As someone who has worked for and led non-profit projects over the past 9 years, it is tough to get respect for the work which I have done. Getting a group of people to work together for no other purpose than the vision and the cause is no easy task. Managing volunteers is vastly different to managing paid staff. In fact, getting people to volunteer in the first place is quite the challenge.
During past job interviews, my volunteer experience was constantly overlooked as nothing more than a hobby. If I cited life lessons or valuable skills I gained from my numerous volunteer experiences, it was easily dismissed and a curt reply asking for a similar experience in a corporate environment was given.
On the flip side, non-profit organisations need to value their volunteers more than just people who can help them achieve their goals. They need to understand that while yes, volunteers are being of service, they also need to be nurtured. Even more critically, this is an excellent way for your organisation to help mould future leaders of our society. Non-profit organisations need to validate the tasks and skills that volunteers get on the job and openly offer to act as references for job openings or academic applications.
It is unacceptable to complain that there are not enough people volunteering. Non-profit organisations need to understand that spending time in your organisation should reap something of value for all stakeholders. But you may ask just by the very nature that they are volunteering shouldn't they just do it out of the goodness of their hearts? Shouldn't the joy of helping your fellow human being or environment be enough?
Does your non-profit offer to give references? The organisation I co-Founded, The Backpack Project is a small organisation but in no way do we discredit the work which we do. I am proud to say that our organisation has recommended past volunteers to international leadership programmes, graduate schools and have given job recommendations.
Do you have an approach for engaging volunteers in a 'fast-paced' world? One time I was appointed the Head of Communications for a project and at that time I didn't have the slightest clue of how to organise a communications team. But what I did know, is that I had a team of competent, intelligent and crazy busy individuals on my team. So I hatched a plan that focused on getting the best work from each member without causing burn out.
I divided up the overall function into specific tasks and charged each member with a specific responsibility. My goal was not to overwhelm any one member of the team and if back up was ever needed I had enough free time to assist anyone in need of it.
Is your nonprofit overwhelming well-intentioned volunteers who get the job done? Is management using an operational or project management approach to execute its work?
Do you have a policy in place for developing your volunteers? When I first wanted a taste at project managing an event, I volunteered to organise a local community talent competition. That turned into me organising a Christmas tour at the malls nationwide, and then being Project Lead for a nationwide talent search competition. While the organisation didn't have an official policy for developing volunteers, I was given the opportunity to challenge and stretch my project management skills. It is important that each non-profit organisation, not only think about sponsors and benefactors of their programme, but to think of themselves as a petri dish for the germination of leadership, team cohesiveness, and communication skills.
Does your non-profit actively record the quantity and quality of volunteer involvement each year? For too long we have allowed accountants to dictate this whole issue, as if the only meaningful way to "account" for volunteers is the dollar value of their service. This is a red herring. It ought to be possible for organizations do provide some basic numbers: how many people volunteered this year? the number of hours? what is the average price per hour? what were their ages, gender, race, etc.? If the argument is that this information is too difficult to collect, we ought to respond that maybe the problem is no one cares.
Volunteering is an overall great experience. It gives us the opportunity to learn about ourselves, to learn about those around us and to build new relationships. But volunteer organisations need to recognise that in order to get more people involved and not to burn out those who give freely, how volunteers are viewed and a new talent management strategy is required.


text:
<!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.01 Transitional//EN">

<meta http-equiv="Content-Type" content="text/html; charset=ISO-8859-1">
<meta name="keywords" content="Erlang, MySQL, client, native, driver." />
<meta name="description" content="The MySQL standalone client for Erlang is using a native, low level MySQL client/server communication protocol." />
<title>Module datasource</title>
<link rel="stylesheet" type="text/css" href="stylesheet.css" title="EDoc">

Overview

Module datasource

Description

Data Types

Function Index

Function Details

  Datasource module implements API to manage datasource process.

Copyright © 2010-2014 Alexei Krasnopolski

Version: 1.2.4

Introduced in: 2012-12-18

Authors: Alexei Krasnopolski (<tt>krasnop@bellsouth.net</tt>) [web site: <tt>http://krasnopolski.org/</tt>].

Description

  Datasource module implements API to manage datasource process. The process keeps information
  about given datasource and controlls a pool of connections to DS. A functions get_connection/1,
  return_connection/2 and invalidate_connection/2 directly delegate calls to the connection pool that creates/reuses/disposes
  a connections. We can think about datasource module as a wrapper for resource_pool_srv module.

Data Types

client_options()

<tt>client_options() = #client_options{}</tt>

The record keeps client options that server is capable to support for this client and this connection.

  -record(client_options, {

charset_number = 33::0..255

- Character set number: default utf-8 (code= 33).

long_password = 1::0|1

- new more secure passwords.

found_rows  = 0::0|1

- Found instead of affected rows.

long_flag = 1::0|1

- Get all column flags.

connect_with_db = 1::0|1

- One can specify db on connect.

no_schema = 0::0|1

- Don't allow database.table.column.

compress = 0::0|1

- Can use compression protocol.

odbc = 0::0|1

- Odbc client.

local_files = 0::0|1

- Can use LOAD DATA LOCAL.

ignore_space = 0::0|1

- Ignore spaces before '('.

protocol_41 = 1::0|1

- New 4.1 protocol.

interactive = 1::0|1

- This is an interactive client.

ssl = 0::0|1

- Switch to SSL after handshake.

ignore_sigpipe = 0::0|1

- IGNORE sigpipes.

transactions  = 1::0|1

- Client knows about transactions.

reserved = 0::0|1

- Old flag for 4.1 protocol.

secure_connection = 1::0|1

- New 4.1 authentication.

multi_statements = 1::0|1

- Enable/disable multi-stmt support.

multi_results = 1::0|1

- Enable/disable multi-results.

trans_isolation_level = default::read_uncommitted | read_committed | repeatable_read | serializable

- transaction isolation level.

read_uncommitted - given session can read uncommitted updates of other session.

read_committed - given session can read only committed updates of other session.

repeatable_read - given session can read only snapshot of committed updates of other session.

serializable - the same as repeatable read but given session blocks own reads.

  }).

connection()

<tt>connection() = #connection{}</tt>

The record keeps information that helps connection server runs.

  -record(connection, {

socket :: port()

- tcp socket linked to MySQL server.

owner :: pid()

- pid of process that borrows this connection from connection pool. undefined if the connection is idle.

ds_def :: #datasource{}

- datasource record.

server_info :: #server_info{}

- server info record.

  }).

datasource()

<tt>datasource() = #datasource{}</tt>

datasource record information describes connections pool.
  The information is using for creating connections of the pool.

  -record(datasource, {

name :: atom()

- datasource name.

host = "localhost" :: string()

- server host name or IP.

port = 3306 :: integer()

- host port number.

database = "" :: string()

- database name.

user :: string()

- user logon name.

password :: string()

- password.

flags = #client_options{} :: #client_options{}

- client options for connections of the datasource.

  }).

eof_packet()

<tt>eof_packet() = #eof_packet{}</tt>

represents end of file (EOF) packet received from server after generic query or command.

  -record(eof_packet, {

warning_count::integer()

- number of warnings.

server_status::#server_status{}

- server status after query.

  }).

error_packet()

<tt>error_packet() = #error_packet{}</tt>

represents ERROR packet received from server after generic query or command.

  -record(error_packet, {

errno::integer()

- MySQL error number.

sqlstate::string()

- MySQL state identifire.

message::string()

- error message.

  }).

field_metadata()

<tt>field_metadata() = #field_metadata{}</tt>

represents field metadata of record set.

  -record(field_metadata, {

catalog::string()

- catalog name (unused in recent versions).

schema::string()

- schema (database) name.

table::string()

- table name from sql query (after AS).

origtable::string()

- original table name from schema.

name::string()

- field name from sql query (after AS).

origname::string()

- original field name from schema.

charsetnr::integer()

- code of character set.

length::integer()

- length of the field.

type::0..255

- type of the field (see mysql_types.hrl).

flags::bitstring()

- flags (see mysql_types.hrl).

scale::0..255

- scale.

default::binary()

- default value.

  }).

metadata()

<tt>metadata() = #metadata{}</tt>

represents metadata of record set.

  -record(metadata, {

field_count=0::integer()

- field count in the query.

param_count=0::integer()

- parameter count in prepared statement.

server_status::#server_status{}

- server status after query.

field_metadata = []::list(#field_metadata{})

- field metadata is a list of metadata
  for each field in the query.

param_metadata = []::list(#field_metadata{})

- parameter metadata is a list of metadata
  for each parameter in the prepared statement..

  }).

mysql_decimal()

<tt>mysql_decimal() = #mysql_decimal{}</tt>

represents value of decimal field.

  -record(mysql_decimal, {

int = ""::string()

- integer part.

fraction = ""::string()

- fraction part.

  }).

mysql_error()

<tt>mysql_error() = #mysql_error{}</tt>

Record represents an exception that is thrown by a client's module.

  -record(mysql_error, {

type:: tcp | connection | sqlquery | statement | transaction

- .

errno = none:: none | integer()

- .

sqlstate = []::string()

- .

source = []::string()

- .

message = []::string()

- .

  }).

mysql_time()

<tt>mysql_time() = #mysql_time{}</tt>

represents value of time/date field.

  -record(mysql_time, {

neg = false::boolean()

- sign of the field.

year = 0::integer()

- year.

month = 0::0..12

- month.

day = 0::0..31

- day of month.

hour = 0::0..23

- hour.

minute = 0::0..59

- minute.

second = 0::0..59

- second.

second_part = 0

- second part.

  }).

ok_packet()

<tt>ok_packet() = #ok_packet{}</tt>

represents OK packet received from server after generic query or command.

  -record(ok_packet, {

affected_rows::integer()

- number affected rows in table(s) during processing query.

insert_id::integer()

- insert query returns generated id for new row.

server_status::#server_status{}

- server status after query.

warning_count::integer()

- number of warnings.

message::string()

- warn/error message.

  }).

ok_stmt_packet()

<tt>ok_stmt_packet() = #ok_stmt_packet{}</tt>

represents OK packet received from server after statement prepare command.

  -record(ok_stmt_packet, {

stmt_handle::integer()

- prepared statement handle for using with following commands.

columns_number::integer()

- number of columns in prepared statement.

params_number::integer()

- number of parameters in prepared statement.

warn_count::integer()

- number of warnings.

  }).

packet()

<tt>packet() = #packet{}</tt>

represents binary packet for low level network connection.

  -record(packet, {

continue = false::boolean()

- indicates it is not last packet in sequence.

seq_number = 0::integer()

- number of the packet in sequence.

uncompressed_size = 0::integer()

- size of packet before compression.

body = <<>>::binary()

- binary body of the packet.

  }).

rs_header()

<tt>rs_header() = #rs_header{}</tt>

represents a header of record set.

  -record(rs_header, {

field_count::integer()

- number of fields in record set.

extra::any()

- extra information.

