Richard Loving, A White Man And Mildred Jeter - 913 Words

ï‚› In 1958, in the District of Columbia, Richard Loving, a white man and Mildred Jeter, a black woman was married. Shortly after the marriage the Loving’s returned to Virginia. Upon returning to Virginia the couple was charged with violating Virginia’s Anti-miscegenation Statue. That bans inter-racial marriages. The Loving’s were found guilty and sentenced to a year in jail but the judge offered to suspend their sentence if the Loving’s were to leave Virginia and not return for 25 years. ï‚› Racial integrity Act of 1924 prohibited interracial marriage and was passed by the General Assembly to protect â€Å"Whiteness† from negative effects of race-mixing. ï‚› What is the Question: ï‚› Did Virginia s anti-miscegenation law violate the Equal†¦show more content†¦In 1959 they were expelled from the state of Virginia for violating the state law that forbids interracial marriage. ï‚› November 6, 1963 - Bernard S. Cohen, representing Richard and Mildred Loving, files a motion in the Caroline County Circuit Court to vacate their 1959 conviction for violating the state law that forbids interracial marriage. He also asks that the two one-year suspended sentences be set aside. ï‚› January 22, 1965 - Leon M. Bazile, a judge for the circuit court of Caroline County, refuses a motion on behalf of Richard and Mildred Loving to vacate their 1959 conviction for violating the state law that forbids interracial marriage. ï‚› March 7, 1966 - In Loving v. Virginia, the Virginia Supreme Court of Appeals upholds the state s antimiscegenation laws. The Court also sets aside the original conviction of Richard and Mildred Loving, finding that a sentence requiring the defendants to leave the state is unreasonable.† ï‚› April 10, 1967 - The U.S. Supreme Court hears oral arguments in the case of Loving v. Virginia. Richard and Mildred Loving, of Caroline County, are appealing their 1959 conviction for violating Virginia s laws prohibiting interracial marriage. ï‚› June 12, 1967 - In Loving v. Virginia, the U.S. Supreme Court unanimously rules that Virginia s antimiscegenation statutes violate the Constitution s Fourteenth Amendment. The decisionShow MoreRelatedThe Life Of Mildred Delores Jeter And Musiel Byrd Jeter1239 Words   |  5 PagesMildred Delores Jeter was born in Central Point, Virginia on June 22, 1939. Mildred’s parents were Theoliver Jeter and Musiel Byrd Jeter. Mildred Loving was of African American, European and Native American origin, specifically from the Cherokee and Rappahannock tribes. Mildred s family had lived in the area around Central Point, Virginia for a long time, where blacks and whites mixed freely with little racial conflict even at the peak of the Jim Crow era. Mildred was a shy woman who became a reluctantRead MoreLoving vs. Virgina783 Words   |  4 PagesLOVING v. VIRGINIA Can you imagine not being able to share your life with the person you love because of the color of your skin? Well, this was the case for those who resided in Virginia decades ago. Interracial marriages were not allowed in Virginia and sixteen other states due to the adoption of the Racial Integrity Act of 1924. The sole purpose of this act was to completely prohibit a white person marrying other than another white person. Marriage licenses were not issued until theRead MoreLoving V. Virginia, Introduction, Facts, Legal Background1567 Words   |  7 PagesLoving v. Virginia Interracial marriage: Respecting the Equal Protection and Due Process Clauses of the Fourteenth Amendment. I. INTRODUCTION This case note will examine the 1967 landmark Supreme Court case of Loving v. Virginia. The Loving v. Virginia case touched on constitutional principles including equality, federalism, and liberty. Just over 30 years ago, it was a crime for interracial couples in Virginia to marry, or to live as husband and wife. Prior to the 1967 case of Loving v. VirginiaRead MoreThe Supreme Court Case Loving V. Virginia1609 Words   |  7 PagesOn June 2, 1958 Mildred Jeter and Richard Loving went to Washington D.C. to get married and they went back to Virginia a few days later. But because Mildred was of African-American and Native American decent, and Richard was white they were arrested for violating the state law that prohibits interracial marriage. At the time, Virginia was one of 17 states, including Texas and Alabama, that had laws prohibiting interracial marriage (Wolfe). The Supreme Court Case Loving v. Virginia is an importantRead MoreEssay on Loving v. Virginia (388 U.S. 1)2059 Word s   |  9 Pagesafter midnight, Richard Loving a white man and Mildred Loving an African American woman were awakened to the presence of three officers in their bedroom. One of the three officers demanded from Richard to identify the woman next to him. Mildred, full of fear, told the officers that she was his wife, while Richard pointed to the marriage license on the wall. The couple was then charged and later found guilty in violation of the states anti-miscegenation statute. Mr. and Mrs. Loving were residentsRead MoreAmerica s Miscegenation Anxiety And The State Of Virginia1383 Words   |  6 Pagescitizenship, or to deny any person â€Å"the equal protection of the laws.† After the Supreme Court ultimately neutralized this amendment through its decision in Plessy v. Ferguson, the southern states, individually, built legal barriers between blacks and whites in practically every aspect of life. America’s miscegenation anxiety, very clearly documented in legislature, can also be found in social thought and theory. Indeed, Thomas Jefferson’s Notes On The State of Virginia must be thought of as one ofRead MoreHow The Virginia State Statue Of Anti Miscegenation And The Fourteenth Amendment Essay2353 Words   |  10 Pagesbiracial marriages and segregation laws that were discriminatory in history. I read the short story about the Loving family and their pursuit to the Supreme Court in 1967, I thought of my own family history and realized that my paternal grandparents would’ve been prosecuted had they lived in Virginia or any other state that prohibited bi-racial marriages in the 1900s. My grandfather was an Irish man and my grandmother was a Native American Crow Creek Dakotah/HoChunk woman, luckily they weren’t victimsRead MoreShould Same Sex Marriage Be Legal?2556 Words   |  11 PagesAlia Thompson Wanosky APUSH, Block F June 10, 2015 US History: Definition of a â€Å"Real Marriage† Can the US government define a marriage in 2015? In the United States, marriage is defined as â€Å"a formal union between a man and woman† [New York Times]. It is estimated that 229 million people currently are legally married in United States [Freedom to Marry]. But at the same time only thirty-six states including the District of Columbia is where same sex marriage is legal [Freedom to Marry]. RecentRead MoreThe Supreme Court and Civil Rights Essay991 Words   |  4 PagesCourt took on more responsibility and started making additional decisions, which in time helped minorities gain their civil rights. It took a couple of years, as a matter of fact till the 1900’s for the Supreme Court to get out of the â€Å"ideology of white supremacy and the practice of racism,† (Smith). Though the decisions of the Supreme Court were not all that appreciated in the beginning, following the 20th century the court really facilitated in the advancements of civil rights. The Supreme CourtRead MoreThe Changes in Society’s Views on Interracial Dating over Time1466 Words   |  6 Pagesinterracial relationships became more relevant. It sparked much controversy after a couple from Virginia was arrested for participating in an interracial relationship. The case, Loving versus Virginia, was taken to the Supreme Court, who found it to be unconstitutional. The couple, Richard Loving, a white man, and his wife, Mildred, a black woman, were married in Washington D.C. Almost immediately after, they went back to their home state of Virginia. However, their happiness was short lived after they

The Effects Of Air Pollution On Hong Kong - 1686 Words

Most people think of the smog that they see that seems to hover over some of the world’s largest cities. Some of the world’s most polluted cities are those in the two countries with the largest populations, China and India. Everybody has seen the pictures of air so thick there is no visibility in Hong Kong, New Delhi and other cities. However, not all air pollution is visible. Air pollution takes place when the air contains gases or fumes and dust. The air pollution that can be seen is called smog, but that which cannot be seen is just as dangerous as the smog that results in dangerous air quality warnings. When there is air pollution, it means that there is a high quantity of dangerous gases in the air that may be injurious to the health†¦show more content†¦That is why individual cities must make every effort to limit the amount of air pollution in their cities as an example to the wider population of their country and of the world. Besides being dangerous t o the overall health of the planet, many of the pollutants found in air pollution can cause harm to humans too. One of those pollutants is aerosols. Secondary organic aerosols are air contaminants released from natural and man-made causes. They are emitted through a complex interaction of sunlight, unstable organic compounds from plants like trees, cars or industrial discharges, and other airborne chemicals. Secondary organic aerosols are a key component of fine particle pollution, which has been found to lead to lung cancer and heart issues and other health implications. Man-made and natural aerosols reaching the air are never a good thing for the macroclimate and microclimate. Created in many ways including everything from volcano explosions to desert winds, aerosol atoms reach the air and can play havoc with the temperature and weather patterns of neighboring areas. Hong Kong, as well as other cities, have this issue worse than other areas around the world. Owing to the massive d eserts of Asia, strong wind carry silt as well as other particles into the air, and the elements collect over the Pacific Ocean. For Hong Kong precisely, recent deforestation that came about as the economy developed has

