116 3rd St SE
Cedar Rapids, Iowa 52401
The Board of Regents on Monday unanimously agreed to shave $50 off a tuition-hike proposal for this fall, decreasing the annual increase for resident undergraduate students from $300 to $250.
The rest of the original tuition proposal - which includes increases as high as $800 for some students - remained untouched during Monday's final reading and approval. But regents said they wanted to address concerns around rising resident undergraduate tuition rates, which had been frozen for an unprecedented three straight years.
'I'm very pleased with our leadership team at the regents here in moving this in this direction, finding this compromise,” said regent Larry McKibben, who proposed the amendment after hearing 'a lot of concern out there about the increase, but also concern about how we maintain the high quality of education at our three universities.”
During the proposal's first reading last month, UI students asked regents to consider an across-the-board $200 tuition increase - regardless of school, student level, and program type - instead of the broad range of increases that included a $300 bump for resident undergraduates. Board President Bruce Rastetter at the time said he would support the original proposal, and UI President Bruce Harreld said decreasing the rate bump would hamper his ability to lead the institution to improved excellence.
But Rastetter on Monday said he's had time to consider more feedback and consult with each university president, and he feels comfortable supporting a $250 increase rather than the $300 hike.
'When going out and asking the three presidents if they can support and manage their budgets with $50 less in in-state tuition … they assured me that they could,” he said.
The original proposal had expected to bring in about $21.1 million, and Rastetter said decreasing it by $50 per resident undergraduate student would cut anticipated revenue by nearly $500,000 at University of Northern Iowa, nearly $700,000 at UI, and just under $1 million at Iowa State University.
Each university had crafted 2016-2017 budgets tied to anticipated tuition-increase revenue, but Rastetter said the presidents will make due with less.
'One of the questions I asked of the three presidents … is can you still accomplish your mission and your goal of what you presented to the board in June with the $50 less?” Rastetter said. 'They went to their people and came back and said they believed they could manage through that.”
The board has tied its need for more tuition revenue to shortcomings in state allocations. Instead of $20.3 million more in general university funds for the budget year that started July 1, per the board's request, lawmakers approved a $6.3 million increase.
During the previously Legislative session, lawmakers similarly approved smaller-than-requested allocations, prompting the board seven months ago to raise tuition 3 percent for resident undergraduates - or $200 a year. That increase went into effect for ISU and UNI students in the spring and will go into effect this fall at UI, meaning resident undergraduate students on all three campuses - with Monday's approved increase - will pay $450 more this fall than they did last.
UNI junior Rachael Johnson, who serves as the board's student regent, said she understands the need to increase tuition with declining state support, but she advocated for a process to make costs more predictable.
'I do believe that is our responsibility to keep costs predictable and sustainable for all Iowans,” she said. 'Therefore, I am asking the board and our institutions to take a deep analytical look into the way we set tuition.”
Rastetter said the board is doing just that and he expects in September, 'at the very least,” to discuss a plan that would bring funding and tuition proposals to the Legislature every two years. University budgets would be reflective of the two-year outlook, according to Rastetter.
'That's what we've asked the universities to think about, in terms of their needs, so we can be more forward-looking for parents and their students on what tuition may look like over the course of two and three years out, rather than just year-by-year,” he said.
Legislative cooperation would be required for such an approach to work, and Rastetter said regents already have instigated some of those discussions.
'Just because we think it's a bright idea doesn't mean that it's going to happen,” he said. 'So we will try and lobby and request that the legislature consider it and the governor consider it going forward.”


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domain: history


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domain: artistic


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Mono Lake Posters
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My name is David Eberhardt, and I live & work in Leipzig, Germany. I started drawing at the age of 3 or 4 and never stopped since. Because I could not decide between a career in graphic arts or literature, I naturally did both: I earned a degree in graphic arts, and later received a university degree in English literature.
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Currently I am working as a freelance graphic artist. I design shirt and poster prints while working on my own publication, a graphic novel named '01.


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U.S. Department of State
Bureau of Arms Control, Verification and Compliance
Key Point: The effort to end nuclear explosive testing has spanned five decades with efforts culminating in the Comprehensive Nuclear Test-Ban Treaty (CTBT), which was opened for signature in 1996.
The first nuclear explosive test was conducted by the United States on July 16, 1945. The Soviet Union followed with its first nuclear test on August 29, 1949. By the mid-1950s, the United States and the Soviet Union were both conducting high-yield thermonuclear weapon tests in the atmosphere. The radioactive fallout from those tests drew criticism from around the globe. The international community’s concern about the effects on health and the environment continued to grow. In 1954, Indian Prime Minister Jawaharlal Nehru proposed a ban on all nuclear testing. The increasing public concern over explosive tests led to the negotiation and entry into force of the 1963 Limited Nuclear Test-Ban Treaty (LTBT). This Treaty banned nuclear testing in the atmosphere, outer space, and under water, but underground tests were still permitted.
When the was being negotiated in 1968, a comprehensive test ban was discussed, but the international community failed to reach agreement on the issue. Advocates for a ban on explosive testing persisted.
In 1974, the United States and the Soviet Union signed the Treaty on the Limitation of Underground Nuclear Weapon Tests, also known as the Threshold Test Ban Treaty (TTBT). It established a nuclear “threshold” by prohibiting the United States and the Soviet Union from conducting tests that would produce a yield exceeding 150 kilotons (equivalent to 150,000 tons of TNT). The mutual restraint imposed by the Treaty reduced the explosive force of new nuclear warheads and bombs, which could otherwise be tested for weapons systems. The TTBT was not intended as a substitute for a comprehensive test ban. Article I of the Treaty states that, “the Parties shall continue their negotiations with a view toward achieving a solution to the problem of the cessation of all underground nuclear weapon tests.”
In 1976, scientists from different countries formed the Group of Scientific Experts (GSE) and began conducting joint research into monitoring technologies and data analysis methods for the verification of a comprehensive test ban.
Almost two decades later, the Cold War ended, bringing with it increased possibilities for progress on disarmament and self-imposed testing moratoriums from the United States and the former Soviet Union. Capitalizing on this momentum, the United Nations’ disarmament body, the Conference on Disarmament (CD) in Geneva, began formal negotiations on the Comprehensive Nuclear Test-Ban Treaty (CTBT) in 1994. Capitalizing on the GSE’s research, the CD was able to reach consensus on the verification regime. Other parts of the negotiations proved more difficult, but members of the CD were able to find common ground and move forward. Australia submitted the Treaty to the U.N. General Assembly, where it was adopted on September 10, 1996 and opened for signature on September 24, 1996.
Since then, 183 nations have signed the Treaty, and 156 have ratified it. Of the 44 nations whose ratifications are specifically required by the Treaty for it to enter into force, 41 have signed and 36 have ratified.

domain: law


text:
Dallas County Health and Human Companies provides testing for sexually transmitted diseases on the division’s central office location and in the cell medical unit. You have been tested for STDs inside the final 3 months. American Family Care (AFC) Centers are walk in STD clinics offering a full range of medical companies and treatments including STD Testing. Reported foremost causes for using public STD clinics amongst clinic users in 5 U.S. cities, 1995. The STD Clinic shall be closed on Wednesday, August 30th.
Through the clinic visit, patients are supplied with info on the best way to keep away from contracting and transmitting STDs and advised that if any of their take a look at results are optimistic they are going to be notified. Research show that almost all managed care organizations provide considerably more comprehensive coverage for reproductive health services than the traditional indemnity insurance coverage (Benson Gold and Richards, 1996).
To be able to provide you with tailor-made testing and vaccination options, questions about age, gender, and sexual orientation help to supply appropriate recommendations in your care. On Wednesday, November 15, 2017, CDPH’s STI Clinics will only offer fast-track providers from eight:00AM – eleven:30AM. Wednesday PHN clinic by appointment 9:00 a.m. – four:00 p.m.
Some clinics, generally these affiliated with tutorial establishments, appear to supply complete, excessive-quality STD-associated companies, whereas different clinics don’t provide either comprehensive or excessive-high quality STD Clinic Singapore care. For example, the lengthy interval between infection and appearance of penalties of STDs may be years; managed care organizations with high turnover rates may have little incentive to emphasise STD-related services.
Associate notification is a process where sexual companions are contacted by the Department of Health and notified that they’ve been exposed to a sexually transmitted illness. 4 The demographics of these discovered to be HIV-positive in the STD clinic are very similar to those who are HIV-constructive in the central Arizona region (see Desk three ), except for the next proportion of Hispanics on the STD clinic.
The STD clinic offers confidential testing for Gonorrhea, Chlamydia, Syphilis, HIV, Herpetic lesions (we do not offer blood testing for herpes), Trichomonas, Chancroid with symptoms and Yeast Infections. These that do not serve populations at high risk for STDs, in particular, might not have a powerful curiosity in offering complete STD-related services.