  }).

server_info()

<tt>server_info() = #server_info{}</tt>

The server_info() contains information that SQL server sent during handshake about itself.
  -record(server_info, {

protocol_version::integer()

- protocol version.

server_version::string()

- server version.

thread_Id::integer()

- current connection thread ID.

server_capabilities::#client_options{}

- server capabilities.

server_status::#server_status{}

- server status.

scramble_buff::binary()

- scrumble buffer for password encryption.

  }).

server_status()

<tt>server_status() = #server_status{}</tt>

The server_status() record contains information about status of SQL server after query.

  -record(server_status, {

inTransaction = false::boolean()

- Transaction has started.

autocommit = false::boolean()

- Server in auto_commit mode.

moreResultExists = false::boolean()

- Multi query - next query exists.

 queryNoGoodIndexUsed = false::boolean()

- .

queryNoIndexUsed = false::boolean()

- .

cursorExists = false::boolean()

- read-only non-scrollable cursor for a query was opened.

lastRowSent = false::boolean()

- read-only cursor is exhausted.

noBackSlashEscapes = true::boolean()

- .

queryWasSlow = false::boolean()

- this query was logged to the slow query log.

psOutParams = false::boolean()

- To mark ResultSet containing output parameter values.

  }).

Function Index

change_user/4	Changes user for the datasource.

close/1	Stops datasource DS_name and disposes all linked resources.

get_connection/1	Provides with connection process pid.

invalidate_connection/2	Sends request to dispose the connection.

return_connection/2	Returns the connection to connection pool.

select_db/2	Selects or changes current database (or schema).

Function Details

close/1

<tt>close(DS_name::atom()) -> ok | {error, Error}</tt>

Stops datasource DS_name and disposes all linked resources.

change_user/4

<tt>change_user(Connection_name::pid(), User::string(), Password::string(), Db::string) -> #connection{} | #mysql_error{}</tt>

Changes user for the datasource. This change concerns all
  connections of the connected datasource/pool.

See also: helper_connection:change_user/4.

select_db/2

<tt>select_db(Connection_name::pid(), DBName::string()) -> Result</tt>

<tt>Result = {Metadata::#metadata{}, [#ok_packet{}]} | #mysql_error{}</tt>

Selects or changes current database (or schema). Change concerns all
  connections of the given datasource/pool.

get_connection/1

<tt>get_connection(DS_name::atom()) -> pid() | #mysql_error{}</tt>

Provides with connection process pid. Connection pool returns idle connection or creates new
  connection process if no idle connection is found.

return_connection/2

<tt>return_connection(DS_name::atom(), Resource::pid()) -> ok | #mysql_error{}</tt>

Returns the connection to connection pool.

invalidate_connection/2

<tt>invalidate_connection(DS_name::atom(), Resource::pid()) -> ok | #mysql_error{}</tt>

Sends request to dispose the connection.

Overview

Generated by EDoc, Mar 19 2014, 17:26:36.

redpajama_set_name: RedPajamaGithub


Description: Add a splash of magical unicorn delight to your party with this brilliantly gold and pink 'We Heart Unicorns' banner. Featuring 13 pennants across 3m of garland, this gorgeous decoration has unicorns with rainbow ribbon unicorn tails and sparkly glitter detail! Shop the range for all the party pieces you need for a truly magical unicorn party.


This 3 bedroom, 2 bathroom home was just completely remodeled. With a spacious fenced in backyard, greenhouse and full size basement this home is move in ready! NEW central air conditioning system, septic system and landscaping! Walk into this home and find a fireplace , new paint and many more updates! Do not wait, this home will not last long!


It's an exciting time to be a Spurs fan, with one of the brightest young managers and most talented squads in the Premier League, plus a new 61,000 stadium due to open in time for the 2018-19 season. Simon Lansley speaks to the Chief Executive of the Tottenham Hotspur Foundation about what this means for the club's community work.
As you approach Spurs' famous old White Hart Lane stadium, the slogans on the boards screening the building work repeatedly scream 'Passionate about Tottenham'. Not unusual, you might think, for a Premier League football club to be shouting about their brand in front of the local community.
That is until, you realise, they are simultaneously referring to the local community.
Next season Spurs will play their Champions League games at Wembley and, during the 2017-18 season, have the option to stage both domestic and European matches at the national stadium, as their iconic old ground is transformed into a state-of-the-art 61,000 all-seater stadium.
As part of the preparations, next summer the club's award-winning charitable Foundation will move into a new 'hub' in a Grade II listed restored residential townhouse on Tottenham High Road, bought and donated by the football club and refurbished with a grant from the Heritage Lottery Fund. According to Grant Cornwell MBE, Chief Executive of the Spurs Foundation, this will serve two purposes – firstly, to provide more 'walk-in' access to the Foundation's services, and secondly to demonstrate that, while the first team may be temporarily based elsewhere, the club's phenomenal community work is unwavering in its commitment to the people of Tottenham.
The new stadium is being seen as the catalyst for the regeneration of east Haringey and south Enfield, with the Foundation's impact across nine wards reported in a 'social audit' published by the social impact research company 'Substance' in late 2015 – itself a first for a Premier League club's charitable foundation.
The report calculated that for every £1 invested by the Foundation, there was a social cost saving of £7. Read it and you realise, this goes way beyond the remit of just another club 'doing good' in the local community. It's a major investment and commitment to the health, wellbeing and future prosperity of some of London's most impoverished wards. The audit specifically looks at the work of the Foundation in nine wards identified in the North Tottenham Regeneration Programme.
"When that report was finished and launched, it was not a time for everyone to sit back and say 'look how great we are'," Cornwell said. "The report highlighted that what we are doing is relevant and is appropriate for the area we represented. It was good to know that the Foundation is having an impact and making a difference.
"The Chairman and Board at Tottenham Hotspur fully buy into the Foundation. There is an acknowledgement that it is an integral part of this football club, where we use what is a valuable brand to go and do some pretty tough work with some of the most hard-to-reach people in our society. The Chairman and the Board have given positive support and encouragement for the Foundation to go out and do what it does.
Cornwell added: "Our stadium is a venue that is available to the local community 365 days a year. We open the executive boxes up on weekdays for people to have health checks, for example. The club is committed to investing the time, energy and commitment into its community work – from first-team player and director engagement, right through the fabric of the club.
"We are in one of the most deprived wards in the country. We have some of the highest rates of obesity and diabetes, unemployment through generations of families, some of the lowest levels of educational attainment – even though we have got some great schools. So building aspirations for young people are a focus for us, for example we are the first club to offer a Foundation degree. We set that up because we want kids who can't usually afford it to still be able to have a University degree.
The audit has also enabled the Foundation to better understand its place in terms of service providers to the local community.
"We have gone through a major re-evaluation and a lot of what we are doing is more than just sport. It's just the Tottenham Hotspur brand is a catalyst for what we do. So for example we have our own Spurs nurse; a qualified health practitioner who is contracted to go out and do health checks mainly targeted at men, because a lot of men don't go to the doctors.
"We were tasked with engaging with 45-year-olds plus, so we send the Spurs nurse in the Spurs tracksuit to Halfords, B&Q and all the other places where men might traditionally go to. She sets up the stand and engages with people, and we offer them a tour of the ground if they have a health check. Now we have done more than 3500 health checks, and we have found a percentage of those did need to go to the doctors and we have identified issues.
Cornwell feels there can sometimes be a "reluctance" outside of football to truly embrace the power of clubs in their communities – perhaps in part because of the omnipotence of the Premier League itself – and he hopes that new strategies from the Government and Sport England over the last six months, which have a greater focus on the social outcomes of sport as opposed to solely driving participation, will open up the sector to more genuine collaboration.
"The skillsets here are far beyond traditional football coaching. All of our staff are mentors and have a deep understanding of the risks associated with young people. We work with all of this borough's looked-after children – at the last count there were 412. We have got some of the highest rates of teenage pregnancy, unemployment, domestic abuse and young people leaving school early. You look at the figures for children in care, they are horrendous and so we have focused on them for the last 10 years – not just the last 10 minutes. They are the most vulnerable in society.
Champions League football, England's best young players, a new stadium taking shape and the Head Coach having just signed a new five-year contract; this is "an incredibly exciting time" for everyone involved with Spurs, both on and off the field, says Cornwell.


Principles of Honda suspension systems, including construction, operation, components, relationships, steering geometry, alignment, rear and front suspension service, repair and tire pressure monitoring systems. 3 Credits (2 Lecture - 3 Lab) Prerequisite(s): HAD120. Corequisite(s): HAD163 and HAD230. Fall Only.


Nature Lover's Paradise! Peace and Tranquility describe this adorable year round Bungalow offering ever changing views of Run Pond Marsh, and privacy seldom found at this price. Bedrooms have French doors which open up to a spacious deck overlooking the backyard and marsh. Open Kitchen, Living, and Dining areas, and tiled Bath with radiant heat flooring. Walkout lower level has full-sized windows for future expansion possibilities, and opens out to backyard and patio with outside shower and irrigation. Three bedroom septic. Excellent Summer Rental History. Stroll to Great Island Bakery for a morning coffee. Close to Windmill Beach, South Middle Beach and the other beaches along South Shore Drive.


text:
define(['hello'], function(hello) {
    hello();
});


text:
Q: How to load .Rdata from an R package? I am developing an R package. In the package, I stored some .Rdata file from the simulation results. Now I would like to load/use those .Rdata in the rmarkdown so that I can write vignettes. I can load those data from my computer. But my question is how do I load data from my own package directory? Because in the future some users of the package may need to follow the same code from the vignette and reproduce the results or plots.

A: Assuming you used devtools::use_data(), you can use library(YOUR_PACKAGE); data("dataset") in your vignette, where "dataset" is the data you're talking about. Running devtools::load_all() will make the data available to you as you are writing the vignette.

redpajama_set_name: RedPajamaStackExchange


You need to send a clean snapshot of this Window Latch and Pull Set – Black – For Phillips window part to make certain it is the replacement window part that you need! Our staff is an expert in identifying your own replacement window parts and we ship hardware country wide by way of USPS.
I highly recommend you publish a pic of your respective Window Latch and Pull Set – Black – For Phillips part prior to placing your order, then again if you fully understand the actual part you need merely publish it in the area down below on the component form our staff members recognizes, responds and ships out components as quickly as feasible if you need rapid assistance just simply give us a call.
Window replacement parts including the Window Latch and Pull Set – Black – For Phillips which we will offer rapid USPS transport on nationwide might require some very distinct hardware that is usually completely about whether or not the house windows can be closed snugly enough to keep out the elements and to remain stable. Our staff distinguishes so you can buy PART online to make positive its the correct one you require.
These are typically window parts for instance the Window Latch and Pull Set – Black – For Phillips that need to offer people added layers of protection considering various climate circumstances. They need to be tough enough, they need to remain covered, and they need to supply people the opportunity and capability to stay warm within complicated circumstances.
Window Latch and Pull Set – Black – For Phillips, for example, functions to make certain that the screens of windows or the sashes of windows are stored very tightly to be certain that the windows are not going to rattle excessively, even in the wake of a great deal of wind, rain, and other outdoor hazards. These Window Latch and Pull Set – Black – For Phillips parts are going to have rust tolerant plating, which can allow them to endure some of the moisture content infiltration that individuals may well experience otherwise.
Window parts will vary per the definition of the kind of hardware you will need for instance storm window hardware or casement window hardware, of course. People are going to ask themselves what qualifies. One example is, some folks would point out that the tough window glass that a lot of storm windows use will meet the criteria as storm window hardware, and this appears like it must be the case. Other folks may say that when the window head or the window sash of tough storm windows is both designed to be strong enough to tolerate a wide variety of various problems, then individuals should also consider these to be storm window hardware.
There is a lot of different parts that could be the one that you need to have, unless of course you are 100% certain that this is the part you require make sure you send a photo of your own hardware so one of our part experts can recognize it and examine if we have it in supply or if we can order it for you, we offer both obsolete and very difficult to locate replacement window parts.
Window Latch and Pull Set – Black – For Phillips Branded aluminium construction; works with 4 inches glass; used on international windows performs with all manufacturers. Printed aluminium design; meets Four in glass. Sticked simple construction; fits 3/4 inch heavy wood slats. part window part is offered right here, still measures 5/16 inch O.D. rather of Prime-Lines 3/8 inch Each set possesses 4 Aluminum groove pin with E ring is used to hook up link end of operator to your Jealousy or Awning window parts.


