The mission of the United States Naval Academy SAPR GUIDE program is to provide guidance, understanding, information, direction and education needed for individuals seeking support regarding incidents concerning equal opportunity, sexual harassment or sexual assault. We work to ensure that all midshipmen are treated with dignity and respect by acting as a trusted peer resource throughout the Brigade. GUIDEs are recognized SAPR resources and sources of support responsible for outreach and education. GUIDEs are equipped through Fleet-relevant training to be sensitive and capable SAPR allies.
The SHAPE program's goal is to foster an environment of dignity and respect, and assist in the development of future officers who will promote and practice proactive leadership to prevent sexual violence.   The program approaches the subject-matter as both a cultural issue, deconstructing myths and accepted behaviors and a leadership issue, providing practical tools to intervene proactively. The curriculum incorporates midshipmen feedback and integrates the unique cultures of USNA, the Navy and the Marine Corps. SHAPE supports the Sexual Assault Response program by reinforcing victim response and support for victims.
USNA SAPRO’s Training and Education programming, through evidenced-based and best-practice techniques of training and education, aims to help foster a culture in which any act of sexual assault or sexual harassment among USNA midshipmen, faculty and staff is not welcome and not tolerated. The programming also aims to train and educate personnel in the institution, in collaboration with USNA SAPRO’s response personnel, about issues regarding survivors and response so that survivors can be optimally supported, advocated for and cared for. We adhere strongly to the research-supported best practice of bystander intervention to encourage proactive leadership and community to prevent sexual assault and harassment.


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The First 60 Minutes Of Need For Speed: Heat
by Paul TamayoI streamed Need For Speed: Heat on Kotaku’s Twitch channel earlier today and got to see the opening moments of the game. Check out the stream archive below and follow Kotaku US on Twitch for more streams.


KISS 💋 News Blog Install KISS Team Guidestones FAQ
Community (NEW) Scrots Package System Testimonials Style
Ports (non-x86-64) Software Contact Donate GitHub ->
+-------------------------------------------------------------------+
| |
| FREQUENTLY ASKED QUESTIONS |
| |
+-------------------------------------------------------------------+
* Why should I use KISS?
That's up to you to decide.
I'm not going to try and sell you something I'm giving away for
free. Have a read of the website and its wealth of information,
try KISS in a virtual machine (a chroot is also an option) and
come to your own conclusions.
* What does KISS mean?
KISS is an acronym for "Keep it simple stupid" (notice _no_ comma).
Stupid does not refer to the user. To quote Wikipedia:
> The principle is best exemplified by the story of Johnson handing
a team of design engineers a handful of tools, with the challenge
that the jet aircraft they were designing must be repairable by
an average mechanic in the field under combat conditions with
only these tools.
> Hence, the "stupid" refers to the relationship between the way
things break and the sophistication available to repair them.
* Is KISS rolling release or stable?
Packages are updated as quickly as possible while at the same time
tested to ensure that no issues arise in the new version. A new
version of a package will be held back if it causes issues.
When a package update brings in a bug during the build process or
at runtime, it will be patched in KISS until it is fixed upstream.
This will typically happen hours after the version is available
and doesn't cause a delay.
Nothing prevents you as a user from choosing the update schedule
you would prefer. Total control is in your hands. KISS merely
provides you with an always up-to-date repository pool.
* Something something BUS factor of ONE.
Every user of the distribution owns their system in its entirety.
The management of the distribution which extends to the management
of updates to the user's system is entirely optional.
All KISS systems contain the full sources for the distribution and
each user has the means of managing and maintaining their machine
on their lonesome (or by forming a collective and secondary
"upstream").
This includes:
- The repositories with full git history.
The repository updates are simply a 'git pull' which results in
each user having a full copy of the git repositories on their
machine at /var/db/kiss/.
- The package manager and the kiss-* scripts.
As these are all simple shell scripts, the installed "binaries"
_are_ the unchanged source code. All of these are self-contained
and separate programs.
The 'kiss-outdated' script allows one to check their system for
outdated packages against the repology.org database independent
of whether or not repology has support for KISS itself.
The 'kiss-export' script allows one to convert an installed
package back into a redistributable and installable binary
tarball. Simply run 'kiss-export pkg_name' and a tarball will be
created in the current directory.
- The distribution's documentation.
As the website sources are merely plain .txt files, the docs are
available in /usr/share/doc/kiss and are readable in less,
vim or the program of your choosing.
- The init scripts.
In addition to the reasoning given for the package manager above,
KISS has no lock-in regarding how the machine boots. Were the
init scripts to disappear, there would be no real loss.
To continue further, the init scripts need not change. There is
no need to update them. Any extensions to them can be made via
the /etc/rc.d directory or the /etc/rc.conf file.
What all of this means is that were the GitHub or website to go
down, there would be no loss in code, documentation or the
distribution itself.
It also extends to each user having everything they will need to
continue the distribution for themselves or for other users. A
change of git remote is all that is needed in the latter case.
KISS was designed to be maintainable by a single person. The
repositories are kept small and focused. There is no requirement
or _need_ for infrastructure of any kind.
A user today could choose to go their own way with their system.
Everything they need is already in the existing system. The power
is in your hands. You are free.
+-------------------------------------------------------------------+
| |
| SOFTWARE |
| |
+-------------------------------------------------------------------+
* Why isn't SOFTWARE packaged?
No one has packaged it yet.
* Can you package SOFTWARE for me?
Do it yourself.
* What init does KISS use?
KISS uses busybox init by default. There is however, no lock-in and
the user may use whatever they like. The distribution's boot up
scripts are written in an init-agnostic way and act as a base for
every init to use.
* What coreutils does KISS use?
KISS uses busybox coreutils by default. There is however, no
lock-in and the user may use whatever they like. All shell code is
written in portable POSIX shell and coreutils invocations.
* One exception is made for 'sed -i' which isn't POSIX but is far
too useful to do away with. Many sed implementations include -i
so this is more of a non-issue.
* The tar command has no standard interface, however support is
included for busybox tar, libarchive tar (bsdtar), GNU tar and
others.
* Some utilities have no standard specification and where their use
is required, we adhere to common options between implementations.
Barring the above exceptions, users are running KISS without the
use of busybox. Popular alternatives are Suckless' sbase/ubase and
GNU's set of core utilities.
+-------------------------------------------------------------------+
| |
| PACKAGE MANAGEMENT |
| |
+-------------------------------------------------------------------+
* How do I remove a package and all of its dependencies?
Short answer: With one command? You can't.
The package manager does not do recursive dependency removal on
removal of a package. This error-prone automation will not be
added to the package manager.
Instead, the workflow is to remove the single package and then look
at the output of the 'kiss-orphans' command to see what can now be
removed. This command will list all packages which have no
relationship with other packages, otherwise known as orphans.
This list may include Firefox and other "end" software so a brain
is required when parsing the list. You'll come to learn the
relationships between packages and their dependencies and this will
eventually become effortless.
+-------------------------------------------------------------------+
| |
| KERNEL |
| |
+-------------------------------------------------------------------+
* Why doesn't my kernel boot?
The kernel not booting can be a variety of issues. This is almost
always related to a configuration issue in the kernel, /etc/fstab
or the bootloader.
KISS doesn't use an initramfs so the configuration of the kernel
may have different requirements to other distributions.
1. The drivers for your disk controller, drives and filesystems
must not be built as kernel modules. They should be set to =y
in your .config or [*] when using make menuconfig.
Essentially, every driver the kernel requires to detect and
mount the drive containing the root filesystem, must be built
as a part of the kernel binary.
2. Multi-drive systems must use PARTUUID or UUID in place of
/dev/sdXX in the bootloader configuration to ensure that the
kernel will find the right drive.
* Why doesn't KISS support initramfs?
KISS technically supports booting via an initramfs, it just doesn't
require or provide one. As a user you have the means to set this up
yourself for your system.
Full disk encryption is also possible without the use of an
initramfs in modern kernels (see dm-mod.create).
The initramfs concept is an ugly, complicated and largely optional
mess. Thank god it isn't a requirement.
* Why must I compile my own kernel?
The kernel must be compiled by the user for a variety of reasons.
1. The user maintains full control over all aspects of their kernel
and further, their entire system. There is no lock-in into a
specific set of kernel sources, version, use of proprietary
firmware, patches, config, etc.
The user decides:
- Which kernel to use (sources and version).
- When to update their kernel.
- Whether to use proprietary firmware blobs.
- Which patches to use (if any).
- How the kernel is configured (endless options).
- How many kernels they'd like to keep.
- Compiler options (-O3, -march=native, etc).
You as a user might actually learn something too. You may come
to understand your hardware, what drivers it needs, how the
kernel works from a configuration perspective, etc.
You should have a better understanding of this part of the
system afterwards and you'll be able fix any issues at this
level with relative ease.
Remember, it's only hard the first time. Once a working config
is created, no further work should need to be done each time you
update your kernel.
If the new kernel version has an issue with your hardware,
simply boot another from your backups. If new hardware was
added, simply run 'make menuconfig' and add what is needed.
I'll say what I always say. Nothing prevents anyone from
providing kernel binaries and an initramfs generation tool for
KISS. Just don't wait on the BDFL to do it for you.
2. Eases distribution maintenance for the BDFL. Shipping a generic
kernel demands a humongous, module heavy kernel with support for
everything under the sun. An initramfs is then a requirement to
boot this damn monstrosity.
There is then a need for a _portable_ initramfs generation tool
which to my knowledge doesn't exist. There can't be a dependence
on bash or anything outside of core. All initramfs tools are
either distribution-specific or non-portable to KISS.
Then there'd be endless support requests for tweaks, additions
and removals to the distribution config and the burden of
updating the kernel on every release.
Does KISS ship the latest kernel? The long term support kernel?
Both? Some users require firmware so we'd need two separate
binaries, one for linux-libre and another for regular linux.
What if the latest kernel has issues on some hardware? New
builds and binaries must be released with backported patches.
It's a large maintenance burden for something which can simply
be solved by the user doing this themselves. The user maintains
full control over every aspect of their kernel and is solely
responsible for it.
* Why doesn't KISS provide kernel sources as a package?
Why should it? See above.
* Must I keep the linux-headers package in sync with my kernel's
version to maintain a working system?
The kernel headers in KISS are pinned to an LTS kernel version
and are only updated when there are changes of interest in the
kernel or headers themselves (usually by users requesting new
features available in the newer headers).
The headers are backwards compatible and are fully usable with a
matching or NEWER kernel version. There are two cases where you'd
be required to update the headers yourself.
1. You'd like to run a kernel _older_ than the default headers.
2. You'd like to make use of features in your _newer_ kernel version.
Here's an excerpt from the kernel's documentation which may
explain the situation better than I.
> Kernel headers are backwards compatible, but not forwards
compatible. This means that a program built against a C library
using older kernel headers should run on a newer kernel (although
it may not have access to new features), but a program built
against newer kernel headers may not work on an older kernel.
https://www.kernel.org/doc/Documentation/kbuild/headers_install.txt
+-------------------------------------------------------------------+
| |
| GCC |
| |
+-------------------------------------------------------------------+
* C compiler cannot create executables.
This is almost always an error in your CFLAGS/CXXFLAGS. Ensure that
you have used -ONUM (CAPITAL O) and not (lowercase o) or (zero 0).
If the above doesn't fix the issue, try building the package with
CFLAGS= CXXFLAGS= LDFLAGS= kiss b pkg. You'll then be able to
discern whether or not this was the issue.
+-------------------------------------------------------------------+
| |
| XORG |
| |
+-------------------------------------------------------------------+
* Why can't I start Xorg as a normal user?
This is likely due to you running 'modprobe amdgpu' or equivalent
post (or late) boot which causes udev to miss setting up the GPU
device in /dev/dri.
There are three separate solutions ranging from easy to hard.
Either of the three will solve the issue.
1. Start the udevd service.
This will keep udevd alive and ensure that it does its thing
when it detects that the GPU driver has loaded.
Enabling the service also turns on device hotplug and other
features. This isn't the default as there's no need for the
daemon to run on a static system.
$ ln -s /etc/sv/udevd/ /var/service
2. Run 'modprobe' earlier in the boot process.
Running 'modprobe' in your inittab or in rc.d will result in it
executing _after_ udevd has finished setting up devices.
By moving 'modprobe' to /etc/rc.conf, it will load the driver
before udevd executes which will allow it to detect and setup
the device.
3. Compile the GPU driver and firmware into the kernel.
This is trickier but the best solution overall. Instead of using
modules, compile the firmware into the kernel.
You'll then not have to worry about udevd/modprobe as the kernel
will automatically load the driver as early as possible.
This may include compiling proprietary firmware into the kernel
as well which is finicky but doable.
* How do I take a screenshot?
Most users will typically use 'ffmpeg' as their screenshot tool as
they'll already have ffmpeg installed for MPV or Firefox. Other
options include 'imlib2' or 'imagemagick' (import).
If you'd like to use another tool, package it yourself. Remember,
if no one has packaged something it means that no one has needed
said package. It is up to you.
The registered trademark Linux(R) is used pursuant to a
sublicense from the Linux Foundation, the exclusive
licensee of Linus Torvalds, owner of the mark on a
worldwide basis.
(C) Dylan Araps 2019-2020
View page source