domain: medical


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«- the nature wonder of the world»(3)
Kazakh land, beautiful nature propaganda for the purpose of presentation this site the Department of form entries, Starting today the vast expanses different regions of for the individual, I decided to conduct a systematic. and, they, in turn,, Depending on the accumulation of educational materials can be reflected in the site a few numbers.
such records to the east of the country, that is, East Kazakhstan region I am from. In this regard,, this this page The nature of the region, tourism potential, most beautiful places on articles and videos given. They were selected articles found on the Internet.
Tamaşalañdar! Friends and colleagues share!
The nature of the East Kazakhstan
The main data
East Kazakhstan region — the Republic of Kazakhstan the north-eastern part of the administrative-territorial and economicografïyalıq zone. 1932 warm 10 Founded in March. 1997 distributed in the region in May Semipalatinsk region Added Territory. land area— 283 300 sq km, The number of people— 1,5 million-dye.
Kazakhstan's Pavlodar, Karagandy and Almaty regions, One region of China, Two Russian border zone. administrative Center— Oskemen City (the basis of 1720 founded in). it Astana the city 1084, but Almaty the city 1100 kilometers.
* * *
the area The climate The sharp continental, seasonal and daily temperature fluctuations can be. The spring— hot and moderately dry, and the winter will be cold and snowy. Cold slopes qoñırajay.
Region Region Nagorny, mountain forest, forests, steppe, the desert, semi-desert, It occupies the landscape. Ore Altai and Southern Altai land of Kalba, Saur-Tarbagatay located in the mountains.
* * *
Water resources are concentrated in Kazakhstan 40 monospetsializatsiya flows from the territory of the East Kazakhstan region. Especially Ore Altai region is rich in surface waters. Most major rivers— black Irtysh, Bukhtarma, Kurchumsky, Kalzhyr, Of, Uba, Ulba.
The main water artery of the three countries in the region (China, Russia, Kazakhstan) trans-boundary rivers passing through the territory - Irtysh river is, It is also the largest river in the country. regional area People's Republic of China crosses the border from the north-west direction. Zaysansky part of the lake black Irtysh is called. Irtysh river the largest branches of: China, Kalzhyr, Qalğuttı, Kurchumsky , Of, birch, Bukhtarma, Ulba, Both, Shulbinsky, cereals, bird, Espe, a name, Bogas, Jüzağaş, antelope, manat, Tainty, Abılaykït, Ulan, Dresvyanka, rock, Kuryk, Kyzylsu, Yellow, Shagan.
Irtysh river and its tributaries are of great importance in the water industry and water supply. so they Bukhtarma, Oskemen and Shulbinsky large dams and hydroelectric power station s built.
* * *
East Kazakhstan one thousand lakes have. Most of the northern and north-eastern part of the area. The region is located - Zaisanskaya and Markakol (reserved), Alakolsky and Sasykkol Kazakhstan is the largest lakes. Ölimyes, Karakul, Turangaköl, Kemïrköl and other small lakes in the south-western side of the region, Shar and often Shagan River and the rivers and Ayaguz Month hummocks, smoothing is located in a small area.
Rivers and lakes (Zaisan, Alakol, Markakol, etc.) Quite significant fishers. Mineral healing of many lakes in Altay region. East Kazakhstan region rich in underground water. It is not only the compensation of the household needs of the population and livestock watering, with a variety of mines and enterprises to provide water.
* * *
Mountains and valleys
Construction of the East Kazakhstan region, the relief is a very complex.
- Irtysh River in the east: ore Altai, Tikyrek, The black, Both, Stanovoy, Koksu, Ivanov, Shopping Cart, Ulba, Qızılqarağay, Of, garlic, Tarbagatai, Southern Altai, Kurchumsky and Azwtaw ridges and spurs
- Irtysh River in the west: Kalba ridge, Families, Arkalyk, Mırjıq, Qaraadır Mountains, Xanşıñğıs, Shyngystau, astragalus ridges
- South of Lake Zaisan: Tarbagatai, Entertainment and Manyrak ridges.
- Irtysh River is located in isolated intervals of the ridges on both sides of the mountains and the hills: Maylıqara, Belterek, Aiyrtau, Arqat, Kökseñgir, Sandyktas, Jağalbaylı, Koktas, Atbasar, Arasantaw, etc..
- South and south-east of the region Sasykkol, Koshkarkoland is located in the lakes Alakol Balkhash - Alakol depression is located in the north-eastern part of the river and hollow.
- Far in the north-western part of the region Beskaragai district covers the whole territory John Balapan It occupies.
At the same time, the total volume of the south-western Altai 99,1 sq.km. 350 There are glaciers.
* * *
Altai slope şımdı-grain grass StYou can, lower than the betegelі-seleulі, and higher birch and aspen mixed şoqtoğay Steppes grass-feather. 700 m and high grain mixed grass meadows in the ancient coniferous forest belt will begin. Ore Altai is dominated by pine and black spruce-fir forests, and the South will see them in the Siberian Altai cedar mixed. Entertainment deciduous forests of the ridge Tien Shan firgrow. ore Altai High subalpine forest belt, Alpine meadows grow, and then transferred to the heights they lichens. Tarbagatai Alma trees and aspen forests spread.
* * *
East Kazakhstan region is very rich in wildlife. Here, due to the different nature of the mountainous pine forests with animals, steppe and desert animals, the most common. Altai and in the forests and healthier places Priirtyshsky squirrel, aqtışqan, Kamshat eltirili expensive, such as animals and birds have a commercial value. Alakol and Zaisan localized hollows reservoirs muskrat. Irtysh and its branches, Markakol, Zaisan and Alakol, Bukhtarma Irtysh, Oskemen many species of fish and Shulbinsky bögenderinde, including sterlet, trout, harius There are a lot of valuable fish, such as. Fishing and fish processing industry, organized on the basis of their.
Beautiful nature of East Kazakhstan
On the region's tourism industry
"The potential in the field of tourism development in the East Kazakhstan, There are of national importance 24 environmental protection zones total volume 1 723 072 hectares. 600-more than historical monumentsand 100-More than tourism sitesand bar.
The most important tourist sites: Katonkaragay National Park, West Altai Nature Reserve, Alakolsky reserve, Katon and Kurchumsky Belukha mountain river pool, Ukok plateau, as well as many historical and cultural monuments.
Its unique tourism opportunities in the region. Depending on the geographical and political features that are available on a large area and clean area remained. The territory of the East Kazakhstan region is rich in natural landscapes range: soup, He gave, taiga, high mountains and many rivers and lakes.
Altai-Sayan ecological zone deserves special attention to the part of the territory of Kazakhstan Altai. Due to its unique landscape and biological diversity of this region, "WWF Living Planet" by the International Organization of two hundred (200) Into the global environmental priorities.
Which is typical for a certain region five types of tourism It should be noted, For example:
- eco-tourism (Katon-Karagay, Kurchumsky in districts and Ridder);
- cultural and educational (Semey City, Abay and Ulan districts);
- Treatment and rehabilitation (Katon-Karagay, Urjar districts);
- beach (Zyryanovsky, Ulan, Kurchumsky areas);
- Chingiz (deep, Zyryanovsky in districts and Ridder);
- rural tourism (Katon-Karagay, Kurchumsky in districts and Ridder);
Kazakhstan, Russia, China and Mongolia is located at the junction of the borders of our ölkemimiz was taken over by the diversity of the Asian desert and the Siberian taygasınıñ. ores, forests and rivers, fish, hammer, herbs, Amazingly beautiful region rich region that has long been attracting people from the business. No wonder the area is located in the large deposits of gold and polymetals also produced coal. Color and large reserves of decorative stones. Field of wheat in the region, sunflower, buckwheat good going fields ... A sheep pastures, cow, Horses graze herds. Altai mountains of the world's largest farms, held where the deer and the white deer. Zaisan, Alakolsky and Sasıqöl fresh water "sea" of hundreds of tons of fish are killed ".
East Kazakhstan 13 amazing places
1. Bukhtarma reservoir
Reservoir, hundreds of kilometers from the city of Oskemen, One of the three major river in Kazakhstan— Stews River built. The world's largest artificial water reservoirs. How convenient propelled ships along the Irtysh River water. Tourist hotels on the shores of the reservoir, sanatoriums, resorts and luxury villas built. Therefore, today the favorite place for locals and foreign tourists, Effectively extreme actions and emotions recreation area is ideal for rafting. Entertainment fans boat on the water, ships and yachts will be able to go. Gets clean water, can relax on the sandy beaches to absorb the sun's rays. fishing, There are good places to go hunting.
* * *
Although very popular in stews such as a water reservoir, this is a great place for tourists. Water reservoir of the city of Semey 70 km away from, Shulbinsky hydropower dam in the Irtysh Riveri was inserted into the. Great views of nature, the warmth of the water storage is a great place for those who want to make a summer vacation at the beach.
Hotels here every years, There is a growing number of sanatoriums, Camp Coast Protected. There are many valuable species of fish is caught.
* * *
3. Nature Lake
Nature Lake — "Pearl of the East Kazakhstan" is. Nature National Wildlife Refuge located on the territory. It is hard to get there, natural beauty and captivating attract tourists. Transparent water of the lake is a period of days, Weather-related changes of many colors.
Many mountain water flows in Cork, There are many other sources of the purest spring and taiga forests. Jean-fauna is also very rich. mammals 55 The type and abundance of healing more than 700 types of plants. Wskwç fish— Only found in this lake in Kazakhstan.
* * *
4. «Katon-Karagay"State Natural National Park
The largest national park in Kazakhstan. UNESCO world heritage sites included in the list of a number of. in the park 3 waterfall, "The necropolis Berel" is located. 400 birds, 1,000 plants, There are more than 100 types of animal life.
Cato State National Natural Park
* * *
Rakhmanov Lake "According to the Kazakh Geographic Society" of tourists arriving in the land of Kazakh 55 included in the list of the most interesting and beautiful places. It is located in the wooded foothills. The depth of the lake 30,6 meters. There are many sources of healing thermal water source in the vicinity of the lake. In this context, natural scenic lake rocky caves, depressions between the two treatment of gastrointestinal diseases and limbs well known procedures Rakhmanov resort "source" is located. The main source of treatment resort— is a natural mineral water underground radon.
* * *
Mfolder in the world, consisting of stones, gravel and sand unique beach. The healing properties of many skin diseases AlakolTicket, as well as, the movement of the focusing action, will have a positive impact on the treatment of nervous system.
* * *
The highest point of the Altai Mountains (4509 m), One of the world's largest şıñnıñ. It is considered sacred among the population. high Sengirbay, vertical rocky reliefs, coincide with all of the glacier -, a perfect complexion of the mountains. Many climbers come from all over the world every year icebergs reached the peak.
* * *
8. Jungar fortress
He XVII century fortified Buddha Lounge монастыры. the founder of 1654 warm within its walls Abylai you want. The monastery was known in scientific circles. Hence, 200 So many years ago, the paper package Abılaykïtte, it is impossible to export shipment of ten horse. unfortunately, he saved the manuscripts to the present day.
* * *
9. Berel mound of kings.
B.e.d. IV ғ. Beware kinäziniñ The burial ground. During excavations of the amazing scientific and archaeological discoveries open-air museum complex. Burial mounds found in the aristocratic and Scythian-Siberian animal style items, organics, clothing, horse rides, carpentry, man leather, As a result of the skin and felt things eternal toñnıñ well preserved relics.
* * *
10. "Berry" Reserve-Museum
The great thinker and poet WARNING "Zhidebay- Wolf "state historical-cultural and literary-memorial museum Is taking place in the center of the city of Semipalatinsk range. Sections of the Museum of Abai district: "Berry" museum-reserve and Börilide M. Auezov House Museum.
* * *
11. Korpesh Bayan Sulu Mazar.
Kazakh people associated with the lyrical name of the heroes of the epic poem Rapkan Present ancient monument. 1982 In the cemetery included in the list of monuments of history and culture of the Kazakh Soviet Socialist Republic, the state protection.
* * *
12. Khalifa Mosque Altai.
The largest in the East Kazakhstan region, One of the three largest mosque in the country. Three thousand people for. Residents of the region mosque, government and commercial enterprises, charitable assistance, and was sponsored by the United Arab Emirates. construction of a mosque 20 About a thousand people took part.
* * *
13. suspension bridge
Hanging bridge over the Irtysh River — One of the attractions of the city of Semey. He ranks 17th in the world and only in Kazakhstan. "IHI" the construction of a bridge built by Japanese firm together with the Turkish firm "Alarko Alsim".
At the end of the page:
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domain: geography


