All shinny black and very new looking lowered into the waiting trench.
The pipe was still uncovered and a huge hole still in the road, but with much daring and great feats of jumping the two oldest (12 and 9) jumped into the open trough, onto the wonderful big shinny pipe, and scrambled up the the other side….I'm sure they went to school dirty but happy. Mom-mom and the little Kindergartner had to walk clear around the huge hole, walk across a smaller white pipe to get to the other side, then RUN—RUN–RUN to the waiting bus down by the mailboxes.
Then last evening the little kids and their Dad stopped by to say HI. Then they were on their way through the fields to their house…Tally (the youngest) wanted to see if she could beat them to her house by running across the fields. You know what? She DID!!
Yesterday Terry started disking up the corn stalks. He worked until nine in the evening and has started the process again today.
We are not fond of large corn stalks creating little dams in the water furrows. Terry also like to have the rotting organic matter in our clay soil…lots of humus significantly influences the density of any soil and contributes to moisture and nutrient retention. We are always, always, always working to protect the land we farm.
That evening I notice a BIG storm playing over the Uncompahgre Plateau. (Un-come-pah-gray).
Today we have wind. Wind always signals Spring in our part of the world.
The January Thaw has arrived! Boom and I love January Thaws…it warms up ya see.
It warms up enough that I don't like sleeping in the house. I go outside (I know how to open the back door and go out, Boomer has NEVER figured it out. HEE HEE) and sleep in my dog house.
Now I have to admit sometimes I go out and want to come back in sooner than when Mom comes looking for me. See I go out when I get hot, that can be when everyone is asleep in the house. Then I have to wait until Sam the cat wants out——he jumps on the bed and runs all over Mom to wake her up so she will let him out — or Mom just gets up. Mom always comes to check on us dogs and if I'm not there she knows I'm in the dog house.
When you are old you can't curl up like you use to be able to do….so I scratch up my dog blankets real good and waller down real deep in them. Even though the dog houses are covered up with lots of blankets and stuff, when the night runs on into morning…well, then, I want to come inside and sleep on my orthopedic bed.
So sometimes I have to wait.
The January Thaw is here!!! The days have warmed up considerable. Boom and I have taken to checking out things, you know…who has been walking on our property, what sort of news is on the corn stalks and maybe just maybe we can find Freddy Fox's house.
Boomer is never still….he runs EVERYWHERE! Up and down and back. He runs off and then comes to find me.
On we went over to the equipment area, then up to the back 40, then over to the Upper End which is where I found it……………………………!!!!
I was clear inside when Mom got there….I didn't want to get out, but she made me. Boom never saw it, so I told him all about it when we were resting in the sun later.
When we got back to the house Mom said we had better enjoy this because it is supposed to snow Saturday and colder weather will follow.
Mom asked if we wanted to come inside, but Boomer and I decided we wanted to hang outside. Even Sam the Cat hung with us. Sam rolled around in the sawdust by the wood pile and Boomer and I lay in the sun next to the house.
Sure is nice, these warmer temperatures.
Now on the bright side, it was just one disk!


Jackson County Jail - HURST-ROSCHE, INC.
The existing Jackson County Jail was a two-story masonry and steel building. It housed 133 beds and consistently reached its limit in housing prisoners, causing personnel to turn away prisoners from other areas. As a result, the likelihood of overcrowding was always imminent, and the County was losing possible revenue that accompanied the housing of prisoners.
The need for an expansion was desired, and a new two-story addition was built. The addition was a 64-bed, two-story masonry and steel building with approximately 10,000 square feet. The building included four cell blocks divided by an 8'-0" wide central corridor resulting in two cell blocks on either side of the corridor.
The design and layout of the building allowed for the separation of different prisoner classifications. The addition was constructed with very little construction related interruptions to daily operations. The existing facility remained in full operation during the construction phase of the project. In the event that additional beds are desired, the building was designed where any future expansions are easily and readily achievable.


text:
by sheep@biosys.net
author's note: the following story contains descriptions of a sexual nature as well as explicit stuff about people being turned into mechanical creatures.
---> it's also mostly about this happening to a *man*. if that freaks you out at all or even sounds the kind of things liable to disgust you, or if you're not supposed to be reading this sort of thing, then rather than writing to tell me how sick I am, please stop reading now. :)
Julia made a few last-minute preparations, checking some screens and meters around the control console before looking up at the subject of her new experiment. He hung lifelessly in the cyborgization cubicle, moaning occasionally. Julia smiled, typed in a control sequence on the keyboard and retreated into the darkness around the edge of the laboratory.
Luke awoke to the dizzying sensation of being completely unable to move his arms or legs, indeed any part of his naked body. As far as he could see, for his scope of vision was limited by the brace that had locked his head tightly in position against the back wall of the cubicle, his limbs and trunk had been fastened in place with reinforced straps and tubes in such a way that no movement at all was possible. Blowing his hair away from his eyes, he struggled slightly and felt the restraints tighten further, accomodating every minute subtlety of his movement. Panic swam into his thoughts as he became convinced that escape was completely impossible. His breathing became faster, and as it did so he noticed an oscillating green meter on the bank of instruments opposite him begin to pulse faster.
"What are you going to do?" he muttered, sure that someone was listening. For a few seconds there was no reply, then Julia's even, soothing voice could be heard coming from a shadowy corner of the chamber:
"The computer is going to take over your brain. You will be enslaved." Before Luke could spit a confused reply, gas began to hiss into the cubicle from jets located in its corners. "The adjustments to your atmosphere I am making will place your mind and body in a state of relaxation, in order to aid the conversion. Do not try to speak." Luke's mouth dropped as he made to say something, but the lethargy creeping over him was making it more difficult every second. He felt a couple of tiny pricks as hypodermic needles punctured his neck and began squirting some alien fluid into his bloodstream. He felt its warmth traverse the length of his body, making him feel numb and helpless.
"W-whhhrrr... wwh......" he gurgled, but the effort was too much. His eyelids drooped. Julia smiled and stepped out of the shadows. She stood in her lab-coat in front of the cubicle.
"Do not become alarmed. You will experience negligible pain. As a cyborg, you will serve me well."
Leaning over to the control console, she entered another control sequence and then strode up towards the cubicle in order to observe the process up close. Luke's eyes were now closed, and the syringes disengaged from his neck smoothly, sliding back into the wall of the cubicle. At the same time, the back wall of the booth came alive, machinery gliding into position around his naked body, and the whole cubicle began to buzz with electricity.
Quietly and efficiently, wiring started coating his arms and legs. The thicker cables wound around him like tendrils. The wires slid under his skin and interfaced with his nervous system. Implants began to fit into his head, while cybernetic components clicked into place all over his chest and limbs. As his brain was equipped with circuitry the computer connected to his cortex and started brainwashing him. Robotic gloves and boots fitted over his own human hands and feet as while sturdy steel armor plating began to cover his trunk. The computer, having finished wiping Luke's memories of his life as a human being, started stripping away his emotions too, replacing them only with logic and obedience. Soon his arms and legs had finished being converted, metal armor snapping shut around them. A gleaming helmet-like device closed around his head, and then it was complete.
There was a slight pause, and the operating machinery slotted back into the sides of the cubicle. The restraints that had held him in place receded also. The cubicle doors also opened. Julia smiled and entered a final sequence into the computer. There was a momentary buzz and Luke's body juddered slightly, then silence. The cubicle was dead. It had done its work.
"Activate." said Julia. The Cyborg-Luke began to come to life. His eyes glowed red and his lips parted slightly, but he did not move. "Diagnostics." There was a slight whirr, and then Luke's voice could be heard. It sounded cold and monotonous, and strangely electronic.
"cyborg-unit---system-integrity-intact." he said.
"Good." murmured Julia, and she stepped forward to admire her new slave. His huge body looked from the outside to be almost entirely robotic, apart from his face. "Are you aware of your past as a human individual, with a free will?"
"yes." said Luke.
"You are no longer that individual. As part of your cyborgization your mind has been totally enslaved to me. You will obey my every command."
"i-will-obey." came the instant reply.
"Good. Step out of the cubicle." Luke's hydraulic limbs whirred into motion as he mechanically walked out of the pod and stood to attention in front of it. "Now..." breathed Julia, moving closer and placing the palm of her hand on Luke's shiny metal chest, "... hold me."
The cyborg which had once been Luke, with surprising gentleness, folded his arms around Julia's back and held her tightly against him. Julia slid her hands around his smooth robotic torso, feeling more and more aroused as the hard, cool metal of his chest touched the soft, warm skin of her breasts and neck. She groaned and smiled. "Obey me." she murmured.
"i-obey-without-question.." came the monotonous, electronic reply from Luke's human mouth.
"Who am I?" asked Julia.
"you-are-my-mistress---
you-are-organic."
"Mmmn..." she moaned.
"you-are-organic---
organic-lifeforms-are-imperfect-and-unstable---
all-organics-must-be-adapted---
you-must-be-converted-into-a-cyborg." Luke concluded.
"Yess...." came the almost inaudible reply as Julia hugged Luke's robotic form tightly. After a pause, Luke disengaged himself and stepped back from Julia. She closed her eyes as Luke began to undress her. He removed her white lab-coat and unbuttoned her blouse, touching the curves of her soft white breasts with his robotic fingers. She took the rest of her clothes off herself and left them in a pile on the floor. Luke opened the door of the conversion cubicle and helped her in. Once inside, with the door shut, Julia leant back against the wall of the cubicle and felt the steel restraints fit around her arms and legs, holding her tightly in position.
Clanking and whirring, Luke jerked over to the control console and interfaced with it directly, feeding it rewritten, efficient control sequences. "you-will-now-be- cyborgized" said Luke as he entered the final commands. "soon-your-brain-will-obey-the-computer-as-mine-does."
Julia smiled happily as the cubicle set to work on her. She knew that she had always wanted to be made robotic and reprogrammed. She was not afraid. She would obey. She made her mind blank so that the computer could take it over easily.
"I will obey. I will be made into a cyborg. I will become a cyborg." she intoned as her body was covered in metal and her brain was adjusted. "I will become a cyborg. I will obey." she repeated, Luke's red eyes watched calmly as his creator underwent her own conversion process. As she became more of a robot, he realised that his something was straining in his groin. His thick steel cock was beginning to grow and stiffen as he watched the beautiful cyborg that was being created inside the cubicle.
Julia's smile faded into a blank look. His arms and legs were entirely encased in metal. Her smooth, sensual chest gleamed in the light of the electricity that buzzed around her. Her eyes began to glow red. Her moist lips parted.
"I am a cyborg.
I - am a - cyborg.
i-am-a-cyborg." Julia's voice sounded monotonous and robotic. Had her computerized brain been capable of emotion, she would have smiled. She was content now that she was being fed instructions from the computer. She obeyed without question.
The cubicle doors slid open with a hiss. The cubicle released its subject from her restraints and she stepped out, her new shiny body clanking and jerking robotically. She looked up at Luke, who was standing in front of her at the door of the pod. They looked at each other's cyborgized body lustfully and knew what they had to do.
Holding each other, the two cyborgs kissed...
Return to the Story Archive

redpajama_set_name: RedPajamaCommonCrawl


United Kingdom Foreign Minister Jeremy Hunt said on Thursday that he is confident London will grant its support to the Brexit agreement if it excludes the Irish backstop arrangement. The official told Polish Gazeta Wyborcza that the backstop would "lock" the UK inside the customs union with Brussels, out of which it won't be able to escape.
"I agree that in December it was difficult to spot a solution. Now, however, it is clear that the parliament will support the divorce deal in the current shape, with one exception- the backstop," Hunt said. "If we change one thing - the entry that could potentially lock the United Kingdom in the customs union without an option to leave- the remaining part of the agreement will gain the support. I am sure this will work out," he added.
As the House of Commons is set for another Brexit debate and vote alter in the day, UK Trade Secretary Liam Fox said earlier today that the EU will closely follow the discussions and decide on future concessions accordingly.


