
Newsgroups: bionet.genome.chrom22
Subject: (none)
Date: 30 Jul 92 03:39:32 GMT
Distribution: bionet

To Chromosome 22 bulletin board users:

Here is the list of 12 PCR polymorphisms we have currently mapped on chromosome
22.  I have also included our latest, but still preliminary, map of these
markers.  Note that the most distal marker, CYP2DP8 is still quite far from
the telomere.  In fact, we probably have most of 22q13.3 left to map.  Hope
this is helpful to somebody.



Locus            Primer1                     Primer2             Het   
----------  ----------------------- ---------------------------  ---- 
CRYB2       CCATGCTACCTTGTAAAGTT      CCGTATGACTTGTATTCACTAC     0.76     
CYP2DP8     CCCTCTTGGCTCTAACAGC       CTCGGTGACAGAGCAAGAC        0.70
D22S156     AGCCTGGGAGTCGAGTGA        AGCTCCAAATCCAAAGACGT       0.78 
D22S257     AGCTGATACACCACTTCTGA      GACAGAAATATCCTTCCCAT       0.67
D22S258     GCCTGAAATTATTCCAGCTG      AATAGTAGAGTTTGCCTTTC       0.82
D22S264     ATTAACTCATAAAGGAGCCC      CACCCCACCAGAGGTATTCC       0.80          
D22S268     TACGTCCTCACAATCCAGCGT     CTGACCTCCCACCATTAC         0.71
IL2RB       CTTGGCTCCTGTGTGCACCT      GCACCCAGGGCCAGATAAAGATCT   0.81          
PDGFB       CGCTGGCTCTGGAGCTCA        GATGTTGGTCATCACAGT         0.57
TOP1P2      GGTTTTCTGTCATTCTTGTTGA    AGTGAGTGGAGATTGCATTG       0.92 
F8VWFP1-1   GATTGCGGGGTCATATGCTAATACC TTACCTCATACGTGCTAGAATGGCC  0.62          
F8VWFP1-2   TTGTAGACAGCACATAGTTG      GGCAACAGAGCCAGACTCCA       0.57          

Locus   # alleles     Range       Repeat  
_____   _________   _________     ______
CRYB2       7       200-212         GT
CYP2DP8    12       108-130         GT 
D22S156     7       96 -110         GT
D22S257     4       125-133         GT
D22S258    10       177-195         GT 
D22S264     7       190-210         GT
D22S268     4       244-252         GT
IL2RB       5       125-135         GT
PDGFB       5       110-120         GT
TOP1P2     22       100-144         GT
F8VWFP1-1   5       190-210         GT
F8VWFP1-2   5       130-150       AAAT
 
 
Latest Genetic Map:
 
pter--F8VWFP--12--D22S264--5--D22S257--1--D22S156--3--TOP1P2--6--D22S258--

--4--CRYB2--14--D22S268--14--IL2RB--9--PDGFB--6--CYP2DP8--lots of space--

--qter.
From: kristoff@genbank.bio.net (David Kristofferson)
Newsgroups: bionet.genome.chrom22
Subject: Re: State of the 22-bulletin board
Date: 27 Jul 92 17:19:19 GMT
Distribution: bionet
Organization: GenBank Online Service

> In response to Nancy's posting, I don't see why Dave K. gets so bent
> out of shape when a usenet new group is not used very often.  What's
> the big deal here?  I love David dearly and have know him for years
> and can only think that threatening to "loose the bbd" will maybe get
> everyone to actually use it!!

Dave K. is not bent out of shape unless you are referring to the fact
that he has bent over backwards to keep this group running.  He was
merely mentioning the rule passed by the BIOSCI readership at the
beginning of this year that newsgroups which averaged less than 52
messages a year would be put on notice of possible closure.  This was
the policy that was adopted:

----------------------------------------------------------------------

Newsgroup Termination Policy
----------------------------

Any group with less than 52 msgs in the previous calendar year will be
put on notice by posting an announcement to the newsgroup (not to
bionet.announce) that it faces cancellation.  It can be reprieved if
80 readers respond within two weeks (this policy will be stated in the
termination announcement).  It then has two months to reach a usage
level of one message per 3 days or else it will be abolished.  Appeals
to the BIOSCI management about high content albeit low volume on the
group will be considered.

