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The juice extraction from fruits and vegetables can be done by size reduction methods as crushing, chopping and comminuting. This is usually employed for fruits that are fully ripened and too soft for whole or diced or sliced packs. The following
Nov 13, 2018 INTRODUCTION Fruits play an important role in human nutrition. Before the 20th century, drinking squeezed fruit juices was the privilege of only a few. Welch was the first to preserve grape juice with heat treatment in America in 1869, followed by Muller-Thurgan in Switzerland in 1896 Thus began the production of preserved fruit juice A wide ...
Juice extraction Filtration (optional) Batch preparation Pasteurisation Filling and bottling. Any fruit can be used to make fruit juice, but the most common ones include pineapple, orange, grapefruit, mango and passion fruit. Some juices, such as guava juice, are not filtered after extraction and are sold as fruit nectars.
percentage of fresh apples (from 55 to 63%) and a lower percentage of juice, canned, dried, and frozen apple products (USDA-NASS, 2003). World apple juice production is expected to remain strong, with record production in China, which in 20022003 accounted for 33% of the world production. This is more than double the
40.1 Introduction. Juice extractionthe elimination of the juice from fibrous, solid particlesis a basic technological step of fruit juice production. The fruit has to be prepared prior to juice extraction, which is then followed by juice clarification and drink completion. Subsequently, the finished drink is packed and preserved.
Fruit can be pulped in a liquidiser. A range of fruit presses are available to extract the juice. Some machines combine pressing with filtration to remove the fine particles. To make fruit squash or cordial, the extracted fruit juice is mixed with sugar syrup to give a final sugar concentration of 12-14%. See added ingredients.
Fruit peeling and extracting machine. This type of fruit juice extractor machine can accomplish fruit peeling, coring and extracting at the same time. It can be applied to oranges, pineapples, lemons, and grapefruits. Extracting process. When the washed fruits are fed into the extractor through the hopper, 2 rollers transmit them to the knife.
The production process of fruit juice and concentrates begins with raw material purchasing. Fruit is verified in terms of quality before unloading. It should look healthy and be ripe. There are two unloading methods wet (known as hydro-unloading) and dry. Both methods are used for unloading and transporting apples to silos of pre-production ...
GOTMORE 4 in 1 Large Juicer Machines with Mixing/Micing/Grinding Function, 1300W, 5-Speed, 84mm Wide Mouth Centrifugal Juicers Easy Clean Juice Extractor for Whole Fruit Vegetable, Titanium Filter, Anti-drip, Non-Slip Feet, BPA-Free. 4.6 out of 5 stars. 3. $199.00.
Apr 06, 2021 The screw type industrial juice extractor uses the change of the gap between the screws to squeeze the fruit to obtain the juice. This type of juicer is the most widely used juicer for fruits, and is suitable for squeezing the juice of fruits and vegetables such as apples, pears, pineapples, tomatoes, coconuts, pomegranates, peaches and all kinds of vegetables.
give juice products the often desired pulpy mouth feel. There is an increasing demand for Brown Extractor quality pulp. This family of Brown extractors is typically installed in lines of 8 to 14 machines of which several are configured to accommodate each fruit size. Each size extractor tolerates some overlap of fruit sizes making ...
The process of jelly making is also similar except that only fruit extract is used for making of jelly and pectin is added for gel formation after boiling. 3.3 Squash Squashes are sweetened juice of fruits containing minimum prescribed quantity of pulp. As per Indian Standards, squash should contain at least 25 per cent (by volume) of fruit juice.
INDUSTRIAL JUICER MACHINE. With the ZMF400 you will be able to cold press fruit and vegetable juices, with or without pasteurizing, to offer always the best quality juice with a machine that keeps pace your business.
Juice can also be extracted by using a screw type juice extractor. Common equipment used for juice extraction are fruit grater or mill, basket or hydraulic press, screw type juice extractor, rosing or burring machine, fruit pulper etc. There are two types of extraction methods i.e., single and double operation system.
Introduction Fruit Juice Making Machine is mainly used for squeezing the fruit and vegetable juices. Widely used for squeezing watermelon, grape, strawberry, pear, apple, lemon, orange, tomato, papaya, carrots, ginger and other fruit and vegetable juices. Suitable for Fruit and vegetable processing equipment, frozen food factory equipment, leisure food factory
Features of Fruit Juice Machine. 1.The juice press machine is made of stainless steel, healthy, durable and easy to clean. 2. The industrial juicer adopts screw extrusion principle, having high juice yield. 3. With crushing device mounted, the machine crushes the materials first before extracting juice. There is no need for the extra crusher.
The Fruit Juice and Fruit Nectars (Scotland) Regulations 2013 SI 2013 No. 305 The Fruit Juice and Fruit Nectars (Northern Ireland) Regulations 2013 SI 2013 No. 253 The main differences in the regulations are with regard to enforcement application. The Regulations lay down compositional standards for fruit juice and certain similar products,
Chapter 8 Fruit specific preservation technologies 8.1 Fruit quality 8.2 Harvesting and preprocessing 8.3 Fresh fruit storage 8.4 Fruit drying and dehydration technology 8.5 Technology of semi-processed fruit products 8.6 Fruit sugar preserves technology jams, jellies, marmalade, fruit paste 8.7 Fruit juice technologies
Electric Fruit Juice Extractor Machine Vegetable Blender Squeezer Juicer Maker. $38.99. Free shipping. Free shipping Free shipping. Centrifugal Electric Juicer Fruit Veg Extractor Juice Maker Machine 3 Speeds. $45.99. $59.99 previous price $59.99 previous price $59.99. Free shipping Free shipping Free shipping. Picture Information.
The aim of the juice extraction process (see Figure 5.4) is to obtain as much juice from the fruit as possible while preventing rag, oil and other fruit components from entering the juice.These may lead to bitterness in taste or other defects during subsequent juice storage. The extraction operation determines product quality and yield and thus has a major effect on the total
Find here online price details of companies selling Fruit Juice Machine. Get info of suppliers, manufacturers, exporters, traders of Fruit Juice Machine for buying in India.
Oct 25, 2016 Jam marmalade The method of preparation for jam marmalade is practically the same as that for jelly marmalade. In this case the pectin extract of fruit is not clarified and the whole pulp is used. Sugar is added according to the weight of fruit, generally in the proportion of 11. The pulp-sugar mixture is cooked till the TSS content reaches 65 ...
As the name implies, fruit juice extractor machine is used to extract juice from different kinds of fruits. with the same working principle, fruit juice extractor machine processes fruits into juice, which is convenient and nutritive. Almost all the fruits can be extracted by fruit juice extractor machine, such as apple, pineapple, pear, orange ...
to prepare and package fruit beverages such as Squashes, RTS, Nectar, Cordial, Crush and Syrup by using appropriate machines such as pulper, juice extractor, autoclave, and corking machine with safety precautions, determine the acidity and
The passion fruit juice machine consists of a hopper, crusher, separator tube, sub-seed tube, transmission, chassis and other components. When start working,the passion fruit first passes through the crusher, and then into the separator tube to separate skin, juice. Pulp and seeds into the seed sub-cylinder, and then the separate the juice,pulp ...
Japan develops rapidly in fruit juice equipments and takes a place in the worlds market. A complete juice processing line is composed of fruit sorter, washer, elevator, crusher, pulping machine, juice extractor, concentrator, sterilizer, filler, etc. As for different fruits, the production processes are varied.
Natural fruit juice (original juice) drink, fruit juice drink or fruit juice drink with pulp, the basic principle and process of their production are roughly the same. Fruit juice production process mainly includes fruit raw material pretreatment, juice extraction, clarification and filtration, homogenization, deoxidization, concentration ...
The fruit juice can be as juice, concentrate or powder, and honey or sweetener may be substituted for sugar. Minimum juice contents are specified for each type of fruit, which for orange nectar in Europe is 50%. Fruit juice regulations do not cover fruit drinks, which fall under general food legislation.
Feb 09, 2011 Red Eye Add a shot of tomato juice to any ale or lager and a splash of Tabasco Skip and go naked Combine beer, lemon juice and gin, with a dash of grenadine. 41. Red wine is the prefered choice of Chinese wine drinkers and is increasingly seen as a status symbol drink among Chinese high society. 80% of wine produced and consumed in ...
May 31, 2017 Juice extraction, clarification, and concentration are critical steps in sugar processing. Below, we discuss what each of these steps entails. Juice Extraction On arrival at the sugar processing plant, sugarcane is loaded onto a conveyor table. It passes through various sugar equipment including a shredder, which uses a series of knives to chop the cane Keep
7 Jam and juice making, syrups, jellies and candied fruit 51 7.1 Making fruit juices 52 7.2 Preparation of other fruit products 58 8 Developing a small-scale food processing enterprise 61 8.1 Marketing a fresh or processed product 62 8.2 Organizing a processing enterprise 64 Further reading 68 Useful addresses 70
Fruit jellies are semisolid, preserved mixtures of fruit juice and sugar. Jelly making is a good way to preserve fruit flavors for enjoyment throughout the year. Fruit jelly is a fairly easy-to-prepare product for the beginning canner and may be made at home without much special equipment. Ingredients Substances essential for fruit jelly making are fruit flavor, pectin,
Passion fruit juice machine is a kind of efficient juicer machine for extracting passion fruit juices and separating passion fruit peels and seeds automatically. With compact structure and high efficiency, this machine can be widely used in many fruit and vegetable juice making and processing fields. And this passion fruit juicer machine has been sold to many countries like
Passion fruit is a vigorous, shallow rooted, perennial vine that climbs by means of tendrils It can grow as tall as 20 ft (6 m or 600 cm) high and a more realistic life expectation is 3 5 years It is mainly grown for fresh market and juice extraction Preferred by
Dec 06, 1996 CONFIGURATION OF A FRUIT JUICE EXTRACTION MACHINE or the fruit juice extracting apparatus of the present invention, in accordance with FIGS. 1-3, comprises a tubular chassis 1, affixed to an extraction box 1 for housing juice, mounted vertically on this box 1 is a gearmotor 2 which drives a crank and rod 3 which provokes the axial movement of ...