The Silver Linings Playbook Chapter 24 Free Essays

Mom’s Handwriting Emerges The sun bursts through the attic window and lands on my face, warming it, until I open my eyes and greet the day with a squint. After a kiss, I return Nikki to my bedroom dresser and find my mother still asleep in my bed. I notice that the glass of water I left her is now empty, and I am glad to have left it there, even if I am mad at Mom now. We will write a custom essay sample on The Silver Linings Playbook Chapter 24 or any similar topic only for you Order Now As I descend the staircase, I smell something burning. When I reach the kitchen, my father is standing in front of the stove. He is wearing Mom’s red apron. â€Å"Dad?† When he turns around, he has a spatula in one hand and a pink oven mitt on the other. Behind him, meat hisses – a thick river of smoke flies up into the exhaust fan. â€Å"What are you doing?† â€Å"Cooking.† â€Å"Cooking what?† â€Å"Steak.† â€Å"Why?† â€Å"I’m hungry.† â€Å"Are you frying it?† â€Å"I’m cooking it Cajun style. Blackened.† â€Å"Maybe you should turn the burner down?† I suggest, but he returns to his cooking, continuing to flip the sizzling cut over and over, so I go down into the basement to begin my workout. The fire alarm goes off for fifteen minutes or so. When I return to the kitchen two hours later, the pan he used is blackened and still on the now greasy stove; a plate and utensils are in the sink. Dad is watching ESPN on his new television, and his surround sound speaker system seems to shake the house. The clock on the microwave reads 8:17 a.m. My mother has forgotten my meds again, so I take out my eight bottles, remove all the caps, and search for the right colors. Soon I have a half dozen pills lined up on the counter, and I confirm that the colors are what I take every morning. I swallow all of my pills, thinking maybe my mother is testing me again, and even though I am technically mad at her, I am also now very worried about Mom, so I climb the steps to my room and see that she is still sleeping. Downstairs, I stand behind the couch and say, â€Å"Dad?† But he ignores me, so I return to my basement gym and continue my workout, listening to the ESPN commentators recap the college games and forecast the upcoming NFL action. Their voices arrive crisply through the floorboards above. I know from reading the paper that the Eagles are favored to win over San Francisco, which makes me excited to watch the game with my father, who will be in a great mood if the Eagles are victorious, and therefore he will also be more likely to speak with me. Midmorning, Mom descends, which is a relief, because I was starting to worry that she was really sick. I am riding the bike, and – after finding the â€Å"Pat† box last night – I just continue pedaling when Mom says, â€Å"Pat?† I do not face Mom, but using my peripheral vision, I see that she is showered, her hair is done, her makeup is applied, and she is wearing a pretty summer dress. Mom also smells really nice – lavender. â€Å"Did you take your pills last night?† she asks. I nod once. â€Å"What about this morning?† I nod again. â€Å"Dr. Patel told me I should have allowed you to take control over your meds when you first came home, that this was a step toward independence. But I was being a mom when you did not need me to be a mom. So congratulations, Pat.† â€Å"Congratulations† is a strange thing for her to say, especially since I have not won a prize or anything, but I am really only thinking about what happened last night, why Mom came home drunk. So I ask her, â€Å"Where were you last night? Did you go out with friends?† Using the corner of my eye again, I see her look down at the old brown rug beneath us. â€Å"I appreciate your putting me to bed last night. The water and the Tylenol helped. It was a bit of a role reversal, eh? Well, I appreciate it. Thanks, Pat.† I realize she has not answered my question, but I don’t know what to say, so I say nothing. â€Å"Your father has been a bear lately, and I’m simply tired of it. So I’m making some demands, and things are going to change a little around here. Both of my men are going to start taking care of themselves a little more. You need to get on with your life, and I’m sick and tired of the way your father treats me.† Suddenly I forget all about the â€Å"Pat† box and face my mother as I continue pedaling. â€Å"Are you mad at me? Did I do something wrong?† â€Å"I’m not mad at you, Pat. I am mad at your father. He and I had a long talk yesterday when you were running. Things might be a little rough around here for a few weeks, but I think we’ll all be better for it in the long run.† A wild thought leaps into my head and terrifies me. â€Å"You’re not leaving us, Mom, are you?† â€Å"No. I’m not,† Mom says, looking me in the eyes, which makes me believe her one hundred percent. â€Å"I would never leave you, Pat. But I am going out today because I’m done with Eagles football. You two are on your own for food.† â€Å"Where are you going?† I ask, pedaling faster now. â€Å"Out,† Mom says, and then kisses the little white scar on my sweaty forehead before she leaves. I am so nervous about what Mom has told me that I do not eat anything all day, but simply drink my water and do my routine. Because the Eagles are playing at 4:15, I get in a full workout. The whole time, I secretly hope my father will come down into the basement and ask me to watch the 1:00 NFL game with him, but he doesn’t. Midafternoon I climb up out of the basement and stand behind the couch for a second. â€Å"Dad?† I say. â€Å"Dad?† He ignores me and keeps watching the 1:00 game, and I don’t even look to see who is playing, because I am so nervous about what Mom told me. I put on my trash bag and hope Tiffany is outside, because I could really use someone to talk to. But after I stretch for fifteen minutes, Tiffany doesn’t show, so I run alone, thinking it funny that when I want to run alone, Tiffany is always there, but today she is not. I am very hungry, and the pain in my stomach increases as I run, which I relish because it means I am losing weight, and well, I feel as though I might have put on some extra fat in the past week, especially after drinking beer with Jake last weekend. This reminds me that I have not spoken with Jake since the Eagles lost to the Giants, and I wonder if he is coming over today to watch the game with Dad and me. Since the pain has sharpened, I decide to run farther than usual, pushing myself. Also, I am sort of afraid to go home, now that my mother has left me alone with my father for the day, and I am not sure what she meant by â€Å"changes† anyway. I keep wishing Tiffany was running with me so I might talk to her and tell her how I feel, which is a strange desire since she usually never says much in response, and the last time I tried to talk to her about my problems, she started cursing very loudly in a public place and said some really awful things about Nikki. Still, I am s tarting to feel as though Tiffany is my best friend, which is sort of strange and scary. At the end of my run, I jog down my street, and Jake’s silver BMW is nowhere to be seen. Maybe he took the train in from Philadelphia, I think. I am hoping not to be left alone with my father for the game, but somehow I know this is exactly what is going to happen. When I enter the house, my dad is still alone on the couch, wearing his McNabb jersey now and watching the end of the 1:00 game. A small collection of beer bottles stand at his feet like bowling pins. â€Å"Is Jake coming over?† I ask my father, but he ignores me again. Upstairs, I shower and put on my Hank Baskett jersey. When I reach the family room, the Eagles game is just coming on, so I sit down at the end of the couch my father is not occupying. â€Å"What the hell is that noise?† Dad says, and then turns down the volume. I realize my stomach is making crazy gurgling noises, but I say, â€Å"I don’t know,† and Dad turns up the volume again. Just as I had hoped, the new television is an experience. The players warming up on the field look life-size, and the sound quality makes me feel as though I am in San Francisco, sitting on the fifty-yard line. Realizing that my brother is not going to make it by kickoff, when a commercial comes on, I jump to my feet and yell â€Å"Ahhhhhhhhh!† but Dad only looks at me like he wants to hit me in the face again. So I sit down and do not say anything else. The announcers state that Donte Stallworth was a late scratch, so I start to hope Baskett will get a few more balls thrown his way, since the Eagles’ number one receiver is out of action. The Eagles set up a nice drive and score on their first possession with a shovel pass to Westbrook, at which point my father’s emotions morph. He reaches across the couch and repetitively claps his hand against my thigh, saying over and over again, â€Å"Touchdown Eagles! Touchdown Eagles!† I start to feel hopeful for my dad, but when the Eagles kick off, he resumes his negative ways and says, â€Å"Don’t celebrate too much. Remember what happened last week.† And it is almost as if he is talking to himself, reminding himself not to be overly hopeful. The defense holds strong, and tight end L. J. Smith scores a touchdown with only a few minutes left in the first quarter, making it 13 – 0. Even though the Eagles have blown big leads before, it seems safe to say the Birds are the superior team today. My thoughts are confirmed after Akers hits the extra point and my father jumps up and starts singing â€Å"Fly, Eagles, Fly.† So I jump up and sing with him, and we both do the chant at the end, spelling the letters with our arms and legs: â€Å"E!-A!-G!-L!-E!-S! EAGLES!† Between quarters, my father asks me if I am hungry, and when I say yes, he orders us a pizza and brings me a Bud from the refrigerator. With the Eagles up 14 – 0, he is all smiles, and as we sip our beer, he says, â€Å"Now all we need is your boy Baskett to get a catch or two.† As if my father’s words were a prayer answered, McNabb’s first completion in the second quarter is to Baskett for eight yards. Dad and I cheer so loudly for the undrafted rookie. The pizza arrives during halftime, and the Eagles are up 24 – 3. â€Å"If only Jake were here,† my father says. â€Å"Then this day would be perfect.† My dad and I have been so happy that I’ve forgotten Jake is not with us. â€Å"Where is Jake?† I ask, but Dad ignores the question. In the third quarter the San Francisco running back fumbles on the Eagles’ one-yard line and defensive tackle Mike Patterson picks up the ball and runs toward the opposite end zone. Dad and I are out of our seats, cheering on the three-hundred-pound lineman as he runs the whole length of the field, and then the Eagles are up 31 – 3. San Francisco scores a few touchdowns late in the second half, but it doesn’t matter, because the game is basically out of reach, and the Eagles win 38 – 24. At the conclusion of the game, my father and I sing â€Å"Fly, Eagles, Fly† and do the chant one last time, celebrating the Eagles’ victory, and then Dad simply turns off the television and returns to his study without even saying goodbye to me. The house is so quiet. Maybe a dozen or so beer bottles on the floor, the pizza box is still on the coffee table, and I know the sink is stacked full of dishes and the pan in which Dad cooked his breakfast steak. Since I am practicing being kind, I figure I should at least clean up the family room so Mom won’t have to do it. I carry the Bud bottles out to the recycle bucket by the garage and throw away the pizza box in the outside garbage can. Back inside, a few used napkins are on the floor, and when I reach down to pick up the mess, I spot a crumpled ball of paper under the coffee table. I pick up the ball, uncrumple it, and realize it is not one but two pieces of paper. Mom’s handwriting emerges. I flatten the papers out on the coffee table. Patrick, I need to tell you I will no longer allow you to disregard the decisions we make together, nor will I allow you to talk down to me any longer – especially in front of others. I have met a new friend who has encouraged me to assert myself more forcefully in an effort to gain your respect. Know that I am doing this to save our marriage. Your options: Return the monstrous television you purchased, and everything will go back to normal. Keep the monstrous television, and you must agree to the following demands: You must eat dinner at the table with Pat and me five nights a week. You must go on a half-hour walk with either Pat or me five nights a week. You must have a daily conversation with Pat, during which you ask him at least five questions and listen to his replies, which you will report to me nightly. You must do one recreational activity a week with Pat and me, such as eating at a restaurant, seeing a movie, going to the mall, shooting baskets in the backyard, etc. Failure to complete either option 1 or 2 will force me to go on strike. I will no longer clean your house, buy or cook your food, launder your clothes, or share your bed. Until you declare which option you wish to take, consider your wife on strike. With best intentions, Jeanie It does not seem like Mom to be so forceful with Dad, and I do wonder if her â€Å"new friend† coached her through the writing of the two-page letter. It is very hard for me to picture Dad returning his new television, especially after watching the Eagles win on the new set. His purchase will be considered good luck for sure, and Dad will want to watch next week’s Eagles game on the same television so he will not jinx the Birds, which is understandable. But the demands Mom made – especially the one where Dad has to talk to me every night – also seem incredibly improbable, although I do think it would be nice to eat dinner together as a family and maybe even go out to a restaurant, but not to the movies, since I am now only willing to watch the movie of my own life. Suddenly I need to speak with my brother, but I do not know his phone number. I find the address book in the cabinet above the stove and place a call to Jake’s apartment. A woman picks up on the third ring; her voice is beautiful. â€Å"Hello?† she says. I know it is not my brother on the other end, but I still say, â€Å"Jake?† â€Å"Who is this?† â€Å"It’s Pat Peoples. I’m looking for my brother, Jake. Who are you?† I hear the woman cover the phone with her hand, and then my brother’s voice comes through loud and clear: â€Å"Did you see that ninety-eight-yard fumble return? Did you see Patterson run?† I want to ask about the woman who answered my brother’s phone, but I am a little afraid of finding out who she is. Maybe I should already know, but forget somehow. So I simply say, â€Å"Yeah, I saw it.† â€Å"Frickin’ awesome, dude. I didn’t know a defensive tackle could run that far.† â€Å"Why didn’t you come over and watch the game with Dad and me?† â€Å"Truthfully?† â€Å"Yes.† â€Å"I can’t lie to my brother. Mom called me this morning and told me not to come, so I went to a bar with Scott. She called Ronnie too. I know because Ronnie called me to make sure everything was okay. I told him not to worry.† â€Å"Why?† â€Å"Should he be worried?† â€Å"No, why did Mom tell you and Ronnie not to come over?† â€Å"She said it would give you a chance to be alone with Dad. She said it would force Dad to talk to you. So did he?† â€Å"A little.† â€Å"Well, that’s good, right?† â€Å"I found a note from Mom to Dad.† â€Å"What?† â€Å"I found a note from Mom to Dad.† â€Å"Okay. What did it say?† â€Å"I’ll just read it to you.† â€Å"Go ahead.† I read him the note. â€Å"Shit. Go Mom.† â€Å"You know he won’t be taking the television back now, right?† â€Å"Not after the Birds won today.† â€Å"Yeah, and I’m worried that Dad won’t be able to meet the demands.† â€Å"Well, he probably won’t, but maybe he’ll at least try, right? And trying would be good for him – and Mom.† Jake changes the subject by mentioning Baskett’s catch in the second quarter, which turned out to be his only catch of the game. My brother doesn’t want to talk about our parents anymore. He says, â€Å"Baskett’s coming along. He’s an undrafted rookie, and he’s getting catches. That’s huge.† But it doesn’t feel huge to me. Jake says he’s looking forward to seeing me next Monday night, when the Eagles will play the Green Bay Packers. He asks me to have lunch in the city before we tailgate with Scott and the fat men, and then we hang up. It’s getting late, and my mother is still not home. I begin to worry about her, and so I do all the dishes by hand. For a good fifteen minutes – with steel wool – I scrub the pan my father burned. And then I vacuum the family room. Dad had splattered some pizza sauce on the couch, so I find some cleaning spray in the hall cabinet and do my best to remove the stain – dabbing lightly and then wiping a little harder in a circular motion, just like it says on the side of the bottle. My mom comes home as I am on my knees cleaning the couch. â€Å"Did your father tell you to clean up his mess?† Mom asks. â€Å"No,† I say. â€Å"Did he tell you about the letter I wrote him?† â€Å"No – but I found it.† â€Å"Well, then you know. I don’t want you to do any cleaning, Pat. We’re going to let this place rot until your father gets the message.† I want to tell her I found the â€Å"Pat† box in the attic, how hungry I was today, that I really don’t want to live in a filthy house, and I need to take one thing at a time – finding the end of apart time first and foremost – but Mom looks so determined and almost proud. So I agree to help her make the house filthy. She says we will be eating takeout, and when my father is not home, everything will be as it was before she wrote the note, but when my father is home, we will be slovenly. I tell Mom that while she is on strike, she can sleep in my bed, because I want to sleep in the attic anyway. When she says she’ll sleep on the couch, I insist she take my bed, and she thanks me. â€Å"Mom?† I say when she turns to leave. She faces me. â€Å"Does Jake have a girlfriend?† I ask. â€Å"Why?† â€Å"I called him today, and a woman answered the phone.† â€Å"Maybe he does have a girlfriend,† she says, and then walks away. The indifference Mom shows regarding Jake’s love life makes me feel as though I am forgetting something. If Jake had a girl friend Mom did not know about, she would have asked me a million questions. Her lack of interest suggests that Mom is keeping another secret from me, maybe something larger than what I found in the â€Å"Pat† box. Mom must be protecting me, I think, but I still want to know from what. How to cite The Silver Linings Playbook Chapter 24, Essay examples