Home Page
Purpose Satement
Everyone strives to be more productive, but most people don’t know how or where to start. This website will provide them the structure they need to be more productive and ensure that productivity is sustainable and motivating as well.
We want users to be informed about the benefits of intermittent study sessions, by providing users scientific literature for their knowledge, and allow users to implement the technique through the in-built tools, such as the timer, and customized study sessions. Allowing users to track their sessions will motivate long-term progress as well.
By providing these tools and information, we are allowing users to achieve their goals, whether that be in their studies, careers, or personal life.
Mission Statement
There is no financial goal with this website; what the website is to be is a tool to allow people to flourish and be at their best and achieve their goals, whether that be personal or career goals.
To enhance discipline is to enhance one’s life. As one achieves more goals and is disciplined in doing so, more of their time is freed up for other important aspects of their lives, while also allowing them to reach their potential.
By helping individuals achieve more of what they want, the benefits will radiate outwards to those around them. Humans are networked individuals and to help one is to help many.
Audience Statement
The main target demographic of this website would be mainly for students of all ages and demographics but, with the customized nature of it, the audience can be anyone who is looking to increase and track their productivity, while also being sustainable as well.
As a student myself and being surrounded by others, students can lack structure and discipline. The website will allow people to give structure to the user’s study/productivity sessions and allowing for sustainable, long-term productivity.
Our demographic will mainly access this via the web app. A mobile application may be implemented at a later date, but a web application allows for universal access across all devices.


|
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| PREV LETTER NEXT LETTER | FRAMES NO FRAMES |
Direction.OUT
: equivalent to outDegree(node)
Direction.IN
: equivalent to inDegree(node)
Direction.EITHER
: equivalent to degree(node)
InspectableGraph.degree(Node)
, i.e. undirected.
InspectableGraph.degree(Node, Direction)
.
Edge
sbetween two Nodes
should have.
|
||||||||||
| PREV LETTER NEXT LETTER | FRAMES NO FRAMES |


Getting Started This unit is to be finished prior to the first day of class (April 25, 2016)
Participant survey Tell us about yourself and why you are taking this course
Pre-course assessment This ungraded assessment will be used to assess course impact.
Participant Post-survey Provide feedback on the course
Post-course assessment This ungraded assessment will be used to assess course impact.


| Interface | Description |
|---|---|
| CompositeEvaluation<T> |
Created by Mike Sobolewski
|
| EvaluationComponent |
Created by Mike Sobolewski on 5/14/16.
|
| Filtration<T> | |
| Getter<T> |
A getter is a generic agent that returns a value calculated in many different
ways.
|
| Model |
An top-level common interface for all service Models in SORCER.
|
| Modeling | |
| ModelingTask | |
| Reference<T> |
A referencing functionality required by context models.
|
| SupportComponent |
Created by Mike Sobolewski on 5/18/16.
|
| Variability<T> |
A variable has id, name and value.
|
| VariabilityModeling |
| Enum | Description |
|---|---|
| Modeling.ParType | |
| Modeling.Type | |
| Variability.MathType | |
| Variability.Type |
Types allow for grouping in the functional model so all variables of a
particular fiType can be retrieved as a functional collection, e.g.,
DESIGN, RESPONSE, DERIVATIVE or GRADIENT.
|
| Exception | Description |
|---|---|
| FilterException | |
| GetterException |


How It Works
SIGN-UP
The first stage is all about
you — the customer. In the very first meeting itself, we
encourage our customers to talk about themselves, the
things that move them, the things that their kids find
excitable, etc. Knowing all this helps our designers to
create a first-cut design presentation that best
resonates with our clients’ likings & sensibilities.
DESIGN
The design phase involves
sitting with your designer through multiple rounds of
design iteration (functional and aesthetic). Your
designer will then design your home for tomorrow by
ensuring it is future-proof and will stand the test of
time.
IMPLEMENTATION
A Project manager and his
team of Implementation experts will be assigned to you
to implement the interiors of your home. Everything will
be delivered to your doorstep, installed, assembled, and
is built to last, to help transform your design dream
into reality.
WARRANTY
Furdo gives you a 5 year
warranty on ‘woodwork’ because we understand how
important it is for your ‘dream home’ to stand the mild
ravages of time. Our materials are therefore sourced
from the very best, giving you peace of mind, and a home
that you can proudly call your own.


oh-my-phd
I am doing my PhD study at ETH Zurich. I try to document what I have been learning here in this Github repo.
Table of Contents
Oh My PhD!
Hi Welcome to Oh My PhD! I will document some interesting and useful things I learn during my PhD study at ETH here.
Writings
orgmode-latex-templates
My org-mode templates for latex-style writing This repo contains different templates as a starter code to write org-mode which can later be exported to latex and pdf.
Coding
programming notes
My programming related notes
oh-my-opengl
My OpenGL practice
Learning
courses at ETH
math foundations
Some math resources I find it very useful online.
optimisation
(discrete) differential geometry
structural mechanics
Time management
phd-org-mode
My phd orgmode to organize my scheduling. Based on this template.


--- layout: post status: publish published: true title: 'Reasons I Could Never Be Mainstream: Separation' author: Ryan Hayden wordpress_id: 844 wordpress_url: http://ryan-hayden.com/?p=844 date: '2013-03-06 06:00:50 -0600' date_gmt: '2013-03-06 12:00:50 -0600' categories: - Fundamentalism - ministry tags: [] ---
[box]
This week, I'm posting the beliefs that I hold that, even if I wanted to be a mainstream evangelical, would keep me from being accepted. These aren't so much the distinctives of being an Independent Fundamental Baptist. But they're close. You can read the introduction here.
[/box]
Ecclesiastical separation, the separation of individuals and churches from professing Christians who espouse false doctrine, is very clearly commanded in the New Testament. Romans 16:17 says:
“Now I beseech you, brethren, mark them which cause divisions and offences contrary to the doctrine which ye have learned; and avoid them.”
“Now we command you, brethren, in the name of our Lord Jesus Christ, that ye withdraw yourselves from every brother that walketh disorderly, and not after the tradition which he received of us.”
“If there come any unto you, and bring not this doctrine, receive him not into your house, neither bid him God speed: For he that biddeth him God speed is partaker of his evil deeds.”
[box]
You can read a past article about separation here.
[/box]
I also believe in personal separation, that Christians are supposed to be separate from the world.
1 Corinthians 6:14 says
“Be ye not unequally yoked together with unbelievers:for what fellowship hath righteousness with unrighteousness? and what communion hath light with darkness?”
“Wherefore come out from among them, and be ye separate, saith the Lord, and touch not the unclean thing; and I will receive you, And will be a Father unto you, and ye shall be my sons and daughters, saith the Lord Almighty.”
These doctrines (particularly the first one) are what make fundamentalists fundamentalists (the term is flawed, but it’s what has been given to us.) The vast majority of those outside of fundamentalism are not consistently practicing any kind of ecclesiastical separation. There are a few who do it sporadicly, but they do not do it consistently.
Also, the majority of those outside of fundamentalism have all but ignored the idea of personal separation from the world. While I personally take issue with the way “standards of separation” have been emphasized in independent Baptist churches, I don’t think we need to swing to the other side of the pendulum and just about ignore the concept of personal separation altogether.
Tomorrow, I'll be writing about my music position.