Michael has over 10 years of experience in design, marketing, and social media. With a degree in industrial design from OCAD University, and having worked with IMM Living, the Children's Own Museum, and the CN Tower, he helps businesses increase their presence online and IRL in creative ways.
In addition to working with the School of Design and Institute without Boundaries, Michael is currently the Marketing Director of DesignTO (formerly Toronto Design Offsite Festival).


text:
Home » SME borrowing rates remain low
SME borrowing rates remain low
New BDRC research has found that in 2017, over 65 per cent of small and medium sized businesses (SMEs) said they would not be happy borrowing money.
The research, which was comprised of 130,000 interviews, found that despite 42 per cent of businesses claiming they needed funds to undertake a business activity, almost 20 per cent of businesses would not utilise an opportunity if there were significant risks with debt.
In fact, as an alternative to borrowing, 29 per cent of owners and directors used some of their personal cash to fund their business.
Moreover, towards the end of 2017, just 27 per cent of small and medium sized businesses (SMEs) said that they had aspirations to expand their business and were prepared to take the risks, including borrowing money, to succeed.
Shiona Davies from BDRC said: "SMEs are as likely to think they would fund a business opportunity themselves as approach a bank for funding.
"For those who do apply for a loan or overdraft, success rates remain high. However, first time applicants' success rates are currently lower than in 2015, albeit still higher than they were in 2012.
"Additionally, fewer SMEs who are not currently using finance show any appetite to do so."
Over 80 per cent of funding applications and renewals have been successful over the past 18 months but not many SME business owners are happy to make use of the options available.
Despite this, 82 per cent of SMEs were profitable and almost half of SMEs reported growth in 2017.
The top three barriers for businesses remain legislation and regulation, political uncertainty and the current economic climate.
Follow Haslers on Facebook for all our latest updates.
So if you want to find out more about what makes us intentionally different us then please contact Haslers today.

redpajama_set_name: RedPajamaCommonCrawl


text:
Shame and Pride: Affect, Sex and the Birth of Self
Author(s): Donald L. Nathanson
This is a revolutionary book about the nature of emotion, about the way emotions are triggered in our private moments, in our relations with others, and by our biology. Drawing on every theme of the modern life sciences, Dr. Nathanson shows how the nine basic affects--interest-excitement, enjoyment-joy, surprise-startle, fear-terror, distress-anguish, anger-rage, dissmell, disgust, and shame-humiliation--not only determine how we feel but shape our very sense of self. For too long there has been a battle between those who explain emotional discomfort on the basis of lived experience and those who blame chemistry. As Dr. Nathanson shows, chemicals and illnesses can affect our mood just as surely as an uncomfortable memory or a stern rebuke. He presents a completely new understanding of all emotion, providing the first link between the exciting affect theory of Silvan Tomkins and the entire world of biology, medicine, psychology, psychotherapy, religion, and the social sciences. Shame is the least understood of the painful emotions, although it affects every phase of life. We have all been made to feel foolish just at the moment we most wanted to appear wonderful; we have all been rebuffed by those we wished to court. Not one of us looks exactly as we might wish. Shame haunts our every dream of love, and influences how we experience ourselves as sexual beings. We react to shame by withdrawing, by making painful alliances with those who humiliate us, by calling attention to what brings us pride, or by attacking whoever has made us feel inferior. The comedian, as Nathanson shows in his discussion of Buddy Hackett, makes us laugh at what we try to keep hidden, transforming shame intoacceptance and even pride. This book explains everything that can possibly make us proud or ashamed. All are in this book; nobody who reads it will be quite the same again.
Shame and Pride is an impressive achievement. Nathanson establishes a solid and systematic foundation for the modern study of affect, much as Erik Erikson accomplished for identity and ongoing development throughout the life cycle. A rich and rewarding experience. -- Richard P. Kluft, M.D., Clinical Professor of Psychiatry, Temple University School of Medicine
Donald L. Nathanson, M.D., is the founder and Executive Director of the Silvan S. Tomkins Institute. He is currently Clinical Professor of Psychiatry and Human Behavior at Jefferson Medical College.
Publisher : WW Norton & Co
Imprint : WW Norton & Co
Illustrations : Ill.


text:
'''
SPDX-License-Identifier: Apache-2.0
Copyright 2017 Massachusetts Institute of Technology.
'''

import sys
import argparse
import os
import shutil

IPSEC_TOOLS_CONF = "flush;\n\
spdflush;\n"

IPSEC_TOOLS_SUBNET = "spdadd 0.0.0.0/0 %s any -P out ipsec esp/transport//require;\n\
spdadd %s 0.0.0.0/0 any -P in ipsec esp/transport//require;\n"

IPSEC_TOOLS_EXCLUDE = "spdadd 0.0.0.0/0 %s any -P out none;\n\
spdadd %s 0.0.0.0/0 any -P in none;\n"

def usage():
    print("ERROR: you must specify a configuration file with one or more subnets for ipsec")
    print("\tExample: generate.py file.txt")
    print("====file.txt format ====")
    print("# Any file starting with # will be ignored")
    print("# All Subnets after ipsec are enabled by default")
    print("ipsec")
    print("192.168.0.0/24")
    print("172.22.2.4/32")
    print("# All subnets after exclude will not use ipsec")
    print("exclude")
    print("192.168.0.1/32")
    sys.exit(1)

def main(argv=sys.argv):
    if len(argv) < 2:
        usage()

    subnets = None
    exclude = None
    with open(argv[1], 'r') as f:
        for line in f:
            line = line.strip()
            if line[0] == '#':
                continue

            if line == 'ipsec':
                subnets = []
                continue

            if line == 'exclude':
                exclude = []
                continue

            if exclude is not None:
                exclude.append(line)
                continue

            if subnets is not None:
                subnets.append(line)

    if subnets is None:
        usage()
    if exclude is None:
        exclude = []

    print("Preparing extra files for ipsec config in directory: ipsec-extra")
    print("enabling ipsec for subnets:  %s" % subnets)
    print("disabling ipsec for subnets: %s" % exclude)

    if os.path.exists('ipsec-extra'):
        shutil.rmtree('ipsec-extra')
    os.mkdir("ipsec-extra")

    with open('ipsec-extra/ipsec-tools.conf', 'w') as f:
        f.write(IPSEC_TOOLS_CONF)
        for subnet in subnets:
            f.write(IPSEC_TOOLS_SUBNET % (subnet, subnet))
        for subnet in exclude:
            f.write(IPSEC_TOOLS_EXCLUDE % (subnet, subnet))

    with open("ipsec-extra/action_list", 'w') as f:
        f.write("local_action_update_crl,local_action_deletesa\n")

    shutil.copy("src/local_action_update_crl.py", "ipsec-extra")
    shutil.copy("src/local_action_deletesa.py", 'ipsec-extra')
    shutil.copy("src/autorun.sh", 'ipsec-extra')
    shutil.copy("src/racoon.conf", 'ipsec-extra')

    print("include directory ipsec-extra when using keylime in cert mode to enable ipsec")

if __name__ == "__main__":
    try:
        main()
    except Exception as e:
        print(e)