Rock Essay Example For Students

Rock Essay In this essay, Im going to introduce to the reader a topic not touched a lot because of its complexity and its avoidance by conservative adults. This topic is, of course, Rock Music. During one week, I looked for information in the library and at my house, and from the information I gathered and my one knowledge about the topic, Im going to lead the reader to a better understanding of Rock n Roll. I chose to do Rock music because I can identify myself with it. Rock musicis very complex. In factIts stylistic scope is to broad to be encompassed by any single definition (Rock Music, Groliers, p.1). Thenearest definition suggests a kind of music that represents and speaks for the teenage society. This music is characterized by using a heavy beat. In this essay, Im going to divide Rock music into four sections: Rock of the 50s, of the 60s, of the 70s and of the 80s. Within these sections Im also going to discuss several sub-topics such as famous composers and groups, and characteristics o f the music. The first section of this essay is Rock n Roll of the 1950s, when Rock n Roll was born. It emerged from rhythm and blues,a music similar to jazz played by blacks. This kind of music started to attract white teenagers. Disc jockey Alan Freed was the one who introduced this music and later gave it the name of Rock n Roll. Record companies distributed records played by whites but composed by blacks. Whites were frustrated because there werent any white artists and they didnt want the blacks to be the stars until Bill Haley appeared with his Rock Around the Clock. In this decade, Elvis Presley introduced amusic that was sexual suggestive and outraged dull adults. In time he changed the style of the music by adopting a country and western style andbecame a national hero. By the end of this decade and the start of the next, Rock n Roll started to decline because it was formula ridden and it was too sentimental. Teenage audiences transferred their allegiance to Folk music. In 1963 the renewal of Rock n Roll came when The Beatles started to play. The Beatles, for some the best rock group ever, were from Liverpool, England. Through the 60s, The Beatles dominated the record industries and with their dominant instrumentation, which included: electric leads, rhythm, and bass guitar, drums and sometimes an electric organ, changed the name of Rock n Roll to just Rock. During the1960s many other styles of music arose from Rock like, Motown, Soul music, Jazz-rock , Folk-rock and others. Folk-Rock the most appreciated of this derivations and was first suggested by Bob Dylan. This kind of music brought to folk music a hard beat and amplification; and to Rock, a new poetic style. California was one of the major centers of rock activity and experimentation during the decade. First it was characterize for its surfing music, a very joyful music that reflected the fun people had while surfing. The Beach Boys were the ones who introduced this kind of music. At the end of the century this happy kind of music changed to a more rebellious style that was designated the name of hippie music. Groups that played this music were Country Joe and The Mamasand The Papas. Along with this hippie ideas popularity of hallucinogenic drugs produced a psychedelic style of music called Acid Rock. By theend of the 60s the distinctions between Rock n Roll and Rock were evident. The early instruments- saxophone, piano, amplified guitar, and drums-had been changed to electric guitar and bass, amplified drums and other electronic devices. Not only did the instruments change but so did the ideas behind the music. For example, to the lyrics of teenage love and adolescent concerns were added social commentary, glorification ofdrugs and free-association poetry(Rock Music, Groliers, p.1). Groups like The Beach Boys, Crew Cuts and The Everly Brothers were replaced bymore imaginative, non-descriptive names groups like The Who, Jefferson Airplane, Big Brother and Holding Company. The Who, the most famousof these groups, were originally from England and were reknowned becauseof their bizarre stage performances, they would destroy their instruments after their performance finished. The Who was one of the first rock groups. In the 70s, the common barriers of rock broke into more divisions, like hard rock and mellow rock. Hard Rock was extremely loud and electronically am plified and Mellow Rock was softer and with acoustic instruments. In 1972, in Jamaica, a new style of music was created called Reggae. Reggae is a mixture of rock, soul, calypsoand Latin music. The king of Reggae was Bob Marley. Other styles more in rocks borders, since Reggae was more latin than rock, were created inthe middle of this century, like: bubble gum rock, a funny playful music directed to the youngest fans, Punk Rock, a loud, hard rock style derived from acid rock and marked by its extremes of costume andstaging (Rock Music, Encarta, p.1), and Heavy Metal, which continued the approach to Acid Rock but with a simpler musical dimension but relying upon the power of repetitiveness, loud volume, and electronic distortion. One famous group of Heavy Metal was Led Zeppelin, a British group that was formed in 1968 by Jimmy Page (lead guitar),Robert Plant(lead singer), John Paul Jones (pianist and bassist) and John Bonham (drummer). Most music of this decade was intended to beli stened to, but not to be danced. But this intention wasnt kept by Disco music that arose in 1977 and was especially for dancing. One great group of Disco music were the Bee Gees. Disco music was described by rock fansas mechanical, commercial and unlyrical (which is true). At the end of this decade, rock, again became a dominant cultural force. The last but not least section of this essay is rock of the 80s. At the start of this last decade, rock groups became more production oriented, mainly because of the sudden explosion of videos. This new sensation was a good way to sell music to the people. Heavy metal bands were greatly pushed by videos, but most helped were the popular performers like Michael Jackson, Prince and Boy George. The influence of British bands of punk, disco, reggae and pop-rock was still big in the U.S. Rock scene. At the same time, there of nostalgia to return to the older pre rock music, like rhythm and blues, which was suggested by played by Elvis Costello. B y the middle of the 80s, almost every country had begun to support indigenous music, and at the end of this a vigorous talk-song style called rap became extremely popular among urban black teenagers (Rock Music, Encarta, p.1.). Rock music taught me to appreciate things in a different way. Ive learned this since rock is not an exact science, it can change. In fact Rock music helps me relax ( I would have been able to finish writing this essay with rock music). After writing this essay I have learned the origins of rock and the branches of it, but that wasnt my intention. I wrote this essay to express myself with it because I feel I can show myself with rock music. I think rock has become social phenomenal.

Investment Decision free essay sample

Tianfu Electronics Ltd is a midsized electronics manufacturer located in Chengdu, China. The company president is Dr. Wang Datong, a graduate from Tsinghua University, who founded the company 20 years ago. The company originally repaired radios and other household appliances. Over the years, the company expanded into manufacturing and is now a reputable manufacturer of various electronic items. Meng Xiaolan, a recent MBA graduate from Fudan University, has been hired by the company’s finance department. One of the major revenue-producing items manufactured by Tianfu is a personal digital assistant (PDA). Tianfu currently has one PDA model on the market, and sales have been excellent. The PDA is a unique item in that it comes in a variety of tropical colors and is preprogrammed to play traditional and modern music. However, as with any electronic item, technology changes rapidly, and the current PDA has limited features in comparison with newer models. Tianfu spent 1,815,000 yuan (RMB) to develop a prototype for a new PDA that has all the features of the existing PDA but adds new features such as cell phone capability. We will write a custom essay sample on Investment Decision or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page The company has spent a further 390,000 yuan for a marketing study to determine the expected sales figures for the new PDA. Tianfu can manufacture the new PDA for 150 yuan each in variable costs. Fixed costs for the operation are estimated to run 4. 5 million yuan per year. The estimated sales volume is 70,000, 80,000, 100,000, 85,000, and 75,000 per each year for the next five years, respectively. The unit price of the new PDA will be 340 yuan. The necessary equipment can be purchased for 19. 5 million yuan and will be depreciated on a sevenyear schedule, with the depreciation percentages for the first five years being. It is believed that the market value of the equipment in five years will be 3. 3 million yuan. As previously stated, Tianfu currently manufactures a PDA. Production of the existing model is expected to be terminated in two years. If Tianfu does not introduce the new PDA, sales will be 80,000 unites and 60,000 units for the next two years, respectively. The price of the existing PDA is 280 yuan per unit, with variable costs of 120 yuan each and fixed costs of 1. 8 million yuan per year. If Tianfu does introduce the new PDA, sales of the existing PDA will fall by 15,000 units per year, and the price of the existing units will have to be lowered to 240 yuan each. Net working capital for the PDAs will be 20 percent of sales and will occur with the timing of the cash flows for the year; for example, there is no initial outlay for NWC, but changes in NWC will first occur in year 1 with the first year’s sales. Tianfu has a 25 percent corporate tax rate and a 12 percent required return. a. Wang Datong has asked Meng Xiaolan to prepare a report and determine the IRR and NPV of the project. What should Xiaolan propose for the decision? b. Tianfu has an idle piece of equipment that can be used for the new project, which, if used, can reduce the purchase of necessary new equipment and its salvage value in five years by 20%. The idle equipment currently has a book value of 4. 05 million yuan, and is to be depreciated to zero in four years in the straight-line method. The current market value of the idle equipment is 3. 735 million yuan and will have no salvage value in five years. What are the IRR and NPV if the idle equipment is used? Compare the difference in your results between (a) and (b) and briefly discuss.