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Apple's New Hire Designed Some Really Beautiful Cars
Aston Martin One-77
By Alex Fitzpatrick
April 19, 2016 10:21 AM EDT
Apple has hired former Tesla executive and Aston Martin engineer Chris Porritt, alternative transportation blog Electrek reports. Porritt is said to have been assigned to Apple's "special projects" division, which is believed to be working on automotive efforts, among other things.
Tesla and Apple have a growing history of swapping staffers. But as Electrek notes, Porritt marks the first Tesla executive poached by the Cupertino, Calif.-based technology firm.
Porritt had a hand in creating some of Aston Martin's most well-received cars while serving as that firm's chief engineer, including the One-77, DB9 and V12 Vantage.
Read Next: Meet the Electric Batmobile That Wants to Take on Telsa
It's unclear exactly what Apple is working on in the automotive space. While some rumors suggest the company is designing its own car, such a project would represent a major shift from Apple's core focus in gadgets and software. Another possibility is that Apple is building prototype cars to better understand vehicles from a user interface perspective. The company has already expressed interest in that approach—its recently introduced CarPlay software, for instance, replaces compatible vehicles' infotainment systems with a more Apple-like interface.
Apple did not immediately return TIME's request for comment.
Theranos Is Under Federal Investigation


Bozeman Police responded to a 911 hang-up call early Tuesday which turned into something much more significant at a Nickles Drive hotel.
UPDATE: At 7:12 this morning the standoff at the Microtel Inn was ended when the male suspect was taken into custody by members of the Special Response Team. We appreciate the professionalism and teamwork of the responding officers, dispatchers, tactical team members and negotiators who worked to successfully resolve this event. The investigation is ongoing and additional information will be made available later today.
At about 1:00 A.M. this morning Bozeman Police responded to a 911 hang up call at the Microtel Inn on Nikles Drive near North 7th Ave and Griffin.
A child caller was on the line and made a statement about being scared and there being a "bad guy" in the room. When Officers arrived they contacted a male suspect who was not compliant with officers and attempted to jump out a window before locking himself in the hotel room with a young male.
During the incident there were possible gun shots heard within the room also. Officers have evacuated affected guests in the hotel and currently have the scene contained to a hotel room.
The Gallatin County – Bozeman Police Special Response Team is on scene and hostage negotiators are in contact with the suspect in the room. Bozeman Police Chief Steve Crawford states that some roads in the area are blocked and asks that the public use alternate routes to avoid delays.
There is no danger to the public in the area at this time. More information will be released later this morning as it becomes available.


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Grand Central shopping centre to host massive virtual reality experience venue - including robotic bartender
Solihull restaurant named the best in the UK for 2023 in SquareMeal's top 100
Man arrested after a laser pen was shone at a police helicopter in the West Midlands
The police helicopter was assisting West Midlands officers on the ground with a vehicle stop
By Adam Dutton, Asmita Sarkar
A man was arrested after being caught in the act of shining a laser pen at a police helicopter during a vehicle investigation in the West Midlands - endangering the lives of the crew on board.
The chopper was assisting officers on the ground with a vehicle stop in Walsall when the bright beam was shone into the cockpit on Tuesday (January 17).
Video footage from the helicopter shows the laser being pointed at the aircraft, dazzling the pilot, before he is quickly detained by police on the residential street.
The 10 richest West Midlands billionaires & millionaires, according to the Sunday Times Rich List 2022 News
Birmingham's 15 richest neighbourhoods based on average income, including Kings Heath & Northfield News
A West Midlands Police spokesperson told BirminghamWorld: "Officers arrested a man in his 30s after a laser was deliberately shone at a police helicopter which was flying over the Walsall area. A laser device was seized. The man was released and will be interviewed at a later date."
The National Police Air Service (NPAS) said the culprit was now facing up to five years in jail for endangering an aircraft. The NPAS shared the footage on social media and said: "We assisted West Midlands Police with a vehicle stop in Walsall.
"Whilst doing so we were targeted with a laser pointer, fortunately force response were nearby and the offender was detained in the act. Offenders can be pin pointed in seconds and can be jailed for five years."
Head of NPAS Flight Operations Captain Paul Watts said previously after a man was jailed for a similar incident: "Laser attacks are a serious concern.
Man arrested after being caught in the act shining a laser pen at a police helicopter in the West Midlands
"The impact on a pilot is, at the very least, distracting, but can be serious enough to cause temporary 'flash' blindness and, in some cases, lasting eye damage. This could be hazardous.
"Those who carry out this dangerous action should expect to be arrested. We will always pursue prosecution with the full support of our police colleagues."
Police helicopter (Photo - SWNS)
PoliceHelicopter


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Home»World»South Africa»'He destroyed my life': Raped and tortured after being sold by family, teen 'bride' recalls ordeal | TIF News
'He destroyed my life': Raped and tortured after being sold by family, teen 'bride' recalls ordeal | TIF News
A 60-year-old man has been sentenced to life in prison for repeatedly raping and torturing a KwaZulu-Natal teenager.
The man raised the defence of the traditional practice of "ukuthwala".
But a Zulu cultural expert said what the man did was "abduction, not ukuthwala".
"He deserves to rot in jail."
These were the words of cultural expert and academic Dr Gugulethu Mazibuko after a 60-year-old man was jailed for life after he repeatedly tortured and raped a KwaZulu-Natal teenager who was forced to marry him.
The man had raised the defence of the traditional practice of ukuthwala, but the court rejected the argument and convicted him.
The justice department defines ukuthwala as a form of abduction involving a man and his friends or peers kidnapping a girl or young woman to compel her family to endorse marriage negotiations.
However, Mazibuko told TIF News that ukuthwala was not abduction.
"Ukuthwala is when two people who are in love feel they are ready to get married and agree that on a specific date… the boy's family pays lobola. Ukuthwala has nothing to do with forcing someone."
READ | KZN man, 60, sentenced to life for torturing, repeatedly raping teen ukuthwala victim
She said the man's actions were criminal and had nothing to do with ukuthwala. She added that he deserved to be punished.
"It is not acceptable. Whoever is involved in that must rot in jail. That is brutality. That man killed everything in that young girl.
"The man should disassociate ukuthwala with [his] criminal actions," she said, adding that the practice was rare.
"I haven't heard about it for quite a while but when people want to do selfish things, they say it is ukuthwala."
The Conversation previously reported that the Western Cape High Court, in a 2015 ruling, "determined features of traditional ukuthwala under customary law" to include consent of both parties to the marriage, the enactment of a "pretend" kidnapping of the bride, and "strict prohibition of sex" during the abduction.
The Greytown Regional Court in KwaZulu-Natal convicted and sentenced the man to life imprisonment for rape, sexual assault and kidnapping on Monday after it heard the ordeal the teenager endured between February and July 2019.
The victim, who was 17 years old at the time, was living with her aunt when she was forced to marry the man, who was a member of the church she attended.
National Prosecuting Authority spokesperson Natasha Kara said the court heard how the teenager was rescued after her harrowing ordeal.
Regional Court prosecutor Premier Naidoo led the evidence of the victim and a friend who had been in her company at the time of the kidnapping, as well as the social worker and police officer who had rescued her.
A medical doctor testified that the victim sustained extensive injuries due to the trauma of the rape over the 10 days she was held captive.
'He has destroyed my life'
In a victim impact statement, the girl said she had isolated herself since the incident "because I am hurt so much emotionally".
She said in the statement:
He has destroyed my life and my future. I have tried suicide a few times but was saved. I just wish the court could do justice for me as I became a laughing stock after being kidnapped and raped.
According to police, the girl's aunt and church elders had agreed to the so-called marriage, with the aunt accepting payment for the arrangement.
"She would allow the man to come to her house and have his way with the girl, and when the girl complained, she was reminded that traditionally she belonged to the man," police spokesperson Constable Thenjiswa Ngcobo said.
After her concerns fell on deaf ears, she attempted to kill herself twice.
Realising she could not flee, the teenager, whose parents had long died, decided to run away from her aunt's house.
The man, however, hired two men to track down his victim.
READ | 'My husband dragged me into a sex trafficking ring' – victims reveal truth behind human trafficking
"She was caught and taken to the accused's house in Greytown, where she was expected to continue with her marital duties.
"He attempted to rape her, and she fought back. When he realised that it was not going to be easy to overpower her, she was taken to Matimatolo (in KwaZulu-Natal), where she was handcuffed and raped repeatedly," said Ngcobo.
The teenager was kept in a room for months.
The local community became suspicious when they heard her crying whenever the man was at home.
Police were alerted and responded swiftly, Ngcobo said.
Meanwhile, Javu Baloyi, spokesperson for the Commission for Gender Equality, welcomed the sentence.
"We have been calling for the judicial system to self-correct in that leniency was sending a wrong message out there because people would be arrested today and tomorrow, they are out on parole, and cases disappear," Baloyi added.
Baloyi said forced marriages were still rife in several provinces.
"The sentence will serve as a message to those who force teenagers, young girls to marry them against their will, let alone raping them."
READ | Foundation sounds alarm on cases of ukuthwala still being practised in rural parts of South Africa
Jason Behrens, the founder of NPO Jason's Angels, which focuses on helping survivors of gender-based violence, described the incident as an example of a "very big challenge we face in South Africa in the fight against gender-based violence".
"South Africa has a diverse population consisting of various races, religions and cultures."
Behrens continued:
What perpetrators need to understand is that the South African law supersedes any cultural or religious belief. We welcome the sanctions imposed as this is an indication that the courts are starting to take GBV more seriously, and especially rape.
Bronwyn Litkie, the founder of SA Women Fight Back, said she was "utterly flabbergasted".
"I wonder how much money the aunt was paid for her niece to marry this man," she said.
"This should be considered human trafficking, and, in my opinion, the aunt should be on trial too. When traditional practices come into play, some men seem to think that tradition means stripping a woman of all human rights, dignity and choice.
"It is our human right to choose who we are with and who we give sexual consent to. Ukuthwala is an ancient tradition that has no place in a country riddled with GBV."
Here are your Daily Lotto results | TIF News
Cricket SA selection head Mpitsang hopes fire-starter Rossouw will come off at World Cup | Sport
Jobs of around 1 000 Putco employees on the line over strike | TIF News


text:
Q: Creating Indexname Dynamically using Spring-Data Elasticsearch with @Document I am trying to get indexname dynamically in
@Document(indexName="Something",type="type")

I have tried using below code:
@Document(indexName="#{systemProperties['databaseName'] }", type = "dbtype")

but at runtime when I am sending two request from UI to get the data from ES from different indexes,it is not working properly.
what should I  do to  solve this issue?