YUCK! I bit into one and threw it away. My sweet kids? These guys?
I can't even make resurrection rolls correctly and I need His help to just make breakfast every day!!! Praise Jesus for the hope He gives us!!!
'Hosanna' is a term used to express adoration, praise or joy.
'Son of David' – There is a prophecy in 2 Samuel 7 that tells of a Messiah coming. Christ is the fulfillment of that prophecy and is our Messiah. You can see geological proof of that in Matthew 1.
'Blessed is He who comes in the name of the Lord' is a direct quote from Psalm 118:26. He is worthy to be blessed and praised because He came to take away the sins of the world. And in scripture, where we meditate today, He was on His journey to do just that. His journey to the Cross was less than a week away. This triumphant entry was the beginning of that journey. He was headed into Jerusalem, where He knew He would eventually be captured, scourged and put to death on a cross for us.
Why? Just so we can know Him and be in relationship with Him. Jesus had to take our sin away so we could be put back into a right relationship with God. Jesus did that for us.
Today, and all this week, let's remember that. Let's think on that and rest in that. What does it mean to you that Jesus endured all that He did for you? You are that important to Him.
So What About All The Bunnies, Chicks, Eggs and Peeps?
Well, honestly, I don't know. I grew up with bunnies, chicks and eggs at Easter. They bring back good memories. I had a really easy time believing in Santa Claus as a child. That was totally believable to me. But the Easter Bunny … now we're stretching it a bit! Isn't that funny?! I totally believed in a jolly fat guy who delivered gifts, via flying reindeer, to all children in one night, but a magical life-sized bunny painting eggs for children … no way.
My mom always did a GREAT job of making every season and holiday something special. We celebrated it and decorated the house for it. I tend to do the same thing. I like wreaths and outside décor for seasons and holidays. I have cute spring and Easter decorations that I enjoy. I have dishtowels with eggs and chicks on them. We dye eggs every Easter with the kids and have an egg hunt. That's always fun … a tradition we all enjoy.
Not because it's bad or evil. Not because I want to be different than everyone else. I'm rethinking it all because God has caused my heart to pause. As Valentine's Day came to a close and I took down my heart-shaped red décor, I started thinking about my Easter décor and when I was going to decorate. It was then that I felt the Holy Spirit create a small tension inside of me about it all. Did you know that's how God speaks to us? There's a beautiful story about it in 1 Kings 19. Elijah was instructed to go and stand on a mountain before the Lord. He was trying to hear from God. There were all these things that happened that made loud noises – a strong wind, an earthquake, a fire – but scripture says "and after the fire a sound of a gentle blowing" 1 Kings 19:12.
Y'all — that was the Lord. That was Him speaking to Elijah. You see, I try to spend time in scripture, prayer, confession and surrender daily JUST FOR MOMENTS LIKE THESE!!!! How exciting it is to have moments of quietness where you are discerning that the Lord is asking you to take pause and think about something.
So that's what I'm doing. I'm reading about it. I'm praying about it. I'm talking to my husband about it. I'm processing with my children about it. I'm NOT throwing the baby out with the bath water and throwing all my Easter decorations away. I have them around my house now. Cute little Eggs on my mantel and chick towels in my bathroom. I'm holding it all with an open hand and asking Jesus to guide Michael, me and our family through it. What is He calling us to do? Why are we doing what we're currently doing?
This Lenten season, I ask you just to pause and think about why you are doing what your doing. When you do that … ask God to show you what HE wants you to do. Ask Him to speak to you, just as He did to Elijah, and give you a heart that discerns His voice. Pray with faith … expecting Him to speak to you. And then let me know about it…I'd love to hear how God is speaking to you!
O God, early in the morning I cry to you. Help me to pray; and to concentrate my thoughts on you; I cannot do this alone.
In me there is darkness, but with you there is light; I am lonely, but you do not leave me; I am feeble in heart, but with you there is help; I am restless, but with you there is peace. In me there is bitterness, but with you there is patience; I do not understand your ways, but you know the way for me. . . .
Restore me to liberty, and enable me to live now that I may answer before you and before men. Lord, whatever this day may bring, your name be praised.
I learned something new at work last week. Catholics do not eat meat on Fridays during Lent. When I inquired about it, no one really knew why, but that's just what they did (or didn't do) on Friday's during Lent. So that left me wondering and doing a little internet research. If you know more about this … please leave comments below. I'd love to learn.
Let's start from a Theological perspective: in the church, we commemorate Jesus' death as being on a Friday. Because of that, people have written of fasting on Friday to remember His sacrifice for us from very early on. That's it. It's just that simple. So in an effort to not forget Jesus' sacrifice, many cut out meat on Fridays. Eating fish is fair game for Fridays because they are cold-blooded animals. Apparently it's only warm-blooded — live off the land sort of animals — that are off-limits.
As the church and Catholicism grew, fasting from meat was not just a Friday during lent sort of thing. Most catholics were supposed to fast from meat every Friday to continually remind themselves of Jesus' death on the cross. When adding in advent and other Holy Days in the christian calendar, Fish Friday's grew in popularity.
I read an interesting USA today article concerning Fish Friday's and a McDonald's in a largely catholic part of Cincinnati. Apparently, the franchise was doing very poorly on Fridays and was going under. The Owner/General Manager knew he needed a non-meat alternative for Friday if his business was going to survive. So he developed the Filet-o-Fish sandwich. The the full article here.
I personally did not adopt the 'no meat Friday' as a part of my fasting this year. I'm going to keep it in my thoughts and see if that is something I think I need to pull in at another time. I will tell you, this would be a hard one for me. I don't really care for fish, so it would definitely be a sacrifice for me. Maybe that is reason enough to pull it in and take part?
Do you eat meat on Fridays during Lent? Was it something you grew up doing or are you doing it just recently as an addition to your Lenten fasting routine? I'm super interested to know!
This month we're focusing on ways to show love. Yesterday, on February 1st we posted our first blog about it … showing love through blueberry muffins. Showing love is something that is not that hard to do, but does take just a little thought and effort; a little bit of time thinking about someone else and what they love. Scripture is pretty specific about this topic.
Nothing. I am to do nothing from selfishness. Well, that's just not going to happen. And that's why I need the grace of God so badly. Jesus is our mediator. He knows we can't live up to that standard. Not living up to that standard shows us our need for a Savior. There's no way we can reconcile the brokenness we have with God because of our sin. So Jesus made the way for us to be brought back to God and to be reconciled in that relationship with Him. Scripture is pretty clear about that, too.
How amazing. Jesus is the one that bridges that gap between us and God so that we might have a personal relationship with Him. Okay, okay, I get it, Holly, but WHY are you going off on this tangent when all I want to do is read about how to 'show love' to those around me. Well, going off on this tangent is SUPER important because God is the author and creator of love. And scripture says that 'we only love because He first loved us'. We can't know true love or even show or give true love unless we know Him first. Knowing God in a personal and intimate way is the BEST way to show love.
Do you know Jesus? Do you have a personal relationship with Him? Have you acknowledged your need for His redeeming work on the cross? I did that for the first time when I was 10. Will you do that for the first time right now? Accept God's love for you and enjoy a whole new perspective on love this month of February.


Delusion was the lead of single from the 'Bring it Back to Basics' album. In addition to 'Delusion' this very limited edition 12" also features the songs 'Make it Hot' and 'Hypnotism' which are exclusive to this EP. The B side of the record features instrumentals of all 3 tracks.