Namibias Independence essays

Namibia's Independence essays Namibia is a country on the southwestern tip of Africa. The geography of Namibia has shaped its course through history. Characterized by two deserts, it was assumed Namibia had little to offer. Despite its low population when Germany took it as a colony in the early 1900s, it still bitterly revolted. Likewise, when South Africa began an attempt at cheap migrant workers for the regions diamond and gold mines, nationalist sentiment caused Namibia to fight for its freedom. Freedom was a long and hard struggle, but after overcoming many hard trials, the country of Namibia gained its independence from external powers in 1990. Namibias earliest occupants were the San people (also known as bushmen). They lived in the area now know as Namibia as early as 8000 BC. Although they were the first inhabitants, they were eventually pushed aside by Bantu-speakers, who, with the advanced technology of iron working for them, pushed the bushmen into the Kalahari. The Bantu-speakers spread throughout the country, and had varied governments in various places. There were villages with chiefs, kingdoms with hereditary succession, and cattle-raisers. These various people occasionally traded goods and had various conflicts with each other. Portuguese sailors placed a cross on the shores of Namibia in 1484 AD, but few Europeans actual began to explore the country until 1650, when the Dutch East Indies Company briefly explored it. The major problem Europeans had was the Skeleton Coast, which took many ships and sailors to their doom. In the 1700s several whaling and sealing ships frequented the area, but there were few European settlers in the area. The early 1800s were when Europeans and Americans stepped up their trading with Namibia for ivory and cattle. The discovery of diamonds in the 1850s brought a veritable flood of miners and traders. Several countries had commercial interests in Namibia by the 1880...

Current Sources of Basic Legislation in the UK Case Study

Current Sources of Basic Legislation in the UK - Case Study Example A company is said to be a private limited liability company if it is incorporated under the laws of either of the following: Scotland, England, Republic of Ireland, and Wales. In addition its shares are strictly limited to a specific group of shareholders but not the general public, consequently cannot be put for trading on a commercial stock exchange.1 The fact that the company is limited by shares is an indication of the presence and ownership of company capital by shareholders who are equally bound by the company's liability to creditors and other third-party institutions and agents. Going by the conventional rules governing the issuance of shares, shareholders and their personal effects are legally insulated from confiscation during insolvency save the value of the premium paid and the nominal value of the shares owned and issued by the company. According to the laws of the United Kingdom, all private limited liability companies are mandated to bear the suffix "Limited" abbreviated as "Ltd." The Case of John and his Partners Suffice to begin this section with a definite reference to the new Company and Companies Act 2006, will serve as the main reference framework of this paper. According to the act which will become functional in the last quarter of this year.

Discussion Board 4-2 Assignment Example | Topics and Well Written Essays - 250 words - 5

Discussion Board 4-2 - Assignment Example Safety plan for intimate partner violence may include assessment of conditions, situation and events that lead to and follow crisis or violence. To ensure treatment of healthy clients, the plan may cover necessity to subject victims of abuse to medical and mental checkups. The plan may further include assessment of history of the violence and manners of handling and addressing the experienced violence (Jackson-Cherry, 2014). Safety plan for intimate partner violence may also include provisions to engage with community, religious or local government leaders to help in establishing frameworks for providing safety for victims of abuse, and offenders in case of vulnerability to community attack. In extreme cases of abuse, safety plan for intimate partner violence include possibility of involving police to initiate arrest and organize for prosecution of abusive partners. The safety plan also has to include promise for shelter, home, work or any necessary resource to comfort the victim of abuse and enable normal running of life routine (Jackson-Cherry, 2014). Another possible component of safety plan for intimate partner violence is increasing accountability level of offenders through measures or ways agreed and approved by the victim, and have to be in manners that do not affect client

Project X Essay Example | Topics and Well Written Essays - 500 words - 2

Project X - Essay Example he form of auditing, internal controls, policies and procedures, budget restrictions, and the like all of which can be enforced at some costs given the assumption of symmetric information (Frame, 2003). The main techniques of risk monitoring include assessment, cross-functional teams, inspection, interviews, reports, observations and reviewing. The most usual ones of these are inspection and control charts, and it is possible to use flowcharting and trend analysis to see whether production facilities and budget to the norm or getting worse. However, these tools are used rather late in the day and are more to do with correcting errors and confirming that what is being delivered is what is required. More emphasis needs to be placed on making sure the correct quality or performance is specified at the outset and clearer recognition of the need for a well-motivated team that clearly understands the project. Some researchers underline that performance monitoring is a part of risk management and control. â€Å"Performance monitoring involves measuring operational activities, analyzing the resulting metrics, and comparing them to internally established standards and industry benchmarks to assess the effectiveness and efficiency of existing operations† (Risk Monitoring and Reporting, n.d.). For project X, the most effective monitoring and control methods will be assessment, cross-functional teams and inspection. Cross-functional teams perform an important role in organizations joining different project areas. To achieve the task requires clear definition, good planning, clear roles and responsibilities, appropriate resources and regular reviews as the project proceeds. Inspection (independent monitoring) will help managers to provide external analysis of the resources, current technological processes and compare them with established standards (applied to the industry in general). Interviews and reports can be identified as internal control methods, which allow a manager