A: A workaround is, to work with NativeSearchQuerries.
There you can set the index to whatever you like:
NativeSearchQuery query = new NativeSearchQueryBuilder().withIndices(indexName)
                .withQuery(QueryBuilders.matchPhraseQuery("_all", request.getSearchTerm().getSearchString()))
                .withPageable(new PageRequest(request.getPaging(), request.getMaxResults()))
....
                .build();
        result =elastic.query(query, query->query2Result(query));

redpajama_set_name: RedPajamaStackExchange


Dentsu Aegis Network has announced that its president of global clients has been appointed as its new CEO, UK & Ireland with immediate effect.
Euan Jarvie, who has experience running large-scale media businesses around the world, will head the UK and Ireland leadership team and run a business with more than 4,000 staff serving over 1,000 clients across 22 media, technology, data and creative agencies.
"I'm looking forward to working with Euan to drive a renewed focus and energy across the UK&I network," said Nick Waters, executive chairman, Dentsu Aegis Network UK & Ireland, who was previously leading the business.
"He is an experienced, considered media professional with strong client relationships. We are fully focused on maximising the significant talent and expertise we have for the benefit of our clients and I am confident that together we will take the business forward to deliver great success."
Prior to Dentsu Aegis, Euan worked at MediaCom for over 18 years, most recently running its operations across the US and then EMEA, and he has experience working with some of the world's biggest global brands.
"I am delighted to return to a UK centric role and thrilled to be leading Dentsu Aegis in the UK & Ireland," said Jarvie.
"The experience I have gained running clients and businesses all over the world has set me up well to lead the wonderful team here. Together we will deliver even more dynamic and innovative solutions to build our clients' brands in the 21st century.
"Success for me is grounded in a culture of trust and bravery where everyone understands how their contribution is helping deliver growth for our clients. I believe it is the most exciting time ever in the industry with many organisations looking to transform and find new growth.
"Our UK&I business is already well down the path of its own transformation and I'm thrilled to be part of it."