domain: food


text:
Nina Simone was Born Eunice Kathleen Waymon in Tryon, NC in 1933
Nina Simone was an instinctive musician, an original, versatile single, pianist, arranger and composer. Her repertoire included jazz standards, gospels and spirituals, classical pieces, folk, blues, pop, African chants, show tunes and her own compositions.
On March 24, 1965, she performed her protest song Mississippi Goddam before 40,000 marchers at the conclusion of the third Selma to Montgomery (AL) march. It was one of many songs she either wrote and/or performed that addressed civil rights, and black liberation and identity.

domain: artistic


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Bible ConcordanceSwelling (14 Occurrences)
2 Peter 2:18 For, uttering great swelling words of emptiness, they entice in the lusts of the flesh, by licentiousness, those who are indeed escaping from those who live in error; (WEB KJV ASV WBS YLT)
Jude 1:16 These are murmurers, complainers, walking after their own lusts; and their mouth speaketh great swelling words, having men's persons in admiration because of advantage. (KJV ASV DBY WBS YLT)
Leviticus 13:2 "When a man shall have a rising in his body's skin, or a scab, or a bright spot, and it becomes in the skin of his body the plague of leprosy, then he shall be brought to Aaron the priest, or to one of his sons, the priests: (See NAS RSV NIV)
Leviticus 13:10 and the priest shall examine him. Behold, if there is a white rising in the skin, and it has turned the hair white, and there is raw flesh in the rising, (See NAS RSV NIV)
Leviticus 13:19 and in the place of the boil there is a white rising, or a bright spot, reddish-white, then it shall be shown to the priest; (See NAS RSV NIV)
Leviticus 13:28 If the bright spot stays in its place, and hasn't spread in the skin, but is faded, it is the swelling from the burn, and the priest shall pronounce him clean; for it is the scar from the burn. (WEB NAS RSV NIV)
Leviticus 13:43 Then the priest shall examine him; and, behold, if the rising of the plague is reddish-white in his bald head, or in his bald forehead, like the appearance of leprosy in the skin of the flesh, (See NAS RSV)
Leviticus 14:56 and for a rising, and for a scab, and for a bright spot; (See NAS RSV NIV)
Psalms 46:3 though its waters roar and are troubled, though the mountains tremble with their swelling. Selah. (WEB KJV JPS ASV DBY WBS NAS)
Psalms 89:9 Thou rulest the pride of the sea: When the waves thereof arise, thou stillest them. (See JPS NAS)
Isaiah 30:13 therefore this iniquity shall be to you like a breach ready to fall, swelling out in a high wall, whose breaking comes suddenly in an instant. (WEB KJV JPS ASV DBY WBS)
Jeremiah 12:5 If thou hast run with the footmen, and they have wearied thee, then how canst thou contend with horses? and if in the land of peace, wherein thou trustedst, they wearied thee, then how wilt thou do in the swelling of Jordan? (KJV DBY WBS)
Jeremiah 49:19 Behold, he shall come up like a lion from the swelling of Jordan against the habitation of the strong: but I will suddenly make him run away from her: and who is a chosen man, that I may appoint over her? for who is like me? and who will appoint me the time? and who is that shepherd that will stand before me? (KJV DBY WBS)
Jeremiah 50:44 Behold, he shall come up like a lion from the swelling of Jordan unto the habitation of the strong: but I will make them suddenly run away from her: and who is a chosen man, that I may appoint over her? for who is like me? and who will appoint me the time? and who is that shepherd that will stand before me? (KJV DBY WBS)
ThesaurusSwelling (14 Occurrences)...
1. (p. pr. & vb. n.) of Swell. 2. (n.) The act of that which swells; as, the swelling
of rivers in spring; the swelling
of the breast with pride. ...SWELLING
. .../s/swelling.htm - 13k
Swell (9 Occurrences)
... 6. (vi) To be turgid, bombastic, or extravagant; as, swelling words; a swelling
style. ... 16. (n.) The act of swelling. 17. (n.) Gradual increase. 18. ...
/s/swell.htm - 13k
Ophel (6 Occurrences)
... and Habakkuk 2:4, where a verbal form is translated "puffed up," seem to imply that
one meaning assigned to the root may be that of "swelling." Recently Dr ...
/o/ophel.htm - 14k
Inflammation (6 Occurrences)
... 1. (n.) The state of being inflamed; redness and swelling in the body. ... It is manifested
outwardly by redness and swelling, attended with heat and pain. ...
/i/inflammation.htm - 9k
... 1. (n.) A morbid swelling, prominence, or growth, on any part of the body; especially,
a growth produced by deposition of new tissue; a neoplasm. ...
/t/tumor.htm - 7k
Swollen (7 Occurrences)
... 1. (pp) of Swell. 2. (p. p) of Swell. 3. (a.) Enlarged by swelling; immoderately
increased; as, swollen eyes; swollen streams. Int. Standard Bible Encyclopedia. ...
/s/swollen.htm - 9k
Assign (22 Occurrences)
... Jeremiah 49:19 Behold, he shall come up like a lion from the swelling of the Jordan
against the strong habitation; for I will make them suddenly run away from ...
/a/assign.htm - 14k
Swelled (5 Occurrences)
/s/swelled.htm - 8k
Quake (23 Occurrences)
... (Root in WEB JPS ASV DBY NAS RSV NIV). Psalms 46:3 though its waters roar and are
troubled, though the mountains tremble with their swelling. Selah. ...
/q/quake.htm - 12k
Vanity (100 Occurrences)
... 2 Peter 2:18 For when they speak great swelling words of vanity, they allure through
the lusts of the flesh, through much wantonness, those that were clean ...
/v/vanity.htm - 39k
Greek5450. phusiosis -- a puffing up ...
Noun, Feminine Transliteration: phusiosis Phonetic Spelling: (foo-see'-o-sis) Short
Definition: a puffing up, pride Definition: a puffing up, pride, swelling
. ... /greek/5450.htm - 6k
5246. huperogkos -- of excessive weight or size
... 5246 (from 5228 , "beyond" and 3591 , "a swelling") -- properly, "oversized,"
("bloated"); used of a who constantly exaggerates, spuing words out from his ...
/greek/5246.htm - 7k
3709. orge -- impulse, wrath
... 3709 (from , "to teem, swelling up to constitutionally oppose") -- properly,
(), ie rising up from an ongoing (fixed) opposition. ...
/greek/3709.htm - 7k
1471. egkuos -- pregnant
... pregnant. From en and the base of kuma; swelling inside, ie Pregnant -- great with
child. see GREEK en. see GREEK kuma. (enkuo) -- 1 Occurrence. << 1470, 1471. ...
/greek/1471.htm - 6k
5445. phurama -- that which is mixed
... From a prolonged form of phuro (to mix a liquid with a solid; perhaps akin to phuo
through the idea of swelling in bulk), mean to knead; a mass of dough -- lump ...
/greek/5445.htm - 6k

domain: literature


Despite the fight between Maharashtra and Andhra Pradesh over the Babli project, Maharashtra has given its nod for the Pranahita Chevella project by agreeing to release the required amount of water to the project. The project irrigates more than 16 lakh acres in eight districts of Telangana including Medak, Nalgonda, Hyderabad, Adilabad, Nizamabad, Warangal Karimnagar and Ranga Reddy.
Out of the proposed 164 tmcft of water for the project anually, around 30 tmcft would go to the Greater Hyderabad area.
After giving its “tacit consent,” the Maharashtra government, in a letter to Aditya Nath Das, Secretary, Irrigation (Telangana), asked Andhra Pradesh to constitute an inter-State committee with officials from both States.
Andhra Pradesh has also offered whatever compensation sought against 1,800 hectares of land in Maharashtra which will be submerged by the barrage’s reservoir.
It is really good to see neighboring states showing such gesture towards each other.
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RALEIGH, N.C. (WNCN) — Advocates for the homeless said housing insecurity is a real problem in Wake County. “Our homeless population is growing like our general population is growing and it’s diverse, ” said Alice Lutz, CEO of Triangle Family Services.
It’s a non-profit organization in Raleigh that started in 1937. The agency focuses on family safety, financial stability, and mental health.
They also work with the homeless population in Wake County. Lutz said some are living in tent cities and encampments. She said for some people it’s by choice, others by unfortunate circumstances.
Lutz said a $1,400 check could be the start to a fresh start. “The stimulus check is a really important piece to have to help provide stability for a long period of time for some of these residents.”
If home is out on the streets without television, internet, or a computer how will they get the information they need to help get their stimulus checks? That’s where Triangle Family Services steps in.
“We have a street outreach team that physically goes out into the street and physically meets people where they are in their homeless encampments as well as on the streets. The whole purpose is to find out what their needs are and get their needs met,” explained Lutz.
But there are still a few barriers that this team is working out.
Last year the IRS recommended that people experiencing homelessness could qualify for a stimulus check by using the non-filers tool, but it has since been removed.
According to the IRS without the portal, you must file your 2020 taxes to claim your stimulus money.
This also goes for the first and second payments if you haven’t received it by now.
You’ll also need a mailing address, which is difficult when you don’t have a home.
“If you’re living in a homeless encampment you don’t have an address. So, we work with the Women’s Center in Wake County and Oak City Cares to have the facilities for mailing. So, then we can have a permanent address for the clients,” continued Lutz.
She said they are also working with banks. “There are banks that are working with the homeless population to help us get access to not having to have a checking account.”
The IRS is also issuing prepaid debit cards called the Economic Impact Payment card for those who may not have access to a checking account.