The Conduct Of Inter Professional Practice Social Work Essay

The Conduct Of Inter Professional Practice Social Work Essay This study aims to investigate the conduct of inter-professional practice in areas of social and health care, with specific regard to the involvement of service users in such practice. The case study prepared by the City and Hackney Local Safeguarding Children Board on Child A and Child B is taken up for analysis and review in this context. The case study is taken as read and is not elaborated for the purpose of this essay. Health and social care in the UK is currently being significantly influenced by a growing commitment towards greater public involvement in the design, delivery and evaluation of services, greater availability and choice of services for all categories of service users, reduction of inequality, greater emphasis on provisioning of services at the local level, (including from the independent and voluntary sectors), the commissioning process, integration of social and health care, and professional roles for delivery of care on the basis of actual needs of service users (Barrett, et al, 2005, p 74). Such reforms call for the blurring of strict boundaries between the different professionals and agencies working in health and social care (Cowley, et al, 2002, p 32). They also call for greater inter-professional and inter-agency working and for significant alterations in organisational cultures in order to enhance the power base of service users and members of the public in different aspects of social care provision (Cowley, et al, 2002, p 32). It is now widely accepted that health and social care professionals need to be more responsive to the rapidly changing needs of service users. Such changes call for the development of health and social care practitioners to improve care for clients and service users (Day, 2006, p 23). Such improvement is required to be brought about by more emphasis on person centred care for clients and service users and the greater involvement of such people in different aspects of planning, delivery and evaluation (Day, 2006, p 23). The increasing contemporary emphasis on user involvement in the policy and practice of social care is however coming in for increasing questioning from disenchanted service users and service user organisations (Branfield Beresford, 2006, p 2). Service users, whilst highlighting the benefits of their involvement in the social and health care process, are raising various questions about their actual participation in social and health care and the continuance of various barriers that prevent their genuine contribution to the process (Branfield Beresford, 2006, p 2). The case study under question details the results of an enquiry into an episode, wherein a mentally disturbed mother killed her two children after (a) being released from institutional surroundings, and (b) being integrated with her children with the full knowledge and approval of an overseeing group of social, health, nursing and mental health professionals. The enquiry raises disturbing issues about the extent of involvement of service users in social and health care processes and in the decision making of the inter-professional group overseeing the care, treatment and rehabilitation of a mentally disturbed and potentially dangerous individual. The essay investigates the involvement of service users in inter-professional practice in the UK, with specific regard to the case study and the enquiry report. Whilst doing so it takes cognizance of (a) identification of sources for evidence based social work practice, (b) the use of enquiry reports as sources of evidence, (c) the relevance of themes that emerge from such enquiries, and (d) the implications of evidenced based practice for the development of practice in social work. The essay is analysed vis-a-vis the Every Child Matters programme and makes use of legal, political and ethical frameworks. Inter-professional Practice Inter-professional practice and inter-agency collaboration aims to ensure the coming together of service providers, agencies, professionals, carers and service users in order to improve the final level of quality of planning and delivery of services (Mathias Thompson, 2001, p 39. Whilst partnership and collaboration are often considered to be interchangeable, collaboration is the actual foundation for joint working and the basis for all successful partnerships (Mathias Thompson, 2001, p 39). The UK has been enacting legislation and policies for the promotion of Inter-professional and inter-agency collaboration (IPIAC) for the last five decades in order to enhance standards and reduce costs in health and social care (SCIE, 2009, p 1 and 2). The development of IPIAC was shaped by the white paper Caring for People in 1989, followed by the enactment of the NHS and Community Care Act 1990. The government has in recent years issued various policy documents for the promotion of collaboration in order to improve efficiency and effectiveness (SCIE, 2009, p 1). Greater emphasis on IPIAC is expected to improve care because different professional groups like social workers, physicians, teachers and police officers will during the course of such working bring their individual perspectives to the collaborative process (SCIE, 2009, p 1and 2). The IPIAC process will aim to ensure the best ways in which such individual and sometimes differing perspectives can be made to come together, as also the ways whereby respective contributions of different professionals and agencies can be utilised to enhance standards of service and experiences of service users and carers (Freeth, 2001, p 38). Consideration requires to be given to collaboration between organisations, as well as professionals, in the course of IPIAC working. It is also important to consider the differences in the working practices and cultures of the various organisations that are required to work together and to take appropriate action to minimise the impact of such differences in order to make inter-professional practice effective (Freeth, 2001, p 38). Policy makers and practitioners agree that adoption of IPIAC will result in greater service delivery despite the existence of various personal, individual and organisational barriers that can practically hinder its efficiency and effectiveness (Day, 2006, p 23). It is however also widely accepted that effective IPIAC working cannot take place in the absence of deliberate involvement of service users and clients in all stages of planning, delivery and evaluation processes (Day, 2006, p 23). The white paper Modernising Social Services, published in 1998 clearly states that people cannot be placed in neat service categories and users will inevitably suffer if partner agencies do not work together (SCIE, 2009, p 1).It is now mandatory that social work programmes, as well as nursing and midwifery, embrace the involvement of patients and service users. Contemporary government reforms are based on public involvement in different aspects of service delivery (SCIE, 2009, p 2). Person centred approaches in health and social care recognise the need for valuing the opinions and experiences of patients and service users and the adoption of person centred approaches by social work practitioners (SCIE, 2009, p 2). Current research however reveals that service users often feel left out of the process of social care, despite the progressive implementation of IPIAC concepts and approaches (Branfield Beresford, 2006, p 2). Service user organisations state that the knowledge of service users is by and large not taken seriously or valued by professionals and service agencies. Many service users find such attitudes from professionals and agencies to be intensely disappointing and disempowering (Branfield Beresford, 2006, p 3). Agencies and practitioners do not appear to be interested in the information provided by service users and do not accord the respect to such knowledge that they otherwise provide to professional knowledge and expertise. Service users also feel that the cultures of social and health care organisations continue to be closed to service user knowledge and reluctant to change (Branfield Beresford, 2006, p 3). The study of the case review of the episode involving the deaths of child A and child B appears to reinforce the impression of service users about their continued exclusion from the working and decisions of different agencies and professionals involved in delivery of social and health care (Henderson, p 261). The Every Child Matters Programme requires social work agencies and professionals like social workers, health care specialists, teachers, nurses, doctors and mental health professionals to constantly ensure the safety, security and protection of children wherever they can. Extant legislation and policies like The Children Act 2004 and the Every Child Matters Programme clarify that it is everyoneà ¢Ãƒ ¢Ã¢â‚¬Å¡Ã‚ ¬Ãƒ ¢Ã¢â‚¬Å¾Ã‚ ¢s job to ensure the safety of children (Henderson, p 261). The report clarifies that various agencies were involved in the assessment and treatment of Ms. C, the wife of Mr. D and the mother of the two children, child A and child B. The report further reveals that agencies, as well as individual practitioners, failed to consider the views, opinions, and experiences of service users, even as it also contains a number of examples of sound agency and inter-agency practice. There is limited evidence of professional contact with Mr. D, the father of the children, after the contact session in October 2006, and it appears likely that professional networks assumed the agreement of Mr. D with arrangements for Ms. C. Professionals also paid inadequate attention during their provisioning of support to Ms. C, in response to her request for re-housing, and did not communicate with Mr. D to ensure that future arrangements would serve the best interests of the children. Interviews conducted with Mr. D and his parents also revealed significant differences b etween their expectations of the roles of social workers roles and what was implied by the records kept in the agency. Mr. Dà ¢Ãƒ ¢Ã¢â‚¬Å¡Ã‚ ¬Ãƒ ¢Ã¢â‚¬Å¾Ã‚ ¢s family members, it appears, were clearly under the impression that they had little choice in the rehabilitation process and were furthermore required to facilitate the contact of the children with their mother. Whilst the report elaborates the role and sincerity of various agencies and professionals in assessing Ms. Cà ¢Ãƒ ¢Ã¢â‚¬Å¡Ã‚ ¬Ãƒ ¢Ã¢â‚¬Å¾Ã‚ ¢s condition and her rehabilitation in society, it specifically refers to (a) the under involvement of Mr. D in the process, (b) the lack of communication with him (Mr D) by social workers and agencies, (c) the differences in perceptions about the role of social workers between Mr. D and his family and the agency, (d) the poor communication of agencies with the parents, (e) the absence of school records of children, and (e) the scope for improvement of involvement of GPs and the police in the social care process. Although the report makes several recommendations, the specific references to involvement of service users calls for detailed and greater involvement of parents and carers of children in planning of discharge and assessment of risk in order to ensure that actions are based on full information. One of the agencies, the East London and the City Mental Trust has been asked to involve family members and carers of children in all processes, even as the Hackney Children and Young Peopleà ¢Ãƒ ¢Ã¢â‚¬Å¡Ã‚ ¬Ãƒ ¢Ã¢â‚¬Å¾Ã‚ ¢s Service has been directed to ensure that decisions are not taken on issues that can affect children without communicating carefully and appropriately with current carers. Emerging Themes and Evidenced Based Practice The revelations of the enquiry into the report reveal a number of themes in different areas of inter-professional practice, inter-agency working and the involvement of service users in planning, delivery, and evaluation of health and social care, which can be beneficially used to inform future social work practice. The report specifically refers to (a) the lack of participation of services users in social and health care processes, and (b) the involvement of different agencies in their exclusion, thereby reinforcing the need for greater emphasis by agencies and practitioners on involvement of service users in their care plans. It also becomes obvious that much of the sentiments and ideas about involvement of service users in social care processes continues to remain in the realm of rhetoric and that it will need determined and deliberate effort by practitioners to truly bring services users into the actual planning, intervention and evaluation functions of social work practice. Enquiry reports serve as important sources of evidence for development of future social work practice. The impact of the enquiry conducted by Lord Laming into the death of Victoria Climbie led to the revelation of evidence on gross inadequacies in the social care system for children and widespread organisational malaise (Roberts Yeager, 2006, p 19). The publication of the report led to radical changes in governmental policy on social care for children and to the introduction of the Every Child Matters Programme and other important policies for the physical and mental welfare of children (Roberts Yeager, 2006, p 19). The utilisation of research evidence for guidance of practice and development of policies in the area of social services and health care is becoming increasingly important for enhancing the effectiveness of social and health care interventions, especially so because of the limited available resources with the government and the pressures to achieve positive outcomes (Johnson Austin, 2005, p 5). Scholars however feel that much of research based evidence is not absorbed by practitioners and have identified five important requirements for research evidence to practically influence practice and policy, namely (a) concurrence on nature of evidence, (b) a strategic approach to the conception of evidence and the progression of an increasing knowledge base, (c) effective distribution of knowledge along with development of useful means for accessing knowledge, (d) initiatives for increasing use of evidence in policy and practice, and (5) a range of actions at organisational level to increase use of evidence (Johnson Austin, 2005, p 5). Conclusions This study investigates the conduct of inter-professional practice in areas of social and health care, with specific regard to the involvement of service users in such practice. The case study prepared by the City and Hackney Local Safeguarding Children Board on Child A and Child B is specifically taken up contextual review. Inter-professional practice aims to ensure the collaborative working of service providers, agencies, professionals, carers and service users in order to improve the planning and delivery of services. Policy makers and practitioners also agree that whilst adoption of inter-professional working is likely to lead to improved care, it cannot occur without the involvement of service users in all stages of the care process. Person centred approaches also recognise the importance of considering the opinions and experiences of service users in planning, intervention and evaluation of care. Contemporary research however reveals that service users feel that their knowledge is not valued by professionals and agencies. The results of the enquiry reinforce the possibility of service users being excluded from the working of agencies and professionals and refer to a number of instances, where the opinions of the service users were not considered for taking of practice and intervention decisions. The report reveals a number of themes in different areas of inter-professional practice that can be beneficially used to inform future social work practice. The use of research evidence for guidance of practice in social work is becoming increasingly important for improving the effectiveness of social and health care interventions. Enquiry reports serve as important sources of evidence for development of future social work practice. Scholars however feel that much of research based evidence is used by practitioners and that certain specific conditions, which have been elaborated in the last section, need to be met for the improvement and application of evidence based practice. Word Count: 2530, apart from bibliography

frost bite :: essays research papers

  Ã‚  Ã‚  Ã‚  Ã‚  Frostbite occurs when skin tissue and blood vessels are damaged from exposure to temperatures below 32 degrees fahrenheit. It mostly affects the toes, fingers, earlobes, chin, cheeks and nose, body parts which are often left uncovered in cold temperatures. Frostbite can occur rapidly or gradually, depending on the temperature conditions and how long it is exposed.   Ã‚  Ã‚  Ã‚  Ã‚     Ã‚  Ã‚  Ã‚  Ã‚  Frostbite has three stages. They are frostnip, superficial frostbite, and deep frostbite. Frostnip occurs when you have this pins and needles sensation and the skin turning very white and soft. This stage has no permanent damage and may be reversed by soaking in warm water or breathing warm breath on the affected area.   Ã‚  Ã‚  Ã‚  Ã‚     Ã‚  Ã‚  Ã‚  Ã‚  Superficial frostbite is the stage of frostbite when blistering occurs. the skin feels numb, waxy, and frozen. There are ice crystals that form in the skin cells and the rest of the skin remains flexible.   Ã‚  Ã‚  Ã‚  Ã‚  Deep frostbite is the most serious stage of frostbite. The blood vessels, muscles, tendons, nerves, and bone all may be frozen. This stage leads to permanent damage, blood clots and gangrene, in severe cases. You have no feeling in the affected area and there usually isn't any blistering. Serious infections and loss of lims accur frequently when frostbite reaches this stage of its development. However, even in deep frostbite, frozen lims may be saved if medical attention is obtained as soon as possible.   Ã‚  Ã‚  Ã‚  Ã‚  If you are in a situation where a patient can't be transported to a hospital immediately, the following rewarming techniques may help until reaching an emergency facility.   Ã‚  Ã‚  Ã‚  Ã‚  - Bring them indoors as soon as possible.   Ã‚  Ã‚  Ã‚  Ã‚  - Apply warm towels or immerse the area in circulating warm water for twenty minutes. However DO NOT   Ã‚  Ã‚  Ã‚  Ã‚   rub or use hot water.   Ã‚  Ã‚  Ã‚  Ã‚  - Do not hold the area near fire since the area may be burned due to the reduced feeling in the area.   Ã‚  Ã‚  Ã‚  Ã‚  - Offer the patient warm coffee or tea, but never alcohol.   Ã‚  Ã‚  Ã‚  Ã‚  - Keep the affected area raised.   Ã‚  Ã‚  Ã‚  Ã‚  After re-warming, a superficial frostbite will reddon and become painful as circulation resumes in the area. Blisters are likely to form within 24 hours.   Ã‚  Ã‚  Ã‚  Ã‚  While a frostbite injury is healing, do the following:   Ã‚  Ã‚  Ã‚  Ã‚     Ã‚  Ã‚  Ã‚  Ã‚  - Avoid infection by leaving the blisters alone.   Ã‚  Ã‚  Ã‚  Ã‚  - Watch for signs of infection such as redness, swelling, fever, oozing pus, and red streaks on skin.   Ã‚  Ã‚  Ã‚  Ã‚  - Take all prescribed medications.   Ã‚  Ã‚  Ã‚  Ã‚  - Don't expose the affected area to cold temperatures until cleared to do so by a physician.