text:
1.A.17.1: The Anoctamin (ANO) Family
1.A.17.1.1
The plasma membrane Ca2 -activated chloride (IClCa) channel, TMEM16A (Anoctamin 1a; ANO1a; ANO1, DOG1, ORAOV2, TAOS2) (Huang et al., 2012; Chen et al. 2011). The mouse orthologue (Q8BHY3), TMEM16A (956aas), is localized to the apical membranes of epithelia as well as intracellular membranes in many cell types. Knockout mice show diminished rhythmic contraction of gastric smooth muscle (Huang et al., 2009). ANO1 is also required for normal tracheal development (Ousingsawat et al., 2009). Expression is upregulated by epidermal growth factor (Mroz and Keely, 2012). Novel 5-substituted benzyloxy-2-arylbenzofuran-3-carboxylic acids are inhibitors (Kumar et al., 2012). TMEM16A channels contribute to the myogenic response in cerebral arteries (Bulley et al., 2012). Membrane stretch activates arterial myocyte TMEM16A channels, leading to membrane depolarization and vasoconstriction. A local Ca2+ signal generated by nonselective cation channels stimulates TMEM16A channels to induce myogenic constriction (Bulley et al., 2012). Ca2+/calmodulin activates bicarbonate (anion) transport (Jung et al. 2012). The protein exists in the membrane as a homodimer where the cytoplasmic N-terminus functions in dimerization (Tien et al. 2013). TMSs 5-6 of the 8 TMSs may comprise parts of the pore-loop that controls Cl- conductance (Adomaviciene et al. 2013). ANO1 confers IClCa in retinal neurons and acts as an intrinsic regulator of the presynaptic membrane potential during synaptic transmission (Jeon et al. 2013). TMEM16A may be a primary driver of the "Grow" (tumor proliferation) or "Go"(metastasis) model for cancer progression, in which TMEM16A expression acts to balance tumor proliferation and metastasis via its promoter methylation (Shiwarski et al. 2014). Regulation of TMEM16A/16B by Ca2+ is mediated by preassociated apo-calmodulin (Yang et al. 2014) as well as CaMKIIδ (Gui et al. 2015). Because the Cl- channel is the only active ion-selective conductance with a reversal potential that lies within the dynamic range of spiral ganglion neurons (SGN) action potentials, developmental alteration of [Cl-], and hence the equilibrium potential for Cl- (ECl), transforms the pre- to the post-hearing phenotype (Zhang et al. 2015). Four basic residues involved in ion selectivity and pore blocker sensitivity have been identified (Peters et al. 2015). Channel activity is required for mucus secretion induced by interleukin-13 (Lin et al. 2015; Zhang et al. 2015). Ano1 may interact cooperatively with TrpV1 (TC# 1.A.4.2.1) to form a thermal sensor (Kanazawa and Matsumoto 2014). Inhibitors have been described (Boedtkjer et al. 2015). The first intracellular loop serves as a Ca2+ binding site and includes D439, E444 and E447 (Pang et al. 2015). It is inhibited by various 4-Aryl-2-amino thiazoles at concentrations as low as 1 mμM (Piechowicz et al. 2016). ANO1 and TRPC6 (1.A.4.1.5) are present in the same macromolecular complex and localize in close spatial proximity in the myocyte plasma membrane. TRPC6 channels probably generate a local intracellular Ca2+ signal that activates nearby ANO1 channels in myocytes to stimulate vasoconstriction (Wang et al. 2016). ANO1 transports bicarbonate which functions in the regulation of pancreatic acinar cell pH (Han et al. 2016). TMEM16A contains two ion conduction pores that are independently activated by Ca2+ binding to sites that are embedded within the transmembrane part of each subunit (Lim et al. 2016). Interactions between the carboxy- terminus and the first intracellular loop in the TMEM16A homo-dimer regulate channel activity (Scudieri et al. 2016). A STAT6-TMEM16A-ERK1/2 signal pathway and TMEM16A channel activity are required for the Interleukin-13 (IL-13)-induced TMEM16A-mediated mucus production (Qin et al. 2016). Angiotensin II elicits a TMEM16A-mediated current, and TMEM16A participates in Ang II-induced basilar constriction via the RhoA/ROCK signaling pathway (Li et al. 2016). 2-acylamino-cycloalkylthiophene-3-carboxylic acid arylamides (AACTs) are inhibitors of TMEM16A, and 48 synthesized analogs (10ab-10bw) of the original AACT compound (10aa) have been synthesized and studied. The most potent compound (10bm), which contains an unusual bromodifluoroacetamide at the thiophene 2-position, had an IC50 ~ 30 nM (Truong et al. 2017). Ano1 plays a role in asthma (Wang et al. 2017). The E143A mutant showed reduced sensitivity to Ca2+ but not to high temperatures, whereas the E705V mutant exhibited reduced sensitivity to both Ca2+ and noxious heat (Choi et al. 2018). Voltage modulation of TMEM16A involves voltage-dependent occupancy of calcium- and anion-binding site(s) within the membrane electric field as well as a voltage-dependent conformational change intrinsic to the channel protein. These gating modalities all critically depend on the sixth transmembrane segment (Peters et al. 2018). TMEM16A in myocytes plays a major functional role in contraction (Mohanakumar et al. 2018). Bile acids activate TMEM16A and thereby increase cholangiocyte fluid secretion (Li et al. 2018). TMEM16A participates in H2O2-induced apoptosis via modulation of mitochondrial membrane permeability (Zeng et al. 2018). Glioma-associate oncogene proteins, Gli1 and Gli2, bind to the promoter and repress ANO1 transcription, dependent on Gli binding to a site close to the ANO1 transcriptional start site (Mazzone et al. 2019). Clarithromycin suppresses IL-13-induced goblet cell metaplasia via the TMEM16A-dependent pathway in guinea pig airway epithelial cells (Hara et al. 2019). TMEM16A is involved in gastric emptying, and TMEM16A inhibition may be effective in treating disorders of accelerated gastric emptying, such as dumping syndrome (Cil et al. 2019). Phosphatidylinositol (4,5)-bisphosphate (PIP2) regulates TMEM16A channel activation and desensitization by binding to a binding site, possibly at the cytosolic interface of TMSs 3-5. The ion-conducting pore of TMEM16A consists of two functionally distinct modules: a Ca2+-binding module formed by TMSs 6-8 and a PIP2-binding regulatory module formed by TMs 3-5, which mediate channel activation and desensitization, respectively (Sui et al. 2020). TMEM16A plays a dual role in LPS-induced intestinal epithelial barrier dysfunction (Sui et al. 2020). Hepatocyte TMEM16A plays a role in nonalcoholic fatty liver disease (NAFLD), and its deletion retards NAFLD progression by ameliorating hepatic glucose metabolic disorder (Guo et al. 2020). Hepatocyte TMEM16A interacts with vesicle-associated membrane protein 3 (VAMP3) to induce its degradation, suppressing the formation of the VAMP3/syntaxin 4 and VAMP3/synaptosome-associated protein 23 complexes (see TC# 1.F.1.1.5). This leads to impairment of hepatic glucose transporter 2 (GLUT2) translocation and glucose uptake (Guo et al. 2020). TMEM16A is a potential biomarker for Lung Cancer (Hu et al. 2019). Allosteric modulation of alternatively spliced Ca2+-activated Cl- channels, TMEM16A by PI(4,5)P2 and CaMKII (TC# 8.A.104.1.11) has been demonstrated (Ko et al. 2020). Signaling through the interleukin-4 and interleukin-13 receptor complexes regulates cholangiocyte TMEM16A expression and biliary secretion (Dutta et al. 2020). A second Ca2+ binding site allosterically controls TMEM16A activation (Le and Yang 2020). A long noncoding RNA (lncRNA), ANO1-AS2, downregulates the ANO1 gene by interacting with the ANO1 gene promoter, which can influence sperm motility and morphology (Saberiyan et al. 2020). Ano1 plays an important role in generating urethral tone (Drumm et al. 2021). Human TMEM16A shows increated expression in many cancers (Chen et al. 2021). TMEM16A is inhibitied by liquiritigenin (Kato et al. 2021) and is activated by the natural product canthaxanthin which promotes gastrointestinal contraction (Ji et al. 2020). TMEM16A ameliorates vascular remodeling by suppressing autophagy via inhibiting Bcl-2-p62 complex formation. It regulates the four-way interaction between p62 (P37198; TC# 1.I.1.1.3), Bcl-2 (TC# 1.A.21.1.10), Beclin-1 (BECN1 or GT197; Q144570; TC# 9.A.15.2.1), and VPS34 (phosphatidylinositol 3-kinase, PI 3-kinase, PIK3C3), and coordinately prevents vascular autophagy and remodeling (Lv et al. 2020). A small molecule inhibitor of TMEM16A (2-bromodifluoroacetylamino-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carbox ylic acid o-tolylamide) blocks vascular smooth muscle contraction and lowers blood pressure in spontaneously hypertensive rats (Cil et al. 2021). Evodiamine and rutecarpine are TMEM16A inhibitors (Zhao et al. 2021). Cepharanthine is a selective ANO1 inhibitor with potential for lung adenocarcinoma therapy (Zhang et al. 2021). Benzophenanthridine alkaloids suppress lung adenocarcinoma by blocking TMEM16A Ca2+-activated Cl- channels (Zhang et al. 2020). The diverse roles of TMEM16A Ca2+-activated Cl- channels in inflammation have been described (Bai et al. 2021). TMEM16A-mediated breast cancer metastasis has been described in which ROCK1 increases TMEM16A channel activity via moesin phosphorylation. An increase in TMEM16A channel activity promotes cell migration and invasion (Luo et al. 2021). Four Ca2+ sensing sites in TMEM16A have been identfied, and the activation properties of TMEM16A by them has been discussed (Ji et al. 2021). Blockade of TMEM16A protects against renal fibrosis by reducing the intracellular Cl- concentration (Li et al. 2021). The TMEM16A/anoctamin 1 inhibitor T16Ainh-A01 reverses monocrotaline-induced rat pulmonary arterial hypertension (Xie et al. 2020). The role of TMEM16A/ERK/NK-1 signaling in dorsal root ganglia neurons in the development of neuropathic pain induced by spared nerve injury has been studied (Chen et al. 2021). Elevated ANO1 (DOG1) expression is frequent in colorectal cancer and is linked to molecular alterations (Jansen et al. 2022). ANO1 plays a role in the occurrence, development, metastasis, proliferation, and apoptosis of various malignant tumors. Guo et al. 2022 reviewed the mechanism of ANO1 involved in the replication, proliferation, invasion and apoptosis of various malignant tumors. Procyanidin (PC) is an efficacious and selective inhibitor of TMEM16A with an IC50 of 10.6 +/- 0.6 muM. The precise sites (D383, R535, and E624) of electrostatic interactions between PC and TMEM16A are known (Li et al. 2022). TMEM16A is a Ca2+activated Cl- channel that plays critical roles in regulating vascular tone, sensory signal transduction, and mucosal secretion. TMEM16A activation also requires the membrane lipid phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) (Tembo et al. 2022). TMEM16A may promote angiogenesis of the heart during pressure-overload (Zhang et al. 2022), and it is an immunohistochemical marker of acinic cell carcinoma (Fiorentino et al. 2022). The progress in understanding solute carrier SLC families in nonalcoholic fatty liver disease has been reviewed (Tang et al. 2022). Arctigenin attenuates vascular inflammation induced by high salt through the TMEM16A/ESM1/VCAM-1 pathway (Zeng et al. 2022). Biologics that inactivate Nav1.7 channels have the potential to reduce arthritis pain over a protracted period of time (Reid et al. 2022).
Anoctamin 1a of Homo sapiens (Q5XXA6)
Anoctamin 1, isoform b (Gnathodiaphyseal dysplasia 1 protein homologue) (39% identical to Anoctamin 1a) (Planells-Cases and Jentsch, 2009). See also Xu et al. 2015.
Anoctamin 1b of Homo sapiens (Q75UR0)
TMEM16B (Anoctamin-2, ANO2) anion channel. Exists in the membrane as a homodimer where the cytoplasmic N-terminus functions in dimerization (Tien et al. 2013). TMSs 5-6 may comprise parts of the pore-loop that controls Cl- conductance (Adomaviciene et al. 2013). TMEM16A and TMEM16B are differentially expressed during development in the olfactory epithelium of the mouse (Maurya and Menini 2014).
TMEM16B of Homo sapiens (Q9NQ90)
Anoctamin-6 (ANO6: TMEM16F) Ca2+-dependent phospholipid scramblase (flippase) (Suzuki et al., 2010; Chauhan et al. 2016). Defects cause Scott syndrome, and promote assembly of the tenase and prothrombinase complexes involved in blood coagulation (Fujii et al. 2015). It is an essential component of the outwardly rectifying chloride channel (Martins et al., 2011; Keramidas and Lynch 2012). It has also been reported to be an anion channel with delayed Ca2+ activation (Adomaviciene et al. 2013) as well as a Ca2+-activated cation channel with activity that is required for lipid scrambing (Yang et al. 2012). However, Suzuki et al. (2013) showed that TMEM16F is a Ca2+-dependent phospholipid scramblase that exposes phosphatidylserine (PS) to the cell surface but lacks calcium-dependent chloride channel activity (see also Segawa et al. 2011). TMEM16C, 16D, 16G and 16J also have Ca2+-dependent scramblase activities but not channel activity (Suzuki et al. 2013). The pore region suggested to be resonsible for Cl- transport in TMEM16A is also responsible for phospholipid scramblase activity (Suzuki et al. 2014). Anoctamin-6 (Ano6) plays an essential role in C2C12 myoblast proliferation, probably by regulating the ERK/AKT signaling pathway (Zhao et al. 2014). It regulates baeline phosphatidyl serine exposure and cell viability in human embryonic kidney cells (Schenk et al. 2016). A single TMEM16F molecule transports phospholipids nonspecifically between the membrane bilayers dependent on Ca2+. Thermodynamic analyses indicated that TMEM16F transports 4.5 x 104 lipids per second at 25 degrees C, with an activation free energy of 47 kJ/mol, suggesting a channel-dependent, facilitated diffusion,"stepping-stone" mechanism (Watanabe et al. 2018). TMEM16F plays roles in platelet activation during blood clotting, bone formation, and T cell activation. Activation of TMEM16F by Ca2+ ionophores triggers large-scale surface membrane expansion in parallel with phospholipid scrambling (Bricogne et al. 2019). With continued ionophore application,TMEM16F-expressing cells undergo extensive shedding of ectosomes which incorporate The T cell co-receptor PD-1. Cells lacking TMEM16F fail to expand the surface membrane in response to elevated cytoplasmic Ca2+and instead undergo endocytosis with PD-1 internalization. This suggests a new role for TMEM16F as a regulator of Ca2+-activated membrane trafficking (Bricogne et al. 2a019). The inner activation gate consists of three hydrophobic residues, F518, Y563 and I612, in the middle of the phospholipid permeation pathway. Disrupting the inner gate profoundly alters phospholipid permeation. Lysine substitutions of F518 and Y563 lead to constitutively active CaPLSases that bypass Ca2+-dependent activation. An analogous lysine mutation to TMEM16F-F518 in TMEM16A (L543K) is sufficient to confer CaPLSase activity to the Ca2+-activated Cl- channel (CaCC) (Le et al. 2019). ANO6, by virtue of its scramblase activity, may play a role as a regulator of the ADAM-network in the plasma membrane. TMEM16F inhibition limits pain-associated behavior and improves motor function by promoting microglia M2 polarization in mice (Zhao and Gao 2019). Polyphenols do not inhibit the phospholipid scramblase activity of TMEM16F (Le et al. 2020). TMEM16F is a ubiquitously expressed Ca2+-activated phospholipid scramblase that also functions as a largely non-selective ion channel with open, closed and intermediate conformations (Jia et al. 2022). An inner activation gate consists of F518, Y563, and I612, and charged mutations of the inner gate residues leads to constitutively active mammalian (m)TMEM16F scrambling. Lysine substitution of F518 and Y563 leads to spontaneous opening of the permeation pore in the Ca2+-bound state of mTMEM16F. Dilation of the pore exposes hydrophilic patches in the upper pore region, greatly increases the pore hydration level, and enables lipid scrambling (Jia et al. 2022).
Anoctamin-6 of Homo sapiens (Q4KMQ2)
Anoctamin-9 (Transmembrane protein 16J) (Tumor protein p53-inducible protein 5) (p53-induced gene 5 protein). It promotes pancreatic tumorigenesis (Jun et al. 2017).
ANO9 of Homo sapiens
Uncharacterized protein of Batrachochytrium dendrobatidis
Anoctamin-like protein
Choanoflagellida
amoctamin-like protein of Dictyostelium purpureum
unchacterized protein of Aureococcus anophagefferens
Anoctamin, Anoh-1 of 822 aas. Functions in a sensory mode-specific manner. Present inamphid sensory neurons to detect external chemical and nociceptive cues (Wang et al. 2013).
Anoh-1 of Caenorhabditis elegans
1.A.17.1.10
Anoctamin, Anoh-2. Present in mechanoreceptive neurons and spermatheca (Wang et al. 2013).
Anoctamin-like protein At1g73020
Ca-ClC Family homologue
Ca-ClC homologue of Paramecium tetraurelia (A0CAP8)
Ciliate CaClC homologue
CaClC homologue of Paramecium tetraurelia (A0CIB0)
Water mold Anoctamin-like protein
Peronosporales
Anoctamin-like protein of Phytophthora infestans (D0NGF4)
Uncharacterized protein of Schizosaccharomyces japonicus
Anoctamin-like protein of Oxytricha trifallax
TMEM16 (Ist2) ion channel/phospholipid scramblase of 735 aas and 8 - 10 TMSs (Malvezzi et al. 2013). Three high-resolution cryo-EM structures of this scramblase, reconstituted in lipid nanodiscs, revealed that Ca2+-dependent activation of the scramblase entails global rearrangement of the transmembrane and cytosolic domains. Activation of the protein thins the membrane near the transport pathway to facilitate rapid transbilayer lipid movement (Falzone et al. 2019).
Ist2 of Aspergillus fumigatus (Neosartorya fumigata)