--- layout: page title: About header-img: "img/about-bg.png" ---
This Page is the full journey of becoming the Full Stack Developer I want to become from start to finish. The ins & out's of my Grind. Giving my advice to the world of what I'm learning here at Learn.co.


Since there is so little information on 2-Aminoindan I decided to take it for a test drive. I was able to obtain a sufficient quantity for experimentation from a source believed to be reliable. The following reports are a bit truncated and represent only a few of several experiments with different dosages and methods of administration. This report represents one of the more thought out and carefully executed series of experiements I have attempted.
Set and setting were always generally the same, at home with time to kill and no responsibilities. Feeling good about my position in life and taking the substance with as few expectations as possible.
With 5mgs insufflated: Nothing note worthy, perhaps a mild stimulation immediately after administration, could just be psychosomatic or nerves. No effects noticable at 20 minutes.
10mgs Insufflated: Mild stimulation, again perhaps just a nervous response, don't feel much of anything after 20-30 minutes.
15mgs Insufflated: I'm willing to say that at this level the is an unmistakable 'something' of a mild stimulation that is probably not nerves, but these effects fade quickly and I am baseline within an hour or so. We're talking minimal, caffeine-like stimulation, though smoother than that chemical, and so almost not present unless I pay attention. I noted that communication was a little weird, not difficult but, just that I was aware that I was altered when I was trying to communicate, though this was not apparent to my partner upon questioning her about it.
15mgs. Insufflated (day after last experiment): No effects outside of anticipation, tolerance?
20mgs. Insufflated: Not much different from 15mgs. Mild caffeine-like stimulation fading fairly quickly from perceptable levels.
25mgs. Insufflated: This is the last time I tried this administration method, the chemical burns like methamphetamine in the sinuses, often bringing a tear to my eye immediately. the 25mg. level seems to not produce much more effect than the 15mg level. This seems a weird dose/response curve to me. Perhaps again this is tolerance? Not sure, have tried to space out experiments by several days to a week each time but perhaps thats not enough.
20mgs. Oral: No tangible effects (although this was taken two hours after a meal).
25mgs. Oral: See above, same circumstances, no effect.
25mgs. Oral, Empty stomach: No tangible effects. (perhaps orally inactive??)
35mgs. Oral: Getting somewhere, a mild caffeine like stimulation, smooth fade in and a relatively quick decline (2 hours total experience from first alert to undetectable)
50mgs. Oral, Empty stomach: Getting a little tired of playing with this substance so I jumped up the dose a little more aggressively with positive results. Defined stimulation, strange, somewhere between methamphetamine and caffeine. Displays qualities of both, perhaps a little like ephedrine. Some cold chills at onset, especially on my scalp, and seem to multiply with physical activity. Thinking is a little scattered like with a good dose of caffeine, but smoother overall, no jitters or palpitations. Relatively quick come-up of about 30 minutes. No euphoria. Very slight visual distortion, no motion but just soft edges, notice colors, VERY mild, perhaps even just a projection on my part. Effects last for 2-3 hours from alert to undetectable, short.
COMCLUSIONS:
Even though I had read of a relatively small dose to produce effects, I found that even at two to three times that level effects were mild. The over-all sensation is pleasant, with no noted side effects. A smooth and mild stimulation. I feel like for someone my size, a dose of 100mgs would be about perfect. I have not taken the experiments any farther than 50mgs, though, and so would still recommend caution as this is a relatively unknown substance and long term or high dose side effects are also unknown.
I would like to add that there may be some addiction potential with this substance, my experiments that were close together produced little or no effects and the instinct to take a larger dose each time seemed apparent and was beyond just my curiosity.
I'm only half way through my supply of this substance. Seems like it could be fun for a club given its mild nature and short term activity. I also think that oral administration is the way to go, no burn, more pleasant effects overall, seems less potent this way but more interesting.
Experience Reports are the writings and opinions of the individual authors who submit them.
Some of the activities described are dangerous and/or illegal and none are recommended by Erowid.


First Arab astronaut urges climate protection on Earth
by AFPDUBAI: Wearing a blue space suit with a United Arab Emirates (UAE) flag on one sleeve and a spaceship on the other, the first Emirati astronaut said on Tuesday his mission highlighted a crucial issue – climate change.
Witnessing Earth and its beauty from space made him realise the importance of preserving it, said Hazzaa al-Mansoori, a 35-year-old former military pilot, who reached the International Space Station in September.
“We have to appreciate the planet and make sure that we save it for the next generations,” said the father-of-four, urging efforts to address “the reasons behind climate change”.
Mansoori became the first Arab astronaut to visit the ISS and returned home to a hero’s welcome after an eight-day mission, during which he participated in scientific experiments including a time-perception study.
His space trip has become a source of great pride in the UAE, a newcomer with ambitions to send an unmanned probe to orbit Mars by 2021.
In a Dubai hall swarming with journalists, Mansoori recounted the magic of his trip.
“When you see our planet from space… it’s really something amazing and a spectacular view,” he said. “I spent a lot of time looking out that window, I didn’t even want to sleep.”
Mansoori said living in space showed him that life on Earth was “a blessing”.
“It is really difficult to live in space, we have to provide a lot of oxygen, air and food… and we have all of this here for free,” he said.
The five-year period ending 2019 is set to be the hottest ever, said a UN report published in September, scientists’ latest grim reminder that climate change is already a reality.
Last month was the hottest October ever recorded worldwide, according to data released by the European Union’s satellite monitoring service.
“We are lucky to live here — let us protect the Earth and its atmosphere,” said Mansoori.
On board the ISS, Mansoori, who was selected from more than 4,000 UAE candidates, donned Emirati dress and treated crew members to his country’s snacks.
He flew to the station after the UAE signed a contract with Russian space agency Roscosmos to make him a “spaceflight participant”, a term used for people from outside the main space agencies who take short trips to the ISS.
As part of its space plans, the UAE has announced its aim to become the first Arab country to send an unmanned probe to orbit Mars by 2021, naming it “Hope”.
“It’s the golden age for space exploration in the UAE,” said Salem al-Marri, of the UAE’s Mohammed Bin Rashid Space Centre.


Trying to get a Job
Customer
Account
Info
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Create an Account
Customer ID:
Account Type:
Checking
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Account nickname:
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Balance:
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submit


Solving Problems By Searching
Node Expansion
To search through a graph, a Search Agent needs to expand nodes. The nodes which can be expanded by the Agent together forms the frontier. Expanding a node refers to marking the node as 'expanded' or 'visited' and adding its immediate neighbors to the frontier.
In the following diagram, 'A' is the initial node and belongs to the frontier. Click on the frontier nodes to see how they are expanded.
- Expanded Nodes
- Frontier Nodes
- Unexplored/Unknown Nodes
Frontier Nodes
Getting in the shoes of a Search Agent
Let's see the prespective of a Search Agent as it searches through the graph.
Remember, the Agent can only see the nodes which are either expanded or currently present in
the frontier.
Now that we know how nodes are expanded and how a search Agent perceives the graph, we are now ready to meet some Search Algorithms. In the above two visualizations, you decided which node to expand next(by clicking on them) but now we shall see some algorithms in action which can decide by themselves which node to expand next.
Breadth First Search
In Breadth First Search, the node which was discovered the earliest is expanded next i.e. the node which joined the frontier earlier, is expanded earlier.
To achieve this, Breadth First Search Algorithm uses a FIFO(First In First Out) Queue. The following graph is explored by a Breadth First Search Algorithm with 'A' as the initial node.
- Nodes waiting to be expanded
- Node to be expanded next from the queue
FIFO QUEUE
Depth First Search
In Depth First Search, the node which was discovered the latest is expanded next i.e. the node which joined the frontier later, is expanded later.
To achieve this, Depth First Search Algorithm uses a LIFO(Last In First Out) Queue. The following graph is explored by a Depth First Search Algorithm with 'A' as the initial node.
- Nodes waiting to be expanded
- Node to be expanded next from the queue
LIFO QUEUE
Uniform Cost Search
Click on the canvas to restart the simulation
Depth Limited Search
Click on the canvas to restart the simulation
Iterative Deepening Search
Click on the canvas to restart the simulation
A Star Search
Click on the canvas to restart the simulation