text:
ILOILO First District Representative Janette Garin said she is not afraid to face the new case filed against her and nine other public health officials for reckless imprudence resulting in homicide over the deaths blamed on anti-dengue vaccine Dengvaxia.
“I have yet to receive the 78-page resolution but this will be the nth case that has been filed against me and the other public health servants who sought to bring a life-saving vaccine in the country,” Garin said.
The Department of Justice (DOJ) on Friday, February 21, indicted Garin and nine other DOH officials, but Garin said she is not afraid of the case.
“Haharapin ko ito ng buong tapang at walang takot dahil alam natin ang katotohanan,” the former Department of Health (DOH) secretary said.
Earlier, Justice Assistant Secretary Neal Bainto said the panel of prosecutors found probable cause to indict Garin and other officials on the second batch of complaints related to Dengvaxia.
“Sa unang batch pa lang out of seven criminal cases for reckless imprudence filed against us, under the direction of PAO Chief Persida Acosta, six have already been dismissed and one remains pending. The dismissals entail the cases’ lack of basis and substance,” Garin said.
Garin added that science has proven over and over again that no deaths have been linked to the Dengvaxia vaccine.
She said almost the whole world is using it now and no case like ours has surfaced.
“Hustisya bang matatawag na hayaan nating mamatay ang ating mga kababayan sa mga sakit na may gamot/bakuna naman? Kung ganunman, sino ang sisingilin natin sa mga namatay sa Dengue nang tanggalin ang bakuna sa bansa? Sino ang sisingilin natin sa mga namatay dahil sa tigdas at mga nagkaroon ng polio dahil natakot sa bakuna?” Garin said.
For Garin, this situation, true justice should listen to science and not to politics.
“Sa dami ng problema at isyu sa bansa pilit pa rin binubuhay ang mga kasong walang basehan para manggipit lang at para pagtakpan ang mga mas importanteng usapin. Hindi dapat bahiran ng pulitika ang hustisya at pampublikong kalusugan,” Garin said. (SunStar Philippines)
Former Department of Health (DOH) and Iloilo First District Representative Janette Loreto-Garin. (Photo by Leo Solinap)
February 22, 2020
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The Spacetoon Media Group & Animation International
The Spacetoon Media Group & Animation International are esteemed multinational companies with extensive experience in the field of kids’ entertainment. The Spacetoon Media Group with its headquarters in Dubai Media City, UAE and Animation International with headquarters in Hong Kong, are currently spreading their base across Asia and the Indian subcontinent via the Distribution of TV Programmes & TV Channel, Children’s Magazines, Videos and allied products and services
Multi Screen Media Pvt. Ltd.
Multi Screen Media Pvt. Ltd., formerly known as Sony Entertainment Television Pvt. Ltd., with continuous delivery of only the best has become one of the most popular entertainment brands in India.We have built a rich library of over 40,000 hours of original programming, which has received national and international acclaim, garnering successful ratings and viewership.
Nickelodeon, Indias leading kids brand and a part of Viacom18, is available in over 34.20 million households in the country. It is a 24-hour pay channel in Hindi, with the option of English, Tamil and Telugu audio feed on DTH. With an approach that puts kids first, Nickelodeon takes pride in encouraging kids to be themselves.
Zing is 24 hour music channel from the Zee bouquet has new shows, vignettes and bigger stars which promise to add more color to your life and have you glued. You won't miss another film preview, music launch, Bollywood party, spicy gossip, or any celebrity anymore.
A healthcare TV channel showing health related information.
Imagine Showbiz, a Turner General Entertainment Networks India Pvt. Ltd endeavour, showcases the best of Bollywood music for a country where music is synonymous with Bollywood Music.Imagine Showbiz is a 24-hour Bollywood music channel catering to the varied tastes of the Indian youth.With its vast library of songs from every genre-soothing & melodious love songs to peppy, foot-tapping dance numbers.
A TV channel offer comedy shows, movies, downloads wallpapers and screen savers.
Sony Entertainment Television
India's leading television channel provide the best in hindi family entertainment to the largest percentage of the indian population-the young adult.
Sanskar Info TV Pvt. Ltd.
Providing hindu bhajans, aartis, aradhanas, program on yoga, satsang samachar and watch live sanskar TV.
BILL PAY POINT
Pay Point India Network Pvt. Ltd. is one of the pioneers in providing a one stop solution for distribution and collection of a world wide range of services. it includes E-recharge,renewal of mobiles,Dish t.v.,and it will also issue transport syatems such as buses,railways,airlines as well as entertainment and cinema shows.
A Hindi language spiritual and religious channel.
ZoomIn accepts all Debit and Credit Cards. If you prefer paying cash – now you can! Order online and pay by cash at any MobileStore location closest to you. You can also use your i-mint loyalty points on ZoomIn. We have a completely vertically integrated infrastructure – we do not outsource.
Multi Screen Media Pvt Ltd.
We at Pix solemnly undertake that every movie shown here features a good story well told. Whether or not our films are crammed with effects, stars and big budgets, we solemnly assure you that they will always contain a good story. May the story take precedence above all else.
An official website of the Cartoon Network India providing information about the cartoon characters, wallpapers & screen savers of cartoons and e-cards.
A hollywood channel showing movies, movie calender, stars, gallery, events, gossip and community.
Sumaria is a customercentric, talent driven organisation that puts the customer in the centre of all its activities.With operations in Mumbai, Thane & Bhiwandi, Sumaria is a retail chain for consumer electronics & home appliances with a commitment to growth and innovation. Sumaria generates more than Rs. 30 crores in annual revenue.


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Did 2020 just loop round to 1999?
Driver software snubbed by macOS 10.15, 10.14, Amazon Music, too
But company also has its eye on Kubernetes at the edge
'That button is there on the app' says minister. How many times have you heard a client make wild claims about functionality?
Snaps vs Flatpaks, losing in mobile, Microsoft and Linux, and avoiding another GNOME 2 to 3 disaster
For everyone else there are splash screens, fixes and iffy anti-malware
Plus: What puts off developers from adopting cloud? Price