TMEM16 of 735 aas and 10 TMSs. Operates as a Ca2+-activated lipid scramblase (Wang et al. 2018). Each subunit of the homodimer contains a hydrophilic membrane-traversing cavity that is exposed to the lipid bilayer as a potential site of catalysis. This cavity harbours a conserved Ca2+-binding site, located within the hydrophobic core of the membrane. Mutations of residues involved in Ca2+ coordination affect both lipid scrambling in N. haematococca TMEM16 and ion conduction in the Cl- channel of TMEM16A. The structure reveals the general architecture of the family and its mode of Ca2+ activation (Brunner et al. 2014). While the cytoplasmic portion of the protein is important for function, it does not appear to regulate scramblase activity via a detectable conformational change (Andra et al. 2018). Dynamic modulation of the lipid translocation groove generates a conductive ion channel in Ca2+-bound nhTMEM16 (Khelashvili et al. 2019) (see family description). Permeation of potassium ions through the lipid scrambling path of nhTMEM16 has been documented (Cheng et al. 2022).
TMEM16 of Nectria haematococca (Fusarium solani subsp. pisi)
Increased sodium tolerance protein, Ist2, of 946 aas and 7 TMSs. Ist2 is an endoplasmic reticulum (ER)-resident transmembrane protein that mediates associations between the plasma membrane (PM) and the cortical ER (cER) in baker's yeast (Kralt et al. 2015).
Ist2 of Saccharomyces cerevisiae
Anoctamin 3, ANO3 or TMEM16C or KCNT1/Slack, of 981 aas and 9 putative TMSs. Has calcium-dependent phospholipid scramblase activity, scrambling phosphatidylcholine and galactosylceramide. Seems to act as a potassium channel regulator and may inhibit pain signaling; can facilitate KCNT1/Slack channel activity by promoting its full single-channel conductance at very low sodium concentrations and by increasing its sodium sensitivity (Scudieri et al. 2012). Mutations cause (i) epilepsy of infancy with migrating focal seizures (EIMFS; also known as migrating partial seizures in infancy), (ii) autosomal dominant nocturnal frontal lobe epilepsy, and (iii) other types of early onset epileptic encephalopathies (EOEEs) (Ohba et al. 2015). TMEM16C/Slack regulation of excitatory synaptic plasticity via GluA1-containing AMPA Receptors is critical for the pathogenesis of remifentanil-induced postoperative hyperalgesia in rats (Li et al. 2021). Specific mutational variants in TMS of ANO3 can be responsible for childhood-onset movement disorders with intellectual disability (Aihara et al. 2022).
ANO3 or KCNT1 of Homo sapiens
Ano5 (GDD1, TMEM16E), of 913 aas and 10 TMSs, is an intracellular calcium-activated chloride channel in the endoplasmic reticulum. It positively modulates bone homeostasis via calcium signaling in GDD (Li et al. 2022), and is associated with bone fragility, limb girdle muscular dystrophy type 2L (LGMD2L), Miyoshi myopathy type 3 (MMD3), and gnathodiaphyseal dysplasia 1 (GDD1) in humans (Jin et al. 2017), but an Ano5 knock-out mutant in mice was not reported to exhibit such symptoms (Xu et al. 2015). The orthologue in mice is TC# 1.A.17.1.2. TMEM16E may function as a phospholipid scramblase in intracellular membranes, promoting sperm motility and function (Gyobu et al. 2016). Dysregulated calcium homeostasis prevents plasma membrane repair in Anoctamin 5/TMEM16E-deficient patient muscle cells (Chandra et al. 2019). Ano5 is involved in familial florid osseous dysplasia (Lv et al. 2019). Pharmacological inhibition of ANO5 or lack of ANO5, prevent Ca2+ uptake into the ER following plasma membrane damage and Ca2+ overload (Chandra et al. 2021). Thus, Cl- uptake into the ER is required to sequester injury-promoted cytosolic Ca2+. Anoctamin 5 regulates the cell cycle and affects the prognosis in gastric cancer (Fukami et al. 2022). Thus, ANO5 may be a key mediator in tumor progression and promises to be a prognostic biomarker for gastric cancer.
Subdued, a calcium-activated chloride channel of 1075 aas. Functions in conjunction with the thermo-TRPs in thermal nociception. Subdued channels may amplify the nociceptive neuronal firing that is initiated by thermo-TRP channels in response to thermal stimuli (Jang et al. 2015). It may also act on phospholipids to transport or hydrolyze them (Le et al. 2019).
Subdued of Drosophila melanogaster
ANO-like protein of 921 aas and 9 predicted TMSs.
ANO-L family protein of Strongylocentrotus purpuratus (Purple sea urchin)
Duplicated full length anoctamin of 2084 aas and an etimated 20 TMSs. The protein has two full length repeats, each of about 1000 aas with a ~500 aas hydrophilic domain followed by the first anoctamin domain, and then another 500 aa hydrophilic domain followed by the second anoctamin domain.
Oomycetes
Dupicated anoctamin of Aphanomyces invadans
TMem16A or Anoctamin-1 (Ano1) Ca2+-activated anion (Cl-) channel of 960 aas and 10 TMSs. Its structure has been solved by cryoEM (Paulino et al. 2017). The protein shows a similar organization to the fungal nhTMEM16, except for changes at the site of catalysis. There, the conformation of transmembrane helices, constituting a membrane-spanning furrow that provides a path for lipids in scramblases, is replaced to form an enclosed aqueous pore that is largely shielded from the membrane (Paulino et al. 2017). It thus provides a pathway for anions such as Cl-. During activation, the binding of Ca2+ to a site located within the transmembrane domain, in the vicinity of the pore, alters the electrostatic properties of the ion conduction path and triggers a conformational rearrangement of an α-helix that comes into physical contact with the bound ligand, and thereby directly couples ligand binding and pore opening (Paulino et al. 2017). The E143A mutant showed reduced sensitivity to Ca2+ but not to high temperatures, whereas the E705V mutant exhibited reduced sensitivity to both Ca2+ and noxious heat (Choi et al. 2018). Loss of TMEM16A resulted in reduced nephron number and, subsequently, albuminuria and tubular damage (Schenk et al. 2018). mAno1 expression is regulated via alternative promoters, and its transcriptional variation results in variation of the N-terminal sequence of the Ano1 protein due to alternative translation initiation sites (Kamikawa et al. 2018). The Ca2+ gating mechanism of TMEM16A, involving a Ca2+-sensing element close to the anion pore, alters conduction and substrate selection. De Jesús-Pérez et al. 2022 studied the gating-permeant anion relationship using mouse TMEM16A, showing that the apparent Ca2+ sensitivity increases with highly permeant anions and SCN- mole fractions, likely by stabilizing bound Ca2+. Conversely, mutations in crucial gating elements, including the Ca2+-binding site 1,TMS 6, and the hydrophobic gate, impaired anion permeability and selectivity. Thus, there is a reciprocal rationship between gating and selectivity (De Jesús-Pérez et al. 2022). Propagation of pacemaker activity and peristaltic contractions in the mouse renal pelvis rely on Ca2+-activated Cl- Channels such as Ano1 and T-type Ca2+ channels (Grainger et al. 2022).
Ano1 of Mus musculus
Anoctamin-10 or TMEM16K of 660 aas and 9 or 10 TMSs. In the presence of Ca2+, TMEM16K directly binds Ca2+ to form a stable complex (Ishihara et al. 2016). In the absence of Ca2+, TMEM16K and TMEM16F (TC# 1.A.17.1.4) aggregate, suggesting that their structures are stabilized by Ca2+. Mutagenesis of acidic residues in TMEM16K's cytoplasmic and transmembrane regions identified five residues that are critical for binding Ca2+. These residues are well conserved between TMEM16F and 16K, and point mutations of these residues in TMEM16F reduced its ability to support Ca2+-dependent phospholipid scrambling (Ishihara et al. 2016). Phosphatidyl serine in the ER of mammalian cells is predominantly localized to the cytoplasmic leaflet, but TMEM16K directly or indirectly mediates Ca2+-dependent phospholipid scrambling (Tsuji et al. 2019). Ano10 plays roles in cell division, migration, apoptosis, cell signalling, and developmental processes (Chrysanthou et al. 2022).
TMEM16K of Homo sapiens
Anoctamin 7, ANO7, or TMEM16G, of 933 aas and 10 TMSs. It has calcium-dependent phospholipid scramblase activity, scrambling phosphatidylserine, phosphatidylcholine and galactosylceramide, but it does not exhibit calcium-activated chloride channel (CaCC) activity. It may play a role in cell-cell interactions (Das et al. 2008). ANO7 is associated with aggressive prostate cancer (Kaikkonen et al. 2018). Insights into the topology and function of Ano7 have been described (Guo et al. 2021). Activation of calcium-activated chloride channels suppresses inherited seizure susceptibility in genetically epilepsy-prone rats (Thomas et al. 2022).
Anoctamin1, Ano1, TMEM16A of 979 aas and 10 TMSs. It is probably an anion (Cl-) cannel. Fertilization activates TMEM16A channels in X. laevis eggs and induces the earliest known event triggered by fertilization: the fast block to polyspermy (Wozniak et al. 2018).
Ano1 of Xenopus laivis
Anoctamin-4, ANO4, TMEM16D, of 955 aas and 10 putative TMSs. 68% identical to ANO4 (TC# 1.A.17.1.20). It has calcium-dependent phospholipid scramblase activity, scrambling phosphatidylserine, phosphatidylcholine and galactosylceramide, and it is a Ca2+-dependent non-selective cation channel (Reichhart et al. 2019). ANO4 is primarily expressed in the CNS and certain endocrine glands, and mutations affecting protein stability have been associated with various neuronal disorders (Reichhart et al. 2021).
Anoctamin-8, Ano8 or TMEM16H, of 1232 aas and 9 TMSs. It tethers the endoplasmic reticulum and plasma membrane for assembly of Ca2+ signaling complexes at the ER/PM compartment (Jha et al. 2019). ANO8 is a key tether in the formation of the ER/PM junctions that are essential for STIM1-STIM1 interaction and STIM1-Orai1 interaction and channel activation at a ER/PM PI(4,5)P2-rich compartment. Moreover, ANO8 assembles all core Ca2+ signaling proteins: Orai1, PMCA, STIM1, IP3 receptors, and SERCA2 at the ER/PM junctions to mediate a novel form of Orai1 channel inactivation by markedly facilitating SERCA2-mediated Ca2+ influx into the ER. This controls the efficiency of receptor-stimulated Ca2+ signaling, Ca2+ oscillations, and the duration of Orai1 activity to prevent Ca2+ toxicity (Jha et al. 2019).
1.A.17.2: The Anoctamin-like (ANO-L) Family
DUF590 family protein
DUF590 protein of Dicyostelium discoideum (Q54BH1)
TMEM16 homologue of 701 aas.
TMEM16 homologue of Naegleria gruberi (Amoeba)
Anoctamin homologue of 689 aas
Cryptophyta
Anoctamin of Guillardia theta
DUF590 homologue of 487 aas
Entamoeba
DUF590 homologue of Entamoeba nuttalli
DUF590 protein of Allomyces macrogynus
Uncharacterized protein of 569 aas and 8 predicted TMSs.
UP of Dictyostelium fasciculatum (Slime mold)
1.A.17.3: The Calcium-permeable Stress-gated Cation Channel-like 1 (CSC-1) Family
Uncharacterized protein of 2464 aas and 11 TMSs. Contains a trypsin-like serine protease domain (residues 100 - 400), a rabaptin (chromosome segregation) domain (residues 900 - 1200), an anoctamin domain (residues 1500 - 2000) and an AAA ATPase-containing von Willebrand factor type A domain (residues 2200 - 2500).
UP of Thalassiosira pseudonana (Marine diatom) (Cyclotella nana)
UP of Phytophthora parasitica (Potato buckeye rot agent)
Uncharacterized protein of 1231 aas and 9 TMSs
Pelagophyceae
UP of Aureococcus anophagefferens (Harmful bloom alga)
Uncharacterized protein of 945 aas and 8 TMSs
UP of Ectocarpus siliculosus (Brown alga)
Uncharacterized protein of 1437 aas
Isochrysidales
UP of Emiliania huxleyi
Ichthyosporea
UP of Capsaspora owczarzaki
DUF590/putative methyltransferase of 1221 aas and 10 TMSs.
DUF490 homologue of Oxytricha trifallax
DUF590 homologue of 1026 aas and 10 TMSs
DUF590 homologue of Paramecium tetraurelia (ciliate)
UP of Ostreococcus lucimarinus
Anoctamin homologue of 1265 aas
Anoctamin homologue of Tetrahymena thermophila
UP of Tetrahymena thermophila
Uncharacterized protein of 10 TMSs in a 3 + 4 +3 arrangement
UP of Paramecium tetraurelia
Uncharacterized protein of 888 aas and 10 TMSs in a 3 + 4 + 3 arrangement
Uncharacterized protein of 958 aas and 11 or 12 TMSs in a 3 or 4 + 5 +3 arrangement.
1.A.17.4: The Transmembrane Channel (TMC) Family
TMC2, like TMC1, plays a role in hearing and gravity detection (Kawashima et al., 2011). Required for normal function of cochlear hair cells, possibly as a Ca2+ channel (Kim and Fettiplace 2013). TMC1 and TMC2 are both components of hair cell transduction channels and contribute to permeation properties (Pan et al. 2013; Kawashima et al. 2014). Both TMC1 and 2 interact with Protocadherin 15 (Maeda et al. 2014). TMC1 and TMC2 are components of the stereocilia mechanoelectrical transduction channel complex (Kurima et al. 2015). While TMC2 is required for mechanotransduction in mature vestibular hair cells, its expression in the immature cochlea may be an evolutionary remnant (Corns et al. 2017). Transgenic Tmc2 expression preserves inner ear hair cells and vestibular function in mice lacking Tmc1 (Asai et al. 2018). Gentamicin and other antibiotics enters neonatal mouse hair cells predominantly through sensorymMechanoelectrical transduction channels, Tmc1 and Tmc2 (Makabe et al. 2020).
TMC2 of Mus musculus (Q8R4P4)
Transmembrane channel-like protein-B, Tmc8 (EVER2). It occurs in the endoplasmic reticulum where it functions to release Ca2+ and Zn2+ and supresses Cl- currents (Sirianant et al. 2014). The functional variant, rs7208422 of the TMC8 gene, has been suggested to have a high impact on susceptibility to beta-papillomaviruses and their oncogenic potential and to also have an influence on alpha-type HPV-related disease (Stoehr et al. 2021).
Tmc8 of Mus musculus (Q7TN58)
Hypothetical protein, HP
HP of Salpingoeca sp. (F2U2C0)
HP of Capsaspora owczarzaki (E9C7I1)
Transmembrane channel-like protein 7, TMC7
TMC7 of Acromyrmex echinatior (F4X8H9)
Transmembrane channel-like protein-1, Tmc1. Also called Transmembrane cochlear-expressed protein-1, Beethoven protein and deafness protein. Required for normal function of cochlear hair cells, possibly as a Na+/K+/Ca2+ channel (Kim and Fettiplace 2013). TMC1 and TMC2 are both components of hair cell transduction channels and contribute to permeation properties (Pan et al. 2013; Kawashima et al. 2014). Channel activity has been demonstrated for the C. elegans orthologue, and the mouse Tmc1. The C. elegans Tmc1 is probably a Na+-activated Na+-selective mechanosensor. The C. elegans Tmc2 may be a Na+/K+ channel. The mouse Tmc1 is functional and replaces Tmc2 when expressed in C. elegans (WR Schafer, personal communication). Ca2+ currents are blocked by the peptide toxin GsMTx-4 (Beurg et al. 2014). Tmc1 and Tmc2, expressed in cochlear and vestibular hair cells, are required for hair cell mechanoelectric transduction (Nakanishi et al. 2014); mutations disrupt mechanoelectric transduction and are a cause of autosomal dominant and recessive forms of nonsyndromic hearing loss (Gao et al. 2015). Using the mutant mouse model (Tmc1; Beethoven) for progressive hearing loss in humans (DFNA36) this mutation has been shown to affect the MET channel pore, reducing its Ca2+ permeability and its affinity for the permeant blocker, dihydrostreptomycin (Corns et al. 2016). Evidence for TMC1 being the hair cell mechanosensitive channel has been evaluated (Fettiplace 2016). The human orthologue (UniProt acc # Q8TDI8) is 96% identical. Mouse LHFPL5 ((HMGIC fusion partner-like protein 5) co-expresses with TMC1 in auditory hair cells (Li et al. 2019). A region within the N-terminus of mouse TMC1 (residues 138 - 168) precludes trafficking from an intracellular location to the plasma membrane (Soler et al. 2019). TMC1 is an essential component of a leak channel that modulates tonotopy and excitability of mouse auditory hair cells (Liu et al. 2019). VRISPER/Cas has been used to correct defects that result in hereditary hearing loss (Farooq et al. 2020). Repair of Tmc1 via genetic engineering in vivo restored inner hair cell sensory transduction and hair cell morphology and transiently rescued low-frequency hearing (Yeh et al. 2020).
Tmc1 of Mus musculus
The sodium sensor/cation conductance channel activated by high extracellular Na+, Tmc-1 (Tmc1) (Chatzigeorgiou et al. 2013). It functions in salt taste chemosensation and salt avoidance and is an ionotropic sensory receptor. Wang et al. 2016 showed that C. elegans TMC-1 mediates nociceptor responses to high pH, not sodium, allowing the nematode to avoid strongly alkaline environments in which most animals cannot survive (Spalthoff and Göpfert 2016). TMC-1 and TMC-2 are required for normal egg laying in C. elegans. Mutations in these proteins cause membrane hyperpolarization and disrupt the rhythmic calcium activities in both neurons and muscles involved in egg laying. Mechanistically, TMC proteins enhance membrane depolarization through background leak currents, and ectopic expression of both C. elegans and mammalian TMC proteins results in membrane depolarization (Yue et al. 2018). TMC-1 is necessary for sodium attraction, but not aversion in the nematode. Dao et al. 2020 showed that TMC-1 contributes to the nematode's lithium induced attraction behavior, but not potassium or magnesium attraction, thus clarifying the specificity of the response. In addition, they found that sodium conditioned aversion is dependent on TMC-1 and disrupts both sodium- and lithium-induced attraction (Dao et al. 2020). The C. elegans Tmc-1 is involved in egg-laying inhibition in response to harsh touch (Kaulich et al. 2021). The initial step in the sensory transduction pathway underpinning hearing and balance in mammals involves the conversion of force into the gating of a mechanosensory transduction channel. Jeong et al. 2022 reported the single-particle cryo-EM structure of TMC-1 from C. elegans. The two-fold symmetric complex is composed of two copies each of the pore-forming TMC-1 subunit, the calcium-binding protein CALM-1 and the transmembrane inner ear protein TMIE. CALM-1 (see TC# 8.A.82.1.1) makes contacts with the cytoplasmic face of the TMC-1 subunits, whereas the single-pass TMIE subunits (see TC# 8.A.116) reside on the periphery of the complex, poised like the handles of an accordion. A subset of complexes includes a single arrestin-like protein, arrestin domain protein (ARRD-6; see TC# 8.A.136.1.15), bound to a CALM-1 subunit. Single-particle reconstructions and molecular dynamics simulations showed how the mechanosensory transduction complex deforms the membrane bilayer to suggest roles for lipid-protein interactions in the mechanism by which mechanical force is transduced to ion channel gating (Jeong et al. 2022).