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[gecode-users] Extending Integer Variable
Christian Schulte
cschulte at kth.se
Mon Apr 7 21:56:12 CEST 2008
Yes, you are right. Also IntVars with unlimited precision using gmp... All
doable, the only thing is that we sort out which occurrence of "int" refers
to variables and which refer to other occurrences.
The problem with 64 bit integers, though is that we have to have a datatype
that is bigger than the data type of variables. Now we use double for that
purpose. With 64 bit integers we might resort to 128 bit integers...
If you implement FloatVars, please keep us posted. If you want to contribute
to Gecode we would be more than happy to help!
All the best
Christian
--
Christian Schulte, www.ict.kth.se/~cschulte/
-----Original Message-----
From: users-bounces at gecode.org [mailto:users-bounces at gecode.org] On Behalf
Of Filip Konvicka
Sent: Monday, April 07, 2008 8:22 AM
To: users at gecode.org
Subject: Re: [gecode-users] Extending Integer Variable
> let me first elaborate a little bit how Gecode's variables and propagators
> are structured before I actually attempt to give an answer.
...
> What do you want to do? Do you think it me be generally useful to be
> included in Gecode anyway?
Hi,
I am currently thinking of implementing floating-point-based variables
(maybe based on Boost.Interval), so thanks for the summary :-)
Anyway, before I decided that I need floating-point numbers, I was
thinking why you can't have "long long"-based variables when you already
have IntVars. On 64bit platforms, long long might be an interesting
choice IMO. But I realize that if that should be a part of Gecode, it
would probably mean adding template parameters to many classes/functions
and maybe it would make things less understandable. But then, if it's
only restricted to integral types, who knows.... the point is that
there's a lot of work done for the IntVars, so it'd be great to reuse it.
Regards,
Filip
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Execution-Executor-Procedure ODP: Description back to ToC
Figure 1 presents an overview of the classes and the properties included in the EEP ODP.
Figure 1. - General overview of the EEP ODP.
Competency Questions back to ToC
This section shows the specification of the EEP ODP requirements in the form of Competency Questions:
- CQ1: What are the observations/actuations performed by a given procedure?
- CQ2: What are the observations/actuations performed by a given sensor/actuator?
- CQ3: What are the procedures implemented by a given sensor/actuator?
- CQ4: What are the features of interest on a given observation/actuation?
- CQ5: What are the properties/qualities sensed/actuated by a given observations/actuations?
- CQ6: What are the features of interest of a given sensor/actuator?
- CQ7: What are the properties/qualities sensed/actuated by a given executor?
Instantiating the EEP ODP back to ToC
This section shows an example of instantiating the EEP ODP. Let us consider a farm scenario where poultry are reared. In this case, a sensor (:sensor36) deployed in the farm (:farm) is in charge of measuring both farm's temperature and humidity (i.e.,
:farmTemperature and :farmHumidity). Furthermore, this sensor implements a
monitoring procedure (:monitoringProc) to make two observations :obs13 and
:obs14. Figure 2 represents this example (for the sake of simplicity, the rdf:type relationships are not shown)
Figure 2. - A farm scenario description example.


{% extends 'base.html' %} {% block content %}
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Hey Gamer welcome to our world where you can explore yor favourite games by ; watching trailers and download them for free.Just scroll down and pik a category of the games you like and explore.


礦ision Build Log
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Rebuild target '智能温湿度计' compiling main.c... compiling User_ADC.c... compiling User_I2C.c... compiling User_Key.c... compiling User_OLED.c... compiling User_SHT30.c... compiling User_Uart.c... compiling User_Timer.c... compiling codetab.c... compiling mcu_api.c... mcu_sdk\mcu_api.c(575): warning C280: 'len': unreferenced local variable mcu_sdk\mcu_api.c(587): warning C280: 'len': unreferenced local variable mcu_sdk\mcu_api.c(599): warning C280: 'len': unreferenced local variable compiling protocol.c... compiling system.c... mcu_sdk\system.c(407): warning C280: 'result': unreferenced local variable linking...  ".\Obj\智能温湿度计" - 0 Error(s), 20 Warning(s). Build Time Elapsed: 00:00:02