text:
Article| Volume 26, ISSUE 8, P1009-1018, April 25, 2016
A Mutation Associated with Stuttering Alters Mouse Pup Ultrasonic Vocalizations
Terra D. Barnes
Department of Neuroscience, Washington University School of Medicine in St. Louis, Campus Box 8108, 660 S. Euclid Avenue, St. Louis, MO 63110-1093, USA
David F. Wozniak
Department of Psychiatry, Campus Box 8134, and the Taylor Family Institute for Innovative Psychiatric Research, Campus Box 8108, Washington University School of Medicine in St. Louis, 660 S. Euclid Avenue, St. Louis, MO 63110-1093, USA
Joanne Gutierrez
National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD 20892, USA
Tae-Un Han
Dennis Drayna
Timothy E. Holy
Open ArchivePublished:April 14, 2016DOI:https://doi.org/10.1016/j.cub.2016.02.068
Previous ArticleCyanobacteria and Eukaryotic Algae Use Different Chemical Variants of Vitamin B12
Next ArticleA Mechanical Feedback Restricts Sepal Growth and Shape in Arabidopsis
Mutations in the lysosomal enzyme-targeting pathway cause stuttering in humans
Mice with a mutation in this pathway also show abnormalities in vocalizations
Other behaviors in the knockin mice appear largely intact
People who stutter with this type of mutation have similar features in their speech
A promising approach to understanding the mechanistic basis of speech is to study disorders that affect speech without compromising other cognitive or motor functions. Stuttering, also known as stammering, has been linked to mutations in the lysosomal enzyme-targeting pathway, but how this remarkably specific speech deficit arises from mutations in a family of general "cellular housekeeping" genes is unknown. To address this question, we asked whether a missense mutation associated with human stuttering causes vocal or other abnormalities in mice. We compared vocalizations from mice engineered to carry a mutation in the Gnptab (N-acetylglucosamine-1-phosphotransferase subunits alpha/beta) gene with wild-type littermates. We found significant differences in the vocalizations of pups with the human Gnptab stuttering mutation compared to littermate controls. Specifically, we found that mice with the mutation emitted fewer vocalizations per unit time and had longer pauses between vocalizations and that the entropy of the temporal sequence was significantly reduced. Furthermore, Gnptab missense mice were similar to wild-type mice on an extensive battery of non-vocal behaviors. We then used the same language-agnostic metrics for auditory signal analysis of human speech. We analyzed speech from people who stutter with mutations in this pathway and compared it to control speech and found abnormalities similar to those found in the mouse vocalizations. These data show that mutations in the lysosomal enzyme-targeting pathway produce highly specific effects in mouse pup vocalizations and establish the mouse as an attractive model for studying this disorder.
Speech disorders affect millions of people worldwide and are usually treated with behavioral therapy [
Bloodstein O.
A Handbook on Stuttering.
Fifth Edition. Singular Publishing Group, 1969
Conture E.G.
Stuttering: Its Nature, Dignosis, and Treatment.
Allyn and Bacon, 2001
]. A better understanding of human speech disorders may lead to a wider array of treatment options and can provide insights into the genetic and neural underpinnings of human speech. To date, there are remarkably few speech disorders with a clearly identified genetic component [
Fisher S.E.
Lai C.S.L.
Monaco A.P.
Deciphering the genetic basis of speech and language disorders.
Annu. Rev. Neurosci. 2003; 26: 57-80
MacDermot K.D.
Bonora E.
Sykes N.
Coupe A.-M.
Vernes S.C.
Vargha-Khadem F.
McKenzie F.
Smith R.L.
Identification of FOXP2 truncation as a novel cause of developmental speech and language deficits.
Am. J. Hum. Genet. 2005; 76: 1074-1080
Fedorenko E.
Morgan A.
Murray E.
Cardinaux A.
Mei C.
Tager-Flusberg H.
Kanwisher N.
A highly penetrant form of childhood apraxia of speech due to deletion of 16p11. 2.
Eur. J. Hum. Genet. 2016; 24: 302-306
]. Developmental stuttering is one of the most common speech disorders, with the persistent form of the disorder affecting three million adults in the United States. The disorder is characterized by frequent repetitions or prolongations of syllables or words or by speech that has frequent hesitations or pauses—known as "blocks"—that disrupt the smooth flow of speech [
]. As an inroad into mechanisms of speech production, stuttering is remarkable because affected individuals are, on average, normal by all other known measures of language, cognitive, and motor function [
]. Recently, it was found that persistent developmental stuttering unaccompanied by other deficits or symptoms can be linked to mutations in the lysosomal enzyme-targeting pathway (LETP) [
Kang C.
Riazuddin S.
Mundorff J.
Krasnewich D.
Friedman P.
Mullikin J.C.
Drayna D.
Mutations in the lysosomal enzyme-targeting pathway and persistent stuttering.
N. Engl. J. Med. 2010; 362: 677-685
]. Mutations in the LETP account for 9%–16% of all cases of persistent nonsyndromic stuttering [
Raza M.H.
Domingues C.E.
Webster R.
Sainz E.
Paris E.
Rahn R.
Gutierrez J.
Chow H.M.
Kang C.S.
Mucolipidosis types II and III and non-syndromic stuttering are associated with different variants in the same genes.
Lysosomes are degradative cellular organelles that contain acid hydrolases. These enzymes are targeted to lysosomes by tagging them with mannose 6-phosphate [
Kornfeld S.
Trafficking of lysosomal enzymes.
FASEB J. 1987; 1: 462-468
]. The mannose 6-phosphate is added to these acid hydrolases in a two-step process carried out by the products of three genes. Mutations in all three of these genes have been found in humans who stutter [
]. However, the mechanisms by which mutations in an apparent "housekeeping gene" produce a deficit with the remarkable specificity of stuttering remain unclear.
While humans are irreplaceable for discovery of the genetic underpinnings of speech disorders, they have a number of disadvantages for analysis of the underlying mechanisms. An animal model would be an invaluable tool for investigations into the circuit and cellular mechanisms of a vocal disorder, despite the obvious difficulties in comparing human speech with any animal vocalization [
]. While no exact correspondence with human speech can be expected, it is possible that one or more "low-level" features—such as planning, initiation, timing, breath control, and/or temporal sequencing—may be shared both in phenotype and underlying neuronal circuitry across mammalian species.
Mouse vocalizations have been extensively studied and have been used successfully in the study of several disorders [
Fischer J.
Hammerschmidt K.
Ultrasonic vocalizations in mouse models for speech and socio-cognitive disorders: insights into the evolution of vocal communication.
Genes Brain Behav. 2011; 10: 17-27
Arriaga G.
Zhou E.P.
Of mice, birds, and men: the mouse ultrasonic song system has some features similar to humans and song-learning birds.
Enard W.
Gehre S.
Hölter S.M.
Blass T.
Somel M.
Brückner M.K.
Schreiweis C.
Winter C.
Sohr R.
A humanized version of Foxp2 affects cortico-basal ganglia circuits in mice.
Scattoni M.L.
Ricceri L.
Crawley J.N.
Unusual repertoire of vocalizations in adult BTBR T+tf/J mice during three types of social encounters.
Ey E.
Torquet N.
Le Sourd A.-M.
Leblond C.S.
Boeckers T.M.
Faure P.
Bourgeron T.
The Autism ProSAP1/Shank2 mouse model displays quantitative and structural abnormalities in ultrasonic vocalisations.
Behav. Brain Res. 2013; 256: 677-689
]. Mice produce ultrasonic vocalizations in a range of social situations. These vocalizations have repeated syllables and complex structure and have been characterized as "songs" [
Holy T.E.
Guo Z.
Ultrasonic songs of male mice.
PLoS Biol. 2005; 3: e386
]. One type of innate mouse vocalization is the isolation call of pups, which has also been extensively studied, especially in disease [
Fujita E.
Tanabe Y.
Shiota A.
Ueda M.
Suwa K.
Momoi M.Y.
Momoi T.
Ultrasonic vocalization impairment of Foxp2 (R552H) knockin mice related to speech-language disorder and abnormality of Purkinje cells.
Gaub S.
Groszer M.
Ehret G.
The structure of innate vocalizations in Foxp2-deficient mouse pups.
Genes Brain Behav. 2010; 9: 390-401
Shu W.
Cho J.Y.
Jiang Y.
Zhang M.
Weisz D.
Elder G.A.
Schmeidler J.
De Gasperi R.
Sosa M.A.G.
Rabidou D.
Altered ultrasonic vocalization in mice with a disruption in the Foxp2 gene.
Grimsley J.M.S.
Monaghan J.J.M.
Wenstrup J.J.
Development of social vocalizations in mice.
Categorical perception of mouse-pup ultrasounds in the temporal domain.
]. Mouse pups, when separated from their mother during the first 2 weeks of life, spontaneously vocalize [
]. The rate of vocalization peaks at around 5–7 days post-natal and decreases thereafter.
This approach is exemplified by studies of the speech disorder associated with mutations of the FOXP2 transcription factor. The Foxp2 gene is conserved across species [
Hurst J.A.
A forkhead-domain gene is mutated in a severe speech and language disorder.
]. In mice, disruption of the Foxp2 gene has been shown to cause changes in pup isolation calls [
Keays D.A.
Deacon R.M.J.
de Bono J.P.
Prasad-Mulcare S.
Baum M.G.
French C.A.
Nicod J.
Coventry J.A.
Impaired synaptic plasticity and motor learning in mice with a point mutation implicated in human speech deficits.
Curr. Biol. 2008; 18: 354-362
] (though not necessarily in adults [
Minge C.
Pääbo S.
A humanized version of Foxp2 does not affect ultrasonic vocalization in adult mice.
Genes Brain Behav. 2015; 14: 583-590
]); in turn, such studies have been used to obtain numerous insights into the molecular and cellular functions of this gene and its role in human speech [
Here, we analyze pup isolation calls and other behaviors of mice with a knockin mutation in the Gnptab gene, for which the equivalent mutation has been shown to cause stuttering in humans [
]. We show that such mice exhibit alterations in timing and sequencing of vocalization that are reminiscent of specific deficits observed in human stuttering.
We engineered mice to carry a homozygous Glu1179Lys mutation in Gnptab (Gnptabmut/mut), homologous to the Glu1200Lys mutation in human GNPTAB well characterized for its role in stuttering [
Fedyna A.
Characterization of a mutation commonly associated with persistent stuttering: evidence for a founder mutation.
J. Hum. Genet. 2011; 56: 80-82
]. Mice were constructed on a pure BALB/c background (Figures 1A and S1), using a heterozygote × heterozygote breeding strategy to ensure the availability of littermate controls with matched pre- and post-natal environment. We recorded pup isolation calls on post-natal day (P) 3, P5, and P8; data from all days are presented in the supplemental figures, with data from day 8 presented in the main figures below.
Figure 1Experimental Scheme
(A) Knockin mice with a Glu1179Lys mutation in exon 19 of the Gnptab gene were constructed on a BALB/c background. Recordings were performed on mice pups separated from dam for recording on post-natal day 3, 5, and 8.
(B) Frequency distribution of mouse pup vocalizations on day 8 in wild-type littermates from the Gnptabwt/wt mice (green, n = 20) and Gnptabmut/mut mice (red, n = 19).
(C) Sonogram of a wild-type mouse pup vocalizing in isolation. Lower window is an expanded version of one portion, showing the repeated structure in the mouse vocalizations. Red bars indicate a detected vocalization.
(D) Sonogram of a Gnptabmut/mut as in (C).
Mouse Vocalizations: Rate, Spectral Characteristics, and Timing
We first asked whether Gnptabmut/mut mice vocalized. Both wild-type littermates (Gnptabwt/wt) and Gnptabmut/mut mice produced vocalizations over the 3.5-min recording session that appeared to have normal amplitude and spectral characteristics (Figures 1B–1D). We found that Gnptabmut/mut mice exhibited significantly fewer vocalizations compared to littermate controls (Gnptabmut/mut 64.2 ± 34.8 vocalizations; Gnptabwt/wt 184.5 ± 39.4 vocalizations; t test, p < 0.034; Figure 2A) during the recording.
Figure 2Differences in the Vocalizations of Mice with a Mutation in the LETP Compared to Wild-Type Littermates
(A) Vocalizations per recording in mouse pup vocalizations in wild-type littermates (Gnptabwt/wt, green, n = 20) and mice homozygous for the Glu1179Lys mutation (Gnptabmut/mut, red, n = 19). Note the significant decrease in the Gnptabmut/mut mice compared Gnptabwt/wt littermates (t test, p < 0.034). Data on a log scale.
(B) Duration of vocalization in Gnptabwt/wt and Gnptabmut/mut mice. No significant difference was found.
(C) Pause duration between vocalizations, with a significant increase in the duration of pauses in the Gnptabwt/wt compared to the Gnptabmut/mut mice (t test, p < 0.020).
(D) Proportion of long pauses with a steadily increasing criterion for long pauses. Gnptabmut/mut mice had significantly more long pauses.
Each dot represents the mean of one subject. Error bars represent SEM across individuals. Color scheme as in panels (A)–(C). See also Figures S2 and S3.
The difference in rate of vocalizing in the homozygous mutants could be due to the length of vocalizations increasing, the length of pauses increasing, or a mixture of these. We found no significant difference in the duration of the vocalizations between the two groups of mice (t test, p > 0.5; Figure 2B). However, there was a significant difference in the mean pause length between vocalizations (Gnptabmut/mut 6.18 ± 1.23 s; Gnptabwt/wt 2.72 ± 0.69 s; t test, p < 0.02; Figure 2C). This increase in the mean pause length in the Gnptabmut/mut could be due to increased duration of all pauses or an increase in a specific subset of pauses. Much like humans, mice exhibit "bouts" of vocalizations, separated by longer pauses (see Figures 1B and 1C). We found that the Gnptabmut/mut showed no difference in the length of intra-bout pauses, those short pauses between vocalizations inside a bout (Figure S2 and Experimental Procedures). Similarly, Gnptabmut/mut had a similar proportion of large pauses when the criterion for large pauses was not stringent (less than 0.8 s). However, using more stringent criteria for large pauses (setting the cutoff to 0.8 s or higher), the proportion of long pauses was increased in the Gnptabmut/mut mice (Figure S3). This result suggests that the key factor contributing to the reduced number of vocalizations in Gnptabmut/mut animals compared to their wild-type littermate controls was an increase in the duration of long pauses between bouts of vocalization.
We analyzed the same features of the vocalizations from heterozygous Gnptabmut/wt littermates and found them to be more similar to the wild-type Gnptabwt/wt mouse vocalizations (Figure S3). We also found that the vocalization anomalies of the Gnptabmut/mut animals became more pronounced from day 3 to day 8 (Figure S3).
To obtain further insight into the changes in number of vocalizations, we also examined the number of bouts per recording and the number of vocalizations per bout in the Gnptabmut/mut mice. As expected from the increase in duration of long pauses, the number of bouts per recording was smaller in the Gnptabmut/mut mice (Gnptabmut/mut 14.74 ± 4.3 bouts; Gnptabwt/wt 45.35 ± 7.8 bouts; t test, p < 0.0023) (Figure 3A). There was no difference in the average number of vocalizations per bout (Gnptabmut/mut 3.0 ± 0.3 vocalizations; Gnptabwt/wt 3.6 ± 0.3 vocalizations; t test, p > 0.65; Figure 3B). In the Gnptabmut/mut mice, there was a small but significant increase in the percentage of bouts that contained only one vocalization compared to their wild-type littermates (Gnptabmut/mut 45.5% ± 0.04%; Gnptabwt/wt 34.2% ± 0.03%; p < 0.029). The percentage of bouts that contained two vocalizations was slightly smaller in the homozygous mutant mice (Gnptabmut/mut 13.0% ± 0.02%; Gnptabwt/wt 20.0% ± 0.03; p < 0.05). There was no difference in the percentage of bouts that contained three, four, or five syllables (Figure 3C).
Figure 3Bout Structure in Gnptabmut/mut Mice Vocalizations Compared to Gnptabwt/wt Mice
(A) Number of bouts per recording in mouse pup vocalizations in wild-type littermates (Gnptabwt/wt, green) and mice homozygous for the Glu1179Lys mutation (Gnptabmut/mut, red). Note there was a significant difference between Gnptabwt/wt and Gnptabmut/mut mice (t test, p < 0.0023). Error bars represent SEM across individuals.
(B) No significant difference was found in the number of vocalizations per bout.
(C) Percentage of bouts containing one (Gnptabmut/mut, red, 45.5% ± 0.04%; Gnptabwt/wt, green, 34.2% ± 0.03%; p < 0.029), two (Gnptabmut/mut, 13.0% ± 0.02%; Gnptabwt/wt, 20.0% ± 0.03; p < .05), three, four, or five vocalizations.
Mouse Vocalizations: Usage and Temporal Sequencing of Syllables
In humans, some of the disfluencies in stuttering consist of silent blocks, but others—such as sound or syllable repetitions—are voiced. To ascertain whether Gnptabmut/mut mice showed such abnormalities in their vocalizations, we analyzed both the usage and temporal sequencing of syllable types as categorized by an established classification scheme based on presence and size of abrupt pitch jumps [