TMC-1 of Caenorhabditis elegans
Tmc2 channel of 1203 aas and 9 - 11 TMSs; functions in touch neurons as a mechanosensitive touch sensor (Chatzigeorgiou et al. 2013; WR Schafer, personal communication). May function as a Na+/K+ channel. TMC-1 and TMC-2 are required for normal egg laying in C. elegans. Mutations in these proteins cause membrane hyperpolarization and disrupt the rhythmic calcium activities in both neurons and muscles involved in egg laying. Mechanistically, TMC proteins enhance membrane depolarization through background leak currents, and ectopic expression of both C. elegans and mammalian TMC proteins results in membrane depolarization (Yue et al. 2018).
Tmc2 of Caenorhabditis elegans
Tmc receptor/channel of 1932 aas and about 10 TMSs. Plays a role in Drosophila proprioception and the sensory control of larval locomotion (Guo et al. 2016). These Tmc channels may be activated by membrane curvature in dendrites that are exposed to strain, possibly explaining how different cellular systems rely on a common molecular pathway for mechanosensory responses (He et al. 2019).
Tmc of Drosophila melanogaster
Transmembrane channel 6, TMC6/EVER1 of 805 aas. Mutations give rise to epidermodysplasia verruciformis (EV), a rare genodermatosis, characterized by increased sensitivity to infection by the beta-subtype of human papillomaviruses (beta-HPVs), causing persistent, tinea versicolor-like dermal lesions (Horton and Stokes 2014). Biallelic mutations in either TMC6 or TMC8 are detected in more than half of the cases of the pre-malignant skin disease epidermodysplasia verruciformis (EV) which together form a complex with CIB1 (TC# 8.A.82.1.9) (Wu et al. 2020).
TMC6 of Homo sapiens
Transmembrane channel 8, TMC8/EVER2/EVIN2 of 726 aas. Mutations give rise to epidermodysplasia verruciformis (EV), a rare genodermatosis characterized by increased sensitivity to infection by the beta-subtype of human papillomaviruses (beta-HPVs) as well as increased incidence of cancer, causing persistent, tinea versicolor-like dermal lesions (Horton and Stokes 2014). This is due to release of Zn2+ and Ca2+ from the endoplasmic reticulum (Sirianant et al. 2014). The channel-like domain has been identified (Miyauchi et al. 2016). It plays a role in several aspects of human pathophysiology, such as ion channel permeability, human papillomavirus infection and skin cancer (Lu et al. 2017). Biallelic mutations in either TMC6 or TMC8 are detected in more than half of the cases of the pre-malignant skin disease epidermodysplasia verruciformis (EV), which together form a complex with CIB1 (TC# 8.A.82.1.9) in lymphocytes (Wu et al. 2020). TMC8 is a prognostic immune-associated gene in head and neck squamous cancer (HNSC) cells (Lin et al. 2021).
Transmembrane channel protein 3, Tmc3 of 1130 aas (Kurima et al. 2003; Beurg et al. 2014). LPS-inducible lncRNA TMC3-antisense-1 (AS1) negatively regulates the expression of IL-10 (Ye et al. 2020). In the brown planthopper, Nilaparvata lugens, TMCs is highly expressed in the female reproductive organ especially in the oviduct (Jia et al. 2020). RNAi-mediated silencing of Nltmc3 substantially decreased the egg-laying number and impaired ovary development.
Tmc1/Tmc2a or Tmc2b/protocadherin 15a (Pcdh15a). The complex is part of a mechanotransduction system (Maeda et al. 2014). Its trafficing to the plasma membrane depends on the transmembrane O-methyltransferase (TOMT/LRTOMT; 259 aas, 1 N-terminal TMS) (Erickson et al. 2017). Water motion is dependent on this complex (Chou et al. 2017). The role of another protein, Tmie (see TC# 8.A.115), in sensory hair cells is to target and stabilize the Tmc channel subunits to the stereocilia, the site of mechano-electrical transduction (Pacentine and Nicolson 2019). Tmc proteins 1, 2a and 2b are essential for zebrafish hearing although Tmc1 is not, probably because they can (at least partially) substitute for each other (Chen et al. 2020). There are two distinct cell types in inner ear hairs, an upper layer of teardrop shaped cells that rely on Tmc2a, and a lower layer of gourd shaped cells that rely on Tmc1/2b (Smith et al. 2020). Tmc reliance in the ear is dependent on the organ, subtype of hair cell, position within the ear, and axis of best sensitivity (Zhu et al. 2020).
Tmc1/Tmc2/Pcdh15 complex of Danio rerio (Zebrafish) (Brachydanio rerio)
Tmc4 (MBOAT7) of 712 aas and 10 TMSs (Mancina et al. 2016). It is a calcium-dependent chloride channel that may play a role in nonalcoholic fatty liver disease (NAFLD) (Sookoian et al. 2018) but is not associated with a risk of hepatocellular carcinoma or persistent hepatitis B infection (Wang et al. 2021). Ibuprofen only minimally inhibits the taste response of the ENaC to NaCl, but it significantly inhibits the TMC4 response to NaCl with an IC50 at 1.45 mM. Thus, ibuprofen interferes with detection of salty taste via inhibition of TMC4 (Kasahara et al. 2021). This agrees with the fact that TMC4 is a chloride channel involved in high-concentration salt taste sensation (Kasahara et al. 2021). TMC4 is involved in pH and temperature-dependent modulation of salty taste (Kasahara et al. 2021). Salt-enhancing peptides were identified based on the allosteric sites in TMC4 (Shen et al. 2022). Mechanisms of salt taste reception and the properties of TMC4 as a salt taste-related molecule have been reviewed (Kasahara et al. 2022). Genetic polymorphisms in TMC4 predispose organisms to a higher risk of liver diseases (Rivera-Iñiguez et al. 2022). Umami peptides bind to the TMC4 receptor to enhance saltiness (Xie et al. 2023).
The mechanoelectric-transduction (MT or MET) complex in auditory hair cells converts the mechanical stimulation of sound waves into neural signals. Tmc1 is of 760 aas and 10 TMSs and is 96% identical to mouse TMC1 (TC# 1.A.17.4.6). Novel TMC1 structural and splice variants are associated with congenital nonsyndromic deafness (Meyer et al. 2005). Variants responsible for hereditary hearing loss have been identified (Wang et al. 2018). There are varying numbers of channels per MET complex, each requiring multiple TMC1 molecules, and together operating in a coordinated, cooperative manner (Beurg et al. 2018). Ballesteros et al. 2018 generated a model of TMC1 based on X-ray and cryo-EM structures of TMEM16 proteins, revealing the presence of a large cavity near the protein-lipid interface that harbors the Beethoven mutation, suggesting that it functions as a permeation pathway. Hair cells are permeable to 3 kDa dextrans, and dextran permeation requires TMC1/2 proteins and functional MET channels (Ballesteros et al. 2018). TMC1 is a pore-forming component of MET channels in auditory and vestibular hair cells (Pan et al. 2018). KCNQ1 rescues TMC1 plasma membrane expression but not mechanosensitive channel activity (Harkcom et al. 2019). A Tmc1 mutation reduces calcium permeability and expression of MET channels in cochlear hair cells (Beurg et al. 2019). Deafness mutation D572N of TMC1 destabilizes TMC1 expression by disrupting LHFPL5 binding (Yu et al. 2020). Homozygous variants in the TMC1 and CDH23 (3354 aas and at least two TMSs, one N-terminal and one near the C-terminus; Q9H251) genes cause autosomal recessive nonsyndromic hearing loss (Zardadi et al. 2020). TMC1 forms a complex with protocadherin 15 (PCDH15, TC# 1.A.82.1.1), lipoma HMGIC fusion partner-like 5 (LHFPL5, TC# 1.A.82.1.1), and transmembrane inner ear protein (TMIE, TC# 8.A.116.1.2). Splicing isoforms of TMC1, LHFPL5, and TMIE have been identified (Zhou et al. 2021). There are four alternative splicing events for the genes encoding these three proteins. The alternative splicing of TMC1 and LHFPL5 is cochlear-specific and occurs in both neonatal and adult (mouse) cochleae (Zhou et al. 2021). Tmc1 deafness mutations impact mechanotransduction in auditory hair cells (Beurg et al. 2021). A TMC1 synonymous substitution is a variant disrupting splicing regulatory elements associated with recessive hearing loss (Vaché et al. 2021). The roles of solute carriers in auditory function have been reviewed (Qian et al. 2022). Autosomal recessive and dominant non-syndromic hearing loss can be due to pathogenic TMC1 variants (Kraatari-Tiri et al. 2022). Mechanical gating of the auditory transduction channel TMC1 involves the fourth and sixth TMSs (Akyuz et al. 2022). Regulation of membrane homeostasis by TMC1 mechanoelectrical transduction channels is essential for hearing (Ballesteros and Swartz 2022). The conductance and organization of the TMC1-containing mechanotransducer channel complex in auditory hair cells has been examined, and it was concluded that each PCDH15 (see 1.A.17.4.13 and 1.A.82.1.1) and LHFPL5 (see 1.A.17.4.15 and 1.A.82.1.1) monomer may contact two channels, irrespective of location (Yu et al. 2020). Homozygous variants in the TMC1 and CDH23 (3354 aas and at least two TMSs, one N-terminal and one near the C-terminus; Q9H251) genes cause autosomal recessive nonsyndromic hearing loss (Zardadi et al. 2020). TMC1 forms a complex with protocadherin 15 (PCDH15, TC# 1.A.82.1.1), lipoma HMGIC fusion partner-like 5 (LHFPL5, TC# 1.A.82.1.1), and transmembrane inner ear protein (TMIE, TC# 8.A.116.1.2). Splicing isoforms of TMC1, LHFPL5, and TMIE have been identified (Zhou et al. 2021). There are four alternative splicing events for the genes encoding these three proteins. The alternative splicing of TMC1 and LHFPL5 is cochlear-specific and occurs in both neonatal and adult (mouse) cochleae (Zhou et al. 2021). Tmc1 deafness mutations impact mechanotransduction in auditory hair cells (Beurg et al. 2021). A TMC1 synonymous substitution is a variant disrupting splicing regulatory elements associated with recessive hearing loss (Vaché et al. 2021). The roles of solute carriers in auditory function have been reviewed (Qian et al. 2022). Autosomal recessive and dominant non-syndromic hearing loss can be due to pathogenic TMC1 variants (Kraatari-Tiri et al. 2022). Mechanical gating of the auditory transduction channel TMC1 involves the fourth and sixth TMSs (Akyuz et al. 2022). Regulation of membrane homeostasis by TMC1 mechanoelectrical transduction channels is essential for hearing (Ballesteros and Swartz 2022). The conductance and organization of the TMC1-containing mechanotransducer channel complex in auditory hair cells has been examined, and it was concluded that each PCDH15 (see 1.A.17.4.13 and 1.A.82.1.1) and LHFPL5 (see 1.A.17.4.15 and 1.A.82.1.1) monomer may contact two channels, irrespective of location (Fettiplace et al. 2022).
TMC5, of 1006 aas and 11 putative TMSs, promotes prostate cancer cell proliferation through cell cycle regulation and could be a target for treatment (Zhang et al. 2019).
Transmembrane channel-like protein 1 of 878 aas and ~ 11 TMSs (Erives and Fritzsch 2019). Mechanosensory transduction (MT) in specialized hair cells of the inner ear may be mediated by TMC1 as the pore component. Other components of the MT complex include protocadherin 15, cadherin 23, lipoma HMGIC fusion partner-like 5, transmembrane inner ear, calcium and integrin-binding family member 2, and ankyrins (Zheng and Holt 2020).
TMC1 of Amphimedon queenslandica
Transmembrane channel-like (TMC7) protein, of 723 aas and 11 TMSs. It probably transports Ca2+, and other cations. It is important for oral tongue squamous cell carcinoma (OTSCC), with rapid local invasion and metastasis. The long noncoding (lnc) RNA MIR4713HG is markedly upregulated in OTSCC. Upregulation of MIR4713HG promotes cell proliferation and metastasis (Jia et al. 2021). Micro RNA let7c5p physically binds MIR4713HG, and knockdown of let7c5p counteracts the effect of MIR4713HG on OTSCC. let7c5p exerted this role by affecting the expression level of TMC7 (Jia et al. 2021). TMC7 also affects other types (rectal and panrecatic) of cancer (Watanabe et al. 2014; Cheng et al. 2019), and may be associated with psychosis proneness (Ortega-Alonso et al. 2017).
1.A.17.5: The Calcium-permeable Stress-gated Cation Channel (CSC) Family
Uncharacterized protein, DUF221, of 703 aas
UP of Zea mays
Uncharacterized protein of 816 aas containe a DUF221 domain
UP of Danio rerio
Uncharacterized transmembrane protein 63B of 832 aas with a DUF221 domain.
UP of Homo sapiens
Longamoebia
UP of Acanthamoeba castellanii
Uncharacterized protein of 853 aas with a DUF221 domain.
UP of Botryotinia fuckeliana
Phosphate metabolism protein 7, Phm7
Phm7 of Saccharomyces cerevisiae
Sporulation-specific protein 75, Spo75
Spo75 of Saccharomyce cerevisiae
RSN-1-like protein of 957 aas
RSN-1-like protein of Saccharomyces kudriavzevii
Early response to dehydrate stress protein, ERD4 of 785 aas. The orthologous channel protein in Dionaea muscipula may play a role in touch-induced hair calcium-electrical signals that excite the Venus flytrap (Scherzer et al. 2022).
The non-rectifying, plasma membrane, calcium-permeable, stress-gated, cation channel 1 (CSC1) of 771 aas (Hou et al. 2014). Activated by hyperosmotic shock. Permeable to Ca2+, K+ and Na+. Inactivation or closure is Ca2+-dependent. The N-terminal region contains 3 TMSs, the first of which may be a cleavable signal peptide., and the C-terminal region contains 6 TMSs corresponding to the DUF221 domain. Arabidopsis contains at least 15 CSCs ((Hou et al. 2014). Some plant homologues are transcriptionally upregulated in response to vaious abiotic and biotic stresses involving mechanical perturbation (Kiyosue et al. 1994).
CSC1 of Arabidopsis thaliana
Osmotically-gated calcium conductance channel of 782 aas. CSC1 (Hou et al. 2014). Activated under hyperosmotic conditions. There are four paralogues in S. cerevisiae.
CSC1 of Saccharomyces cerevisiae
The osmosensitive calcium-permeable cation channel, CSC1 or Tmem63c, of 806 aas and ~10 TMSs. It is activated by hyperosmolarity and Ca2+ (Hou et al. 2014). Tmem63c is a potential pro-survival factor in angiotensin II-treated human podocytes (Eisenreich et al. 2020). It is regulated by microRNA-564 and transforming growth factor beta (TGFβ) in human renal cells, and is therefore a potential target for albuminuria development (Orphal et al. 2020). TMEM63C mutations cause mitochondrial morphology defects and underlie hereditary spastic paraplegia (Tábara et al. 2022).
CSC1 of Homo sapiens
Hexamitidae
UP of Spironucleus salmonicida
Uncharacterized protein of 1267 aas and 12 TMSs
UP of Dictyostelium discoideum (Slime mold)
Uncharacterized protein of 1548 aas and 12 TMSs.
UP of Phytomonas sp. isolate EM1
UP of Agaricus bisporus (White button mushroom)
Csc1 homologue of 866 aas and ~ 11 TMSs. Deletioin of this gene causes C. albicans to become senstive to cations and SDS, tolerant to antifungal agents and produce filamentation (Jiang and Yang 2018).
Csc1 homologue of Candida albicans
OSCA1.2 of 772 aas and 11 TMSs. It is a dimer containing eleven TMSs per subunit, similar to other TMEM16 proteins. Jojoa Cruz et al. 2018 located the ion permeation pathway within each subunit by demonstrating that a conserved acidic residue is a determinant of channel conductance. Molecular dynamics simulations revealed membrane interactions, suggesting a role of lipids in gating. The high resolution structure of this hyperosmolality-gated calcium-permeable channel has been determined (Liu et al. 2018). It contains 11 TMSs and forms a homodimer. The pore-lining residues were clearly identified. Its cytosolic domain contains an RNA recognition motif and two unique long helices. The linker between these two helices forms an anchor in the lipid bilayer and may be essential to osmosensing. Genome-wide analyses of OSCA gene family members in Vigna radiata have revealed their involvement in the osmotic response (Yin et al. 2021).
OSCA1.2 of Arabidopsis thaliana (Mouse-ear cress)
Dimeric OSCA1.2 of 766 aas and 11 TMSs. The 3-D structure has been determined (K. Maity et al., PNAS, in press). This protein is 69% identical to the A. thaliana ortholog (TC# 1.A.17.5.10). It is a putative early stress-responsive osmolality-sensing ion channel protein. A model has been proposed by which it may mediate hyperosmolality-sensing and consequent gating of ion transport. It has a cytosolic domain structurally related to RNA recognition proteins that includes helical arms paralell to the plane of the membrane. They may sense lateral tension in the inner leaflet, caused by changes in turgor pressure, allowing gating of the channel via coupling of the two domains.
OSCA1.2 of Oryza sativa subsp. japonica (Rice)
TMEM63A or CSC1-like protein of 807 aas and ~10 TMSs. Heterozygous variants in TMEM63A have been identified as the cause of infantile-onset transient hypomyelination. TMEM63A variants are thought to cause transient hypomyelination with favorable developmental progress, but identification of a novel TMEM63A variant showed that the TMEM63A-related clinical spectrum is broad and includes severe developmental delay with seizures (Fukumura et al. 2021).
TMEM63A of Homo sapiens
1.A.17.6: The Transmembrane Channel-like (TMC-L) Family
Uncharacterized protein of 878 aas and 7 putative TMSs.
UP of Oxytricha trifallax
TMC-like protein 8 of 890 aas and 8 TMSs
TMC homologue of Oxytricha trifallax
UP of Guillardia theta
TMC protein of 890 aas and 10 TMSs
TMC protein of Tetrahymena thermophila
UP of Saprolegnia diclina
Plasmodiophoridae
UP of Plasmodiophora brassicae
1.A.17.7: The Calcium-permeable Stress-gated Cation Channel-like 2 (CSC-L2) Family
UP of Giardia intestinalis (Giardia lamblia)
Distant Anoctamin homologue of 718 aas and 14 TMSs in a 4 + 1+1+1+2+2+2+1 arramgement.
Anoctamin homologue of Spironucleus salmonicida
Uncharacterized Anoctamin homologue of 502 aas and 8 putative TMSs
Uncharacterized anoctamin homologue of 823 aas and 8 predicted TMSs in a 3 + 2 + 3 arrangement.