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Text source name = 11604102.txt
[12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to
Text source name = 11604102.txt
should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16]. Prrp binds
Text source name = 11604102.txt
estes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family an
Text source name = 11604102.txt
eated here, should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16].
Text source name = 11604102.txt
with polyribosomes in mouse testes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the funct
Text source name = 11604102.txt
ZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to understanding the mechan
Text source name = 11532192.txt
he insertion allele. The other allele was identical to the published sequence [22]. The insertion allele also had a previously reported substitution (A to G, Thr164Ala) in exon 1 and several other single base changes in the promoter region [22]. The presence or absence of this allele does not associa
Text source name = 11319941.txt
y down-regulate synaptic activity in the striatum [45]. Another gene that falls into the Bsc10a interval is Macs, the gene encoding the myristoylated alanine-rich C kinase substrate (MARCKS protein). This molecule is important in cerebral development. MARCKS-deficient mice have a high incidence of exenceph
Text source name = 11604102.txt
e proteins were mixed with Freund's adjuvant and injected into rabbits to generate the anti-DAZAPl-C antibody. An oligopeptide containing the last 19 ammo acid residues of the mouse DAZAP1 was synthesized in vitro using the services of Bethyl Laboratories (Montgomery, TX). The peptide was conjugated to KLH as carrier
Text source name = 11604102.txt
reading frame and results in a shorter Prrp with a different C-terminal end sequence. Figure 1 Evolutionary conservation of the DAZAP1 proteins. The amino acid sequences of the human and mouse DAZAP1s and the Xenopus Prrp are compared. The two RNA binding domains are boxed. Differences between the human and
Text source name = 11597317.txt
ein are, like many cancer cells, genetically unstable and accumulate gross chromosomal rearrangements [5,6]. The sequence of this large protein (3418 amino acids) offers very little clue to its function, although there are eight repeated segments (termed BRC repeats) in the middle of the protein that are highl
Text source name = 11604102.txt
nant protein containing the C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against an oligopeptide containing the last 19 amino acid residue at the C-terminus. Both antibodies recognized in vitro synthesized DAZAP1 in an immunoprecipitation assay (data not shown). Western blotting of mouse
Text source name = 11597317.txt
bination in the human pancreatic tumor cell line CAPAN-1. CAPAN-1 cells carry a deletion of BRCA2 on one homolog and codes for a protein truncated at amino acid 1981 on the other homolog. The authors indicate that the level of I-SceI-induced recombination in CAPAN-1 is over 100-fold less than that seen using
Text source name = 11604102.txt
was isolated by 5' RACE [15]. The near fall length cDNA consists of a 53 bp 5' untranslated region (UTR), an open reading frame for a protein of 405 amino acid residues, and a 362 bp 3' UTR (GenBank Accession No: AF225910). The coding region shares 89% similarity with that of the human orthologue. The 3' UTR sequence
Text source name = 11604102.txt
BDS) and a C-terminal portion that is rich in proline (Figure 1). It is highly conserved evolutionarily. The mouse and the human proteins differ in 9 amino acids only, with 7 substitutions and two deletions/insertions. The mammalian proteins shares 89% similarity and 81% identity with Xenopus Prrp (for proline
Text source name = 11532192.txt
tely thirty 3 to 4 month old mice were analyzed at each dose and time B-D. Scatter-plots demonstrating no change in IOP over the 12 minutes following anesthetic administration in three relatively unrelated mouse strains. All strains consisted of males and females that ranged from 3 to 6 months of age. The sex
Text source name = 11532192.txt
me). The greater the dose the greater the decrease in IOP. At the 5 minute measurement, however, IOP was the same using all doses suggesting that the anesthetic effect on IOP had not yet occurred. To identify any early window when it may be possible to assess IOP without an obvious anesthetic effect, 195 mice
Text source name = 11532192.txt
tic dose within 12 minutes of administration. A Mean IOP ± SD is shown for C57BL/6J mice at 5 and 25 minutes after administration of various doses of anesthetic. The 1X dose consisted of 99 mg/kg ketamine and 9 mg/kg xylazine. All doses decreased IOP by 25 minutes. At all doses the IOP at 5 minutes was the sa
Text source name = 11532192.txt
he mice to terpenes. Terpene administration or environmental exposure to terpenes in wood shavings alters drug resistance and decreases the effect of anesthetic agents in both rats and mice [40-45]. Risk factors for increased IOP Some epidemiological studies implicate factors such as diabetes, vascular hypertensio
Text source name = 11532192.txt
do not immediately identify them as likely glaucoma candidates. No effect of anesthetic protocol on IOP during a 12 minute measurement window Many anesthetic agents including xylazine lower IOP. Ketamine usually appears to increase IOP [31-33], but there are reports of ketamine having no effect on IOP or even red
Text source name = 11532192.txt
ronments may contribute to these differences. The relationship between the IOPs we measure and those in conscious mice depends upon the effect of our anesthetic protocol (intraperitoneal injection of 99 mg/kg ketamine and 9 mg/kg xylazine). An anesthetic effect could potentially alter IOP and mask genetically
Text source name = 11532192.txt
s (Figure 6). In these experiments, IOP measurements were made between 5 and 12 minutes after administration of anesthesia. Figure 5 No effect of 1X anesthetic dose within 12 minutes of administration. A Mean IOP ± SD is shown for C57BL/6J mice at 5 and 25 minutes after administration of various doses of ane
Text source name = 11532192.txt
those in conscious mice depends upon the effect of our anesthetic protocol (intraperitoneal injection of 99 mg/kg ketamine and 9 mg/kg xylazine). An anesthetic effect could potentially alter IOP and mask genetically determined differences in IOP. Therefore, it is very important to understand this effect. Her
Text source name = 11532192.txt
minutes after administration. Thus, to avoid effects of anesthesia on IOP, all measurements should be made within a window of up to 12 minutes after anesthetic administration. Similarities and differences to rat studies Our results agree with the time course of cardiovascular depression caused by intraperi
Text source name = 11532192.txt
own functions do not suggest that they affect aqueous humor dynamics or do not immediately identify them as likely glaucoma candidates. No effect of anesthetic protocol on IOP during a 12 minute measurement window Many anesthetic agents including xylazine lower IOP. Ketamine usually appears to increase IOP
Text source name = 11604102.txt
12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to understan
Text source name = 11604102.txt
BMC Genomics. 2001; 2: 6. PubMed ID: 11604102 Copyright © 2001 Dai et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. -------------------------------------------------------------------- Characterization of the mouse Dazap1 gene encoding an RNA-binding protein that interacts with infertility factors DAZ and DAZL Abstract Background DAZAP1 (DAZ Associated Protein 1) was originally identified by a yeast two-hybrid system through its interaction with a putative male infertility factor, DAZ (Deleted in Azoospermia). In vitro, DAZAP1 interacts with both the Y chromosome-encoded DAZ and an autosome-encoded DAZ-like protein, DAZL. DAZAP1 contains two RNA-binding domains (RBDs) and a proline-rich C-terminal portion, and is expressed most abundantly in the testis. To understand the biological function of DAZAP1 and the significance of its interaction with DAZ and DAZL, we isolated and characterized the mouse Dazap1 gene, and studied its expression and the subcellular localization of its protein product. Results The human and mouse genes have similar genomic structures and map to syntenic chromosomal regions. The mouse and human DAZAP1 proteins share 98% identity and their sequences are highly similar to the Xenopus orthologue Prrp, especially in the RBDs. Dazap1 is expressed throughout testis development. Western blot detects a single 45 kD DAZAP1 protein that is most abundant in the testis. Although a majority of DAZAP1 is present in the cytoplasmic fraction, they are not associated with polyribosomes. Conclusions DAZAP1 is evolutionarily highly conserved. Its predominant expression in testes suggests a role in spermatogenesis. Its subcellular localization indicates that it is not directly involved in mRNA translation. Background Spermatogenesis is a complex developmental process in which male germ cells progress through mitotic proliferation, meiotic division and dramatic morphological changes to form mature sperm. This process is vital for the propagation of a species, and involves a large portion of the genome of an organism to ensure the quality and quantity of the final products. It is estimated that mutations in up to 11% of all genes in Drosophila might lead to male sterility [1]. This is likely to be true for humans also, considering the extremely high incidence (4–5%) of infertility in men [2]. Among the genes associated with male infertility is the DAZ (Deleted in Azoospermia) gene family. The family includes the Y-linked DAZ genes that are present only in great apes and old world monkeys [3], and the autosomal DAZL1 (DAZ-like 1) and BOULE genes [4,5] in all mammals. Deletion of the DAZ genes is found in about 10% of infertile males with idiopathic azoospermia [2], and disruption of Dazl1 causes infertility in both male and female mice [6]. Mutations in the DAZ family members of Drosophila[7], C. elegans[8], and Xenopus[9] also affect the fertility in either males, females, or both sexes. The DAZ gene family encodes RNA binding proteins that are expressed specifically in germ cells. DAZ and DAZL are expressed in the nucleus and cytoplasm of primordial germ cells and spermatogonia, and in the cytoplasm of meiotic spermatocytes [6,10]. BOULE is expressed later, in the cytoplasm of pachytene spermatocytes [5]. Genetic and biochemical studies suggest a role for the DAZ family in the regulation of mRNA translation. Drosophila Boule mutants was defective in the translation of the meiosis-specific CDC25 homologue, Twine [11], and DAZL was found to be associated with polyribosomes in mouse testes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to understanding the mechanisms of its action, we used a yeast two-hybrid system to isolate two human genes encoding DAZ associated proteins (DAZAPs) [14]. One of them, DAZAP1, is expressed predominantly in testes. It encodes a protein with two RNA binding domains and a proline rich C-terminal portion. The DAZAP1 protein interacted with both DAZ and DAZL in vitro. It also bound to RNA homopolymers. We now report our characterization of the mouse Dazap1 gene and its protein product. The subcellular localization of DAZAP1 suggests that it is not involved directly in mRNA translation. Results Characterization of the mouse Dazap1 cDNA Mouse Dazap1 cDNA clones were isolated by library screening, and the 5' end of the cDNA was isolated by 5' RACE [15]. The near fall length cDNA consists of a 53 bp 5' untranslated region (UTR), an open reading frame for a protein of 405 amino acid residues, and a 362 bp 3' UTR (GenBank Accession No: AF225910). The coding region shares 89% similarity with that of the human orthologue. The 3' UTR sequence is remarkably conserved. It contains three segments of 35 bp, 133 bp and 90 bp that share 85%, 90%, and 97% similarity with segments in the human 3' UTR, respectively. These segments probably contain regulatory elements. The DAZAP1 protein contains two RNA-binding domains (RBDS) and a C-terminal portion that is rich in proline (Figure 1). It is highly conserved evolutionarily. The mouse and the human proteins differ in 9 amino acids only, with 7 substitutions and two deletions/insertions. The mammalian proteins shares 89% similarity and 81% identity with Xenopus Prrp (for proline-rich RNA binding protein) [16]. The two RBDs are especially highly conserved. They share 98% and 97% similarity and 97% and 92% identity, respectively, between DAZAP1 and Prrp. These proteins may therefore have a similar RNA binding specificity. The C-terminal proline-rich portions of DAZAP1 and Prrp are less conserved (81% similarity and 71% identity). There is an insertion of a 58 bp segment in Prrp cDNA that causes a change of reading frame and results in a shorter Prrp with a different C-terminal end sequence. Figure 1 Evolutionary conservation of the DAZAP1 proteins. The amino acid sequences of the human and mouse DAZAP1s and the Xenopus Prrp are compared. The two RNA binding domains are boxed. Differences between the human and the mouse sequences, and between the mouse and Xenopus sequences are marked by #'s and *'s, respectively. Genomic structure of Dazap1 and chromosomal mapping Several overlapping lambda clones containing mouse Dazap1 genomic sequences were isolated. The locations of exons were determined by PCR amplification across exon-intron boundaries following by sequencing. All but the first exon were isolated and mapped. The genomic structure of the human DAZAP1 gene was also determined by blasting the human genome sequence at National Center for Biotechnology Information with the human DAZAP1 cDNA sequence. The mouse and the human genes have very similar structures, consisting of 12 exons spanning about 28 kb. All intron insertion sites are conserved (Table 1). The two RBDs are encoded by exons 1–4 and 5–8, respectively. Table 1 Exon-intron Organization of the Mouse and Human DAZAP1 genes A pair of PCR primers was designed from Dazap1 intronic sequences that amplified mouse but not hamster genomic sequences. Using a panel of mouse-hamster radiation hybrids, the mouse Dazap1 gene was mapped to chromosome 10 placed 27.84 cR from D10Mit260 (lod > 3.0) (data not shown). This region is syntenic to human 19p13.3 where the human DAZAP1 gene is located [14,17]. It contains no known mutant alleles that are associated with infertility. Expression of Dazap1 Northern analyses of adult mouse tissues showed the presence of two Dazap1 transcripts of 1.75 kb and 2.4 kb, respectively (Figure 2). Only the shorter transcript has been isolated in cDNA clones. Dazap1 was expressed most abundantly in the testis, much less in liver, heart and brain, and even less in other tissues. This pattern of expression is similar to that of the human DAZAP1 (14). RT-PCR analyses showed that Dazap1 mRNA was already present in fetal testes at embryonic day 15, similar to Dazl1 mRNA (Figure 3). The expression of both Dazl1 and Dazap1 persisted throughout testes development, in both the prenatal and postnatal periods. Dazl1 and Dazap1 transcripts were also present in the testes of Wv/Wv mutant mice which contained diminished number of germ cells [18]. However, only Dazap1 was expressed in a mouse germ cell line GCl-spg [19] and a Sertoli cell line MT4. The results suggest that Dazap1 is expressed in both somatic and germ cells in the testis. Figure 2 Expression of Dazap1 in adult mouse tissues. A mouse multiple-tissue Northern blot was hybridized with a Dazap1 cDNA probe, stripped, and rehybridized with a β-actin probe. Dazap1 is expressed most abundantly in the testis. Figure 3 Developmental expression of Dazap1 and Dazl in mouse testes. RT-PCR was performed on total testicular RNAs isolated from day 15 (El 5) and day 17 (El 7) embryos, new born mice (Day 0), and mice at various days after birth. Wv/Wv testes contain diminished germ cell population due to a mutated W (White spotted) gene. GC1 and MT4 are mouse germ cell and Sertoli cell lines, respectively, and gDNA is mouse genomic DNA. The PCR primers span over introns and produce much larger (if any) fragments from genomic DNA. To study the expression of the DAZAP1 protein, two antibodies against mouse DAZAP1 were generated. The anti DAZAP1-C antibody was raised against a recombinant protein containing the C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against an oligopeptide containing the last 19 amino acid residue at the C-terminus. Both antibodies recognized in vitro synthesized DAZAP1 in an immunoprecipitation assay (data not shown). Western blotting of mouse tissue extracts detected a 45 kD protein that was present most abundantly in the testis, and to a lesser degree in spleen, liver, lung and brain (Figure 4). The protein was also present in the ovary. The expression of DAZAP1 during germ cell development paralleled that of DAZL (Figure 5). It is present at a low level in the testes