] (Figure 4A). We chose this scheme because it was initially introduced based on quantitative observations of clustering, which makes it easy to automate and therefore unbiased. Several other syllable classification schemes have been used [
Grimsley J.M.
Gadziola M.A.
Automated classification of mouse pup isolation syllables: from cluster analysis to an Excel-based "mouse pup syllable classification calculator".
Front. Behav. Neurosci. 2012; 6: 89
]; these alternative schemes differ primarily in the classification of syllables lacking pitch jumps, but they also exhibit many points of commonality. Wild-type Gnptabwt/wt and knockin Gnptabmut/mut mice were indistinguishable in the syllable types and their usage (see Experimental Procedures, p > .39, Figure S4). This indicates that these mutations do not affect the ability to produce different syllable types.
Figure 4Mouse Vocalizations from Gnptabwt/wt Compared to Vocalizations of Gnptabmut/mut
(A) Syllable identification scheme showing examples of each type of syllable.
(B) Percentage of each type of syllable in wild-type and knockin mice. Differences were not statistically significant. Each color represents one syllable type.
(C) Percentage of times that one syllable type was followed by the same syllable type. Differences were not statistically significant.
(D) Percentage of syllables that were of the same type as the mode of each bout, for bout sizes ranging from 1 to 20. Data are from Gnptabwt/wt and Gnptabmut/mut. Shaded areas represent 95% confidence intervals.
(E) Diversity of vocalization sequences as quantified by the entropy of the corresponding first-order Markov process (t test, p < 0.022).
Error bars indicate the SEM. See also Figures S4 and S5.
We next looked at syllable repetition. Wild-type mice normally exhibit a strong tendency toward syllable repetition, with doublets (defined as two of the same syllable type in a row, in bouts of two or more syllables) occurring 38% of the time. We found no significant increase in the presence of doublets (Figure 4C) in the vocalizations of the Gnptabmut/mut mice. However, within bouts, Gnptabmut/mut mice exhibited a higher percentage of syllables of a single type, suggesting that Gnptabmut/mut mice might be more stereotyped in their vocalizations (Figure 4D). We tested this possibility using a first-order Markov process model to compare the entropy present in the temporal structure of these vocalizations and found a tendency toward increased repetition in the vocalizations of the Gnptabmut/mut animals (t test, p < 0.022; Figure S5), indicating an overall reduction in temporal diversity. Consequently, Gnptabmut/mut mice use all syllable types in proportions similar to wild-type but exhibit higher stereotypy in temporal sequencing.
Non-vocal Phenotypes
We also asked whether phenotypic differences between wild-type Gnptabwt/wt and homozygous knockin Gnptabmut/mut mice were specific to vocal behavior. Gnptabmut/mut pups exhibited normal weights (Figure S5).
We next examined adult wild-type and homozygous knockin mice using a wide variety of behavioral tests. Mice were evaluated first on a 1-hr locomotor activity test. There was a significant decrease in ambulatory activity (total ambulations) on the part of the Gnptabmut/mut mice across the 60-min test session (Figure 5A; p = 0.0498). In contrast, no differences were found between groups with regard to vertical rearing frequency, or time spent, entries made, or distance traveled in the central zone of the test field.
Figure 5Non-vocal Behavioral Test Results from Gnptabwt/wt and Gnptabmut/mut Mice
(A) Ambulatory activity was significantly decreased (∗p < 0.05) in mice homozygous for the Glu1179Lys mutation (Gnptabmut/mut, red, n=15) compared to wild-type (Gnptabwt/wt, green, n=15) mice with differences being greatest during block 6 (∗∗p < 0.018).
(B–D) The two groups of mice did not differ on several sensorimotor tests such as the time spent on an elevated ledge (B), the time to climb down a pole (C), or the time spent hanging upside down on an inverted screen before falling (D).
(E and F) The Gnptabwt/wt and Gnptabmut/mut mice also exhibited similar magnitudes of the acoustic startle response (arbitrary units) (E) and PPI (F).
(G) Time spent on accelerating rotorod before falling. No significant differences were found.
(H) Path length on Morris water maze for place trials. No significant differences were found.
(I and J) The performance of the Gnptabmut/mut mice was not impaired during the spatial learning (place) trials (I) nor in terms of retention performance during a probe trial where each group showed spatial bias for the target quadrant (Tgt) by spending significantly more time in it versus the times spent in each of the other quadrants (p < 0.00005). (J) The Gnptabwt/wt and Gnptabmut/mut mice showed comparable levels of sociability in that each group spent significantly more time in an investigation zone surrounding a stimulus mouse that remained inside a withholding cage compared to time spent near an empty cage (Emp) (p < 0.00005), although there were no differences between the two groups for either of the two conditions.
(K) For each group, the duration of hole pokes was significantly greater for the odorant-containing (Odr) versus the empty (Emp) holes (p < 0.024), although the Gnptabwt/wt mice had significantly higher poke durations on average across the two hole types (∗p < 0.029).
(L) The two groups did not differ in levels of anxiety-related behaviors in the elevated plus maze as exemplified by the lack of significant effects involving genotype on the percentage of distance traveled in the open arms out of the total distances traveled in all of the arms.
(M) The absence of any significant effects involving genotype suggested that there were no differences in contextual fear conditioning between the groups.
Error bars indicate the SEM.
The mice were then tested on a battery of sensorimotor measures to assess possible functional deficits. No performance differences were observed between groups on the ledge (Figure 5B), pole (Figure 5C), inverted screen tests (Figure 5D), or on the walking initiation, platform, or inclined screens tests. These results suggest that balance, strength, coordination, and initiation of movement were not grossly disturbed in the Gnptabmut/mut mice. These animals and their wild-type littermates also performed equivalently on tests of acoustic startle (Figure 5E), prepulse inhibition of startle (PPI) (Figure 5F), and rotarod tests (Figure 5G).
Spatial learning and memory capabilities of the Gnptabmut/mut mice were assessed using the Morris water maze task with a computerized tracking system as previously described [
Wozniak D.F.
Hartman R.E.
Boyle M.P.
Vogt S.K.
Brooks A.R.
Tenkova T.
Young C.
Olney J.W.
Muglia L.J.
Apoptotic neurodegeneration induced by ethanol in neonatal mice is associated with profound learning/memory deficits in juveniles followed by progressive functional recovery in adults.
Neurobiol. Dis. 2004; 17: 403-414
]. Cued (visible platform, variable location) and place (submerged, hidden platform, constant location) trials were conducted. No significant effects involving genotype were found (Figures 5H and 5I).
We next examined the sociability of the mice as assessed by measuring the time investigating a conspecific stimulus mouse that was loosely sequestered in a small withholding cage compared to the investigation times observed for an empty cage and found no significant differences between genotypes [
Dougherty J.D.
Maloney S.E.
Rieger M.A.
Sonnenblick L.
Coppola G.
Mahieu N.G.
Zhang J.
Cai J.
Patti G.J.
The disruption of Celf6, a gene identified by translational profiling of serotonergic neurons, results in autism-related behaviors.
] (Figure 5J).
Mice were next evaluated for differences in exploratory behaviors and olfactory preference using a previously published procedure [
Diggs-Andrews K.A.
Conyers S.
Yuede C.M.
Dearborn J.T.
Brown J.A.
Tokuda K.
Izumi Y.
Zorumski C.F.
Gutmann D.H.
Motivational disturbances and effects of L-dopa administration in neurofibromatosis-1 model mice.
], where hole poking with the nose served as the main behavioral response. The apparatus contained four corner and four side holes in the floor. One corner hole contained a familiar odorant, and the diagonally opposite corner hole contained a novel odorant. Gnptabmut/mut and control mice performed similarly on several test variables including the frequency of nose pokes made into the side or corner holes, pokes made into odorant-containing versus empty corner holes, and pokes made into the corner holes containing a novel versus a familiar odorant. However, wild-type mice showed a significant preference for the hole containing the familiar odorant (p = 0.006), while the Gnptabmut/mut mice did not show a significant preference for the familiar odorant, though there was a trend in that direction (p = 0.11). In addition, the Gnptabmut/mut mice had decreased poke durations on average across empty and odorant-containing holes as indicated by a significant genotype effect (Figure 5K; p = 0.028). The Gnptabmut/mut and wild-type groups each had significantly longer poke durations for the odorant-containing holes versus the empty ones (p = 0.024 and p = 0.012, respectively).
People who stutter (PWS) have been shown to have a higher incidence of anxiety disorders [
Menzies R.G.
Onslow M.
Packman A.
Anxiety and stuttering: exploring a complex relationship.
Am. J. Speech Lang. Pathol. 1999; 8: 3
]. Therefore, we assessed anxiety-like behaviors using the elevated plus maze (EPM) and fear conditioning. Analysis of the classic open arm variables in the EPM including time spent, entries made, and distance traveled in the open arms did not yield any significant effects involving genotype (Figure 5L). We also evaluated the performance of Gnptabmut/mut mice on a fear-associated Pavlovian conditioning test and found no genotype differences (Figure 5M).
Human Vocalizations
Given that the deficits of mice were primarily vocal in nature, we compared these vocal deficits with those of PWS. Stuttered and normal speech have been compared using a wide variety of language-based and acoustic analyses [
Czyzewski A.
Kaczmarek A.
Kostek B.
Intelligent processing of stuttered speech.
J. Intell. Inf. Syst. 2003; 21: 143-171
Healey E.C.
Ramig P.R.
Acoustic measures of stutterers' and nonstutterers' fluency in two speech contexts.
J. Speech Hear. Res. 1986; 29: 325-331
Howell P.
Sackin S.
Glenn K.
Development of a two-stage procedure for the automatic recognition of dysfluencies in the speech of children who stutter: II. ANN recognition of repetitions and prolongations with supplied word segment markers.
J. Speech Lang. Hear. Res. 1997; 40: 1085-1096
Lustyk T.
Bergl P.
Cmejla R.
Evaluation of disfluent speech by means of automatic acoustic measurements.
J. Acoust. Soc. Am. 2014; 135: 1457-1468
Van Riper C.
The Nature of Stuttering.
Prentice-Hall, 1982
Robb M.
Blomgren M.
Formant frequency fluctuation in stuttering and nonstuttering adults.
J. Fluency Disord. 1998; 23: 73-84
Seery C.H.
Watkins R.V.
Mangelsdorf S.C.
Shigeto A.
Subtyping stuttering II: contributions from language and temperament.
J. Fluency Disord. 2007; 32: 197-217
]. To ensure optimal comparability, we applied our mouse-centric analyses to the vocalizations of PWS. Moreover, we were able to analyze vocalizations from normally fluent human controls, PWS, and PWS who carry a well-characterized mutation in one of the three genes encoding the LETP (PWS-L) (Table S1). We thus tested whether the vocal abnormalities found in the Gnptabmut/mut mice could be seen in the more complex vocalizations of humans. We obtained recorded speech samples using a standard 500-word stuttering diagnostic reading passage containing balanced representation of all speech sound classes [
Webster R.L.
Empirical considerations regarding stuttering therapy.
in: Gregory H.H. Controversies about Stuttering Therapy. University Park Press, 1979: 209-239
]. To ensure comparability with mouse data, no language-specific criteria were used to determine vocalization segments. Computer-assigned vocalization segments could therefore include more than one syllable or word.
We compared the rate of vocalizations in PWS versus controls (CT). Consistent with previous studies [
de Andrade C.R.
Cervone L.M.
Sassi F.C.
Relationship between the stuttering severity index and speech rate.
Sao Paulo Med. J. 2003; 121: 81-84
Johnson W.
Measurements of oral reading and speaking rate and disfluency of adult male and female stutterers and nonstutterers.
J. Speech Hear. Disord. 1961; : 1-20
Ryan B.P.
Articulation, language, rate, and fluency characteristics of stuttering and nonstuttering preschool children.
], stuttering case subjects had significantly fewer vocalizations per minute (PWS, 90.2 ± 3.0 vocalizations; CT, 124.9 ± 3.1; mean ± SEM; t test, p < 10−11; Figure 6B). Like the Gnptabmut/mut mice, the PWS-L group also displayed significantly fewer vocalizations per minute than controls (PWS-L, 81.1 ± 4 vocalizations per minute; t test, p < 10−11; Figure 6B). These differences were comparable to the difference found between the Gnptabmut/mut mice and Gnptabwt/wt mice.
Figure 6Vocalizations of Human Controls, PWS, and PWS with a Known Mutation in the LETP, as Measured by a Language-Agnostic Analysis
(A) Sonogram of a person saying "(Breath) One of big…the…one of the big advantages of nylon over" while reading from a standard diagnostic script. Red bars indicate detected vocalization intervals.
(B) Vocalizations per minute in the control subjects reading the script (CT, green, n = 51), PWS (PWS, gray, n = 74), and PWS with a known mutation in the LETP reading the same script (PWS-L, black, n = 26). Each dot represents the mean of one subject. Error bars represent SEM across individuals. Brackets and stars indicate significance. There was a significant decrease in the number of vocalizations in the PWS group (t test, p < 10−11) and in the PWS-L (t test, p < 10−11) group compared to the CT group. Data on log scale.
(C) Duration of vocalizations. Note there was no significant difference in the duration of vocalizations between the CT and PWS groups, while there was a significant difference between the CT and PWS-L groups (t test, p < 0.02).
(D) Mean pause duration between vocalizations, with a significant increase in the PWS group (t test, p < 10−12) as well as in the PWS-L group (t test, p < 10−9) compared to the CT group.
(E) Proportion of long pauses, with a steadily increasing criterion for long pauses, in the recordings of the subjects reading the script using the same color scheme as in (B)–(D), showing that over a range of criteria, PWS in both categories had more long pauses.
Bars indicate SEM. See also Table S1.
An analysis of the durations of vocalizations revealed no significant difference between the general stuttering cases and controls (PWS mean 0.335 ± 0.020 s; CT mean 0.313 ± 0.012 s; t test, p > 0.3). However, there was a significant increase in the duration of the vocalizations in stuttering cases carrying an identified mutation in the LETP (PWS-L mean 0.385 ± 0.033 s; t test, p < 0.02; Figure 6C).
A primary feature of stuttering is involuntary pauses known as blocks, and such pauses have been found to be important in assessing fluency of speech [
]. Like the differences in the pause lengths in the Gnptabmut/mut mice and Gnptabwt/wt mouse vocalizations, the speech of PWS had significantly longer pauses between vocalizations compared to that of control subjects (PWS, 0.385 ± 0.021 s; CT, 0.170 ± 0.005 s; t test, p < 10−12; Figure 6D). This was also true for case subjects carrying an LETP mutation (PWS-L, 0.375 ± 0.038 s; t test, p < 10−9). Moreover, we found a significantly greater proportion of long pauses in both case groups compared to the control subjects over a wide range of cutoff values for defining long pauses (Figure 6E). These data indicate that we can detect a statistically significant difference in specific features of vocalization in PWS using the same methods used to analyze the mouse data. Such measures are "language agnostic" because they do not depend on detailed features (formant structure, vowels and consonants, etc.) of human speech. Using just two such features, number of vocalizations per minute and length of pauses, a k-nearest neighbor classifier could distinguish a PWS from a control subject 79% of the time, significantly better than chance (shuffled data, p < 10−5).
To determine whether these same phenomena apply to stuttering during extemporaneous speech, we also examined the length and number of speech intervals in 80 3-min samples obtained from publicly available podcasts, for both controls and PWS. Consistent with the results from people reading a script, we found that in extemporaneous speech, PWS had fewer vocalizations per unit time compared to controls. This difference was significant (PWS, 274.7 ± 8.4 vocalizations; CT, 434.8 ± 6.5 vocalizations; t test, p < 10−12; Figure 7A). The average length of vocalization also differed significantly between groups (PWS 415 ± 17 ms; CT 258 ± 6; t test, p < 10−10; Figure 7B). We also found that the mean length of pauses between speech intervals was significantly longer in PWS compared to controls (PWS 255 ± 12 ms; CT 160 ± 3; t test, p < 10−8; Figure 7C).