EVOC’s Edinburgh Climate Forum is a place for communities, groups and voluntary organisations to discuss the climate and nature emergency and specifically the transformation that needs to happen in Edinburgh and how we can play a part in delivering that change. This is a platform for community groups and voluntary organisations to come together and discuss shared actions, plan and initiate collaborations, exchange resources learning and skills, coordinate on shared funding bids, access materials and resources, as well as inform strategic decision making.
The forum is open to all community groups and voluntary organisations in Edinburgh and you are invited to join the Organising Group where your group can take part in leading on the activities of the forum.
Find out more and how to get involved: firstname.lastname@example.org
Join the Edinburgh Climate Forum Whatsapp Group.
This Whatsapp group is for community, voluntary and social enterprise groups to share important information , news and updates with other groups in Edinburgh interested in climate and nature emergency activities.
The Edinburgh Climate Forum is now operating a hub and spokes approach to climate direction with 14 new thematic Forums which have been created for specific policy influencing, shared projects creation and strategic planning. These forums will be led and organised by community groups and most will have the opportunity to interface with City of Edinburgh Council officers and key stakeholders/budget holders in the city. In the group description, you will see theme links below on Policy Influencing, Projects and Planning Forums.
The Community Climate Forum is focused on making sure community and voluntary groups across the City play a part in Edinburgh’s journey to becoming a net zero, climate ready city by 2030.
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text:
The extraembryonic ectoderm (ExE) is essential for mammalian placental formation and survival of the embryo in utero. We have obtained a mouse model lacking the ExE, by targeted deletion of the transcription factor Elf5. Although Elf5 mutant embryos implant and form an ectoplacental cone, no trophoblast stem (TS) cells can be derived, indicating that the absence of ExE is a result of the lack of TS cell maintenance. Embryos without ExE tissue are able to form the anterior visceral endoderm but fail to undergo gastrulation, demonstrating an essential role for the ExE in embryonic patterning during a defined window of development.
Two of the earliest lineages that can be distinguished during mammalian embryogenesis are the inner cell mass (ICM) and the trophectoderm cells of the blastocyst stage (Rossant et al.,2003). Whereas the ICM segregates into epiblast and primitive endoderm, and gives rise to embryonic as well as extraembryonic tissues, the trophectoderm cells will contribute solely to extraembryonic lineages. In particular, the polar trophectodermal (pTE) cells, which overlie the ICM, will form the extraembryonic ectoderm (ExE) and the ectoplacental cone (EPC). In the mouse, the ExE makes up the proximal half of the egg cylinder after implantation.
The proliferative potential of the pTE and ExE is dependent on a population of trophoblast stem (TS) cells that can be isolated from these tissues at blastocyst to gastrula stages (Tanaka et al., 1998; Uy et al.,2002). FGF signalling from the ICM, and subsequently from the epiblast, is required for maintaining these TS cells. Removal of Fgf4 in TS cell cultures leads to the differentiation of TS cells into giant cells and the loss of early ExE-specific markers, such as Eomes and Cdx2 (Tanaka et al.,1998). Eomes and Cdx2 appear to be crucial to the formation and maintenance of TS cells within the pTE as no TS cells can be derived from null mutants of Eomes(Russ et al., 2000) or Cdx2 (Rossant et al.,2003), in accordance with the in vivo trophoblast defects in these embryos. The EPC, which overlies the ExE, is devoid of TS cells(Uy et al., 2002). It contains differentiated trophectodermal cells that are thought to form the spongiotrophoblast (Cross et al.,2003).
Towards the end of gastrulation, the ExE forms a bilayer with extraembryonic mesoderm and becomes separated from the epiblast by the exocoelomic cavity (Kaufman,1995). This bilayer is termed the chorion and is deflected towards the proximal pole of the conceptus. At around embryonic day (E) 8.5, the allantoic mesoderm attaches and fuses with the basal layer of the chorion. Chorionic trophoblast cells begin to differentiate into syncytiotrophoblast cells and villi progenitors, and, in conjunction with allantoic cells, will form the chorioallantoic placenta essential for maternal-foetal nutrient and gaseous exchange and therefore embryonic survival(Cross et al., 2003; Rossant and Cross, 2001).
However, the ExE also fulfils an earlier inductive function by signalling to the subjacent epiblast during germ cell formation(Yoshimizu et al., 2001) and embryonic patterning (Beck et al.,2002). These roles of the ExE are mediated by at least two distinct pathways involving the TGFβ superfamily members Bmp4 and Nodal. Bmp4 is expressed at gastrulation stages in the ExE adjacent to the epiblast, and, in chimeric loss-of-function mutants in which extraembryonic Bmp4 expression is selectively ablated, neither primordial germ cells nor extraembryonic mesoderm is formed(Lawson et al., 1999).
Secondly, Nodal activity in the epiblast is necessary for both mesoderm and anterior visceral endoderm (AVE) formation(Brennan et al., 2001). The AVE is formed by migration of a group of distal visceral endoderm cells to one side of the egg cylinder well before gastrulation commences. It secretes antagonists into the adjacent epiblast thereby restricting Nodal activity and thus mesoderm formation to the opposite (posterior) side of the egg cylinder(Lu et al., 2001). However, Nodal translation generates Pro-Nodal, which has to be cleaved by endoproteases to generate the fully active Nodal signalling molecule. This cleavage is performed by Furin/Spc1 and Spc4/Pace4, which are expressed in,and secreted from, the ExE (Beck et al.,2002). Double loss-of-function mutants for these two proteases closely resemble Nodal-deficient embryos and do not form AVE or mesoderm (Beck et al.,2002).
We describe here the identification of a novel key gene involved in maintaining the polar trophectoderm/ExE lineage. This gene is Elf5(ESE2 in humans), which encodes a transcription factor belonging to the Ets superfamily. It is characterised by a DNA-binding Ets domain, is able to bind to a subset of Ets-binding sites and can transactivate constructs containing Ets-binding sites upstream from a minimal promoter(Oettgen et al., 1999). Elf5 has previously been shown to be expressed in foetal and adult epithelial cells of organs such as the mammary and salivary glands, kidney,prostate and lung in mice and in humans(Oettgen et al., 1999; Zhou et al., 1998). Significantly, placenta of pregnant mice at E9.5 and later exhibited Elf5 expression, as assayed by northern blots(Zhou et al., 1998). Presently no in vivo role for Elf5 has been demonstrated though Elf5 expression appears to be increased significantly in mouse mammary tumors relative to in normal mammary tissue (Galang et al.,2004).
We report here on the early expression of Elf5 in the ExE lineage and demonstrate an essential function of Elf5 for the generation of this tissue. We discuss our findings in relation to trophoblast stem cell maintenance and epiblast-ExE interactions.
Materials and methods
Elf5 genomic clones were isolated from a 129 mouse λlibrary (Stratagene) using a 380 bp 5′ probe of the published cDNA sequence. The targeting vector was constructed by ligating the 3′ short arm of 660 bp commencing within exon 2 and isolated by PCR adding SalI sites, and inserted into the XhoI site after the polyadenylation signal of the phosphoglycerate kinase promoter-driven, puromycin-containing, PGKPuroA cassette(Ridgeway et al., 2000). The dipthera toxin gene (DT-A)(McCarrick et al., 1993),allowing negative selection, was inserted (blunt/SalI) upstream of puromycin. The PCR-derived 5′ flank of 11.3 kbp extending into exon 2 was inserted into the SalI site between DT-A and puromycin. The NotI linearised construct was electroporated into thirteenth passage R1 ES cells (Nagy et al., 2003) (Jackson Laboratory). Puromycin resistant colonies were screened for correct targeting using PCR, and by Southern analysis using a probe external to the short arm. This probe was isolated by PCR with the primers 5′-AATCACCTCCTACCCCCTAC and 5′-GCAAACATTAGCATAGCATCAG. The correct targeting event resulted in insertion of the puromycincassette into exon 2 of Elf5 76 bp downstream of the ATG. Clones 33 and 59 produced germ-line chimeras and were maintained on a 129 background. Diploid embryo-tetraploid embryo aggregation chimeras were produced as described (Nagy et al., 2003). This work was performed under ERMA permit GMD002183 and animal ethics approval RAEC4307.
Genotyping by PCR
PCR genotyping was performed using the two allele, three primer PCR strategy. The common primer A (5′-GAGCAATGGGAATAAACAGGG) was located within the long arm at the 3′ end of intron 1 and could give a 410 bp product only on the targeted allele with primer B(5′-TGGATGTGGAATGTGTGCGA) located within the puromycin cassette. On the wild-type allele, primer A gave a 578 bp product in combination with primer C(5′-GGAGAAAGGTGGGGAGGATAA). The inserted puromycin cassette prevented the primer A-C combination from yielding a product with the targeted allele as template. Tail tips were digested in Proteinase K buffer [100 mM Tris (pH 8.0), 5 mM EDTA, 0.1% SDS, 200 mM NaCl] at 55°C, with shaking,for 2 hours to overnight, boiled for 5 minutes, centrifuged and then 0.25-1μl used in a standard 25 μl PCR reaction containing 1 Unit Taq polymerase, 1.5 mM Mg2+, 10 pmol of primer A, and 5 pmol of primers B and C. PCR conditions were 94°C for 4 minutes, followed by 35 cycles of 94°C for 30 seconds, 62°C 45 seconds and 72°C for 1 minute.
Whole-mount in situ hybridisation
The whole-mount in situ hybridisation protocol has been described(Nagy et al., 2003). Staining reactions were carried out for 2 hours up to 5 days, with littermates always treated in the same vessels in the same way. Embryos were genotyped after photography by PCR. Mouse Elf5 cDNA clones were isolated by screening a mammary gland cDNA library with a bovine Elf5 fragment. The Elf5 antisense probe used covered nucleotides 9 to 334 of the reference sequence NM_010125, excluding the conserved ETS domain. A 530 bp mouse Hex fragment was cloned into pGEM-Teasy (Invitrogen) in the T7/sense orientation using the primers 5′-CCCTCTGTACCCGTTCCC and 5′-CCGATGACTGTCATCCAGC and a Ta of 50°C.
Trophoblast stem (TS) cell culture
E6.5 embryos from Elf5+/– matings were separated into proximal and distal halves. The distal half was used for genotyping. The proximal half was stripped of visceral endoderm(Nagy et al., 2003) and treated at 37°C for about 10 minutes with 0.25% pronase in Tyrodes Ringer saline. Cells were dispersed by brief pipetting and plated on primary feeder cells (Nagy et al., 2003; Tanaka et al., 1998; Uy et al., 2002). We used 70%conditioned medium (Tanaka et al.,1998) from the outset. Several colonies were observed in wild-type and Elf5+/– cultures by 3 days after dissociation. After the third passage, TS cultures were grown in the absence of feeder cells. For the Fgf4/heparin withdrawal experiment, sixth passage TS cultures were grown in four-well dishes to 30% confluency, whereupon growth was continued for 5 days in conditioned medium with or without Fgf4 and heparin. Cells were pelleted and subjected to real-time PCR as described below.
RNA was isolated using TRIZOL (Invitrogen) and reverse transcribed with Superscript3 (Invitrogen) and oligo-dT, according to the manufacturer's instruction. Real-time PCR was performed using SYBR-Green master mix (Applied Biosystems) and the following primers (introns spanned; amplicon size in base pairs):
Bmp4, 5′-GAGTTTCCATCACGAAGAACA-3′ and 5′-GCTCACATCGAAAGTTTCCC-3′ (1; 301);
Cdx2, 5′-CCAAGTGAAAACCAGGACAAAA-3′ and 5′-AACGAAGAAGCCCCAGGAA-3′ (1; 669);
Elf5, 5′-CTTGTCTTCACGGTGATGTTGGA-3′ and 5′-CATTCTTCTTCCTTTGTCCCC-3′ (4; 640);
Eomes, 5′-GCAAACAACAACAACACACA-3′ and 5′-GGGGCAAGGACTTAATACCA-3′ (3; 583);
Fgfr2, 5′-ACCAAATACCAAATCTCCCAAC-3′ and 5′-ATTCATTCTCCACCAGGCA-3′ (3; 592);
Furin, 5′-TGCCAGACCACATGACTAC-3′ and 5′-CAAGGACTTGGGGGATGAA-3′ (3; 320);
Otx2, 5′-AAACAGCGAAGGGAGAGGA-3′ and 5′-AGGAGGAGGAAGTGGACAA-3′ (1; 400);
Spc4, 5′-GGCATCAGACCCAACTACA-3′ and 5′-ATCCACCAAGCCAAATCCA-3′ (4; 570).
PCR conditions were 95°C for 5 minutes, followed by 40 cycles of 95°C for 10 seconds, 56°C for 30 seconds, 72°C for 30 seconds, and 78°C for 10 seconds, followed by dissociation curve analysis. RT-minus controls were run routinely and representative PCR products analysed on agarose gels to ensure spcecificity of reactions. Amplification efficiencies were monitored by standard curves using serially diluted samples and ranged from 1.6 to 1.9. Relative copy numbers were calculated and normalised against actin.
Elf5 is expressed in the extraembyonic ectoderm lineage
Elf5 is transcribed in adult and foetal epithelial tissues in both humans and mice, but has not been reported to be expressed during early embryogenesis. We noticed using RT-PCR that Elf5 mRNA is present before somitogenesis, and performed whole-mount in situ hybridisation to determine the spatiotemporal pattern of transcripts. Interestingly, Elf5 is already expressed before gastrulation in the ExE(Fig. 1A). This pattern of expression is maintained throughout gastrulation with Elf5transcripts marking the ExE lineage, which, by E7.5, has formed the chorionic ectoderm (Fig. 1B-D). The chorion still exhibits Elf5 transcription at E8.5(Fig. 1E), when chorioallantoic fusion is occurring. Until the start of somitogenesis, we did not detect expression in epiblast derivatives or other extraembryonic tissues, such as ectoplacental cone (EPC) and parietal or visceral endoderm. Low levels of Elf5 transcripts could be seen by RT-PCR in E3.5 pre-implantation blastocysts (data not shown). This exquisitely specific expression pattern suggested a role for Elf5 in the ExE lineage.
Elf5 deficient embryos are embryonic lethal
To determine whether this novel extraembryonic-specific factor does have a function in early development, we inactivated the Elf5 gene by homologous recombination. The puromycin gene was inserted into the second exon of Elf5, at the start of the N-terminal pointed domain,leaving only 26 amino acid residues of the Elf5 protein(Fig. 2A-C). Heterozygous Elf5+/– mice derived from two different ES-cell targeting events appeared normal and were fertile. However, no homozygous pups from numerous heterozygous crossings were ever retrieved on either 129 or BALB/c backgrounds, or on mixed 129:BALB/c genetic backgrounds, suggesting embryonic lethality.
Analysis of genotypes of embryos derived from Elf5+/–intercrosses on the 129 background revealed normal Mendelian ratios up to E8.5. Thereafter, an increasing number of embryos were resorbed, as manifested by the retrieval of only dead or resorbing Elf5-/- embryos at E9.5 (Table 1). We noticed an effect of genetic background. BALB/c Elf5-deficient embryos showed a more variable phenotype, with the odd developmentally delayed embryo still present at E10.5. Such survivors could not develop much further, as by E12.5 no Elf5-/- embryos were recovered(Table 1). We restricted our further analyses to the more severe phenotype seen on the 129 background.
|Age .||Strain .||Wild type .||Heterozygous .||Mutant .||Total .|
|E6.5||129||19 (26%)||36 (49%)||17 (23%)||73|
|E7.5||129||11 (25%)||24 (55%)||9 (20%)||44|
|E8.5||129||10 (26%)||22 (56%)||7 (18%)||39|
|E9.5||129||9 (30%)||15 (50%)||6 (20%)*||30|
|E10.5||BALB/c||7 (25%)||14 (50%)||7 (25%)†||28|
|E12.5||BALB/c||5 (26%)||14 (74%)||0||19|
|Age .||Strain .||Wild type .||Heterozygous .||Mutant .||Total .|
|E6.5||129||19 (26%)||36 (49%)||17 (23%)||73|
|E7.5||129||11 (25%)||24 (55%)||9 (20%)||44|
|E8.5||129||10 (26%)||22 (56%)||7 (18%)||39|
|E9.5||129||9 (30%)||15 (50%)||6 (20%)*||30|
|E10.5||BALB/c||7 (25%)||14 (50%)||7 (25%)†||28|
|E12.5||BALB/c||5 (26%)||14 (74%)||0||19|
Four dead/extremely underdeveloped; two resembling E8.5, one E9.5 embryo
Morphological and patterning defects in Elf5 mutant embryos
Elf5-deficient embryos cavitated normally. However, from early streak stages, mutant embryos were consistently smaller than their littermates and exhibited no signs of primitive streak formation(Fig. 3A). The constriction marking the extraembryonic-embryonic ectoderm (ExE-EmE) border was absent. Mesoderm had not formed by E7.5, and mutant embryos still resembled egg cylinders, composed of ectoderm surrounded by visceral endoderm and no ExE-EmE constriction (Fig. 3B,C). The EPC was present. By E8.5, Elf5-/- embryos were severely reduced in size, consisting of a sack of ectoderm which often had continued to proliferate with folding of the excess epithelium(Fig. 3D,E). This ectoderm displayed an anterior neuroectodermal character in the form of patchy Otx2 expression, whereas more posterior genes, such as the mid-and/or hindbrain markers Pax2 and Krox20, were not transcribed (Fig. 3E, and data not shown). T/brachyury, marking mesoderm from the onset of gastrulation, was seen in half of the E8.5 mutant embryos(Fig. 3D). These T-positive embryos, which we term type II Elf5-mutants,occasionally contained a mass of mesenchymal cells located at the proximal end of the embryo, indicative of delayed aberrant mesoderm formation (data not shown). Elf5-/- embryos did not form a chorion(Fig. 3C), thus precluding chorioallantoic placenta formation. The resulting foetal-maternal loss of nutritional and waste exchange would be expected to lead to the observed midgestational lethality (Cross et al.,2003; Rossant and Cross,2001).
Elf5 deficient embryos are devoid of extraembryonic ectoderm