Eastern religion Essay

Transcendentalism denotes an abstract thought composed of several layers of meaning. The Oxford Companion to Philosophy states, â€Å"Transcendentalism is belief in the existence of things that transcend sense-experience, or more reflectively, belief in the possibility of transcendent metaphysics† (pp-878).   In the religious sense, it can be defined as the quest for reality through spiritual intuition and/or those qualities unique to the creator of all natural things (God). There are many transcendental traits of available in the Upanishads, the Vedas, the Gita and remarkable contribution of great persons like Maharishi ji, Chaitanya Mahaprabhu.   Lord Krishna reveals transcendental knowledge in Bhagwat Gita as under. Transcendental knowledge – the spiritual knowledge of the soul, of God, and of their relationship is both purifying and liberating. Such knowledge is the fruit of selfless devotional action (karma-yoga) (Bhagawat Gita, chapter 13 to 15). 1) Lord Chaitanya instructed the mass of people in the Sankhya philosophy of acintya-bhedabheda-tattva, which maintains that the Supreme Lord is simultaneously one with and different from His creation. Lord Chaitanya taught this philosophy through the chanting of the holy name of the Lord. He taught that the holy name of the Lord is the sound incarnation of the Lord and that since he Lord is the absolute whole, there is no difference between His holy name and His transcendental form. Thus by chanting the holy name of the Lord one can directly associate with the Supreme Lord by sound vibration. As one practices this sound vibration, he passes through three stages of development: the offensive stage, the clearing stage and the transcendental stage. In the offensive stage one may desire all kinds of material happiness, but in the second stage one becomes clear of all material contamination. When one is situated on the transcendental stage, he attains the most coveted position – the stage of loving God. Lord Chaitanya taught that this is the highest stage of perfection for human beings. 2) Maharishiji contributed a great Transdental meditation tequenique. The Transcendental Meditation (TM) technique is a simple, natural, effortless procedure whereby the mind easily and naturally arrives at the source of thought, the settled state of the mind — Transcendental Consciousness — pure consciousness, self-referral consciousness, which is the source of all creative processes. Transcendental meditation technique, the individual’s awareness settles down and experiences a unique state of restful alertness. As the body becomes deeply relaxed, the mind transcends all mental activity to experience the simplest form of awareness, Transcendental Consciousness, where consciousness is open to itself. This is the self-referral state of consciousness. The experience of Transcendental Consciousness develops the individual’s latent creative potential while dissolving accumulated stress and fatigue through the deep rest gained during the practice. Reference: Bhagawat Gita; Chapter 13 – 15 Gyan; Maharishi Sanwatsar-51; 2006

Free Essays on Images In William Shakespeares Macbeth

of Macbeth. Just after Macbeth murders Duncan, he begins to show his feelings of guilt and remorse when he says: With all great Neptune’s ocean wash this blood Clean from my hand? No; this my hand will rather The multitudinous seas incarnadine, Making the green one red. (2.2, 60-64) Macbeth is saying that the sea does not contain enough water to wash the blood from his hands without turning the seas themselves red with blood. He is illustrating to the viewer ... Free Essays on Images In William Shakespeares Macbeth Free Essays on Images In William Shakespeares Macbeth Imagery may be defined as a collection of mental pictures or thoughts, formed in an individual’s mind, that appeals to any of one their five senses. In the play Macbeth, written by William Shakespeare, various images are used. Macbeth is a play about an ambitious young man who goes to great lengths in order to become king of Scotland. In the play, the dominant images are of clothing, blood, animals, and sleeplessness. The use of imagery creates an effect in the readers’ minds and enhances their understanding of the play through helping to create a moral, mental, and physical atmosphere in a work. Firstly, images of blood and animals play a prominent role in establishing the moral atmosphere. In Lady Macbeth’s statements to Macbeth before the murder of Duncan, an animal image helps to convey moral atmosphere. Lady Macbeth says to her husband, â€Å"†¦look like the innocent flower, / But be the serpent under’t†. (1.5 65-66) Lady Macbeth is telling Macbeth that to escape suspicion he needs to act innocent, while acting like the devilish fiend to accomplish his murderous goals. The serpent has often been used as a representation of evil or a representative of the devil. Thus, although Lady Macbeth is arguing to Macbeth that the murder of Duncan is the righteous thing to do, her use of the image of the serpent suggests that the murder of Duncan is wrong. Another image that particularly helps to establish moral atmosphere is found in the words of Macbeth. Just after Macbeth murders Duncan, he begins to show his feelings of guilt and remorse when h e says: With all great Neptune’s ocean wash this blood Clean from my hand? No; this my hand will rather The multitudinous seas incarnadine, Making the green one red. (2.2, 60-64) Macbeth is saying that the sea does not contain enough water to wash the blood from his hands without turning the seas themselves red with blood. He is illustrating to the viewer ...

Overview of Minimum Wage in Canada

Overview of Minimum Wage in Canada When Canadas federal minimum wage laws governing all 10 provinces and three territories were eliminated in 1996, the minimum hourly wage rates for experienced adult workers were set by the provinces and territories themselves. These minimum wage rates have periodically changed, and the new minimum wage laws usually take effect in either April or October.   Exceptions to Canadas Minimum Wage Some circumstances circumvent the general minimum wage, applying different minimums to some workers. In Nova Scotia, for example, employers can pay an inexperienced minimum wage to workers for the first three months of employment if they have less than three months prior experience in a field; that wage is 50 cents lower than the general minimum wage. Similarly, in Ontario, the minimum wage for students is 70 cents less than the general minimum wage. Different work situations affect the minimum wage in some provinces, too. In Quebec, the minimum wage for all workers who receive tips is $9.45, which is $1.80 less than the minimum wage of general workers, and the minimum wage for liquor servers in British Columbia is $9.60, more than $1 lower than the general minimum wage. Manitoba has separate minimum wages for security guards ($13.40 per hour in October 2017) and construction workers, whose pay depends on the type of work and experience. Liquor servers in Ontario earn $1.50 less than the minimum wage but home workers earn $1.20 more. Minimum Weekly and Monthly Wages Not all occupations are covered by the general hourly minimum wage. Alberta, for example, passed a three-stage wage increase for sales workers, from $486 per week in 2016 to $542 per week in 2017 and $598 per week in 2018. The province did the same with live-in domestic workers, raising the 2016  wage from $2,316 per month to $2,582 per month in 2017, and to $2,848 per month in 2018. Examples of Minimum Wage Increases in Canada Most provinces have periodically revised minimum wage rates since Canadas federal mandates were eliminated. For example, in 2017 Saskatchewan tied its minimum wage to the Consumer Price Index, which adjusts for the costs of goods and services, and plans to announce on June 30 each year any change to the minimum wage, which will then take effect on Oct. 1 of the same year. In the first fiscal year of this plan, the 2016 minimum wage of $10.72 was raised to $10.96 in 2017. Other local governments have scheduled similar increases based on other criteria. Alberta scheduled its $12.20 rate to rise to $13.60 on Oct. 1, 2017, the same date Manitoba ($11 to $11.15), Newfoundland ($10.75 to $11) and Ontario ($11.40 to $11.60) scheduled minimum wage rate hikes. Province General Wage More Employment Standards Alberta $13.60 Alberta Human Services BC $10.85 B.C. Ministry of Jobs, Tourism and Skills Training Manitoba $11.15 Manitoba Family Services and Labour New Brunswick $11.00 New Brunswick Employment Standards Newfoundland $11.00 Labour Relations Agency NWT $12.50 Education, Culture and Employment Nova Scotia $10.85 Labour and Advanced Education Nunavut $13.00 Ontario $11.60 Ministry of Labour PEI $11.25 Environment, Labour and Justice Quebec $11.25 Commission des normes du travail Saskatchewan $10.96 Saskatchewan Labour Standards Yukon $11.32 Employment Standards

Challenges faced (Cultural, perceptional and religious perspective ) Research Paper

Challenges faced (Cultural, perceptional and religious perspective ) and acceptance of Islamic Finance in western (Non Islamic C - Research Paper Example Also discussed is the concern over Islamic finance allegedly supporting terrorism, and why some western countries equate Islamic finance with supporting terrorism. Some implications are highlighted and recommendations are then made based on the research as to how to deal with such issues and overcome the barriers to making Islamic finance more acceptable in Western countries. Although most Islamic banks are concentrated in Muslim countries, they are also to be found in many non-Muslim countries, especially in Europe and the U.S.). In addition, some conventional banks have also begun to offer Islamic financing schemes such as the HSBC Amanah division of HSBC Group established in 1998. Also, although several studies have been conducted on attitudes towards Islamic banking and the patronization of Islamic banks with reference to IFIs located in Muslim countries, some studies, albeit very few have also been conducted to gather the views and preferences of Western customers. A selection o f these IFIs located in Western countries is also the focus of attention and the few studies referred to above are mentioned. General perceptions of Islamic finance It proved to be difficult to ascertain the perceptions of Westerners towards Islamic finance due to a lack of studies in this area. Most studies have examined customers from Muslim and other developing countries. To give an example of one significant study, Erol & El-Bdour (1989) studied attitudes towards Islamic banking in Jordan. They used a nine-part question/statement instrument and showed that religious motivation was not such an important factor as a fast and efficient service, reputation and image, and confidentiality. Nonetheless, a general awareness of Islamic banks and their methods was evident. Sudin et al. (1994) conducted a more extensive study among both Muslims and non-Muslims in Malaysia. The three most important criteria for non-Muslims were firstly, friendliness of staff, secondly a fast and efficient s ervice, and thirdly the bank’s image and reputation. Another study on Malaysian customers showed that although most of them did not have a complete understanding of Islamic financial products, they did not differentiate between products from Islamic and conventional banks (Hamid & Nordin, 2001). In another study, Gerrard & Cunningham (1997) surveyed the attitude towards Islamic banking among Singaporeans where Muslims are in a minority. It was found that non-Muslims were generally lacking in awareness of Islamic banking. Furthermore, whereas Muslims were mainly motivated by religious reasons besides profitability, and had little interest in getting a high interest rate on savings, it was the opposite situation for non-Muslims. It is a similar situation in Turkey (Okumus, 2005). Even in non-Muslim countries like India where Muslims form a significant proportion of the country’s population, awareness of IFIs was low at the turn of the present century (Munawar & Llewellyn , 2002: 188). Less than half of the 720 persons interviewed knew that they even existed. This general finding of non-Muslims being more motivated by reasons other than religious ones could be the case in Western countries as well among non-Muslims that do use Islamic finance. However, during the past decade there has probably been an increasing awareness

King Arthur Essay Example | Topics and Well Written Essays - 500 words

King Arthur - Essay Example Geoffrey also translated an ancient book titled â€Å"History of the Kings of Britian†, which was most likely highly elaborated upon in his hands. This was the first work to cover the life of King Arthur in much detail. It was taken as truth until around the 17th century. Modern historians trace much of the content of Geoffreys â€Å"History† to Celtic mythology and other Breton writings, as well as some historical works tying the content back to actual events of the time period. â€Å"Life of Merlin†, another of Geoffreys writings, was both written and placed into the timeline after â€Å"History of the Kings of Britian†. However, since Merlin appeared in the original â€Å"History† as well, his role was made more mythical by extending his lifespan to an impossible degree. Geoffrey did this in order to make the events in â€Å"Life of Merlin† and â€Å"History† agree with each other, even though it is most likely that the Merlin from â€Å"History† and the Merlin in â€Å"Life of Merlin† were two different people. As a reward for his work, Geoffrey was first named Bishop of St. Asaphs and then Archbishop Theobald. However, he was unable to fill this role well due to the Welsh revolution that was taking place. In addition, he died shortly after being named Archbishop, and was never really able to enjoy being elected to the position. I selected this reading because it is more interesting for me to learn about real historical figures and the background of the stories than it is to study the life of characters that never existed. Geoffrey of Monmouth is the man responsible for the popularity of the Arthur stories we know and love today; this alone makes him worthy of further study. This reading attempts to be as historically accurate as is possible. There are probably errors, given the scarcity of records that remain about Geoffrey of

Annotated list of resources Bibliography Example | Topics and Well Written Essays - 750 words