Figure 7Extemporaneous Vocalizations of Human Controls and PWS
(A) Vocalizations per minute in podcast segments from controls (CT, green, n = 68) and PWS (PWS, black, n = 28; t test, p < 10−21). Each dot represents the mean of one subject. Error bars represent SEM across individuals.
(B) Duration of vocalization periods. Note the significant increase in PWS group (t test, p < 10−12) compared to the CT group.
(C) Pause duration between vocalizations, with a significant increase in the duration of pauses in the PWS group (t test, p < 10−17) compared to the CT group.
(D) Proportion of long pauses with a steadily increasing criterion for long pauses, in the recordings of the control subjects reading the script (CT, green) and PWS (ST, black). Bars indicate SEM. These results are similar to that of reading a script Figure 6.
Vocal Alterations: Mouse
Identification of some of the genetic underpinnings of human stuttering has created an opportunity to study whether homologous mutations could affect vocalization in a genetically tractable animal model. We found that the vocalizations of Gnptabmut/mut mice were significantly different than those of their Gnptabwt/wt littermates. Gnptabmut/mut mice had fewer vocalizations per unit time and showed an increased incidence of long pauses. Gnptabmut/mut mice produced all the same syllables as their wild-type littermates and used them in similar proportions. We note that these conclusions are specific to the syllable classification scheme adopted here, and it remains possible that other classification schemes would come to a different conclusion. From manual inspection of the sonograms, at present, we have not seen evidence leading us to suspect any kind of difference in syllable repertoire.
However, by two measures, mice with an LETP mutation showed more temporal stereotypy in their vocalizations. First, during bouts of vocalization, a higher percentage of syllables were of a single type. Second, the entropy of the first-order Markov process was reduced, indicating an overall reduction in temporal diversity.
Vocal Alterations: Human
The techniques used to analyze mouse vocalization were validated by applying them to human speech from PWS, PWS with a mutation in the LETP, and controls. Stuttered speech has been extensively analyzed using a variety of measures including words per minute, voice onset time, formant frequencies of consecutive glottal periods, and other language-based acoustic techniques [
]. Previous studies have demonstrated that stuttering is strongly correlated with fewer vocalizations per unit time and an increase in pause length [
]. We confirmed and extended these results using language-agnostic measures designed to be generalizable to mice. In addition, we found that using just three such language-agnostic features (number of vocalizations per minute, length of syllables, and length of pauses), we could classify a recording as coming from a PWS or control.
Stuttering is heterogeneous disorder, and there have been many proposed subtypes of stuttering [
Yairi E.
Subtyping stuttering I: a review.
]. Here, we have analyzed the speech of a subset of PWS with a genetically identified etiology and compared it to the speech of a general group of PWS. Our results show that using our language-agnostic analyses, PWS with a known mutation in the LETP showed similar abnormalities as those displayed by PWS of an unknown etiology.
Vocal Alterations: Comparison between Mouse and Human
Human speech is produced by air flow-induced vocal fold vibrations [
Honda M.
Human speech production mechanisms.
NTT Tech. Rev. 2003; 1: 24-29
]. In contrast, mouse vocalizations are thought to be a form of whistle [
Johnson A.M.
Ciucci M.R.
Russell J.A.
Hammer M.J.
Connor N.P.
Ultrasonic output from the excised rat larynx.
J. Acoust. Soc. Am. 2010; 128: EL75-EL79
]. Many types of comparisons between human speech and mouse vocalization are unlikely to be informative due to anatomical differences and the complex and communicative nature of human speech. In the case of stuttering, it is important to note that while stuttering may manifest as deficiencies in the control of vocal tract movements, there is no evidence that these are the proximal cause of the disorder. In fact, there is evidence that persons who are deaf or hearing impaired stutter when using sign language, suggesting the cause of stuttering resides elsewhere [
Montgomery B.M.
Fitch J.L.
The prevalence of stuttering in the hearing-impaired school age population.
J. Speech Hear. Disord. 1988; 53: 131-135
Snyder, G. (2006). The existence of stuttering in sign language and other forms of expressive communication: Sufficient cause for the emergence of a new stuttering paradigm? In International Stuttering Awareness Day Online Conference, J.M. Kuster, ed. (Minnesota State University).
]. In addition, mouse vocalizations require control of airflow, subglottal pressure, and glottal adduction, and these share some common neural pathways with human vocal production [
]. Moreover, there is evidence linking these more cognitive pathways to stuttering [
Fox P.T.
Ingham R.J.
Ingham J.C.
Zamarripa F.
Xiong J.-H.
Lancaster J.L.
Brain correlates of stuttering and syllable production. A PET performance-correlation analysis.
Brain. 2000; 123: 1985-2004
Rosenbek J.
Messert B.
Collins M.
Wertz R.T.
Stuttering following brain damage.
Brain Lang. 1978; 6: 82-96
GNPTAB mutations exhibit several common features in both organisms. PWS, and specifically subjects carrying an LETP mutation (PWS-L), showed fewer vocalizations per unit time and as fluency blocks, which acoustically correspond to large pauses. Also like the mouse, the tendency toward syllable repetition is a defining characteristic of human stuttering. That some of the prominent features of human stuttering are recapitulated by the mouse model suggests that such a model may be useful in discovering the circuit and neurochemical underpinnings of at least some aspects of stuttering. One interpretation of these results is that a primary deficit in stuttered speech is the inability to initiate vocalization sequences and that this specific aspect of the behavior is shared between organisms and can be disrupted by similar genetic means.
It is worth noting that mice are deaf during the post-natal period until at least P10 depending on the frequency [
Development of absolute auditory thresholds in the house mouse (Mus musculus).
J. Am. Audiol. Soc. 1976; 1: 179-184
]. Auditory feedback in humans has been alternatively shown to reduce or increase stuttering [
Kalinowski J.
Armson J.
Roland-Mieszkowski M.
Stuart A.
Gracco V.L.
Effects of alterations in auditory feedback and speech rate on stuttering frequency.
Lang. Speech. 1993; 36: 1-16
]. However, with humans, being able to hear is not necessary for stuttering behavior. As mentioned previously, it has been found that people who are hard of hearing and people who are deaf stutter, both in oral speech and sign language [
We also noted some differences between human stuttering and the mouse model. There have been several cases of PWS who are heterozygous for a mutation in the LETP; in contrast, we found that the vocalizations of heterozygous mice were more similar to those of wild-type controls. In humans, repetition of a single syllable is unusual and strongly indicative of stuttered speech; in mice, syllable repetition is commonplace, with the consequence that only longer sequences of repetition exhibited significant differences. Finally, an important outstanding question is whether the Gnptabmut/mut mice were attempting to vocalize during the abnormal pauses or simply remaining quiet.
Non-vocal Behaviors
We examined sensory-motor function and learning in a series of non-vocal behavioral paradigms. We ran an exhaustive behavioral screening on these animals, including 50 statistical comparisons. We found that Gnptabmut/mut mice were normal over the large majority of these measurements. Two measures that did exhibit differences were general ambulatory activity (p = 0.007 [corrected for six bins, p = 0.042] during the final time bin, or p = 0.049 over the entire time interval) and exploratory behaviors related to hole pokes (p = 0.01–0.03). This is not significantly better than chance (one would expect 2.5 tests to pass this criterion by chance). Also, no single p value was sufficiently strong to survive Bonferroni correction for multiple comparisons. With such an extensive battery of tests, we therefore conclude that our finding appears to be a specific abnormality in vocalization. An alternative interpretation of these findings is that Gnptabmut/mut mice may show reduced rate of certain spontaneous behaviors.
Our data show that mutations in the LETP affect mouse pup isolation calls and human speech in similar ways, suggesting that there is a commonly disrupted vocalization function in both. We suggest that this animal model could allow the large array of genetic and neurobiological tools that exist in mice to be applied to the study of the cellular and molecular features of stuttering and perhaps normal speech.
Studies were conducted under institutionally reviewed and approved animal study protocols (NIDCD/NIH #1318-13 [D.D., Principal Investigator] and Washington University protocols 20130179 [T.E.H., Principal Investigator] and 20120110 [D.F.W., Principal Investigator]). Knockin mice were constructed on a BALB/c background using the strategy shown in Figure S1. To generate test subjects, heterozygous mice were mated. Females were separated from all other mice before they gave birth. Dams were checked for pups daily, and the first day that a litter was discovered was considered P0. Mice came from a total of 18 litters. We recorded from mice on P3, P5, and P8. Mouse data were collected blind to genotype.
Analysis of Vocalizations
All analyses were done using in-house MATLAB software, available online at http://holylab.wustl.edu/. Analysis code implemented a fully automated algorithm and was therefore blind to genotype. For mice, waveforms were pre-processed and band-pass filtered (25–110 kHz), and vocalizations were identified using mean frequency, "spectral purity" (fraction of total power concentrated into a single frequency bin), and the "spectral discontinuity" (the change in the allocation of power across frequencies between two adjacent time bins) [
]. Stored acoustical waveforms were processed using MATLAB to compute the sonogram (512 samples/block, half-overlap, resulting in a time resolution of 1.02 ms and a frequency resolution of 0.98 kHz). We also examined broad band "clicks" in the mouse vocalizations that were defined as non-contiguous milliseconds where less than 200 of the 512 sampled half frequencies were empty. No difference in the number of clicks was detected between groups. Stored acoustical waveforms were processed in MATLAB to compute the sonogram (512 samples/block, half-overlap, resulting in a time resolution of 1.02 ms and a frequency resolution of 0.98 kHz).
For humans, vocalizations were identified in terms of supra-threshold power in either of two frequency bands (0.6–1.6 kHz and 5.6–9.6 kHz). Vocalization periods separated by gaps shorter than 20 ms were merged. Comparison with manual scoring demonstrated that breathing and other non-speech noises were excluded (Figure 5A). We examined the length and number of speech intervals and non-speech intervals (pauses between speech) in the speech samples. These intervals did not necessarily have only one word; many times words were run together without an appreciable non-speech interval.
To calculate the number of syllables and the duration of syllables and pauses, each vocalization or pause contributed to the mean for each animal or subject; each individual's mean was then averaged to obtain the group average. A t test was then performed to compare groups with each individual's mean. An alternative analysis, where each syllable contributes to the overall mean, was also performed and produced comparable results, not shown.
For the syllable analysis, the definitions set forth in Arriaga et al. were used, with the exception of classes "i, j, k," which were grouped into the category "other" [
]. This classification was done using a fully automated algorithm. To determine whether the usage frequency differed significantly between genotypes, we first picked a random pair of recordings from the Gnptabmut/mut mice. We performed a bootstrap analysis of the differences in syllable usage between two individual mice, measuring the difference by
D=∑s=19(a˜s−b˜s)2
where s is the syllable type index, and a˜s and b˜s are the normalized numbers of observed or predicted syllables of type s. The normalization was performed in terms of A=∑as and B=∑bs (where variables without ∼ denote raw counts), scaling a˜s=(B/max(A,B))as and b˜s=(A/max(A,B))bs. As a consequence, the recording with more syllables was scaled to a number equal to that of the recording with fewer syllables, and the one with fewer was unmodified. The bootstrap was calculating the mean difference D among randomly selected pairs of Gnptabmut/mut mice and then comparing this mean against the distribution of mean D obtained from an identical number of wild-type/mut pairs, with wild-types resampled to match the number of syllables produced by mut animals.
We used a t test to compare the number of doublets and found no difference between experimental groups. Bout-level analyses defined bouts based on histograms of pause lengths for all groups of mice in each day we recorded. Histograms were constructed with a range of bin sizes (50 ms to 300 ms). The middle of minimum bin in the range of 0.15 to 0.33 s was averaged across all bin sizes to determine the criteria for an inter-bout pause. The resulting minimum intra-bout/inter-bout cutoff was determined to be 0.33 s in day 3 recordings, 0.28 s in day 5 recordings, and 0.22 s in day 8 recordings. To analyze bout-level repetition, the syllables in each bout were analyzed to determine how many of the syllables in each bout were the same type as the mode of that bout. In Figure 4D, the confidence intervals were calculated by bootstrapping bouts; bout sizes with fewer than three exemplars are depicted only in terms of their mean. Entropy of syllable usage was calculated from the proportion of different syllable types; entropy for the temporal sequence (modeled as a first-order Markov process) was given by H2 = −∑ p(X) ∑p (X|Y) log2p (X|Y), with X and Y being each syllable type [
For the classification analysis, the number of vocalizations per minute, the average duration of each vocalization, and the average duration of each pause were normalized to a range of 0–1 and plotted in three-dimensional space. We then categorized each recording as being either from a control or person who stutters based on nearest neighbors. Choices of one, three, and five closest neighbors all yielded very similar results. We analyzed three cases: (1) all recordings from controls and PWS; (2) controls and PWS with an unknown cause; and (3) controls and PWS with a known mutation in the LETP. Data in all cases were significant compared to empirically calculated p value of 0.05 (data labels [e.g., controls, person who stutters] shuffled 10,000 times, 95% confidence intervals).
T.D.B. designed experiments, gathered mouse vocalization data, analyzed human and mouse vocalization data, and wrote the paper. D.F.W. directed the non-vocalization behavioral studies, which involved statistical and graphical analyses of the results, and wrote the paper. T.-U.H. provided comments on the manuscript and made Figure S1. D.D. had the original idea for the paper, recruited human subjects, designed knockin mice, supervised experiments, and wrote the paper. J.G. recruited subjects and obtained speech recordings from scripts. T.E.H supervised mouse experiments, analyzed data, and wrote the paper.
We thank the Hollins Communication Research Institute for assistance with subject recruitment and stuttering diagnosis. We thank Peter Rietzes of StutterTalk for help with speech samples from PWS. We thank Grace Liao and Cassandra Roberts for help with vocal recordings of mice; Wayne Barnes and Thanh Loan Nguyen for their help with mouse genotyping; and Sara Conyers for her work involving the non-vocalization behavioral testing. This work was supported by the NIH Director's Pioneer Award (DP1 OD006437), Intellectual and Developmental Disabilities Research Center grants (P30 HD062171; U54 HD087011), and NIDCD Intramural grant Z1A-000046-15.
Document S1. Supplemental Experimental Procedures, Figures S1–S5, and Table S1
Received in revised form: February 18, 2016
Received: December 16, 2015
© 2016 Elsevier Ltd.