of 6 days old mice which contained only primitive type A spermatogonia. The expression of DAZAP1 increased afterward, as the testes contained increasing number of proliferating and meiotic germ cells. Figure 4 Expression of the DAZAP1 protein in adult mouse tissues. Equal amounts of total protein from various tissue extracts were applied to a 10% SDS-polyacrylamide gel and western blotted with the anti-DAZAP1-P antibody. Figure 5 Western blot analyses of the expression of DAZAP1 and DAZL in mouse testes during postnatal development. Subcellular localization of DAZAP1 Our previous fractionation of mouse testis extracts showed that most DAZL were present in the post mitochondrial fraction, and some of them were associated with polyribosomes [12]. Similar analyses showed that a majority of DAZAP1 in adult mouse testes was also present in the cytoplasmic fraction (data not shown). However, sucrose gradient analyses of the post- mitochondria fraction showed that, unlike DAZL, DAZAP1 did not co-sediment with polyribosomes (Figure 6). Figure 6 Sucrose gradient analyses shows that DAZAP1 is not associated with polyribosomes. The post-mitochondrial supernatant of mouse testis extracts was analyzed on a 15–45% sucrose gradient. Sedimentation was from left to right. The presence of DAZAP1 and DAZL in each fractions was analyzed by Western blotting. Discussion RNA-binding proteins have been found to participate in many cellular functions, including RNA transcription, pre-mRNA processing, mRNA transport, localization, translation and stability [20]. A role for the DAZ family in the regulation of mRNA translation is supported by lines of circumstantial evidence, including the association of DAZL with polyribosomes [12]. The absence of DAZAP1 from polyribosomes indicates that it is not directly involved in protein synthesis. This finding is different from two RNA-binding proteins, FXR1P and FXR2P, that were identified through their interaction with another polysomal-associated RNA-binding protein, the fragile X mental retardation protein [21]. Both FXR1P and FXR2P are associated with the polyribosomes [22]. The significance of the interaction between DAZAP1 and DAZL/DAZ remains to be defined. These proteins may act together to facilitate the expression of a set of genes in germ cells. For example, DAZAP1 could be involved in the transport of the mRNAs of the target genes of DAZL. Alternatively, DAZL and DAZAP1 may act antagonistically to regulate the timing and the level of expression. Such an antagonistic interaction between two interacting RNA-binding proteins is exemplified by the neuron-specific nuclear RNA-binding protein, Nova-1. Nova-1 regulates the alternative splicing of the pre-mRNAs encoding neuronal inhibitory glycine receptor α2 (GlyR α2) [23]. The ability of Nova-1 to activate exon selection in neurons is antagonized by a second RNA-binding protein, brPTB (brain-enriched polypyrimidine tract-binding protein), which interacts with Nova-1 and inhibits its function [24]. DAZAP1 could function in a similar manner by binding to DAZL and inhibiting its function. Comparing the phenotypes of Dazl1 and Dazap1 single and double knock-out mice may provide some clues to the significance of their interaction. Dazl1 knock-out mice have already been generated and studied [6]. The spermatogenic defect in the male becomes apparent only after day 7 post partum when the germ cells are committing to meiosis (H. Cooke, personal communication). The genomic structure of Dazap1, delineated here, should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16]. Prrp binds to a 340 nt sequence in the 3' UTR of Xenopus Vg1 mRNA. This Vg1 localization element (VLE) is sufficient for the migration and clustering of Vg1 mRNA to the vegetal cortex of mature oocyte. Prrp also interacts through its proline-rich domain with two microfilament-associated proteins profilin and Mena, which may facilitate the anchoring of Vg1 mRNA to the vegetal cortex. The Vg1 RNA encodes a member of the transforming growth factor-β family that is required for generating dorsal mesoderm at the blastula stage of Xenopus embryogenesis [25]. Sequence conservation between the RBDs of DAZAP1 and Prrp suggests that these proteins may bind to similar RNA sequences. However, a BLAST search of the GenBank for the 340 nt VLE sequence failed to identify any mammalian sequences with significant homology. Further mapping of the RNA sequence within VLE that binds Prrp, and possibly DAZAP1, may help to identify the natural substrates of DAZAP1. Conclusions DAZAP1 is an evolutionarily conserved RNA-binding protein. It is present at variable levels in many tissues. Its predominant expression in testes suggests a role in spermatogenesis. In mouse testes, DAZAP1 was found both in the nuclei and in the cytoplasm. Its absence from polyribosomes indicates that it is not directly involved in mRNA translation. Materials and methods Isolation of mouse Dazap1 cDNA clones Dazap1 cDNA clones were isolated from a mouse testis cDNA library (#937308, Stratagene, La Jolla, CA) using a human DAZAP1 cDNA as a probe. The 5' end of the cDNA was isolated by 5' RACE [15], using prdap11 (ttgcgggccatatccttg, #749–732) as the primer for cDNA synthesis from mouse testis RNA, and prdap37 (ttgttgccacgtgggcg, #734–718) and an adaptor primer as the primers for PCR amplification. The PCR products were cloned into a TA cloning vector pCR2.1-TOPO (Invitrogen, Carlsbad CA). Dazap1 clones were identified by colony hybridization and sequenced. The 5' RACE clone with the longest 5' UTR region and the cDNA clone P21 were ligated together through a unique PmlI site at # 722 to generate a cDNA clone (Dazap1-C) with a near full-length insert. Chromosomal mapping of Dazap1 Dazap1 genomic clones were isolated from a mouse 129SV genomic library (#946305, Stragagene), and sequences flanking each exons were determined. PCR primers (prdap25: cacctccaggatgtgttagc and prdazp26:gtcaccaagggtgtctgaag) were designed from intronic sequences flanking Dazap1 exon 8. These primers amplified a 271 bp fragment from mouse but not hamster genomic DNA. DNA samples of a panel of 100 radiation hybrids containing mouse chromosome fragments in a hamster background were purchased from Research Genetics (Huntsville, AL). The presence of mouse Dazap1 in the radiation hybrids was determined by PCR and the results were sent to the MIT server for computerized physical mapping of the gene. Expression of Dazap1 transcripts Northern hybridization was carried out according to standard procedures [26] using a mouse Multiple Tissue Northern Blot #7762–1 from Clontech (Palo Alto, CA). The blot was hybridized sequentially with DAZAP1 and β-actin cDNA probes, with stripping of the bound probes in between. Reverse transcription-polymerase chain reaction (RT-PCR) was carried out as preciously described using an annealing temperature of 54°C [27]. The primers were prdap35: agctcagggagtacttcaaga and prdap24 :ggagcttgattcttgctgtcc for Dazap1 which generated a product of 211 bp, and prdaz71: atcgaactggtgtgtcgaagg and prdaz72: ggaggctgcatgtaagtctca for Dazl1 which generated a product of 245 bp. Both primer pairs annealed across intron insertion sites. Generation of anti-DAZAP1 antibodies Antibodies were generated against both a recombinant protein produced in E. coli and an oligopeptide synthesized in vitro. The insert of a Dazap1 cDNA clone P21, which encoded the C-terminal portion of DAZAP1 (starting from aa #197), was cloned in-frame into the EcoRI/XhoI sites of an expression vector pET32b (Novagen, Madison, WI). Sequences at the junctions were verified by DNA sequencing. Milligrams of fusion proteins between thioredoxin and DAZAP1 were prepared and purified on His-Bind metal chelation resins (Novagen, Madison, WI). The proteins were mixed with Freund's adjuvant and injected into rabbits to generate the anti-DAZAPl-C antibody. An oligopeptide containing the last 19 ammo acid residues of the mouse DAZAP1 was synthesized in vitro using the services of Bethyl Laboratories (Montgomery, TX). The peptide was conjugated to KLH as carrier and injected into a goat. The anti-DAZAP1-P antibody thus produced was purified on an affinity column containing the oligopeptide antigen. Western blotting Mouse tissues were homogenized in the RIPA lysis buffer (150 mM NaCl, 1.0% NP-40, 0.5% DOC, 0.1% SDS, 50 mM Tris, pH 8.0) at a concentration of 0.2 g tissue per ml of buffer. The homogenized samples were cleared of debris by centrifugation at 10,000 × g for 10 minutes. Protein concentration of the tissue extracts was determined by the Bradford method using the Bio-Rad Protein Assay system (Bio-Rad, Hercules, CA). About 50 μg of tissue extracts were separated on 10% SDS-polyacrylamide gels and blotted with either the anti-DAZAP1-C antibody (at a 1/2,000 dilution) or the anti-DAZAP1-P antibody (at a 1/5,000 dilution). After incubation with horseradish peroxidase-conjugated secondary antibodies, the binding of antibodies was detected using the ECL Western Blotting System (Amersham Pharmacia Biotech, Piscataway, NJ). Fractionation of mouse testicular extracts Adult mouse testes were homogenized in a buffer containing 20 mM Tris, pH 7.5, 100 mM KCl, 5 mM MgCl2, 0.3% NP-40, 40 U/ml of Rnasin ribonuclase inhibitor (Promega, Madison, WI), and a mixture of 10 protease inhibitors provided in the Protease Inhibitors Set (Roche Molecular Biochemicals, Indianapolis, IN). Homogenates were centrifuged at 1,000 × g for 10 minutes to pellet cell debris and nuclei. After an additional centrifugation at 10,000 × g for 10 minutes to pellet the mitochondria, aliquots of the supernatant were applied to 15–45% sucrose gradients in 20 mM Tris, 100 mM KCl and 5 mM MgCl2 and centrifuged in a Beckman SW41 rotor at 39,000 rpm for 2 hours at 4°C. Fractions of 0.5 ml were collected from the bottom of the tubes and analyzed by western blotting. Acknowledgments We thank Gary Kuo for his involvement in the construction of Dazap1 expression vectors, and Ron Swerdloff's group for helpful discussion. The work was supported by grants from the National Institutes of Health (HD28009 and HD36347). Y. Vera was supported by an NIH grant (GM56902) on Initiative for Minority Student Development. References 1. Hackstein JHP, Hochstenbach R, Pearson PL. Towards an understanding of the genetics of human male infertility.. TIG. 2000;16:565-572. doi: 10.1016/S0168-9525(00)02140-5 2. De Kretser DM. Male infertility.. Lancet. 1997;349:787-790. doi: 10.1016/S0140-6736(96)08341-9 3. Reijo R, Lee T-Y, Salo P, Allagappan R, Brown LG, Rosenberg M, Rozen S, Jaffe T, Straus D, Hovatta O, de la Chapelle A, Silber S, Page DC. Diverse spermatogenic defects in humans caused by Y chromosome deletions encompassing a novel RNA-binding protein gene.. Nat Genet. 1995;10:383-393. doi: 10.1038/ng0895-383 4. Cooke HJ, Lee M, Kerr S, Ruggiu M. A murine homologue of the human DAZ gene is autosomal and expressed only in male and female gonads.. Hum Mol Genet. 1996;5:513-516. doi: 10.1093/hmg/5.4.513 5. Xu EY, Moore FL, Pera RAR. A gene family required for human germ cell development evolved from an ancient meiotic gene conserved in metazoans.. PNAS. 2001;98:7414-7419. doi: 10.1073/pnas.131090498 6. Ruggiu M, Speed R, Taggart M, McKay SJ, Kilanowski F, Saunders P, Dorin J, Cooke HJ. The mouse DAZLA gene encodes a cytoplasmic protein essential for gametogenesis.. Nature. 1997;389:73-77. doi: 10.1038/37987 7. Eberhar CG, Maines JZ, Wasserman SA. Meiotic cell cycle requirement for a fly homologue of human Deleted in Azoospermia.. Nature. 1996;381:783-785. doi: 10.1038/381783a0 8. Karashima T, Sugimoto A, Yamamoto M. Caenorhabditis elegans homologue of the human azoospermia factor DAZ is required for oogenesis but not for spermatogenesis.. Development. 2000;127:1069-1079. 9. Houston DW, Zhang J, Maines JZ, Wasserman SA, King ML. A Xenopus DAZ-like gene encodes an RNA component of germ plasm and is a functional homologue of Drosophila boule.. Development. 1998;125:171-180. 10. Reijo RA, Dorfman DM, Slee R, Renshaw AA, Laughlin KR, Cooke H, Page DC. DAZ family proteins exist through male germ cell development and transit from nucleus to cytoplasm at meiosis in humans and mice.. Biol Reprod. 2000;63:1490-1496. 11. Maines JZ, Wasserman SA. Post-transcriptional regulation of the meiotic Cdc25 protein Twine by the Daz1 orthologue Boule.. Nat Cell Biol. 1999;1:171-174. doi: 10.1038/11091 12. Tsui S, Dai T, Warren ST, Salido EC, Yen PH. Association of the infertility factor DAZL1 with actively translating polyribosomes.. Biol Reprod. 2000;62:1655-1660. 13. Venables JP, Ruggiu M, Cooke HJ. The RNA-binding specificity of the mouse Daz1 protein.. Nucl Acids Res. 2001;29:2479-2483. doi: 10.1093/nar/29.12.2479 14. Tsui S, Dai T, Roettger S, Schempp W, Salido EC, Yen PY. Identification of two novel proteins that interact with germ-cell specific RNA-binding proteins DAZ and DAZL1.. Genomics. 2000;65:266-273. doi: 10.1006/geno.2000.6169 15. Frohman MA, Dush MK, Martin GR. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.. PNAS. 1988;85:8998-9002. 16. Zhao W, Jiang C, Kroll TT, Huber PW. proline-rich protein binds to the localization element of Xenopus Vg1 mRNA and to ligands involved in actin polymerization.. EMBO J. 2001;20:2315-2325. doi: 10.1093/emboj/20.9.2315 17. DeBry RW, Seldin MF. Human/mouse homology relationships.. Genomics. 1996;33:337-351. doi: 10.1006/geno.1996.0209 18. Fleischman RA. From white spots to stem cells: The role of the Kit receptor in mammalian development.. Trends Genet. 1993;9:285-290. doi: 10.1016/0168-9525(93)90015-A 19. Hofmann MC, Narisawa S, Hess RA, Millan JL. Immortalization of germ cells and somatic testicular cells using the SV40 large T antigen.. Exp Cell Res. 1992;201:417-435. doi: 90291-F 20. Siomi H, Dreyfuss G. RNA-binding proteins as regulators of geneexpression.. Cur Opin Genet Develop. 1997;7:345-353. doi: 10.1016/S0959-437X(97)80148-7 21. Zhang Y, O'Connor JP, Siomi MC, Srinivasan S, Dutra A, Nussbaum RL, Dreyfuss G. The fragile X mental retardation syndrome protein interacts with novel homologs FXR1 and FXR2.. EMBO J. 1995;14:5358-5366. 22. Siomi MC, Zhang Y, Siomi H, Dreyfuss G. Specific sequences in the fragile X syndrome protein FMR1 and the FXR2 proteins mediate their binding to 60S ribosomal subunits and the interactions among them.. Mol Cell Biol. 1996;16:3825-3832. 23. Jensen KB, Dredge BK, Stefani G, Zhong R, Buckanovich RJ, Okano HJ, Yang YY, Darnell RB. Nova-1 regulates neuron-specific alternative splicing and is essential for neuronal viability.. Neuron. 2000;25:359-371. doi: 10.1016/S0896-6273(00)80900-9 24. Polydorides AD, Okano HJ, Yang YY, Stefani G, Darnell RB. A brain-enriched polypyrimidine tract-binding protein antagonizes the ability of Nova to regulate neuron-specific alternative splicing.. PNAS. 2000;97:6350-6355. doi: 10.1073/pnas.110128397 25. Thomson GH, Melton DA. Processed Vg1 protein is an axial mesoderm inducer in Xenopus.. Cell. 1993;74:433-441. doi: 10.1016/0092-8674(93)80045-G 26. Sambrook J, Fritsch EF, Maniatis T. null. Molecular Cloning: A Laboratory Manual 2nd ed, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,. 1989;null:null-null. 27. Salido EC, Yen PH, Mohandas TK, Shapiro LJ. Expression of the X-inactivation-associated gene XIST during spermatogenesis.. Nat Genet. 1992;2:196-199. doi: 10.1038/ng1192-196
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ins. Most tested strains exhibit a diurnal rhythm with IOP being the highest during the dark period of the day. Homozygosity for a null allele of the carbonic anhydrase II gene (Car2n) does not alter IOP while homozygosity for a mutation in the leptin receptor gene (Leprdb) that causes obesity and diabetes
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evant to glaucoma. Car2 deficiency does not alter IOP Bicarbonate formation is important for aqueous humor secretion from the ciliary processes and carbonic anhydrase (CA) facilitates this secretion. There are multiple forms of CA and the CAII isoform is reported to be the predominant form in the ciliary
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rified and sequenced as described [22]. Abbreviations Intraocular pressure (IOP), C57BL/6J (B6), chromosome (Chr), polymerase chain reaction (PCR), carbonic anhydrase (CA) Acknowledgments We thank Norma Buckley, Felicia Farley and Jennifer Smith for their assistance with data entry, references, and figu
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± SD; db/db 422 ± 110 mg/dl ; db/+ 147 ± 19 mg/dl; Anthony Nicholson, The Jackson Laboratory, personal communication) Car2 To test if deficiency of carbonic anhydrase II leads to decreased IOP, we analyzed mice of a B6 background that were genetically similar but with normal or mutant alleles of the Car2
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nst both a recombinant protein produced in E. coli and an oligopeptide synthesized in vitro. The insert of a Dazap1 cDNA clone P21, which encoded the C-terminal portion of DAZAP1 (starting from aa #197), was cloned in-frame into the EcoRI/XhoI sites of an expression vector pET32b (Novagen, Madison, W