List of resources - Annotated Bibliography Example I think that it would be a reliable source for parents or educators seeking such programs. Raz-Kids is, essentially, an online library and reading program all in one, intended for young readers in the range of K through 6. They offer to improve children’s reading through reading for practice, recording the student reading, listening for modeled fluency, and checking comprehension through quizzes assigned to them. Children are encouraged to read, â€Å"†¦The right books for the right students.† ("Raz-kids.com: Online leveled," 2012) This can give the instructors the opportunity to properly track progress. By having the opportunity to have the library of the books in the â€Å"level† with a subscription, children can log in and read from anywhere that they have internet access. This side is adequate in its presentation. It is intended to be kid-friendly with bright colors, but internal access beyond the overview and sample pages is denied without opening an account with the site. This, in my opinion, makes this resource, by no means less credible, but less likely my first choice when reviewing such sites. So with no disrespect intended it is just less essay to give a truly genuine opinion of the overall potential of this particular program The San Diego Zoo website offers a section dedicated to kids, of all ages, that have games, activities, and articles that designed for the young. However, they do not offer a reading program per say, but they have â€Å"kits† available that educators can purchase filled with interactive, hands on activities, as well as, reading and science activities. The overall intention is to teach children those vital skills but, also, inspire children toward science and conservation. The lessons plans provided include skills tests for each section in the form of language arts, arts, and early science skills.("San

Song of Myself Essay Example for Free

Song of Myself Essay ‘Song of Myself’ is one of the most representative poems of Walt Whitman. It reveals what Walt Whitman is and what he stands for. In this poem, the poet discovers himself and gives the boldest expression to his true and ever expanding poetic-self defying all the limitations. In fact, it is a voyage of the self into the realms of imagination in an attempt to find its true nature. The poem celebrates his joy of finding his limitless and unbounded self identifying with the universal spirit. It is a journey from the individual self to the universal self. The poet expresses himself joyfully with the ‘original energy of nature which is unchecked and in exhaustive. The opening lines prepare the readers for an unprecedented flight of imagination. â€Å"He says, I CELEBRATE myself, and sing myself, And what I assume you shall assume, For every atom belonging to me as good belongs to you. † (Section-1) Whitman directly expresses the universality of the self and indirectly suggests the equality of all. He expresses the true nature of the self. He feels it is common to all and everyone has the same self. Everyone in the world has an equal claim on this world. He identifies his physical self and makes a distinction between the physical self of the poet and universal self of the poet. In the third stanza he says, †¦. form’d from this soil, this air, Born here of parents born here from parents the same, and their parents the same, I, now thirty-seven years old in perfect health begin, Hoping to cease not till death. (Section-1) The identity of the poet is revealed here. He was born to his parents and he was thirty seven years old enjoying robust health. These lines throw light on the personal life of the author. With this identity he starts his journey into the realms of freedom and equality where he finds himself perfectly in tune with the universe. He enjoys himself in his physical self and feels contented. He has neither worries nor any anxieties. He does not consider this life as preparation for the next. He is absolutely happy about his life and happy about what he is. The mention of soil and air clearly reveal that he is conscious of the world where he has come from. The poet is very happy with this beautiful world. The fresh air breathes new spirit in him. He identifies himself with nature and he wants to be as close to nature. He does not like anything to come in between himself and nature. He wants to be honest with nature. Here, we find the poet equating nakedness with honesty. He says, I will go to the bank by the wood and become undisguised and naked, I am mad for it to be in contact with me. (Section-2) He rejoices himself being close to nature. The proximity with nature gives him vitality and strength. He totally identifies himself with the soil, water and air. He says, My respiration and inspiration, the beating of my heart, the passing of blood and air through my lungs, The sniff of green leaves and dry leaves, and of the shore. (Section-2) The poet celebrates himself for being a part of such wonderful nature. He is neither worried nor concerned about any thing that may happen in future. He is very much concerned with the present. He is down to the earth practical, and at the same time, he expresses the wisdom of living in the present instead of thinking about future and worrying about the past. He expresses his happiness saying, â€Å"I am satisfied—I see, dance, laugh, and sing. † He has no complaints about the present and his presence in this world. He says he is not blind to the problems and evil in this world. But he does not allow them to spoil his happiness and his celebration. His reference to â€Å"myself† refers to his soul. It is the essence of his personality. He identifies the soul of the speaker is the soul in everyone. It is the universal self. It is quite untouched by the sufferings of the world. In fact, Whitman identifies the speaker of the poem with the birth of his poetic self. The newly born poet finds himself joyful and happy. The poet mentions that the spirit of the poet is a brother of god. He says, â€Å"And I know that the hand of God is the promise of my own, And I know that the spirit of God is the brother of my own, And that all the men ever born are also my brothers, and the women and my sisters and lovers, And that a kelson of the creation is love. † (Section-5) Whitman says the new self of the poet identifies itself with everything in nature. It identifies itself with a blade of grass. He feels the grass represents the same what a man represents. The awakened self of the poet strikes a common cord with the creative forces of the world. He says: A child said What is the grass? fetching it to me with full hands, How could I answer the child? I do not know what it is anymore than he. The grass stands as symbol for equality. It also represents the creative spirit in the world. He then goes on listing the things he has seen in American life, the joys, the worries, the celebration of the human race and celebration of everyday life. He describes the people he has met on his travels. He accepts them all without any complaints against them. Here, the poet shows his maturity of understanding and his modesty in accepting everything that life offers him. He does not question anything nor suspect any one. He describes himself as a lover of life. He says in the section 13 of the poem, he is a caresser of life wherever moving, backward as well as forward sluing. Absorbing all to myself and for this song. (Section-13) He strikes a beautiful comparison between his nature and the grass. He says: This is the grass that grows wherever the land is and the water is, this the common air that bathes the globe. As the grass grows every where he also respects everything and everyone in this world. He says, I play not marches for accepted victors only, I play marches for conquerd and slain persons, and I will not have a single person slighted or left away. (Section-18) Thus, the speaker in the poem, the new born poet shows a compassionate spirit and rejoices himself. The intensity in the search for the true meaning increases as the poem progresses. He asks in a philosophical tone, â€Å"Who goes there? Hankering, gross, mystical, nude: How is it I extract strength from the beef I eat? What is a man anyhow? What am I? What are you? † He is not dazzled by these questions nor withdraws himself to silence. He expresses himself and answers the questions he has raised. He says, In all people I see myself, none more and not one a barley-corn less, And the good or bad I say of myself I say of them. (Section-20) He also adds saying, I am the poet of the Body and I am the poet of the Soul, the pleasures of heaven are with me and pains of hell are with me. (Section-21) He rightly expresses that his poetry is a combination of both body and soul. There is something for the soul and there is something for the body. He mixes these things as naturally as a forest mixes different trees. His poetry is not like a cultivated garden. It is like a jungle. It is full of nature’s bounty. It is full of variety and it is vast. Just like the poem, the poet’s self grows to be the universal self and identifies itself with multitude of people and the creation in the world. The poet’s self grows beyond any limits and reaches the borders of vast expanses. The true self of the poet is no different from the vast and expanding world of his poetry. He grows as a man; his self grows to be a universal self and his soul ripens with wisdom and understanding, having made the entire tumultuous journey. He makes it clear that his poetic voice and his realization of his true self are not two different things. They are like the twins. He says, â€Å"My voice goes after what my eyes cannot reach, With the twirl of my tongue I encompass world and volumes of worlds. Speech is the twin of my vision, it is unequal to measure itself, It provokes me forever, it says sarcastically, Walt you contain enough, why dont you let it out then? † (section-25) The poet’s identity of his voice with his poetic vision is a major leap in the journey. He realizes that the two senses reveal the same reality that he finds. He goes on exploring further and he says, I believe a leaf of grass is no less than the journey work of the stars. (Section-31) Here, he realizes the some living force behind all the things in the world which makes them equal. He feels he is perfectly at home with everything in the world. He has no complaints nor any plans for improvement. He enjoys the sight of animals which are placid and self-contained. They are in absolute peace. He wants to live with them in that condition. He spends his time looking at them for a long time. He denounces the worries and frustrations of people as meaningless. The poet realizes the uniqueness of this realization and treats it with all the specialty and respect it commands. In the section 39 he begins speaking about himself in the third person. It is an indirect reference to what he has realized in the course of his journey. In that state of mind, wherever the poet goes he is respected and admired. He becomes the most liked person. He says, Wherever he goes men and women accept and desire him, They desire he should like them, touch them, speak to them, stay with them. He assumes the role of a guide and wants to lead the people with his realization. He says, â€Å"I launch all men and women forward with me into the Unknown. † Thus, the poet finds himself in a new role after the journey. He feels the realization he has, made him unique and this experience will be useful to the people who need guidance amid their turbulent lives.