text:
Were Nutz - Delicious And Crunchy Praline Pecans Jar Of 2.3 Lbs
Click Image for Gallery
Shellers - Delicious and crunchy Praline Pecans jar of 2.3 lbs (37 oz) Very fresh and sweet candy coated snacks and/or toppings
- Includes 1 jar with 2.3 lbs of delicious praline pecan halves
- We listened to your reviews!! We are using a new sturdier jar that withstands carriers and fulfillment centers!
- Different brandname, better presentation, sturdier jar but it's our same original recipe inherited from grandmothers and aunts!
- Delicious as a sweet topping for your salad.
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domain: food


text:
Staff and management of Indigenous Conference services wish our friends, delegates and supporters of Indigenous Conferences a Merry Christmas and a prosperous New Year 2016!
2015 was a very successful year for us with almost a thousand delegates for the year. We consider ourselves to be privileged to meet such talented, diverse and professional individuals and dynamic groups. We wish to say thank you and our heartfelt gratitude to one and all. With your support, we look forward to making 2016 and 2017 even more successful!
Listed below are the conferences for 2016 and 2017. As we lead up to the beginning of the New year, calling for papers & are now open and sought for. Please submit your entry online.
Exciting upcoming 2016-2017 Conferences
· National Indigenous Youth Justice Conference
· National Indigenous Board of Director’s Conference
· Global Indigenous Domestic Violence Conference
· International Indigenous Men’s Conference
· International Indigenous Women’s Conference
· International Indigenous Allied Health & Chronic Diseases Conference
· National Culturally and Linguistically Diverse Worker’s Conference
· National Indigenous Aged Care Conference
· National Indigenous Drug & Alcohol Conference
· National Indigenous Children’s Conference
· National Stolen Generation Bringing Them Home Conference
· National Mental Health Conference
· Save Our Mob, Stop the Closure of Our Communities Conference
· National Indigenous Grief & Loss Conference
ICS-MEES Pty Ltd success is based upon our principle of trusting of our culture and listening and adhering to our grassroots, community controlled grounding and as part of this our future planning is open to input from communities and grassroots individuals. With this in mind, we are extremely keen to receive input and ideas of topics of concern from around Australia which may not have the chance to be address this year.
We look forward to working with individuals, community controlled organizations, governments and non-government organizations in a not too distant future. Once again, may your holiday season be filled with laughter, family fun and above have a safe celebration to you all!
Guidelines in Submitting Paper:
For more information, please visit the conference website at www.indigenousconferences.com or contact us by email at [log in to unmask]
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domain: sociology