The lack of chorion formation in Elf5-/- embryos suggested defects in the ExE lineage. We therefore analysed the Elf5mutant embryos using a panel of molecular markers that are expressed in the ExE. The genes coding for Cdx2 (Beck et al., 1995), Eomes (Ciruna and Rossant, 1999; Russ et al.,2000), the fibroblast growth factor receptor Fgfr2(Haffner-Krausz et al., 1999),Bmp4 (Lawson et al., 1999) and the endoproteases Furin and Spc4 (Beck et al., 2002) are primarily or exclusively expressed in the ExE during early gastrulation stages. None of these genes were transcribed in the proximal ectoderm of E6.5 to E7.5 Elf5 deficient embryos(Fig. 4C-J; Fig. 8D,G), molecularly confirming the absence of ExE. Notably, Cdx2, the earliest known marker for the undifferentiated trophectoderm lineage(Rossant et al., 2003), is expressed in the polar trophectoderm of Elf5-deficient blastocysts,but no expression is evident by E5.5 (Fig. 4A,B). Thus from pregastrula stages, Elf5 deficient embryos do not contain ExE.
It should be pointed out that the absence of the ExE does not affect the formation of the EPC, as judged histologically(Fig. 3C) and by the expression of Furin, Spc4 and Fgfr2. At E5.5, Spc4 and Fgfr2 expression marks both EPC and ExE tissue(Fig. 4E,I)(Beck et al., 2002). In Elf5 mutants, these genes were robustly expressed only in the EPC overlying the forming egg cylinder (Fig. 4E,I). Similarly, Furin, which is normally expressed in the ExE, EPC, proximal epiblast and visceral endoderm at early gastrula stages(Beck et al., 2002), could be detected only in the EPC region of Elf5-deficient embryos(Fig. 4H).
The absence of ExE and a visible ExE-EmE constriction suggested that the ectodermal layer of the Elf5 mutant egg a marker for undifferentiated embryonic ectoderm (Scholer et al.,1990). We found that Oct4 expression in mutants extended to the proximal end of the egg cylinder(Fig. 4K,L), confirming the absence of ExE tissue well before the commencement of gastrulation. Similarly Otx2, normally restricted to the anterior embryonic epiblast(Perea-Gomez et al., 2001),was expressed across the entire mutant ectoderm, reaching the EPC region(Fig. 4M,N). We conclude from these marker studies that in the absence of Elf5 activity, the ExE is not formed, resulting in embryos composed of EPC directly abutting the embryonic ectoderm.
Elf5 is required for the maintenance of trophoblast stem cells
Why were Elf5-/- embryos depleted of ExE by E5.5? The ExE is a direct derivative of the polar trophectoderm (pTE) of the blastocyst. Yet unlike Cdx2 and Eomes deficient embryos, which die around the implantation stage as a result of defects in trophectoderm and pTE cells, respectively (Chawengsaksophak et al., 1997; Russ et al.,2000), Elf5-/- embryos implant and develop at expected Mendelian ratios to E6.5 (Table 1). Furthermore, Cdx2 marking the pTE was still expressed in E4.5 Elf5-/- mutants(Fig. 4A). This suggests that pTE formation is not impaired in Elf5 deficient embryos.
However, both pTE and ExE contain trophoblast stem (TS) cells(Tanaka et al., 1998; Uy et al., 2002), and a failure to maintain these stem cells after implantation would be expected to affect ExE formation. Indeed, our observations of the absence of expression of the undifferentiated TS cell markers Cdx2, Eomes and Fgfr2in Elf5-/- embryos suggested that TS cells are not maintained in mutant embryos. We therefore attempted to isolate TS cells from Elf5-/- embryos by culturing E6.5 dissociated proximal ectoderm tissue on primary feeder cells in the presence of Fgf4 and heparin(Tanaka et al., 1998; Uy et al., 2002). Notably,TS-like colonies were only formed from wild-type and Elf5+/– proximal ectoderm (n=18/18),whereas Elf5-/- tissue formed no colonies (n=0/4; Fig. 5A,B).
Analysis of TS cell colonies by RT-PCR indicated that these cells expressed, not only Cdx2, Eomes, Fgfr2, Errb (Esrrb –Mouse Genome Informatics), Bmp4, Furin and Spc4, but also Elf5 (Fig. 5C). Fgf4 withdrawal causes the differentiation of TS cells into secondary giant cells and spongiotrophoblasts, both found in the EPC(Tanaka et al., 1998; Yan et al., 2001). We observed that Fgf4 withdrawal resulted, within 5 days, in a downregulation not only of the TS cell markers Cdx2, Eomes, Fgfr2 and Errb but also of Elf5. Concomitantly, the spongiotrophoblast and EPC marker Tpbp/4311 (Lescisin et al., 1988) was upregulated, indicative of TS cell differentiation(Fig. 5C). We conclude that Elf5 is a marker for undifferentiated TS cells. In Elf5-/- embryos, TS cells no longer self renew but differentiate into EPC progenitors, leading to the observed absence of the ExE by E5.5.
Extraembryonic ectoderm is required for patterning of the embryo proper
Elf5 deficient embryos not only exhibit defects in the ExE lineage but also display severe patterning defects in the embryo proper. Although we observed expression of Elf5 solely in the ExE and chorion using whole-mount in situ hybridisation, low levels of expression in other regions might have escaped our detection and be partly or wholly responsible for the phenotype seen in the embryo proper. We therefore wished to determine whether patterning defects occurred in embryos lacking Elf5 function only in the epiblast. To this end, we performed a tetraploid rescue experiment. Wild-type eight-cell-stage tetraploid cells known to contribute only to extraembryonic tissue were aggregated with diploid cells derived from four- to eight-cell-stage embryos of Elf5+/–× Elf5+/– matings and allowed to develop to E10.5, when Elf5-/- embryos are either dead or severely retarded and malformed. Statistically, one quarter of the chimeric embryos would be expected to contain epiblast with an Elf5-/-genotype. Genotyping revealed that four out of 15 embryos were composed solely of Elf5-/- cells (Fig. 6A). These four embryos were morphologically normal(Fig. 6B,C), proving that Elf5 is not required in embryonic tissues for development up to this developmental stage.
The extraembryonic ectoderm is not required for AVE formation
The ExE has been shown to be involved in reciprocal interactions with the subjacent embryonic ectoderm, activating Nodal, which, in turn, orchestrates AVE development and axis formation in the epiblast(Beck et al., 2002; Brennan et al., 2001; Lu et al., 2001). We therefore investigated whether AVE formation was impaired in Elf5 mutants. The AVE is formed from a group of distal visceral endoderm cells expressing the homeobox gene Hex, which migrate towards the future anterior side between E5.7 and E6.5 (Yamamoto et al.,2004). We found that, in E5.8 Elf5-/- embryos, Hex expression was confined to the visceral endoderm on one side of the embryo, similar to in controls (Fig. 7A). Furthermore, the Nodal/BMP inhibitor Cer-1(Cerl) was expressed in the presumptive AVE of Elf5-/- embryos (Fig. 7B). Similarly, the AVE/anterior neurectoderm marker Hesx1 could still be detected in the anterior endodermal region of mutants by E7.5 (Fig. 7C). In addition, we could confirm Nodal expression in Elf5deficient embryos at E5.5 (Fig. 7D), a prerequisite for AVE formation. These results indicate that the AVE can be specified and established in the absence of the ExE.
Extraembryonic ectoderm is required for gastrulation
In marked contrast to AVE formation, mesoderm formation is impaired in Elf5 null mutants. At the start of gastrulation, Nodalexpression marks the epiblast where the primitive streak will form. In Nodal mutants, posterior epiblast/mesoderm markers, such as Cripto, Eomes, T and Fgf8, fail to be expressed and no primitive streak is formed (Brennan et al.,2001). Elf5-/-embryos either showed no expression of Nodal and its downstream markers (type I mutants), or exhibited ectopic expression abutting the EPC (type II)(Fig. 8A-E: see Table S1 in supplementary material for statistics). Conversely, transcription of Otx2, normally showing a reciprocal pattern to Nodal, was not restricted in Elf5 deficient embryos(Fig. 4J,K). Cdx2 and Bmp4 transcription, characteristic of nascent extraembryonic mesoderm, was not seen in mutant embryos(Fig. 8F,G). Thus the lack of ExE leads to a secondary defect in mesoderm formation, the severity of which can vary and may be due to heterogeneity in the 129 background(Simpson et al., 1997).
Elf5 as a lineage-determining gene for the extraembryonic ectoderm
The polar trophectoderm (pTE) overlying the inner cell mass in the implanted blastocyst at E4.5 proliferates and differentiates in the following 24 hours to yield the morphologically distinct extraembryonic ectoderm (ExE)and overlying ectoplacental cone (EPC) regions of the egg cylinder stage embryo. We report here the expression of the transcription factor Elf5 specifically in the ExE from E5.5 onward. Furthermore, in our Elf5loss-of-function mouse model this tissue is specifically lost. This phenotype of normal implantation followed by the loss of the ExE but not the EPC region is unique. Several other genes have been implicated in the formation or maintenance of trophectoderm-derived lineages but show quite distinct phenotypes when disrupted by homologous recombination(Cross et al., 2003; Rossant and Cross, 2001). Cdx2 loss-of-function mutants die before implantation, rarely forming expanding blastocysts (Chawengsaksophak et al., 1997). Cdx2 is already differentially expressed at the morula stage and is believed to be one of the earliest lineage determining genes for the trophectoderm (Kunath et al., 2004; Rossant et al.,2003). Eomes appears to be required at a slightly later stage, as mutants do implant but arrest shortly thereafter, failing to undergo trophoblast differentiation in vitro (Russ et al., 2000). Whereas mural trophectoderm generates only primary giant cells, which are essential for implantation, further development requires the pTE which gives rise to the ExE, the EPC, as well as secondary giant cells (Copp, 1979). Eomes could thus be involved in the mural versus polar trophectoderm lineage decision (Fig. 9A).
Little is known about the molecular events leading to the generation of the ExE and the EPC region from the pTE. However, it has emerged that trophoblast stem cells can be isolated from the ExE but not the EPC(Rossant and Tamura-Lis, 1981; Uy et al., 2002). These TS cells require FGF as well as Nodal/Activin signalling for their maintenance(Erlebacher et al., 2004; Guzman-Ayala et al., 2004; Tanaka et al., 1998). Upon FGF withdrawal, TS cells differentiate either into polyploid giant cells or spongiotrophoblast cells. This is reminiscent of the EPC, the outer edge of which differentiates into secondary giant cells, whereas the main part generates spongiotrophoblast cells (Cross et al., 2003). The requirement of FGF signalling is also supported by in vivo results. Fgf4 and Fgfr2 loss-of-function embryos die shortly after implantation (Arman et al., 1998; Feldman et al.,1995), and inactivation of Erk2, an important downstream effector of FGF receptors, leads to a specific loss of both the ExE and EPC, indicating a primary proliferative defect at the level of the pTE(Saba-El-Leil et al., 2003). As Fgf4 is expressed in the ICM of the blastocyst, as well as by the epiblast of the egg cylinder, current models suggest that the tissues in close proximity to the FGF signals, namely the pTE and later the ExE, maintain TS cells, whereas the more distant mural trophectoderm and EPC form differentiated trophoblast cell types(Kunath et al., 2004; Tanaka et al., 1998). Thus the`lineage decision' between ExE and EPC at post pTE stages may simply reflect differences in the ability to maintain a population of undifferentiated proliferative TS cells. This interpretation places the emphasis on the maintenance of TS cells. In line with this, TS cells cannot be isolated from Cdx2 nor Eomes defective blastocysts(Rossant et al., 2003; Russ et al., 2000). Both genes can be considered to be trophoblast lineage determining genes by virtue of specifying or maintaining TS cells at the morula and blastocyst stages,respectively.
We suggest that Elf5 acts at the next step, being required for maintaining trophoblast stem cell potential beyond the implanted blastocyst stage(Fig. 9A). Thus in the absence of Elf5, TS cells within the pTE are no longer maintained, instead differentiating into EPC precursors that will differentiate finally into sphongiotrophoblasts and giant cells. By egg cylinder stages, the ExE is absent whereas the EPC trophoblast is still seen. Several lines of evidence support this scenario. The normal implantation rates of Elf5deficient embryos implies that Elf5 is only required after the formation of trophectoderm. The presence of an EPC in mutants means that the pTE must have formed correctly, as it gives rise to this lineage. This is supported by the correct pTE-specific expression of Cdx2 at E4.5 in Elf5-/- embryos. That TS cells are no longer maintained in Elf5 mutants past the pTE stage is supported by the absence of the TS-cell containing ExE and the observation that TS cells could not be derived from the proximal half of the egg cylinder of Elf5 mutant embryos. Moreover, TS cell markers such as Cdx2, Eomes and Fgfr2 were no longer expressed in mutant embryos. Lastly, Elf5 is expressed in TS cell lines and lost upon their differentiation after FGF removal. Elf5 can therefore be considered to be a lineage-determining factor, required for the formation of the ExE lineage by virtue of maintaining TS cells.
The role of the extraembyonic ectoderm in patterning the embryo
The absence of ExE from its inception in Elf5 deficient embryos has provided a novel mouse model allowing an examination of the role of this tissue in the development of the embryo proper. Patterning of the embryo involves two distinct steps, the first being the establishment of the AVE signalling centre thereby preparing the adjacent epiblast for anterior induction by subsequent mesendodermal signalling. Thereafter, the primitive streak with its associated organiser activities is formed on the opposite side of the egg cylinder. Both events require Nodal signalling within the epiblast(Brennan et al., 2001; Lu et al., 2001). In turn,Nodal activity has been shown to be dependent on the ExE, which secretes the proteases Furin and Spc4 to cleave and thereby activate the Nodal precursor in the epiblast. In the absence of both proteases, mesoderm and AVE formation are abolished (Beck et al., 2002). Why do Elf5-/- embryos, which do not have an ExE, still form the AVE? We propose (Fig. 9B) that in the absence of ExE, Nodal cleavage is mediated by secretion of Spc4, which is strongly expressed in the EPC located adjacent to the epiblast in Elf5-/- pregastrula embryos. Once proteolytically activated, Nodal can then amplify its own expression(Fig. 7D, Fig. 9B) via its autoregulatory intronic enhancer, leading to AVE formation and migration(Brennan et al., 2001; Norris et al., 2002).
Although the EPC may substitute for the ExE at stages preceding gastrulation, this is not the case at later stages. The loss of Nodaltranscription at E6.5 in type I Elf5 mutants explains the absence of expression of Eomes, Fgf8, T and Cripto, all of which are downstream targets of Nodal activity and required for posterior patterning and mesoderm formation. Significantly, the absence of transcription from the Nodal locus at E6.5 in Elf5 mutants differs from ExE-containing Nodal null mutants and Furin/Spc4 double mutants, which do exhibit transcripts in the proximal epiblast(Beck et al., 2002; Brennan et al., 2001). These mutants differ from Elf5 deficient embryos in two fundamental ways– they have no AVE and they do contain ExE. Could the continued presence of the AVE in Elf5 mutants result in the absence of posterior markers? We consider this to be unlikely as the Nodal-repressive AVE is restricted to only one side of the mutant egg cylinders. We would thus favour the alternate hypothesis (Fig. 9C), that a protease independent signal emanating from the ExE(absent in Elf5 but present in Nodal mutants) is required for Nodal transcription at E6.5. Potential candidates for such signalling are Bmp4 and/or Bmp8b, which are expressed in the ExE, but not the EPC (Fujiwara et al., 2002; Ying et al., 2000). ExE-derived Bmp4 has been shown to induce posterior genes in the epiblast and is required (in a mouse background strain-dependent fashion) for the generation of a normal primitive streak(Beck et al., 2002; Fujiwara et al., 2002; Winnier et al., 1995). In Nodal-null mutants, Bmp4 is present in the ExE at E6.5(Brennan et al., 2001) and thus could theoretically contribute to transcription from the Nodal locus,as it does in other contexts (Fujiwara et al., 2002; Piedra and Ros,2002; Schlange et al.,2002).
Whatever the identity of the signals emanating from the ExE are, our ExE-deficient mouse model and tetraploid rescue experiments strongly support the proposed inductive role of this extraembryonic tissue in primitive streak formation, thus suggesting that this tissue is equivalent to the avian posterior marginal zone (Bachvarova et al.,1998).
We have found a novel factor exquisitely restricted to and required for the formation of the ExE, and have created a mouse model that clearly separates two temporally distinct requirements for the ExE in instructing the patterning of the epiblast. Whereas the early ExE function in anterior patterning via AVE establishment can be replaced presumably by Spc4 secreted from the EPC, there is an essential requirement for ExE in initiating gastrulation and posterior patterning of the embryo proper. Furthermore, we now can identify three genes that consecutively function to determine cell choices in the maintenance of TS cells. First, Cdx2 for the trophectoderm/ICM choice at the morula stage; second, Eomes for the polar/mural trophectoderm choice at the blastocyst stage; and third, Elf5 in the ExE/EPC decision at the implanted blastocyst stage.
We thank Drs Daniel Bachiller, Daniel Constam, Chuxia Deng, Jacqueline Deschamps, Brigid Hogan, Juan Carlos Izpisua-Belmonte, Michael Kuehn, Gail Martin, Roel Nusse, Elizabeth Robertson, Andreas Russ, Hans Scholer and Michael Shen for kind gifts of probes; Jan Oliver for help with electrofusion;and Drs Goetz Laible and Richard Wilkins for critical reading of the manuscript. This work was supported by the Foundation for Research, Science and Technology, New Zealand, and by AgResearch CRI.