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text:
The navigation conditions at the Port of Aabenraa are particularly good and the harbour can be sailed 24 hours a day. Approach takes place from Aabenraa Fjord via a 1000-metre long and 11-metre deep channel to the harbour. With water depths of up to 11 metres, both large and small vessels can call at the port – safely and efficiently.
The Port of Aabenraa's location is 55⁰ 02' 5N - 0⁰25' 7E. The harbour consists of 3 basins (Nyhavn, Gammelhavn and Sydhavn), as well as a quay (Sønderjyllandskajen), Ro-Ro facilities and 2 tanker jetties.
Approach to the harbour takes place via a 1000-metre long channel. The largest bulk carriers that can navigate the Port of Aabenraa have a length of 250 metres and no width restrictions. Tankers can have a maximum length of 210 metres and no width restrictions, though max. 50,000 displacement TS. The highest permitted speed in the harbour basins is 4 knots.
Lighthouse of Sønderstrand is located South of Port of Aabenraa. The lighthouse has White, red and green lights with occlusions. The White sector leads through the Aabenraa fjord from Varnæs Hage. (Ses map above)
Nyhavn 6,5-9,3 m.
Gammelhavn 7,5 m.
Sydhavn 3,0 m.
Sønderjyllandskajen 11,0 m.
Ro-Ro terminal 11,0 m.
Tank Jetty I 11,0 m.
Tank Jetty II 7,0 m.
Hight of tided
There is no noticeable difference in terms of tide levels. Strong north-easterly/easterly winds can result in high water of up to 1.5 metres, whilst strong south-westerly winds can result in low water of up to 1 metre.
Further information about the Port of Aabenraa can be found at Den Danske Havnelods.
ISGOTT
The Port of Aabenraa has drawn up a handbook, Port Information and Safety Regulation, for tankers that call at the port.
In connection with the implementation of rules and regulations, the Port of Aabenraa has drawn up the attached handbook, Port Information and Safety Regulation, which contains information in connection with tankers calling at the port.
Port Information and Safety Regulations