text:
India logs 2,706 new COVID-19 cases, 25 more deaths
India has so far conducted a total of 85,00,77,409 COVID-19 tests.
Updated 30 May 2022, 7:30 am
India on Monday recorded as many as 2,706 new COVID-19 cases and 25 COVID-19 related deaths, according to the Union Health Ministry. India's cumulative number of confirmed COVID-19 cases has reached 4,31,55,749 while the death toll stood at 5,24,611.
As per the Health Ministry, India also recorded 2,070 new COVID-19 recoveries, taking the cumulative tally of recovered cases to 4,26,13,440. India’s recovery rate stands at 98.74 per cent.
Consequently, India’s active COVID-19 caseload stands at 17,698, which constitute 0.04 per cent of the country's total positive cases.
India has so far conducted a total of 85,00,77,409 COVID-19 tests, including 2,78,267 COVID-19 tests done in the last 24 hours.
The weekly positivity rate in the country currently stands at 0.58 per cent and the daily positivity rate is reported to be 0.97 per cent, the Health Ministry stated.
Meanwhile, India’s COVID-19 vaccination coverage has reached 1,93,31,57,352 as per provisional reports till 7 am Monday.
First published:30 May 2022, 7:30 am

domain: news


text:
When Lev Raphael’s debut story collection, Dancing on Tisha B’Av, was published 25 years ago, I was enthralled. For me, it was the first book of fiction that truly brought together my gay and Jewish identities. Reviewing the book for the Washington Blade, I called it “a dazzling collection of short fiction” and noted that it “ties together a dual identity into a cohesive and logical whole.”
What has stayed with me the most over the years was the very first story in the collection: the title story, which Tablet has republished today, just in time for Tisha B’Av. That story, in which a young gay man faces ostracism from his Orthodox minyan, had originally appeared in Shmate magazine in 1985, and was later included in Men on Men 2, a 1988 collection of gay short stories edited by George Stambolian.
“New American Library did not want this story in Men on Men 2 because it was too Jewish,” Raphael told me in a recent phone interview. “George never told me that until the book came out, and then he took me aside at an Outwrite conference and said that he fought very hard to keep the story in—which is ironic since that’s one of the stories that was singled out by reviewers as being one of the best in the collection.”
Raphael credits Stambolian with helping him shape the story, and punching up its ending. But the initial inspiration came from his own life. He recalled attending Orthodox services and talking over lunch afterward to a couple who were “casually mentioning that a lesbian came out in their parents’ home congregation, and she was asked to leave. I said, how horrible. And, that would be a great story.”
That first story, he told me, “put me on the map in gay writing… It’s an old story, but I see it as something that marked an important stage in my career.”
After his debut collection came out in 1990 from St. Martin’s Press, it won a Lambda Literary Award. In my Washington Blade review at the time, I wrote: “Raphael has begun a dialogue too long unspoken.” Raphael has gone on to write two dozen more books across a wide variety of genres. And while the dialogue around gay and Jewish issues has since broadened to include a wide array of authors, he remains at the center of the discussion he began 25 years ago.
Related: Dancing on Tisha B’Av

domain: library


POLICE are probing claims that animal rights activists are responsible for two racoons escaping from a zoo.
Racoons Bowser and Wookie are thought to have been freed through a hole made by the unknown activists.
The one-year-old rodents who disappeared from Tropiquaria Zoo in Washford, Somerset,UK, were spotted two miles away.
Authorities said the animals can carry disease – as well as having a nasty bite – and freeing them deliberately is extremely dangerous.
Chris Moise, Thropiquiaria Zoo director said: “I believe animals were released by activists looking to cause trouble before a badger cull begins in Somerset.
He added: “I have no evidence to say it is, but I have circumstantial evidence to say it might be. The wire was folded back in a way the raccoons wouldn’t have pulled it back.”
Under the EU laws it is illegal to import, breed, sell or release raccoons into the wild as they are classed as an invasive species.
The zoo spokesperson warned people not to approach the raccoons, but to report any sightings of them to the Animal and Plant Health Agency.


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Ask almost any adult what their favorite childhood memory is, and they will tell you it is getting a puppy. It is undeniable that today’s dogs are not just animals to us. They have become family members and a source of great joy. Dogs simply make life better. They are great and loyal companions and protective of their family, and it doesn’t hurt that they are absolutely adorable and fluffy. However, there is another side to this that is not all roses. Dogs are also a big responsibility. It’s why some parents have a no-pet rule in their homes. You need to put effort into training a dog; not everybody has enough time for that. If you want you and your puppy to have the best life possible, then you can’t underestimate the importance of socialization for puppies.
Caption: A well-socialized puppy will play with other dogs instead of fighting.
Socialization for puppies makes them safer in the future
There are many reasons why equipping puppies with social skills is essential, but one of the main ones is that it will make the dog and the owner safer. While dogs are lovely and caring creatures, they can also be aggressive and dangerous. That will mostly depend on how well the dog was socialized as a puppy. Dogs that don’t have proper social skills are more likely to be aggressive and challenging to handle. If you put the unsocialized dog in an environment with another dog or dog, you risk a terrible outcome. When dogs are not used to being around other animals, sometimes they instinctively growl, snap, or in the worst cases, attack. And unfortunately, sometimes that results in tragedy. That is why social skills will make your dog safer.
Other people will be safer as well
It is not just your dog’s safety. You should worry about other people’s safety as well. When you get a puppy, slowly introducing it to your extended family and friends is incredibly important. He will need to get used to the other people around him. You can give family and friends treats to offer to your dog when they come over so they build a good relationship. That will make him not just used to other people but also excited. Slowly introduce him to the outside world and teach him not to get spooked by all the strangers around. Because if you don’t do this early, you risk your puppy becoming anxious and scared. And dogs like that can potentially bite someone because they will feel threatened. Imagine you are walking your dog, and you meet your friend who wants to say hello, but your dog sees this as a threat and instinctively reacts. That is why socialization for puppies is so important.
You will be able to leave your pet at daycare
As we previously said, dogs are a big responsibility, and they can’t be left alone for a long time. Thankfully there are pet daycares that will take care of your dog while you are away if you need it. However, this can be difficult if you don’t socialize with your puppy. A badly behaved dog is not likely to be a good match for pet daycare. And if you can’t leave your dog here, you will be forced to stay home, which will be greatly limiting. So, if you want to be able to leave your dog at daycare, you will have to socialize with them while they are a puppy. Also, here is a quick tip regarding dog daycares. Before leaving your dog in one of these places, you should research how pet daycares keep their turf clean. Dogs risk getting a disease, so it's important to keep it clean at all times. So before choosing a particular pet daycare, ask them about this first.
Caption: Proper socialization for puppies will make vet visits more pleasant
Vet visits will be easier
As a dog owner, you are responsible for regularly taking your dog to the vet for emergencies or regular checkups. However, if you ignore the importance of socialization for your puppy, these visits will likely become an anxiety-ridden nightmare. Poorly socialized puppies will be highly stressed during vet visits, and they might act out or make the exam impossible. That is a massive issue with larger dog breeds, which can be dangerous for you and the vet. You need to teach your dog that the vet is not a scary place. When your puppy is tiny, always bring treats to the vet. Also, buy him a toy or other goodies while you are there. That will help the puppy associate vet visits with something fun and joyous instead of a scary experience. It might take some time, but with enough effort, you will succeed.
Caption: Well-socialized dog will have a happier life Alt text: A white dog
A socialized puppy is a happy puppy
And finally, if you socialized your puppy on time, you have contributed significantly to your dog’s overall happiness. Simply put, a socialized puppy is a happy puppy. When you equip a dog with the necessary social skills, you open them up to a whole new world of fun experiences, like going to dog parks or sitting in pet-friendly restaurants with you. You wouldn’t be able to do all these things with a poorly behaved dog. Also, your puppy will be much less anxious and scared if socialized well. They won’t fear loud noises, cars, other people, vets, etc.
The importance of socializing for puppies is enormous. It makes life so much easier for both you and your furry friend. You can easily engage in everyday activities with your dog, such as walks in the park, vet visits, leaving him in the daycare, etc. Also, you are more likely to avoid tragic incidents if you socialize with your dog early on. Doing this would make you a responsible dog owner and do a huge favor to your dog. His life will be so much fuller, richer, and less stressful.
Meta Description: Before making a big commitment and getting a dog, you should learn about the importance of socialization for puppies.
✍🏼 Natalie Taylor