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With a base MSRP of $845,000, the 918 Spyder was the most expensive Porsche production car ever, and the options list included the $84,000 Weissach Package.Jul 26, 2021
Since Porsche stopped producing this model in 2015, the asking price for a used Porsche 918 Spyder has skyrocketed — it can easily exceed $1.6 million.
With prices originally starting in the US at $845,000 in 2013, the last Porsche 918 for sale had been sold by December 2014. Now the only way to procure a used Porsche 918 Spyder is from private owners or firms when the automobiles do periodically come up for sale.
1955 Porsche 550 Spyder — Estimated Price- $6,000,000.
The Bugatti La Voiture Noire is the most expensive car in the world. It’s a part of Bugatti’s plans to build two custom cars every year.
The Koenigsegg Gemera is the world’s quickest production car that hits the 0-60 mph mark within 1.9 seconds. It’s the very first four-seater by Koenigsegg and the world’s first Mega-GT that weighs 4,079 pounds.
Porsche 918 Spyder GCC Spec
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1’42.627 is the record-breaking time clocked by Porsche’s 918 Spyder on the Le Mans Bugatti circuit. The new time breaks the previous record of 1’45.26 held by the Radical SR3, making the Porsche 918 Spyder the fastest production car to drive this circuit.
How many Porsche 550 Spyders are there left? According to TopSpeed, around 90 Porsche 550 Spyder examples left the factory in total. Owners of these cars bought and raced them successfully throughout the 1950s.Apr 18, 2021
A 550 Spyder sold in 2012 for $3.685 million. Expected to sell for between $5 and $6 million, Jerry Seinfeld’s 550-060 changed owners for $5.35 million at a Gooding & Company auction in March 2016.
2005 Maybach Exelero
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8 Aston Martin DBR1
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As he was walking to a math class during his freshman year at Ohio State University, R. J. Kromer spotted a poster for a student-run team designing a fuel-cell-powered car.
The SSC Tuatara is the fastest production car in the world.
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Black History Month 2023, Protect and Preserve
Black History Month should a time to embrace, celebrate and most of all to learn about the untold stories and immense contributions African Americans have made to our nation from its inception. It’s often been pointed out that there shouldn’t need to be a Black History Month because Black History is American history, and for a while it seemed as if we were making progress towards that realization.
But anyone who has been paying attention over the past few years in conservatively controlled statehouses and governorships should be acutely aware the progress made in Black History finding is rightful full place in our story is being actively and legislatively suppressed. It began with laws banning books from libraries and topics forbidden from classrooms. Now the Ron DeSantis has banned an entire Advanced Placement course on African American Studies from Florida’s high schools because it “inexplicably contrary to Florida law and significantly lacks educational value.”
Related article: “Florida Rejects A.P. African American Studies Class”
Florida claims the course “significantly lacks educational value” despite it being designed by some of the foremost scholars and educators in the country. Not to mention, it’s already been approved for implementation in 2024 by numerous State’s Boards of Education. What Gov. DeSantis and Florida are trying not to say is the course significantly undermines White American mythology with facts.
First, DeSantis and his Florida Republicans passing, “Don’t Say Gay;” now it’s “Don’t Say Black.” Makes one wonder who will be next? If this sounds familiar it should; we seen it before. In 1935 Hitler and the NAZIs passed the Nuremberg Laws, stripping Jewish people of their citizenship, their rights, and centuries of culture in Germany.
We can’t let what’s happening in Florida and elsewhere spread. This Black History Month we must preserve and protect all that it is from those who are doing their best to erase all it should be. To that end, as we did last year we will again be celebrating Black History Month with a daily Twitter series @TheOUTFront . In light of current events, this one seems especially appropriate to begin with.
(Main Image: “Letter from a Birmingham Jail” 1963)