Almost Most of people likes and loves movies and Best Hd Movies Point is the place where users can reach and download their favorite Full HD 720p Movies for Mobile, Tablet PC's and PC's. We always try to provide our best to deliver the best rated and best quality collection.
The Most of our Provided movies include, Bollywood 720p Movies, Hollywood Movies and Hindi Movies.
Moreover BesthdMoviesPoint always provide very safe to use download links to Download Free HD Movies.


A beautifully presented and architecturally designed three bedroom garden apartment providing excellent lateral space over the entire ground floor of this elegant white stucco fronted building in an enviable position. The apartment has its' own private entrance providing in excess of 1,900 sq. ft. with stylish kitchen/dining room with underfloor heating and beautiful reception room which both lead out to a lovely south facing mature garden. The master bedroom benefits from a luxury en suite bathroom, fitted wardrobes and an additional walk in wardrobe which provides ample space for a study or nursery, two further good size double bedrooms have use of a family bathroom and there is also an additional guest WC. This apartment is simply outstanding throughout and is offered unfurnished from the beginning of July.


Orange white green stripes mango wood rustic nightstand. Wood and metal bedside tables rustic white nightstand. Pink wooden c shaped nightstands for brown lamp shade. Nightstands interesting wooden nightstands high resolution. Woodbury night stand farmside wood bedroom furniture. 90% off rustic wood nightstand / tables. Belcourt cherry nightstand nightstands dark wood.
Nightstand friendly white nightstand ikea decor: white. Red nightstands, pair. Nightstand mini fridge bedroom galerry. #hat @home accents today magazine ty for including emma. Fairfield lane cherry nightstand nightstands dark wood. Ethan allen georgian court cabinet nightstand 11 5216. Purple nightstands antique chalk painted side table end. Purple nightstands foter. Reclaimed barn wood furniture at the galleria. Captivating colorful nightstands also new colorful bedroom. Mirrored nightstands and dressers, mirrored dresser pier.
Santa cruz cherry nightstand nightstands dark wood. 247 best images about purple painted furniture on. Black wood nightstand steal a sofa furniture outlet los. Talbot wood two drawer nightstand in driftwood humble abode. Great large night stand tables black nightstand walmart. 28quot; transitional three drawer nightstand in driftwood mathis brothers furniture. Black nightstand set maralynchaseorg.