domain: pet


text:
Transfer of type-1 helper T-conditioned (Th1-conditioned) cells promotes functional recovery with enhanced axonal remodeling after spinal cord injury (SCI). This study explored the molecular mechanisms underlying the beneficial effects of pro-inflammatory Th1-conditioned cells after SCI. The effect of Th1-conditioned cells from interferon-γ (ifn-γ) knockout mice (ifn-γ−/− Th1 cells) on the recovery after SCI was reduced. Transfer of Th1-conditioned cells led to the activation of microglia (MG) and macrophages (MΦs), with interleukin 10 (IL-10) upregulation. This upregulation of IL-10 was reduced when ifn-γ−/− Th1 cells were transferred. Intrathecal neutralization of IL-10 in the spinal cord attenuated the effects of Th1-conditioned cells. Further, IL-10 is robustly secreted from Th1-conditioned cells in an ifn-γ-dependent manner. Th1-conditioned cells from interleukin 10 knockout (il-10−/−) mice had no effects on recovery from SCI. These findings demonstrate that ifn-γ-dependent secretion of IL-10 from Th1 cells, as well as native MG/MΦs, is required for the promotion of motor recovery after SCI.
In mammals, once the central nervous system (CNS) is injured, in contrast to peripheral nervous system, neural regeneration and functional recovery are very limited.1 Until recently, immune reactions have been considered detrimental after CNS injuries. For example, infiltrating macrophages (MΦs) and resident microglia (MG) are considered deleterious to recovery after CNS injuries2, 3, 4 and T cells are pathogenic in the injured CNS.4, 5, 6 Hence, massive dose of glucocorticoid has been infused to patients after spinal cord injury (SCI) to attenuate immune reactions in the CNS, although there are no convincing evidences that prove significant efficacy of the treatment. However, in some conditions, T cells are beneficial for repair of the injured CNS, although this notion remains controversial.4, 7, 8, 9 These contradictory findings regarding the role of T cells after CNS injuries could be explained by differences in the subsets of helper T cells. In support of this hypothesis, a previous study demonstrated that adoptive transfer of interferon-γ (IFN-γ)-producing type-1 helper T-conditioned (Th1-conditioned) cells promote functional recovery after SCI in mice.10 Enhanced axonal remodeling of corticospinal tract (CST) and serotonergic fibers are also observed in mice that undergo Th1-cell transfer. Although type-2 helper cells (Th2 cells) are considered neuroprotective in some situations,11, 12 this previous study did not demonstrate enhanced recovery from SCI after transfer of Th2-conditioned cells.10 Furthermore, transfer of IL-17-producing helper T (Th17) cells, pathogenic in multiple sclerosis (MS) and in an animal model of MS,13 limits recovery in the acute phase after SCI.10 Importantly, upregulation of the anti-inflammatory/neuroprotective M2 subtype of MG/MΦs was observed in the injured spinal cord after transfer of Th1 cells that had previously been considered to be involved in the onset and progression of autoimmune diseases such as MS.14, 15 In the current study, we explored the mechanism underlying the restorative effects of transferred Th1 cells after SCI in mice.
Therapeutic effects of Th1 cells after SCI depend on IFN-γ
The objective of this study was to explore the mechanism behind the effects of adoptively transferred Th1-conditioned cells on the functional recovery after SCI. We first assessed whether IFN-γ, a major cytokine secreted by Th1 cells,16 was required for the effects. We isolated cluster of differentiation 4 (CD4)+ T cells from spleens of wild-type (WT) and interferon-γ (ifn-γ)-knockout (ifn-γ−/−) mice. The cells were then cultured with IL-2/IL-12 and anti-IL-4 neutralizing antibody to induce differentiation into Th1 cells. We confirmed with enzyme-linked immunosorbent assay (ELISA) that IFN-γ secretion from Th1 ifn-γ−/− cells was totally abrogated compared with WT Th1 cells and Th17 cells (Figure 1a). In addition, abundant expression of T-bet, a T-box transcription factor of the Th1 master gene,17 was observed in cultured WT Th1-conditioned cells by flow cytometry (Figure 1b). T-bet in ifn-γ−/− Th1-conditioned cells was expressed at an intermediate level between Th0 and Th1 cells (Figure 1b).
Mouse vertebrae were laminectomized and the exposed spinal cords were contused at the level between thoracic vertebrae 9 (T9) and 10 (T10) by an impactor with a force of 60 kdyn. On the first day after contusion, the hindlimbs of all mice were completely paraplegic and gradually displayed partial recovery of motor function after day 1. At 4 days after contusion SCI, 1.0 × 107 WT and ifn-γ−/− Th1-conditioned cells were transferred intraperitoneally (i.p.) into the mice and recovery of motor function was assessed using the Basso Mouse Scale (BMS).18 Enhanced motor recovery was observed in mice that received Th1 ifn-γ−/− cell transfer compared with control mice, but the extent of the improvement was less than that observed in mice receiving a transfer of WT Th1-conditioned cells (Figure 1c).
Consistent results were obtained when the grid-walk test was performed, which assesses voluntary movement control mediated by the corticospinal and the rubrospinal systems.19 Transfer of Th1 ifn-γ−/− cells led to a significantly greater frequency of hindlimb dropping (mistakes) compared with that of WT Th1 cells (Figure 1d). Thus, IFN-γ in Th1-conditioned cells contributes to improved functional recovery.
ifn-γ-dependent activation of MG by adoptive transfer of Th1-conditioned cells
Previous observations with flow cytometry after SCI showed that transfer of Th1-conditioned cells led to an increase in MG/MΦs and to upregulation of the neuroprotective M2 subtype: CD206+ cells among the MΦs and arginase 1+(Arg1) cells among the MG.10 Morphological analysis by immunohistochemistry for ionized calcium-binding adapter molecule 1 (Iba1) in the contused spinal cord revealed that with Th1 transfer, MG/MΦs increased, enlarged, and were more round compared with those in control mice (Figures 2a and b). This observation is consistent with the previous data that showed both activation of MG/MΦs with CD11b and CD45 immunoreactivity using flow cytometry and upregulation of the M2 subtype, which has characteristics including anti-inflammatory effects, phagocytic behavior, and a round shape compared with the pro-inflammatory, destructive, and ramified M1 subtype.10 MG/MΦs were assessed quantitatively using flow cytometry20 and microglial activation by Th1-cell transfer after SCI was shown to depend on IFN-γ (Figures 2c and d).
IL-10 produced from MG/MΦs contributes to repair after SCI
It was hypothesized that some molecules secreted from MG/MΦs have a role in the enhanced functional recovery induced by Th1-cell transfer after SCI. Intracellular cytokine staining of MG/MΦs indicated that interleukin 10 (IL-10), which is considered to be cerebroprotective after an ischemic stroke21 and is one of the M2 anti-inflammatory markers,22 was upregulated in MG/MΦs in the spinal cord after transfer of Th1-conditioned cells (Figure 3a). This upregulation was reduced if Th1 ifn-γ−/− cells were adoptively transferred (Figure 3a), demonstrating that IFN-γ secreted from Th1-conditioned cells contributes to IL-10 production from MG/MΦs. We previously demonstrated that neutralization of IL-10 from Th1 cells attenuates the functional recovery initiated by the transfer of Th1 cells after SCI.10 However, it remains to be determined whether IL-10 secretion in the CNS after Th1 transfer has neuroprotective effects, because anti-IL-10-neutralizing antibody could block not only IL-10 secretion from transferred Th1 cells but also from native cells in the injured CNS, as shown in our previous experiment. To assess whether IL-10 produced by MG/MΦs in the CNS is required for the in vivo effects, anti-IL-10-neutralizing antibody or control IgG was injected into the cerebral ventricles (i.c.v.) using an osmotic pump catheterized to the right lateral ventricle for 7 days after SCI in mice. Th1-conditioned lymphocytes were injected i.p. to these mice on day 4 after SCI. Compared with treatment with control IgG, antibody infusion significantly suppressed motor recovery at time points later than 4 weeks after SCI, as assessed by BMS scoring (Figure 3b). Although there were no significant differences observed in the grid-walk test (Figure 3c), the ameliorated recovery of motor functions by Th1-cell transfer was attenuated in the inclined-plane test (Figure 3d). Therefore, IL-10 secreted from the cells in the CNS is a partial contributor to enhanced motor recovery after SCI.