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text:
from django.contrib.auth.models import Permission
from django.core.urlresolvers import reverse
from django.utils.six.moves import http_client
from django_webtest import WebTest
from purl import URL

from oscar.core.compat import get_user_model

User = get_user_model()

def add_permissions(user, permissions):
    """
    Grant permissions to the passed user

    :param permissions: e.g. ['partner.dashboard_access']
    """
    for permission in permissions:
        app_label, __, codename = permission.partition('.')
        perm = Permission.objects.get(content_type__app_label=app_label,
                                      codename=codename)
        user.user_permissions.add(perm)

class WebTestCase(WebTest):
    is_staff = False
    is_anonymous = False
    is_superuser = False

    username = 'testuser'
    email = 'testuser@buymore.com'
    password = 'somefancypassword'
    permissions = []

    def setUp(self):
        self.user = None

        if not self.is_anonymous:
            self.user = self.create_user(
                self.username, self.email, self.password)
            self.user.is_staff = self.is_staff
            add_permissions(self.user, self.permissions)
            self.user.save()

    def create_user(self, username=None, email=None, password=None):
        """
        Create a user for use in a test.

        As usernames are optional in newer versions of Django, it only sets it
        if exists.
        """
        kwargs = {'email': email, 'password': password}
        if 'username' in User._meta.get_all_field_names():
            kwargs['username'] = username
        return User.objects.create_user(**kwargs)

    def get(self, url, **kwargs):
        kwargs.setdefault('user', self.user)
        return self.app.get(url, **kwargs)

    def post(self, url, **kwargs):
        kwargs.setdefault('user', self.user)
        return self.app.post(url, **kwargs)

    # Custom assertions

    def assertIsRedirect(self, response, expected_url=None):
        self.assertTrue(response.status_code in (
            http_client.FOUND, http_client.MOVED_PERMANENTLY))
        if expected_url:
            location = URL.from_string(response['Location'])
            self.assertEqual(expected_url, location.path())

    def assertIsNotRedirect(self, response):
        self.assertIsOk(response)
        self.assertTrue(response.status_code not in (
            http_client.FOUND, http_client.MOVED_PERMANENTLY
        ))

    def assertRedirectsTo(self, response, url_name, kwargs=None):
        """
        Asserts that a response is a redirect to a given URL name.
        """
        self.assertIsRedirect(response)
        location = response.headers['Location']
        for unwanted in ['http://localhost:80', 'http://testserver']:
            location = location.replace(unwanted, '')
        self.assertEqual(reverse(url_name, kwargs=kwargs), location)

    def assertNoAccess(self, response):
        self.assertContext(response)
        self.assertTrue(response.status_code in (http_client.NOT_FOUND,
                                                 http_client.FORBIDDEN))

    def assertIsOk(self, response):
        self.assertEqual(http_client.OK, response.status_code)

    def assertContext(self, response):
        self.assertTrue(response.context is not None,
                        'No context was returned')

    def assertInContext(self, response, key):
        self.assertContext(response)
        self.assertTrue(key in response.context,
                        "Context should contain a variable '%s'" % key)

redpajama_set_name: RedPajamaGithub


text:
OpenXR comes to multiple PC VR headsets, HoloLens 2, and Oculus Quest
Jeremy Horwitz@horwitz
Image Credit: Khronos Group
One year ago, the Khronos Group released the royalty-free mixed reality standard OpenXR in an effort to make cross-platform software coding easier for VR and AR developers. Today, Khronos is trumpeting OpenXR's availability across a collection of well-known headset platforms, some with full support and others in developer preview form, with additional backing from key chipset and XR accessory makers.
As of now, OpenXR apps can run on Microsoft's Windows Mixed Reality headsets and the HoloLens 2, as well as the Oculus Rift and Quest platforms — a feat more impressive than that brief list might suggest. Supporting all of these headsets meant Microsoft and Oculus had to release OpenXR-conformant runtimes for both Intel and ARM architectures. For businesses and developers, that means the same app will be able to run across tethered PC headsets or standalone Android-based and HoloLens mixed reality goggles without custom modifications, saving money and development time while creating more universal user experiences.
Other major PC platforms have also launched OpenXR developer previews, including Valve's SteamVR and the high-end XR headsets from Varjo, which focuses on ultra-high-resolution displays for industrial and military applications. "Now is the time for developers and engine vendors to start looking at OpenXR as the foundation for their upcoming content," said Valve's Joe Ludwig, noting that the standard will let "one build of their VR title … 'just work' across the entire PR VR ecosystem." Ultraleap, perhaps best known for Leap Motion's hand-tracking accessories, has also released an OpenXR developer preview for its trackers, while Collabora's Monado open source runtime is "approaching conformance" with the standard.
Several key apps and games are expected to start benefitting from OpenXR, notably including Microsoft's flagship title Minecraft, which the company announced today is using OpenXR in its new RenderDragon graphics engine for desktop PCs. This will enable one build of the popular game to run on Oculus, Windows Mixed Reality, and SteamVR PC headsets, as well as future OpenXR-compatible PC hardware. On the browser front, Google's Chromium 81 uses OpenXR for WebXR content, so Chrome and Microsoft's Edge will run on OpenXR-ready hardware. Mozilla's Firefox Reality supports OpenXR for HoloLens 2.
Industry support for OpenXR continues to be significant, with additional backing from Epic's Unreal Engine team, HTC's Vive group, and chip developers ranging from Arm to Qualcomm. Based on the standard's momentum, additional announcements of OpenXR-conformant platforms are likely to follow in the coming months.


FORDYCE, Ark. — As the only congressman in Arkansas seeking re-election, U.S. Rep. Mike Ross should have plenty of reason to fret as he runs when many Democrats and incumbents are endangered.
But Ross is one of the few Democratic candidates his party isn't worried about as it tries to prevent GOP gains in blue-leaning Arkansas.
Ross is distancing himself from President Barack Obama and House Speaker Nancy Pelosi. He's also making clear that he's no Blanche Lincoln, the Arkansas senator fighting to win re-election.
Ross is running ads touting that he never moved to Washington. It's a contrast to criticism Lincoln has received for moving to suburban Washington.
His Republican challenger, Beth Anne Rankin, is trying hard to undercut Ross' image.