domain: news


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PICAYUNE — Picayune Police caught a strong arm robber minutes after the crime occurred.
Thursday afternoon officers were sent to the area of Hancock Bank on Memorial Boulevard in reference to a strong arm robbery.
Upon arriving to the area officers quickly located the suspect, 18-year-old Drin Shaw of Waveland, and took him into custody.
Officers also took his girlfriend, Shavonne Honeycutt of Picayune, into custody on an unrelated charge of possession of marijuana.
Check Friday's edition of the Picayune Item for a full story on the crime.


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The kids wanted to swim, so we went to a little place called Firehole Canyon. Getting there was a treat on the big rig, because the sign said "RVs not advised" - but who reads those signs anyways? We saw the beautiful Firehole Falls (I do like those waterfalls) before getting wet.
The swimming was one of the more challenging tasks of the trip - but a huge adventure for the brave ladies. Rocks, currents, and the unknown made for crazy fun!

domain: geography


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The Renaissance of Lesbianism in Early Modern England
Lieferbar innert 2 Wochen
BeschreibungTraub analyzes the representation of female-female love and eroticism in early modern literature and drama.
InhaltsverzeichnisAcknowledgements; List of illustrations; Introduction: 'practicing impossibilities'; 1. Setting the stage behind the seen: performing Lesbian history; 2. 'A certaine incredible excesse of pleasure': female orgasm, prosthetic pleasures, and the anatomical Pudica; 3. The politics of pleasure; or, queering Queen Elizabeth; 4. The (in)significance of Lesbian desire; 5. The psychomorphology of the clitoris; or, the reemergence of the Tribade in England; 6. Chaste femme love, mythological pastoral, and the perversion of Lesbian desire; 7. 'Friendship so curst': Amor Impossibilia, the homoerotic lament, and the nature of Lesbian desire; 8. The quest for origins, erotic similitude, and the melancholy of Lesbian identification; Notes; Index.
PortraitValerie Traub is Professor of English and Women's Studies at the University of Michigan, Ann Arbor. She is the recipient of a National Endowment for the Humanities Fellowship, and author of numerous works on gay/lesbian studies, including the book Desire and Anxiety: Circulations of Sexuality in Shakespearean Drama and coeditor of Feminist Readings in Early Modern Culture: Emerging Subjects.
Pressestimmen'Breathtaking in its knowledge and dazzling in its analysis, The Renaissance of Lesbianism shows us how female same-sex desires become not only legible but pivotal during the course of the English Renaissance. Traub's brilliant readings of sexual representation, both in early modern texts and in the scholarly imagination today, turn the screw of interpretation on figures from Queen Elizabeth to Katherine Philips and on topics from terminology to teleology. This synthesis of 'theory' and 'history' sets a new standard for lesbian scholarship and opens new paths for studying both sexuality and early modernity.' Susan Lanser, Brandeis University 'The Renaissance of Lesbianism in Early Modern England is a dazzling achievment. Analyzing an extraordinary range of texts and visual artifacts, Traub changes the very terms through which it is possible to read forms of female intimacy in Early Modern England. Her work on the tribade, on female friendship, and on the psychomorphology of the clitoris is pathbreaking. Theoretically deft and immensely learned, The Renaissance of Lesbianism is without question one of the most important books of the decade in Early Modern studies.' Jean Howard 'One of the most significant books to be published in 2001 ... A tour de force of theoretically informed inquiry, this is cultural history at its most powerful and elegant - completely rewriting the history of sexuality in the 17th century.' Recent Studies in the English Renaissance '...[F]rank and enabling engagement with large methodological debates about sexuality...'. Times Literary Supplement
Untertitel: 'Cambridge Studies in Renaissan'. New. Sprache: Englisch.
Verlag: CAMBRIDGE UNIV PR
Erscheinungsdatum: Juni 2002
Seitenanzahl: 512 Seiten

domain: library


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The holiday season is here, and it's the perfect time to start doing activities to celebrate the holidays. No matter what holidays you celebrate, it's important to find an activity that you enjoy. As someone who has done each of the following four activities, I can personally attest that some will become your new favorites, as they have all become mine.
- Visit Celebration’s Winter Wonderland
Each year, the town of Celebration puts on a Winter Wonderland event that offers carriage rides, nightly random snowfalls and great views of the town decorated for Christmas. Going to this event is a favorite activity of mine because seeing snow (even if it's fake) is a rarity for me as a born and raised Floridian, and it puts me in the holiday spirit like nothing else! It's also fun to take pictures in the town, eat at the local restaurants and just enjoy the atmosphere.
- Experience Dazzling Nights at Harry P. Leu Gardens
During the holidays, Harry P. Leu Gardens is transformed into a holiday wonderland with lights, music and interactive elements for its annual event, Dazzling Nights. Walking through the gardens during the day is a truly amazing experience, but at this event, I think the experience is even more so. The music is great, and the lights light up the garden beautifully.
- View holiday displays and events at theme parks
Theme parks like Disney and Universal have fantastic holiday displays that are perfect for viewing to get you in the holiday spirit. Whenever I'm there during the holidays, I love taking a photo in front of the holiday decor, eating a holiday-inspired treat, and watching whatever holiday event is going on in the park!
- Walk through movie scenes with the "I Love Christmas Movies" exhibit at Gaylord Palms
Gaylord Palms Orlando has an exhibit this year where you can walk through 13 scenes from iconic Christmas movies including Elf, The Polar Express, National Lampoon’s Christmas Vacation, and more. I love the surreal feeling of being transported into some of my favorite holiday movies when I walk through this exhibit. It's definitely been my favorite holiday activity I've done this year! If you like any of the movies featured, I highly recommend you try it.
Doing these activities are some of my favorite ways to celebrate the holiday season. They serve as a way for me to relax, immerse myself in the holidays and have fun with friends and family. The holidays are surely different this year, but these events (running with safety guidelines in place) are great ways to keep you in the holiday spirit!