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e C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against an oligopeptide containing the last 19 amino acid residue at the C-terminus. Both antibodies recognized in vitro synthesized DAZAP1 in an immunoprecipitation assay (data not shown). Western blotting of mouse tissue e
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oteins (DAZAPs) [14]. One of them, DAZAP1, is expressed predominantly in testes. It encodes a protein with two RNA binding domains and a proline rich C-terminal portion. The DAZAP1 protein interacted with both DAZ and DAZL in vitro. It also bound to RNA homopolymers. We now report our characterizatio
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AZAP1 protein, two antibodies against mouse DAZAP1 were generated. The anti DAZAP1-C antibody was raised against a recombinant protein containing the C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against an oligopeptide containing the last 19 amino acid residue at the C-t
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identity). There is an insertion of a 58 bp segment in Prrp cDNA that causes a change of reading frame and results in a shorter Prrp with a different C-terminal end sequence. Figure 1 Evolutionary conservation of the DAZAP1 proteins. The amino acid sequences of the human and mouse DAZAP1s and the Xe
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similarity and 97% and 92% identity, respectively, between DAZAP1 and Prrp. These proteins may therefore have a similar RNA binding specificity. The C-terminal proline-rich portions of DAZAP1 and Prrp are less conserved (81% similarity and 71% identity). There is an insertion of a 58 bp segment in P
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any) fragments from genomic DNA. To study the expression of the DAZAP1 protein, two antibodies against mouse DAZAP1 were generated. The anti DAZAP1-C antibody was raised against a recombinant protein containing the C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against a
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system (Bio-Rad, Hercules, CA). About 50 μg of tissue extracts were separated on 10% SDS-polyacrylamide gels and blotted with either the anti-DAZAP1-C antibody (at a 1/2,000 dilution) or the anti-DAZAP1-P antibody (at a 1/5,000 dilution). After incubation with horseradish peroxidase-conjugated secon
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h both the Y chromosome-encoded DAZ and an autosome-encoded DAZ-like protein, DAZL. DAZAP1 contains two RNA-binding domains (RBDs) and a proline-rich C-terminal portion, and is expressed most abundantly in the testis. To understand the biological function of DAZAP1 and the significance of its interac
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he human 3' UTR, respectively. These segments probably contain regulatory elements. The DAZAP1 protein contains two RNA-binding domains (RBDS) and a C-terminal portion that is rich in proline (Figure 1). It is highly conserved evolutionarily. The mouse and the human proteins differ in 9 amino acids
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[12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to
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[12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to
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d as they contain most ocular structures. Analysis of Myoc Exons and the proximal promoter of the mouse Myoc gene were amplified from mouse genomic DNA using the following combinations of primers: Exon 1: 5'-cttgcaggagaactttccagaa-3' and 5'-atctcgaaggagattgttatagg-3' 5'-gaccagctggagacccaaaccag-3' a
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ure was .81 (P < .05), which is similar to the reliability seen in previous estimates of neuron number [39, 40]. Genotyping and QTL Mapping Genomic DNA was extracted from spleens of F2 animals using a high-salt procedure [63]. A set of 82 microsatellite loci distributed across all autosomes and the X
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al forebrain [27, 50]. Brain volume and neuron number It has previously been shown that differences in brain weight are proportional to total brain DNA content and consequently to total CNS cell number [51, 52]. For this reason, brain weight has been suggested to be a good surrogate measure for total
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ng this period in any analyzed strain. Environmental differences may be important. Cage cleanliness, changing frequency and housing density can alter drug metabolism and the effect of anesthesia in rats [38,39]. The type of bedding used also may be important. We use wood shavings for bedding and wood sh
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vings for bedding and wood shavings expose the mice to terpenes. Terpene administration or environmental exposure to terpenes in wood shavings alters drug resistance and decreases the effect of anesthetic agents in both rats and mice [40-45]. Risk factors for increased IOP Some epidemiological studies
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l portion of Chr 10 in proximity with a number of genes known to affect brain development. One of these is Grk2, a member of the family of ionotropic glutamate receptor genes that is thought to play a role in modulating Huntington disease [44]. In the mouse, members of this receptor type act to indirectly do
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l portion of Chr 10 in proximity with a number of genes known to affect brain development. One of these is Grk2, a member of the family of ionotropic glutamate receptor genes that is thought to play a role in modulating Huntington disease [44]. In the mouse, members of this receptor type act to indirectly do
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should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16]. Prrp binds
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estes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family an
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sertion allele. The other allele was identical to the published sequence [22]. The insertion allele also had a previously reported substitution (A to G, Thr164Ala) in exon 1 and several other single base changes in the promoter region [22]. The presence or absence of this allele does not associate wi
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estes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family an
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should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16]. Prrp binds
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ptide consisting of one of the eight BRC repeats from human BRCA2. Expression of a single BRC repeat (BRC4) had previously been reported to act as an inhibitor of DNA repair by Chen et al [27]. These investigators showed that expression of constructs containing the BRC4 repeat in MCF-7 cells enhances the rad
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cts Adult mouse testes were homogenized in a buffer containing 20 mM Tris, pH 7.5, 100 mM KCl, 5 mM MgCl2, 0.3% NP-40, 40 U/ml of Rnasin ribonuclase inhibitor (Promega, Madison, WI), and a mixture of 10 protease inhibitors provided in the Protease Inhibitors Set (Roche Molecular Biochemicals, Indianapolis,
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at the junctions were verified by DNA sequencing. Milligrams of fusion proteins between thioredoxin and DAZAP1 were prepared and purified on His-Bind metal chelation resins (Novagen, Madison, WI). The proteins were mixed with Freund's adjuvant and injected into rabbits to generate the anti-DAZAPl-C antib
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hese risk factors. IOP is increased during the dark period of the day Diurnal variation in IOP is common in humans and laboratory animals [53]. The molecular mechanisms underlying the diurnal rhythm are not defined but increased aqueous humor production or flow occurs during the period of increased IOP in
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d Not genes. Vax1 is localized during development to the anterior ventral forebrain, and is expressed in the striatum during embryogenesis [28]. This molecule also has an important role in axon guidance: both the anterior portion of the corpus callosum and the optic chiasm are malformed or absent in Vax1 kn
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rior portion of the corpus callosum and the optic chiasm are malformed or absent in Vax1 knockout mice [49]. In addition, Vax1 interacts with several molecules including sonic hedgehog,Pax2, Pax6, and Rx that are known to be important during development of the basal forebrain [27, 50]. Brain volume and neur
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accurate and rapid enough to allow large scale genetic studies of factors determining IOP. Mice have great potential for helping to characterize the molecular mechanisms affecting IOP. Materials and Methods Animal husbandry All experiments were performed in compliance with the ARVO statement for use of a
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OP in mouse strains, we analyzed the gene in an assortment of strains with different IOPs. Two alleles were identified. One of these alleles had a 12 nucleotide insertion in the promoter region (ccagagcagggt, between positions -340 and -341) compared to the previously published sequence and is called the inse
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ral substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16]. Prrp binds to a 340 nt sequence in the 3' UTR of Xenopus Vg1 mRNA. This Vg1 localization element (VLE) is sufficient for the migration and clustering of Vg1 mRNA to the veg
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etween the RBDs of DAZAP1 and Prrp suggests that these proteins may bind to similar RNA sequences. However, a BLAST search of the GenBank for the 340 nt VLE sequence failed to identify any mammalian sequences with significant homology. Further mapping of the RNA sequence within VLE that binds Prrp, an
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ormalin fixative) processed, paraffin embedded and sectioned as previously reported [15,79], except that the paraformaldehyde was buffered with 0.1 M phosphate buffer. All strains other than CBA/CaHN and C57BLKS/J strains were evaluated. The eyes of CBA/CaJ and BALB/cJ mice were also fixed and processed for
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nt with this result, mice with albino eyes that are homozygous for tyrosinase or pink eye dilution mutations have altered diurnal rhythms compared to pigmented mice [75]. Albinism by itself is either not sufficient to alter the diurnal rhythm of IOP or alters it in different ways depending upon genetic backg
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these abnormalities affect mammalian IOP. Here, we show increased IOP in mice lacking tyrosinase activity compared to otherwise genetically identical pigmented B6 mice. Additionally, IOP differences between the light and dark period of the day were detected in the pigmented but not the albino B6 mice. Thus,
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to otherwise genetically identical pigmented B6 mice. Additionally, IOP differences between the light and dark period of the day were detected in the pigmented but not the albino B6 mice. Thus, albinism can affect the diurnal pattern of IOP changes. In agreement with this result, mice with albino eyes that a
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tion element (VLE) is sufficient for the migration and clustering of Vg1 mRNA to the vegetal cortex of mature oocyte. Prrp also interacts through its proline-rich domain with two microfilament-associated proteins profilin and Mena, which may facilitate the anchoring of Vg1 mRNA to the vegetal cortex. The V
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interacts with both the Y chromosome-encoded DAZ and an autosome-encoded DAZ-like protein, DAZL. DAZAP1 contains two RNA-binding domains (RBDs) and a proline-rich C-terminal portion, and is expressed most abundantly in the testis. To understand the biological function of DAZAP1 and the significance of its
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e segments probably contain regulatory elements. The DAZAP1 protein contains two RNA-binding domains (RBDS) and a C-terminal portion that is rich in proline (Figure 1). It is highly conserved evolutionarily. The mouse and the human proteins differ in 9 amino acids only, with 7 substitutions and two deleti
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and 97% and 92% identity, respectively, between DAZAP1 and Prrp. These proteins may therefore have a similar RNA binding specificity. The C-terminal proline-rich portions of DAZAP1 and Prrp are less conserved (81% similarity and 71% identity). There is an insertion of a 58 bp segment in Prrp cDNA that cau
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in, two antibodies against mouse DAZAP1 were generated. The anti DAZAP1-C antibody was raised against a recombinant protein containing the C-terminal proline-rich portion, and the anti DAZAP1-P antibody was raised against an oligopeptide containing the last 19 amino acid residue at the C-terminus. Both ant
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o acids only, with 7 substitutions and two deletions/insertions. The mammalian proteins shares 89% similarity and 81% identity with Xenopus Prrp (for proline-rich RNA binding protein) [16]. The two RBDs are especially highly conserved. They share 98% and 97% similarity and 97% and 92% identity, respectivel
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associated proteins (DAZAPs) [14]. One of them, DAZAP1, is expressed predominantly in testes. It encodes a protein with two RNA binding domains and a proline rich C-terminal portion. The DAZAP1 protein interacted with both DAZ and DAZL in vitro. It also bound to RNA homopolymers. We now report our characte
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in interacted with both DAZ and DAZL in vitro. It also bound to RNA homopolymers. We now report our characterization of the mouse Dazap1 gene and its protein product. The subcellular localization of DAZAP1 suggests that it is not involved directly in mRNA translation. Results Characterization of the mouse Dazap1
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nteraction with DAZ and DAZL, we isolated and characterized the mouse Dazap1 gene, and studied its expression and the subcellular localization of its protein product. Results The human and mouse genes have similar genomic structures and map to syntenic chromosomal regions. The mouse and human DAZAP1 proteins sha
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eated here, should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16].
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with polyribosomes in mouse testes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the funct
Text source name = 11604102.txt
ZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to understanding the mechan
Text source name = 11604102.txt
eated here, should facilitate the generating of Dazap1 null mutation. DAZAP1 was shown to bind RNA homopolymers in vitro, with a preference for poly U and poly G. Its natural substrates have not been identified. Recently, the Xenopus orthologue of DAZAP1, Prrp, was identified and characterized [16].
Text source name = 11604102.txt
ZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the function of the DAZ gene family and to understanding the mechan
Text source name = 11604102.txt
with polyribosomes in mouse testes [12]. More recently, DAZL was shown both in vitro and in a yeast three-hybrid system to bind specifically to oligo(U) stretches interspersed by G or C residues, including a U-rich segment in the 5' UTR of mouse Cdc25C mRNA [13]. In an attempt to elucidate the funct
Text source name = 11532192.txt
ariation in IOP is common in humans and laboratory animals [53]. The molecular mechanisms underlying the diurnal rhythm are not defined but increased aqueous humor production or flow occurs during the period of increased IOP in both rabbits and humans [63-65]. Small changes in the resistance to aqueous hum
Text source name = 11532192.txt
important approaches as they may allow the association of genes with IOP and glaucoma whose currently known functions do not suggest that they affect aqueous humor dynamics or do not immediately identify them as likely glaucoma candidates. No effect of anesthetic protocol on IOP during a 12 minute measure
Text source name = 11532192.txt
sed aqueous humor production or flow occurs during the period of increased IOP in both rabbits and humans [63-65]. Small changes in the resistance to aqueous humor drainage may also contribute to diurnal differences in IOP [66,67]. We first suspected IOP changes with time of day when the IOP of a group of