ifn-γ-dependent secretion of IL-10 from Th1 cells have a pivotal role in improved recovery after SCI
We next performed in vitro experiments to confirm whether we can recapitulate IL-10 upregulation from MG and MΦs by Th1-conditioned lymphocytes. First, we cultured MG from neonatal mouse brains and mouse femoral bone-derived MΦs. Secretion of IL-10 was measured using ELISA after MG (Figure 4a), or MΦs (Figure 4b) cultured in 96-well plates were treated with conditioned media (CM) of Th1 cells, Th1 ifn-γ−/− cells, and Th17 cells or recombinant IFN-γ (Figure 4 upper schematic). Even at the starting point, high levels of IL-10 were observed in Th1-cell group (Figures 4a and b), demonstrating that CM of Th1 cells themselves contained abundant IL-10 compared with Th1 ifn-γ−/− cells and Th17 cells. Although IL-10 expression from MG/MΦs unexpectedly did not surge, IL-10 levels from MG or MΦs treated with CM of Th1 cells were significantly higher than only CM of Th1 cells at 3 h after the treatment (Figures 4a–c). It is suggested that comparatively small amounts of IL-10 were secreted from cultured MG/MΦs in the early phase after treatment of Th1 CM. As Th1 cells robustly secreted neuroprotective IL-10 (Figures 4a and b), we examined whether genetic deletion of il-10 in transferred Th1-conditioned cells decreased the effects of the Th1 cells in SCI mice. As a result, Th1 interleukin 10 knockout (il-10−/−) cells transfer completely diminished the ameliorative effects of Th1 cells transfer on hindlimb locomotion after SCI, as assessed by BMS scoring (Figure 4d).
Th1-conditioned lymphocytes secrete neurotrophic factors
Candidate neurotrophic factors in Th1-conditioned cells that promote neuroprotection were investigated after contusion SCI and the expression of neurotrophic factors, such as glia-derived neurotrophic factor (GDNF) and neurotrophin 3 (NT-3), in Th1-conditioned cells was shown to be higher than that in Th17 cells, which limited functional recovery after SCI.10 It has been shown that GDNF is neuroprotective after SCI23 and that NT-3 promotes growth and differentiation of neurons and synapses.24 In the current study, the expression levels of GDNF and NT-3 among Th1 cells, Th1 ifn-γ−/− cells, and Th17 cells were compared using ELISA. GDNF expression was higher in Th1 cells than that in Th1 ifn-γ−/− cells or Th17 cells (Figure 5a), although expression of NT-3 was not significantly different between Th1 cells and Th1 ifn-γ−/− cells (Figure 5b).
The current study demonstrated that ifn-γ-dependent secretion of IL-10 from Th1 cells as well as from MG/MΦs is required for the enhanced recovery from SCI induced by Th1 transfer (Figure 6). It is surprising that expression of the neuroprotective cytokine IL-10 is upregulated from MG/MΦs in an ifn-γ-dependent manner (Figure 3a), although pro-inflammatory Th1 cells were transferred after SCI. However, effects of these cytokines especially IL-10 upregulation in the CNS are partial (Figures 3b and d). Th1 cell-transferred spinal cord after SCI showed accumulation of activated round MG and MΦs around the injured dorsal and lateral CST, although activated and ramified MG/MΦs are also observed without treatment after SCI (Figures 2a and b). It is suggested that the increased proportion of neuroprotective M2 MG/MΦs induced by Th1 cells transfer10 may protect and repair injured neuronal fibers, including the CST. To elucidate the progression of IFN-γ from Th1 cells leading to IL-10 upregulation in MG/MΦs, we treated cultured MG/MΦs with Th1 CM or recombinant IFN-γ and measured IL-10 levels (Figures 4a–c). The result suggests that Th1 cells themselves secrete abundant IL-10 and, comparatively, an unexpectedly small amount of IL-10 was secreted from MG/MΦs when treated with Th1 CM. It is possible that the cell state of the cultured MG/MΦs in our system may be different from in vivo MG/MΦs after SCI (Figure 3a) and that other micro-environmental factors, such as astrocyte activity, may be required to robustly elevate IL-10 secretion from MG/MΦs. Whereas experiments neutralizing IL-10 in the CNS only attenuated the effects of Th1-conditioned cells transfer (Figures 3b and d), transfer of Th1-conditioned cells derived from il-10−/− mice showed complete abrogation of the restorative effects of Th1 cells (Figure 4d). Thus, ifn-γ-dependent secretion of IL-10 from transferred Th1 cells (Figures 4a and b) may contribute to the entirety of the effects by directly shifting the CNS into a neuroprotective state25 in addition to increasing the proportion of the neuroprotective M2 subtype of MG/MΦs observed and upregulation of Arg1, CD206,10 and IL-10 (Figure 6). Indeed, transfer of Th1-conditioned cells has enhanced restorative effects compared with just inducing viral expression of IL-10 in the CNS after SCI.25 This may be caused by shifting systems in the periphery, such as MΦs in the lymph nodes into a neuroprotective state, as well as the CNS with Th1-cell transfer. On the other hand, IFN-γ did not have direct effects on IL-10 upregulation from MG/MΦs (Figures 4a–c). We conclude that ifn-γ is required to maintain transferred Th1 cells to abundantly secrete IL-10 rather than directly upregulating IL-10 from MG/MΦs in the lesion site after SCI. Our result is consistent with a report that ifn-γ mediates IL-10 reactivation by parasite-reactive Th1 cells.26 The notion may be consistent with our previous report that IFN-γ is necessary for maintaining Th1 cells to promote neurite outgrowth from cortical neurons rather than directly elongating neurites.27 On the other hand, other mechanism independent of ifn-γ may be partially responsible for IL-10 secretion from Th1 cells, because comparatively very small amounts of IL-10 were still secreted from Th1 ifnγ−/− cells (Figures 4a and b). This residual secretion of IL-10 may be attributed to ifn-γ-independent activation of signal transducer and activator of transcription 4, as previously reported.28
In general, inflammation is considered to be detrimental to the CNS injury, but there are some reports demonstrating that pro-inflammatory T cells are beneficial to the CNS injury. Those T cells may be attributed to the functions of Th1 cells in ifn-γ-dependent manner. Indeed, IFN-γ promotes neuronal differentiation29 and neurogenesis.30 Secretion of IFN-γ by the transferred Th1-conditioned cells has a role in MG activation (Figures 2c and d), leading to functional recovery from SCI. It has been suggested that activated MG induce neuronal differentiation31 and are neuroprotective.22 In addition, the beneficial role of MΦs in the injured CNS is known.32, 33