domain: entertainment


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Research & Publications
We work on the molecular basis of a number of physiological phenomena related to the hearing and balance organs.
Phenomenon of electrical resonance.
Electrical tuning is a phenomenon by which certain vertebrates discriminate between different frequencies of sound. Electrical resonance results when the inherent oscillation in the membrane potential of hair cells corresponds to sound of a particular frequency. This gives rise to a resonance and amplification of signal with consequent transmitter release from these cells. The inherent oscillation in membrane potential in a hair cell is brought about by an inward Calcium current and an outward Potassium current (calcium dependent). Inherent to this view is that the two proteins are physically proximate.
We had previously erroneously believed that the range in BK channel currents was brought about by alternative splicing. We now hypothesize that this variation in current is brought about by association with other proteins. We have isolated several binding partners using the yeast two hybrid technique and are in the process of evaluating their ability to alter BK kinetics and bring about channel clustering and co-localization. The role of Prestin (in Collaboration with Dr. Joseph Santos-Sacchi). Prestin is a recently described protein in outer hair cells that is responsible for the sharp tuning seen in the hearing organ of mammals. It is critical for normal hearing. Knocking out of this protein results in the loss of hearing in mice.
Specialized Terms: Hearing and Balance Organs
Extensive Research Description
Phenomenon of electrical resonance
Electrical tuning is a phenomenon by which certain vertebrates discriminate between different frequencies of sound. Electrical resonance results when the inherent oscillation in the membrane potential of hair cells corresponds to sound of a particular frequency. This gives rise to a resonance and amplification of signal with consequent transmitter release from these cells.
The inherent oscillation in membrane potential in a hair cell is brought about by an inward Calcium current and an outward Potassium current (calcium dependent). The systematic variation in the frequency of such an oscillation in hair cells that occurs across the tonotopic axis is brought about primarily by a variation in the kinetic properties of the Potassium current. Previously we had shown that some of this variation in kinetic properties of this current was brought about by alternative splicing of this BK potassium channel. However, a large part of this variation in kinetics cannot be explained by alternative splicing alone.
Our present hypothesis is that this variation in current is brought about by association with other proteins. Towards this end we have made yeast two hybrid libraries from the cochlea organ and are screening these libraries for binding proteins using the purported intracellular domains of the BK channel as “bait”. In addition we have also demonstrated by fluorescence immunohistochemistry that the calcium channel and the potassium channel are in fact in close physical approximation in these hair cells.
There is considerable theoretical reasons to support this observation. We have preliminary data that these two proteins are physically linked to one another. We are presently attempting to duplicate these results using a more sensitive method of detection (tandem mass spec at the Keck facility). We are also seeking to identify proteins that might serve as scaffolding linking both these channels. The role of Prestin (in Collaboration with Dr Joseph Santos Saatchi, Prof in Otolaryngology see www.yaleearlab.org) Prestin is a recently described protein in outer hair cells that is responsible for the sharp tuning seen in the hearing organ of mammals.
It is critical for normal hearing. Knocking out of this protein results in the loss of hearing in mice. Prestin is a “motor” protein that gives rise to electromotility in outer hair cells. Its an unusual phenomenon in that motility in outer hair cells can be rapid, upto 20KHz, which is orders of magnitude faster than conventional motor proteins like myosin. We are attempting to determine the functional domains within this protein by mutagenesis. We have made over a dozen mutants of this protein and are presently evaluating how it affects the motor function of this protein.
Regeneration in the auditory epithelium
Hearing loss in the elderly population affects upto 60% of this age group. This hearing loss is largely brought about by a loss in hair cells in the auditory epithelium. In previous experiments we had demonstrated that increasing levels of the second messenger cyclic AMP results in a cascade of events giving rise to cell division and regeneration in hair cells.
These results have since been duplicated in mice. We are attempting to identify the molecular events that underlie such regeneration and we have undertaken a subtractive hybridization experiment to determine the identity of mRNA transcripts that are upregulated with increasing cAMP. We have identified several molecules.
One such molecule is Id 2 a protein that has a binding domain but lacks the activation domain of BHLH transcription factors. Id2 is thought to repress other BHLH transcription factors by preventing their dimerization. To determine the effects of Id2 on regeneration we have used antisense oligos to suppress the expression of this protein in cochleas treated with forskolin (which increases cAMP levels).
Our preliminary data suggests that suppressing Id2 results in a reduced number of cells entering the cell cycle (measured by incorporation of BrDU). We are presently attempting to further corroborate this finding and will undertake experiments to determine the binding partners of Id2.
Cholinergic synaptic function
Acetylcholinesterase in cholinergic synapses removes acetylcholine. Acetylcholine, the neurotransmitter in cholinergic neurons, is rapidly removed from cholinergic synapses by hydrolysis by the enzyme acetylcholinesterase.
Failure to remove acetylcholine in a timely fashion gives rise to a sustained depolarization of the postsynaptic membrane and a failure to respond to further neurotransmitter release. Thus the timely removal of acetylcholine brought about by the localization of acetylcholinesterase in these synapses is critical for normal neurotransmission to occur. We have identified two proteins that likely serve to localize the protein to these neuronal synapses. One of these proteins is related to the bacterial Cu transporter CUTA1.
The other is a novel protein which however has a proline rich domain that has been shown to be important in interactions between acetylcholinesterase and the colq protein that localizes the enzyme to the synapse in the neuromuscular junction. We have made several mutants of the latter protein and are presently attempting to relate the ability of these mutants to alter the localization of acetylcholineterase to synaptic membranes.
Postural Balance; Hearing; Neurobiology; Neurology; Neurosciences

domain: medical


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Jedi: Survivor is a strong entry in the modern Star Wars canon, part of a new subsect of adventures in this universe finding ways to be sci-fi fantasy without ignoring the innate horror and banal evils inherent in the premise. The story paints an impressively dire picture of the new status quo in the galaxy, and it weaves in elegantly with the interactivity of the game, tying it directly into the fact that Cal is still powerful but only one Jedi in a galaxy that fell even when there was an entire army of them.
The game’s dedication to graphical fidelity feels like a blinder to thinking outside the box in every other regard. It can’t help but feel like intensive overcompensation for inconsistent, tension-less stealth, one-note combat, level design that doesn’t reward exploration, generically fleshy enemies, upgrades that don’t reward experimentation, and ineffective jump scares, from enemies that get cheap hits in on Jacob every single time, regardless of how well-prepared the player is. Much has been made of the fact that this was meant as the heir apparent to beloved survival horror series Dead Space, a game that, 12 years later, can still induce goosebumps just from its terrifying attract sequence. By contrast, if not for its graphics, The Callisto Protocol feels like a relic from 1998, undone creatively even by the decaying likes of Shadow Man.
Gollum just feels so shockingly old hat—a disheartening collection of mechanics that, at best, bring to mind one of the lesser pre-2013 Tomb Raider games and, at worst, suggest leftovers from the N64 bargain bin. Every success involves wrestling the loose controls, unhelpful camera, and iffy collision detection into submission against an ever-increasing wave of bugs and glitches, only some of which have been fixed by the game’s Day One patch.
The fallout of that ending makes what had been a wafer-thin murder mystery with a gimmick into an exercise in psychological sadism, where the player is nauseatingly complicit. Despite the immense pool of talent giving their all to breathe life into these characters, Twelve Minutes is a game thoroughly lacking in humanity, in any sense of the word.
Battlefront II is actually a rather fitting sequel to its immediate predecessor, which was itself a fun, visually phenomenal but woefully shallow and convoluted experience. Everything that was right with the original game is exactly as it was before. Everything that wasn't, however, has mutated into something more craven and significantly uglier.
Rockstar's remastered trilogy is, appropriately, an absolute car wreck of creative neglect.


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Home Posts tagged Ireland Visit
The US President Joe Biden’s visit to Ireland continues on Thursday with set-piece engagements, including an address to the Irish parliament. He will spend most of the day in the company of leading Irish politicians and meet Irish President Michael D Higgins. His visit to the Republic of Continue Reading
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As this blog has already mentioned, the past few months have seen new guidelines for sunscreen makers to consider when making labels for their creams and lotions. Terms must be clarified, with the phrase "broad spectrum protection" denoting the range of radiation the product shields against (known as UVA and UVB).
As a recent New York Times article suggests, however, the chemicals or other ingredients that form the base of these products may present difficulties for customers looking to integrate them into their standard habits, and cosmetics producers can put corresponding information on their cosmetic labels in order to clarify.
The article describes the dilemma that makeup users may encounter in terms of getting sufficient protection from the sun, as some options reportedly don't offer the same kinds of defense as creams. Checking the SPF level on cosmetic labels is considered a necessity, as achieving the right level of total SPF (recommended as somewhere between 15 and 50) may require use of both kinds of products. But that doesn't mean that a product can't be preventative and cosmetic.
The Times interviewed makeup veteran Sylvie Chantecaille on this subject, as she defended mineral-based sunscreens over makeup products that may use chemicals like oxybenzone to add this element.
"Mineral sunscreens don't penetrate the bloodstream," she said. "That was top of my mind when we started developing it."
With conflicting information out in the market, it can be hard to communicate to consumers which types of products are in their best interest. This is part of why making labels that keep customers well-informed is such an important task, and cosmetic labels in particular can work to provide the types of information shoppers are most likely to be looking for.