----------------------------------------------------------------------
Newsgroups: bionet.genome.chrom22
From: chipper@welchgate.welch.jhu.edu (Mike Chipperfield)
Subject: Re: (none)
Organization: Johns Hopkins Univ. Welch Medical Library
Distribution: bionet
Date: Fri, 31 Jul 1992 14:49:24 GMT

In article <01GMYU1Z3SFM8WW0BQ@HELIX.MGH.HARVARD.EDU> HAINES@HELIX.MGH.HARVARD.EDU ("Jonathan L. Haines") writes:
>To Chromosome 22 bulletin board users:
>
>Here is the list of 12 PCR polymorphisms we have currently mapped on chromosome
>22.  I have also included our latest, but still preliminary, map of these
>markers.  Note that the most distal marker, CYP2DP8 is still quite far from
>the telomere.  In fact, we probably have most of 22q13.3 left to map.  Hope
>this is helpful to somebody.
>
>
>
>Locus            Primer1                     Primer2             Het   
>----------  ----------------------- ---------------------------  ---- 
>CRYB2       CCATGCTACCTTGTAAAGTT      CCGTATGACTTGTATTCACTAC     0.76     
>CYP2DP8     CCCTCTTGGCTCTAACAGC       CTCGGTGACAGAGCAAGAC        0.70
>D22S156     AGCCTGGGAGTCGAGTGA        AGCTCCAAATCCAAAGACGT       0.78 
>D22S257     AGCTGATACACCACTTCTGA      GACAGAAATATCCTTCCCAT       0.67
>D22S258     GCCTGAAATTATTCCAGCTG      AATAGTAGAGTTTGCCTTTC       0.82
>D22S264     ATTAACTCATAAAGGAGCCC      CACCCCACCAGAGGTATTCC       0.80          
>D22S268     TACGTCCTCACAATCCAGCGT     CTGACCTCCCACCATTAC         0.71
>IL2RB       CTTGGCTCCTGTGTGCACCT      GCACCCAGGGCCAGATAAAGATCT   0.81          
>PDGFB       CGCTGGCTCTGGAGCTCA        GATGTTGGTCATCACAGT         0.57
>TOP1P2      GGTTTTCTGTCATTCTTGTTGA    AGTGAGTGGAGATTGCATTG       0.92 
>F8VWFP1-1   GATTGCGGGGTCATATGCTAATACC TTACCTCATACGTGCTAGAATGGCC  0.62          
>F8VWFP1-2   TTGTAGACAGCACATAGTTG      GGCAACAGAGCCAGACTCCA       0.57          
>
>Locus   # alleles     Range       Repeat  
>_____   _________   _________     ______
>CRYB2       7       200-212         GT
>CYP2DP8    12       108-130         GT 
>D22S156     7       96 -110         GT
>D22S257     4       125-133         GT
>D22S258    10       177-195         GT 
>D22S264     7       190-210         GT
>D22S268     4       244-252         GT
>IL2RB       5       125-135         GT
>PDGFB       5       110-120         GT
>TOP1P2     22       100-144         GT
>F8VWFP1-1   5       190-210         GT
>F8VWFP1-2   5       130-150       AAAT
> 
> 
>Latest Genetic Map:
> 
>pter--F8VWFP--12--D22S264--5--D22S257--1--D22S156--3--TOP1P2--6--D22S258--
>
>--4--CRYB2--14--D22S268--14--IL2RB--9--PDGFB--6--CYP2DP8--lots of space--
>
>--qter. 

Additonal details about these data are available in the Genome Data Base. We 
will be receiving some band size information for a few loci from Dr. Haines in 
the next few days. The GDB has additional information of interest to 
investigators working on chromosome 22:

	Genes: 			68
	Polymorphic Loci: 	140
	PCR primers: 		20


To get an account with the Genome Data Base, please contact:


	GDB User Support
	3rd Floor
	1830 E. Monument St.
	Baltimore, MD 21205
	Phone: (410) 614-0436
	FAX:   (410) 614-0434
	Email: help@welchgate.welch.jhu.edu


Sincerely,

Michael A. Chipperfield
DNA Data Coordinator
Genome Data Base
From: shephens@esvax.dnet.dupont.com
Newsgroups: bionet.genome.chrom22
Subject: State of the 22-bulletin board
Date: 23 Jul 92 21:48:30 GMT
Distribution: bionet