IFN-γ contributes to the upregulation of GDNF in Th1 cells (Figure 5a), which may have positive effects on neuroprotection after SCI. Thus, the in vivo effects of IFN-γ secreted by Th1-conditioned cells are important for functional recovery after SCI. However, it should be noted that Th1-conditioned cells lacking IFN-γ still had limited beneficial effects (Figures 1c and d). These remnant effects may be mediated by neuroprotective IL-10 (Figures 4a and b) and trophic factors, such as NT-3 (Figure 5b).
Taken together, pro-inflammatory Th1 cells have restorative effects on functional recovery after SCI in ifn-γ- and il-10-dependent manner. There are many side effects after massive dose of glucocorticoid treatment such as increased susceptibility to infection, gastroinstestinal ulcer, diabetes mellitus, and mental disorder. Therefore, our study provides new insights into future immunomodulatory treatment after CNS trauma (for example, transfer of Th1 cells or skewing to Th1), which not only promotes beneficial immune reactions but also attenuates harmful ones such as devastating Th17 cells.
Materials and Methods
C57BL/6j mice were purchased from Charles River Laboratories (Wilmington, MA, USA). As previously reported, ifn-γ−/− mice34 and il-10−/− mice35 on the C57BL/6 background were used. All mice used in this study were housed in specific pathogen-free conditions and were treated and cared for in accordance with the guidelines of Osaka University pertaining to the treatment of experimental animals.
Animal model of SCI
Adult (7–9 weeks old) female C57BL/6 mice were anesthetized with sodium pentobarbital (50 mg/kg; Kyoritsu Seiyaku, Tokyo, Japan). Following dorsal laminectomy (at the T9–T10 level), the spinal cord was contused with a force of 60 kdyn using an Infinite Horizon Impactor (Precision Systems & Instrumentation, Fairfax Station, VA, USA), as previously described.36 The muscle and skin layers were then sutured. The bladder was expressed daily using manual abdominal pressure until 10 days after injury. Food and water were provided ad libitum.
Adoptive transfer of cultured T cells
The procedure was performed according to a previously described procedure.10 The procedure is briefly indicated as following: spleens were collected from the mice. CD4+ T cells were isolated using magnet sorting with anti-CD4 magnet beads (Miltenyi Biotec, Cologne, Germany). CD4+ T cells were stimulated with anti-CD3-ɛ/anti-CD28 antibodies (5 μg/ml, respectively, eBioscience, San Diego, CA, USA) coated on 24-well plates (Greiner Bio-One, Kremsmünster, Austria). Th1-conditioned cells were differentiated by the addition of recombinant IL-2 (25 U/ml; R&D Systems/TECHNE, Minneapolis, MN, USA), IL-12 (10 U/ml; R&D systems), and anti-IL-4 antibodies (25% culture supernatant of hybridoma; clone HB-188; American Type Culture Collection, Manassas, VA, USA). Th17-conditioned cells were differentiated by the addition of recombinant IL-6 (20 ng/ml; R&D systems), IL-23 (20 ng/ml; R&D systems), TGF-β1 (3 ng/ml; R&D systems), anti-IL-4 antibodies (25% culture supernatant of the hybridoma), and anti-IFN-γ antibodies (1% culture supernatant of hybridoma; clone HB-170; American Type Culture Collection) in the presence of anti-IL-2 antibody (10 μg/ml; R&D Systems). Th0 cells were cultured in the presence of only IL-2 (25 U/ml; R&D Systems). On day 4, the helper T cells were collected. The cells were centrifuged at 1000 × g for 20 min with Lympholyte-M (Cedarlane Laboratories, Burlington, Ontario, Canada), and the intermediate layer was collected to eliminate the dead cells. The cells were restimulated for 3 h with anti-CD3-ɛ and anti-CD28 antibodies. On day 4 after SCI, helper T cells (1.0 × 107) suspended in 500 μl phosphate-buffered saline (PBS) were injected i.p. into mice. As a control, 500 μl PBS was injected into mice after SCI.
Hindlimb motor function was evaluated using the BMS open-field locomotor test, in which scores range from 0 to 9.18 BMS scores were recorded on days 1, 3, and 7 after injury, and once weekly thereafter for a total of 6 weeks.
The grid-walk test was also performed to evaluate hindlimb locomotion.19 Deficits in descending motor control were examined by assessing the ability of mice to step on a square framework, 26 cm on a side, consisting of 1-cm2 grids made of round metal bars. Analysis was performed by counting the number of errors (that is, dropping the hindlimb from the grid) in 50 steps for each hindlimb. The mean percentage of errors was then calculated. Error percentages were not calculated before 2 weeks after SCI because, before this time, animals were unable to move their hindlimbs owing to insufficient recovery.
A subset of animals was analyzed using the inclined-plane test,10 which evaluates the animal’s ability to maintain body position on a board raised incrementally to increasing angles. Performance on the inclined plane correlates with the integrity of the rubrospinal tract (and other nonpyramidal pathways) after SCI. Animals were tested at days 1, 3, 7 after injury, and once weekly thereafter for a total of 6 weeks. Mice were tested in each position, after which the angle was increased incrementally by 5°. The maximum angle at which the animal could maintain a stationary position on the board for 10 s was recorded. Investigators were blinded to the treatment protocol and recorded behavioral scores.
Tissue preparation and histochemistry
For histochemistry, animals were perfused with 4% paraformaldehyde and preserved spinal cord tissues were collected. The entire spine was dissected out and post-fixed in 4% paraformaldehyde for 3 h at 4 °C. The spinal cord was removed from the vertebral column and stored for 12 h in 15% sucrose in 0.2 M PBS at 4 °C. The spinal cord was then placed in 30% sucrose in 0.2 M PBS for 24 h at 4 °C. The spinal cord was embedded in Tissue Tek OCT compound (Sakura, Tokyo, Japan) and immediately frozen on dry ice at −80 °C. A series of 20-μm transverse sections were cut on a cryostat and mounted on Matsunami Adhesive Silan-coated glass slides (Matsunami Glass, Osaka, Japan). After washing three times with PBS, all sections were blocked in PBS containing 5% BSA and 0.3% Triton X-100 for 1 h at room temperature. The sections were then incubated with polyclonal anti-Iba1 antibody (1 : 200; Wako, Osaka, Japan) overnight at 4 °C, washed three times with PBS, and incubated with fluorescein-conjugated secondary antibodies (1 : 500; Life Technologies, Carlsbad, CA, USA) for 1 h at room temperature. Sections were then rinsed three times in PBS and mounted. The sections were viewed under an inverted light microscope equipped with epifluorescence optics and a dry condenser for phase-contrast microscopy (DP70, Olympus, Tokyo, Japan).
Preparation of leukocytes from injured spinal cords
The extraction of leukocytes from the spinal cord was performed according to a previously described procedure.37 First, mice were transcardially perfused with ice-cold PBS. The spinal cord was then dissected and suspended in Hank’s Balanced Salt Solution (Life Technologies) supplemented with 3% fetal bovine serum (FBS), 100 IU/ml penicillin, and 100 μg/ml streptomycin. The spinal cords were digested with collagenase D (5.0 mg/ml; Roche Diagnostics, Tokyo, Japan) plus 2.5 mM calcium chloride. After filtration with a 70-μm Cell Strainer (BD Biosciences, Franklin Lakes, NJ, USA), lysis of red blood cells with RBC Lysis Buffer (BioLegend, San Diego, CA, USA) and washing, the pellet, isolated by 38% Percoll centrifugation at 1500 × g for 20 min, was collected for flow cytometry.
Cell-surface and intracellular antigen staining
Cultured lymphocytes or collected leukocytes from mouse spinal cords were stimulated with 50 ng/ml phorbol 12-myristate 13-acetate (Sigma-Aldrich, St. Louis, MO, USA) and 750 ng/ml ionomycin (Calbiochem/EMD Millipore, Billerica, MA, USA) for 4 h and with 10 μg/ml brefeldin A (Sigma-Aldrich) for the last 2 h. The cells were suspended in Fixation Buffer (eBioscience) and surface stained with anti-CD45-Alexa647 (BioLegend) and anti-CD11b-PE (BioLegend). Intracellular staining was performed according to the manufacturer’s protocol, using anti-T-bet-PE (eBioscience) and anti-IL-10-FITC (BioLegend).