Bhubaneswar to host India’s home matches in Hockey Pro League
After becoming a hockey hub in recent past, Bhubaneswar will host the Netherlands first in the second edition of the Hockey Pro League on January 18.
by PTIBhubaneswar will host India’s home matches during the 2020 Hockey Pro League, the sport’s governing body announced on Monday.
The second edition of the Hockey Pro League will be played between January 11 and June 28, the International Hockey Federation (FIH) said in a statement.
Bhubaneswar has become a hockey hub in India and has been hosting a majority of tournaments, the most recent being the Olympic qualifiers. The first team that India will host is the Netherlands on January 18.
The Pro League will feature 144 matches in the coming season.
Australia’s home matches will be played in Perth and Sydney, while England will be playing its share in London.
The first match will feature the Netherlands and China on January 11, 2020 at the Wujin Hockey Stadium in Changzhou.
The venues:
Argentina: Buenos Aires, San Miguel de Tucuman
Australia: Perth, Sydney
Belgium: Antwerp
China: Changzhou
Germany: Mönchengladbach, Hamburg, Berlin
Great Britain: London
India: Bhubaneswar
The Netherlands: Utrecht, Rotterdam, Hertogenbosch, Amsterdam
New Zealand: Christchurch, Auckland
Spain: Valencia
USA: North Carolina


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The Hope spacecraft will be a compact, hexagonal-section spacecraft. It will be built from aluminium in a stiff but lightweight honeycomb structure and surfaced with a strong composite face-sheet. Its overall size and weight are comparable to a small car: it will weigh approximately 1,500 kg including fuel, and measure 2.37m wide by 2.90m tall.
The UAE aims to accomplish key science goals by integrating with the global Mars science community on key questions that no other mission has addressed.