As was mentioned earlier in a memo from Dave Kristofferson, the chromosome 
22 community is likely to loose this bulletin board if the use does not 
increase to an average of one message per week.  We are well into 1992 and 
I am fairly certain that this will not be met unless traffic increases 
dramatically.  Maybe it was simply just an idea before its time, but I am 
sincerely astonished at the lack of use.  For example, other than a reply 
from Peter Pearson of GDB, there was no bulletin board response to Jonathan 
Haines request for chromosome 22 GT repeats to map.  Why?  I know there is 
an up-coming chromosome 22 meeting, but I haven't seen any bulletin board 
traffic about it (or have I???).  I suggested that everyone give a brief 
comment about their more recent research efforts and interest on the 
chromosome, but there was again only a few responses.  From my inquiry 
concerning who is READING the board, I had approximately 20 responses... so 
someone IS out there.  Finally, in February of this year, Johnathan Haines 
mentioned that he hoped to have primer sequences, map locations etc. in a 
little database, and hopefully others are doing the same.  Will there be 
any effort to alert the chromosome 22 community about new submissions to 
GDB for the chromosome or will others have any inclination to use the 
bulletin board for dissemination of such?  
     Finally, I would just like to mention that my username has changed to 
"SHEPHENS@esvax.dnet.dupont.com" (rather than Shepherd), and that as part 
of a pharmaceutical company my research efforts are changing with the goals 
of the company (not uncommon).  I therefore will not be attending the 
meeting in Philadelphia and hope to see the results of the meeting on THIS 
bulletin board! Also, we did make some progress on getting chromosome 22
clones in the P1 vector system, but will not at this time pursue the
project further. Nancy  P.S.  What makes the Arabidopsis 
newsgroup so successful?
From: HAINES@HELIX.MGH.HARVARD.EDU ("Jonathan L. Haines")
Newsgroups: bionet.genome.chrom22
Subject: access
Date: 30 Jul 92 14:26:25 GMT
Distribution: bionet

I seem to have lost my membership in the chr22bb.  I send messages, but
get nothing back from the BB.  HELP!
From: shephens@esvax.dnet.dupont.com
Newsgroups: bionet.genome.chrom22
Subject: RE:
Date: 30 Jul 92 13:20:24 GMT
Distribution: bionet

Congratulations on being the first person to actually put data on the 
bulletin board.  Did you also submit this to GDB?  Nancy
From: JIM2@PSUVM.PSU.EDU ("June Medford")
Newsgroups: bionet.genome.arabidopsis
Subject: B-ATPase
Date: 21 Jul 92 14:57:00 GMT
Distribution: bionet

We're quantitating the expression of some shoot meristem genes in the
juvenile and adult meristem with PCR and would like to use B-ATPase as a
control in our experiments.  The sequence has been published for tobacco.
Does anyone have a sequence for the Arabidopsis gene?  Thanks for the help.
June Medford, JIM2@PSUVM.
From: brwnng@CCWF.CC.UTEXAS.EDU (@ccwf.cc.utexas.edu, Karen Browning)
Newsgroups: bionet.genome.arabidopsis
Subject: (none)
Date: 21 Jul 92 22:48:07 GMT
Distribution: bionet

I am looking for an Arabidopsis cDNA in lambda ZAP.  Does anyone have a
library to share?  Thank you.
Karen S. Browning                      (email) brwnng@ccwf.cc.utexas.edu
Dept. of Chemistry and Biochemistry    (fax)   512-471-8696
The University of Texas at Austin      (phone) 512-471-4562
Austin, TX  78712
From: bafp3@uk.ac.sussex.central (Neil Butt)
Newsgroups: bionet.genome.arabidopsis
Subject: check
Date: 27 Jul 92 13:56:37 GMT
Distribution: bionet
Original-To: arab-gen@uk.ac.daresbury
X-Mailer: ELM [version 2.3 PL11]

This is just to check our new network is running
From: pierce@esvax.dnet.dupont.com (John Pierce, phone: 302-366-5392, fax: -5236)
Newsgroups: bionet.genome.arabidopsis
Subject: Position Announcement
Date: 23 Jul 92 01:05:20 GMT
Distribution: bionet


     Position Announcement: Du Pont Agricultural Products

     We're looking for a Ph.D.-level scientist who will take a biochemical
     and/or genetic approach to investigating herbicide action and
     selectivity.  A  plant physiologist with a strong background in
     genetics, biochemistry, or analytical chemistry would be ideal.  
     Initial areas of study could include analysis of mode of action of
     new herbicides, identification of metabolic targets for rational
     design approaches, and evaluation of potential resistance mechanisms
     in model and target organisms. 

     The successful candidate will be interested in participating in a
     multi-disciplinary research team and in applying modern techniques to 
     the discovery of novel approaches to crop protection.

     Please provide a CV, names of references, and a brief summary of
     research interests to 

     John Pierce  
     Du Pont Agricultural Products 
     Stine-Haskell Research Center 
     Box 30, Elkton Road 
     Newark, DE 19714

     Bitnet: 	pierce@esvax.dnet.dupont.com 	
     Fax:	302-366-5236