Flow cytometric analysis
Flow cytometry was performed with the FACSCalibur flow cytometer (BD Biosciences), and was analyzed using ProQuest (BD Biosciences) and FlowJo software (TreeStar, Ashland, OR, USA). Control experiments using isotype-matched immunoglobulins (BioLegend) were performed to determine the specificity of the signals of antibodies against specific antigens.
Following SCI, the right lateral ventricle of the mouse was stereotaxically perforated with a brain infusion kit 3 (ALZET, DURET Corporation, Cupertino, CA, USA; coordinates: 0.5 mm posterior to the bregma, 1.0 mm lateral to the bregma, 2 mm deep) and connected to an osmotic minipump (ALZET; pump model: 1007D) filled with 1 mg/ml anti-IL-10-neutralizing antibody (eBioscience; clone: JES5-16E3) or rat IgG (eBioscience). The anti-IL-10 antibody was infused for 7 days.
Culture of MG/MΦs and treatment of T-cell culture media
Culture of MG and MΦs was performed according to a previously described procedure.38 MG were obtained from the brain of C57BL/6j mice on postnatal day 1–2. The cerebral cortex of each mouse was dissected out and the meninges were eliminated. The brains were minced with surgical knives, and then digested with 0.25% trypsin for 15 min and 0.5 mg/ml DNase for the last 1 min at 37 °C. The cells were passed through a 70-μm Cell Strainer (BD Biosciences). The resultant cell suspension was diluted with Dulbecco’s Modified Eagle Medium (DMEM; Life Technologies) and spun down. The pellet was suspended with 10% FBS/1% penicillin and streptomycin/DMEM (DMEM growth medium) and plated on poly-L-lysine-coated 75-cm3 culture flask (Greiner Bio-One). On day 1 and every 3 days thereafter, DMEM growth medium was replaced. After >7 days, the flask was shaken for 30 min so that floating MG could be collected from the astrocyte monolayer sheet and cultured at the density of 3.3 × 104 cells in 100 μl growth medium for 1 well of 96-well plate (Greiner Bio-One). More than 90% of the cells were CD11b+ MG.
Bone marrow-derived MΦs were obtained from bilateral femoral bones of 9-week-old C57BL6/j mice. Marrow cores were flushed using 10-ml syringes filled with RPMI1640 (Invitrogen)/10% FBS. After trituration and lysis of red blood cells with RBC Lysis Buffer (BioLegend), the cells were washed in the medium, and then plated and cultured in RPMI1640 with 10% FBS, penicillin/streptomycin (RPMI growth medium), and M-CSF (50 ng/ml; BioLegend). Non-adherent cells were collected at day 4. On day 6, the cells were replated on 96-well plates at the density of 3.3 × 104 cells in 100 μl of RPMI growth medium for one well. On day 7, the cells were treated with the T-cell supernatant. More than 90% of the cells were CD11b+ MΦs.
One hundred microliters of CM of cultured T cells (collected 4 days after T-cell culture) or 40 ng/ml IFN-γ (BioLegend) dissolved in 100 μl medium were applied to MG or MΦs cultured as above. At 0, 1, 3, 6, and 24 h after treatment of T-cell CM, supernatant of the cultured cells were collected to detect IL-10 with ELISA.
Enzyme-linked immunosorbent assay
Th1, Th1 ifn-γ−/−, and Th17 cells were prepared as described above. Three hours after restimulation with anti-CD3-ɛ/anti-CD28 antibodies, cells were centrifuged and their supernatants were collected and stored at −80 °C until use. Concentrations of IFN-γ, IL-10, NT-3, and GDNF in the supernatants of each cell were analyzed using the ELISA kits, according to the manufacturer’s protocol (IFN-γ and IL-10: R&D Systems; NT-3: Emax Immunoassay System; GDNF: Promega, Fitchburg, WI, USA). Absorbance values were read at 450 nm on a plate reader (SpectraMax M2; Molecular Devices, Sunnyvale, CA, USA).
All values are expressed as mean±S.E.M. Motor function scores were analyzed using two-way ANOVA with repeated measures with a Bonferroni post-test for 2–3 groups. For all other data, a one-way ANOVA with a Bonferroni post-test collection was used. Values of P<0.05 were considered statistically significant.
central nervous system
spinal cord injury
- Th1 cells:
type-1 helper T cells
- Th2 cells:
type-2 helper T cells
IL-17-producing helper T cells
cluster of differentiation 4
glia-derived neurotrophic factor
enzyme-linked immunosorbent assay
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We thank Drs. T Nakayama, M Yamashita, and M Kuwahara at Chiba University for their technical advice on T-cell culture; Drs. K Morimoto and S Lee at Osaka University for their help in histological assessment; Ms Nakamura at Osaka University for her advise on surgical procedure; Dr. Y Souma at the National Institute of Biomedical Innovation for taking care of the IFN-γ knockout mice; and Drs. H Iijima, A Mukai, H at Osaka University for advice on knockout mice. This work was supported by a research grant from a Grant-in-Aid for Young Scientists (S) from the Japan Society for the Promotion of Science (JSPS).
The authors declare no conflict of interest.
Edited by A Verkhratsky
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Cite this article
Ishii, H., Tanabe, S., Ueno, M. et al. ifn-γ-dependent secretion of IL-10 from Th1 cells and microglia/macrophages contributes to functional recovery after spinal cord injury. Cell Death Dis 4, e710 (2013). https://doi.org/10.1038/cddis.2013.234
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- microglia macrophages
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The Colorado Legislature just convened yesterday, with four months of hard work and tough budgeting choices ahead. So it's no surprise that in their off-hours, citizen lawmakers would rather ignore this new year altogether and go directly to 2010, which promises plenty of political surprises.
Already, Troy Eid, outgoing U.S. Attorney, is talking about running for Attorney General, since John Suthers is eying the governor's seat -- or maybe the Senate slot soon to be occupied by Michael Bennet. Governor Bill Ritter's surprise appointment of Bennet to replace Ken Salazar has given politicos plenty to yak about -- including not just how Bennet will fare in a race in 2010, but whether Ritter himself might be challenged within his own party.
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Ritter will deliver his State of the State speech at the Capitol at 11 a.m. today. But it's no secret that right now, many ambitious pols are in a state of readiness. -- Patricia Calhoun


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In pictures: 2017 Australian International Air Show, featuring F-35 Joint Strike Fighter debut
The F-35A Joint Strike Fighter landed in Australia for the first time earlier this week and today it took center stage at the Australian International Air Show at Avalon, with two of the US$100 million jet fighters making their first public touchdown some 15 years after the Government first announced it would participate in the JSF program. With Australia set to spend AU$17 billion on 72 F-35s, the first of which should be operational in 2020, the public was eager to get a glance at its tax dollars in action.
Running concurrently at the site is the Aerospace and Defence Expo, where all manner of flight, weapons and military tech is on display. Multirotors are certainly playing a larger and larger part in any air show, and they're out in force here.
The air show and expo are open to the public this weekend, featuring a ton of aerial and static displays of both civilian and military aircraft.
Jump into the photo gallery for a visual tour of 2017 Australian International Air Show.