Sperm Assessment Using Flow Cytometry

Sperm Assessment Using Flow Cytometry State of the art in sperm assessment using flow cytometry Abstract Flow cytometry is emerging as an important tool in the field of modern andrology for routine analysis of spermatozoa. Recently, application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, semen sample analysis was routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. The last decade, several new flow cytometric techniques have been introduced for farm animal semen assessment that enable a more detailed evaluation of several sperm characteristics. Here in this paper, an initiative has been taken to focus on a number of recent flow cytometry developments important for addressing questions in andrological tests. After the invention of flow cytometry, sperm evaluation by traditional microscopic means became questioned due to the robust advantages of flow cytometry over the microscopic method. Due to the recent development of large number of fluroscence probes, flow cytometry is now capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gamets based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the possibility of working with small sample sizes, increases the repeatability of assessment, removes the subjectivity of assessment and allows simultaneous assessment of multiple fluorochromes. Flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of experiments that a re not possible with any other technique. Nowadays, semen evaluation using laboratory assays is extremely important to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen parameters that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Semen evaluation is the single most important laboratory test that has helped us to identify clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Methods are available that can sometimes estimate the potential fertilizing capacity of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005). The methods routinely used for evaluation of the quality of a semen sample involved an evaluation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm concentration, viability, morphology and motility. Most of these techniques are microscopic analyses that only measure a small number of spermatozoa within a population, are time-consuming, can be subjective and generally measure sperm attributes individually. Recently, limitations of semen evaluation methodology have been brought into sharp focus by controversies raised in the epidemiological literature. It should also be noted that such conventional measurements are prone to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed se men, and IVF. Such handling implies semen extension, fluorophore loading, ultraviolet and laser illumination, high-speed sorting, cooling and cryopreservation, procedures that impose different degrees of change in sperm function following damage to sperm membranes, organelles or the DNA. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed that buck of these procedures are incomplete, time consuming and laborious. Flow cytometry in different technical applications offers many advantages for the analysis of sperm quality. Flow cytometry allows the simultaneous measurement of multiple fluorescences and light scatter induced by illumination of single cell or microscopic particles in suspension, as they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers can acquire data on several subpopulations within a sample in a few minutes, making it ideal for assessment of heterogenous populations in semen sample. Initially developed in the 1960s, flow cytometry made automated separation of cells based on the unique recognition of cellular patterns within a population feasible (Hulett et al., 1969). Using such a separation approach, cellular patterns can be identified by as sessing, in individual cells within a population, protein expression using fluorescently labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). Flow cytometry was first developed for medical and clinical applications such as haematology and oncology. These areas still account for the vast majority of publications on this technique, but during the past few years it has been used in other areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. FCM is increasingly used for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream. In other word, a flow cytometer will be defined as ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are made while the cells or particles pass, preferably in single file, through the measuring apparatus in a fluid stream. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other analytical tools, where a single value for each parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives objective and accurate results (Bunthof et al., 2001; Shleeva et al., 2002), overcoming the problems with the manual methods described above. Function and types of flow cytometry Fluidics, optics and electronics are the three main systems that make up a flow cytometer. In a few minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was used for measuring their DNA content (Evenson et al., 1980) and its application to semen analysis has gradually increased over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. New fluorescent stains and techniques are continuously being developed that have potential application to the flow cytometric evaluation of spermatozoa. Flow cytometry permits the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters. Sorters have the ability not only to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function During the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective nature of the manual method commonly used for semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with manual techniques. Because of time and cost restraints, most laboratories analyze only 50 to 100 sperm to compute the percentage of each cell population and the viability rate. This small sample from a population of millions probably results in a statistical sampling error (Russel and Curtis, 1993). The conventional methods used are limited to microscopic determination of sperm concentration using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and morphology (Keel et al., 2002). These methods usually involve a subjective asses sment of a few hundred sperm, and quality assurance is rarely implemented in the laboratories performing such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in terms of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been used on human sperm to determine a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method. While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the expense and difficulties of operation associated with the requirement of a skilled operator. In addition, a flow cytometer is quite large and cannot withstand shocks associated with movement, meaning it requires a dedicated position in the laboratory. However, the development of more affordable ‘‘bench-top flow cytometers has recently increased the potential application to semen analysis. If we consider flow cytometric analysis further, we can see that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. Comparing these assays to the more widely used epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of fertilization with acrosome reactivity potential (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). Using this information, the World Health Organization stated that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further data—for example, from Tomlinson and colleagues—showed that 2 proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower spe rm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, numerous reports document unacceptable discrepancies between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Several reports emphasize the need for improvement in overall quality of semen testing within and between laboratories (Neuwinger et al., 1990; Jorgensen et al., 1997; Keel et al., 2000). However, the subjective nature of conventional semen analyses, combined with their relatively low precision due to the low number of cells assessed, leads to poor intra- and interlaboratory reproducibility; therefore, the introduction of standardized or quality controlled procedures will probably have a limited effect. The conventional analyses are used to determine whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only unclear cut-off values when used for the prediction of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems found in conventional semen analysis. In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to determine the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to advise a patient about his fertility and the prognosis for the treatment of the couple. To use established cut-off values and ensure uniform diagnosis, within and between laboratory variations should be determined and followed closely. Accurate determination of sperm cell concentration is critical to the AI industry because it provides assurance both to bull studs and to customers that straws of extended semen contain the sperm numbers indicated. An accurate measure of sperm concentration is particularly important in export markets in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly accurate; because of inherent errors in the technique, Freund and Carol (13) found that mean differences of 20% were not uncommon between duplicate sperm count determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19). Currently, the primary method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration. New, more accurate methods for sperm count determinations are being sought to replace the older ones. Some laboratories are trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. It may be argued that when comparing fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence rather than fluorescence intensity, i.e., the flow cytometer is not capable of discriminating sperm which have a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Viability The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 1997), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit (Molecular Probes Inc., OR, USA). When used in combination, the nuclei of living sperm fluoresce green (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration [11]. Due to its compact size and its relatively inexpensive purchase price, this instrument could be useful for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and found a negative correlation between the percentage of damaged cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified: live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an effective indicator of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not find a relationship between bull sperm viability detected by CFDA/PI and fertility. Flow cytometry for assessment of sperm viability appears to be a valuable tool for the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating betwee n the base pairs (Krishan, 1975). Viability stains have also been used in association with fluorescently labeled plant lectins to simultaneously assess the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of flushed porcine embryos in culture. Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. An intact plasma membrane will prevent these products from entering the spermatozoa and staining the nucleus. Commonly used examples include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility. Changes in sperm membrane permeability Detection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993; Ormerod et al., 1993; Castaneda and Kinne, 2000; Reber et al., 2002). Sperm plasma membrane status is of utmost importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stability of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose purpose is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen during sperm capacitation, thus shorten ing their lifetime (Perez et al., 1996; Cormier et al., 1997; Maxwell and Johnson, 1997; Green and Watson, 2000; Schembri et al., 2000; Watson, 2000). During the freezing process, cells shrink again when cooling rates are slow enough to prevent intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. Though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity. Classical combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson RodrÄ ±Ã‚ ´guez-MartÄ ±Ã‚ ´nez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a boar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to track changes in sperm membrane permeability, has been developed recently by our group (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only after destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al. , Sperm Assessment Using Flow Cytometry Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as an important tool in the field of modern andrology for routine analysis of spermatozoa. Recently, application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, semen sample analysis was routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. The last decade, several new flow cytometric techniques have been introduced for farm animal semen assessment that enable a more detailed evaluation of several sperm characteristics. Here in this paper, an initiative has been taken to focus on a number of recent flow cytometry developments important for addressing questions in andrological tests. After the invention of flow cytometry, sperm evaluation by traditional microscopic means became questioned due to the robust advantages of flow cytometry over the microscopic method. Due to the recent development of large number of fluroscence probes, flow cytometry is now capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gamets based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the possibility of working with small sample sizes, increases the repeatability of assessment, removes the subjectivity of assessment and allows simultaneous assessment of multiple fluorochromes. Flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of experiments that a re not possible with any other technique. Nowadays, semen evaluation using laboratory assays is extremely important to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen parameters that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Semen evaluation is the single most important laboratory test that has helped us to identify clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Methods are available that can sometimes estimate the potential fertilizing capacity of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005). The methods routinely used for evaluation of the quality of a semen sample involved an evaluation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm concentration, viability, morphology and motility. Most of these techniques are microscopic analyses that only measure a small number of spermatozoa within a population, are time-consuming, can be subjective and generally measure sperm attributes individually. Recently, limitations of semen evaluation methodology have been brought into sharp focus by controversies raised in the epidemiological literature. It should also be noted that such conventional measurements are prone to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed se men, and IVF. Such handling implies semen extension, fluorophore loading, ultraviolet and laser illumination, high-speed sorting, cooling and cryopreservation, procedures that impose different degrees of change in sperm function following damage to sperm membranes, organelles or the DNA. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed that buck of these procedures are incomplete, time consuming and laborious. Flow cytometry in different technical applications offers many advantages for the analysis of sperm quality. Flow cytometry allows the simultaneous measurement of multiple fluorescences and light scatter induced by illumination of single cell or microscopic particles in suspension, as they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers can acquire data on several subpopulations within a sample in a few minutes, making it ideal for assessment of heterogenous populations in semen sample. Initially developed in the 1960s, flow cytometry made automated separation of cells based on the unique recognition of cellular patterns within a population feasible (Hulett et al., 1969). Using such a separation approach, cellular patterns can be identified by as sessing, in individual cells within a population, protein expression using fluorescently labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). Flow cytometry was first developed for medical and clinical applications such as haematology and oncology. These areas still account for the vast majority of publications on this technique, but during the past few years it has been used in other areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. FCM is increasingly used for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream. In other word, a flow cytometer will be defined as ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are made while the cells or particles pass, preferably in single file, through the measuring apparatus in a fluid stream. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other analytical tools, where a single value for each parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives objective and accurate results (Bunthof et al., 2001; Shleeva et al., 2002), overcoming the problems with the manual methods described above. Function and types of flow cytometry Fluidics, optics and electronics are the three main systems that make up a flow cytometer. In a few minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was used for measuring their DNA content (Evenson et al., 1980) and its application to semen analysis has gradually increased over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. New fluorescent stains and techniques are continuously being developed that have potential application to the flow cytometric evaluation of spermatozoa. Flow cytometry permits the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters. Sorters have the ability not only to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function During the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective nature of the manual method commonly used for semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with manual techniques. Because of time and cost restraints, most laboratories analyze only 50 to 100 sperm to compute the percentage of each cell population and the viability rate. This small sample from a population of millions probably results in a statistical sampling error (Russel and Curtis, 1993). The conventional methods used are limited to microscopic determination of sperm concentration using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and morphology (Keel et al., 2002). These methods usually involve a subjective asses sment of a few hundred sperm, and quality assurance is rarely implemented in the laboratories performing such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in terms of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been used on human sperm to determine a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method. While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the expense and difficulties of operation associated with the requirement of a skilled operator. In addition, a flow cytometer is quite large and cannot withstand shocks associated with movement, meaning it requires a dedicated position in the laboratory. However, the development of more affordable ‘‘bench-top flow cytometers has recently increased the potential application to semen analysis. If we consider flow cytometric analysis further, we can see that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. Comparing these assays to the more widely used epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of fertilization with acrosome reactivity potential (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). Using this information, the World Health Organization stated that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further data—for example, from Tomlinson and colleagues—showed that 2 proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower spe rm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, numerous reports document unacceptable discrepancies between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Several reports emphasize the need for improvement in overall quality of semen testing within and between laboratories (Neuwinger et al., 1990; Jorgensen et al., 1997; Keel et al., 2000). However, the subjective nature of conventional semen analyses, combined with their relatively low precision due to the low number of cells assessed, leads to poor intra- and interlaboratory reproducibility; therefore, the introduction of standardized or quality controlled procedures will probably have a limited effect. The conventional analyses are used to determine whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only unclear cut-off values when used for the prediction of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems found in conventional semen analysis. In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to determine the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to advise a patient about his fertility and the prognosis for the treatment of the couple. To use established cut-off values and ensure uniform diagnosis, within and between laboratory variations should be determined and followed closely. Accurate determination of sperm cell concentration is critical to the AI industry because it provides assurance both to bull studs and to customers that straws of extended semen contain the sperm numbers indicated. An accurate measure of sperm concentration is particularly important in export markets in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly accurate; because of inherent errors in the technique, Freund and Carol (13) found that mean differences of 20% were not uncommon between duplicate sperm count determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19). Currently, the primary method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration. New, more accurate methods for sperm count determinations are being sought to replace the older ones. Some laboratories are trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. It may be argued that when comparing fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence rather than fluorescence intensity, i.e., the flow cytometer is not capable of discriminating sperm which have a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Viability The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 1997), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit (Molecular Probes Inc., OR, USA). When used in combination, the nuclei of living sperm fluoresce green (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration [11]. Due to its compact size and its relatively inexpensive purchase price, this instrument could be useful for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and found a negative correlation between the percentage of damaged cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified: live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an effective indicator of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not find a relationship between bull sperm viability detected by CFDA/PI and fertility. Flow cytometry for assessment of sperm viability appears to be a valuable tool for the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating betwee n the base pairs (Krishan, 1975). Viability stains have also been used in association with fluorescently labeled plant lectins to simultaneously assess the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of flushed porcine embryos in culture. Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. An intact plasma membrane will prevent these products from entering the spermatozoa and staining the nucleus. Commonly used examples include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility. Changes in sperm membrane permeability Detection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993; Ormerod et al., 1993; Castaneda and Kinne, 2000; Reber et al., 2002). Sperm plasma membrane status is of utmost importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stability of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose purpose is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen during sperm capacitation, thus shorten ing their lifetime (Perez et al., 1996; Cormier et al., 1997; Maxwell and Johnson, 1997; Green and Watson, 2000; Schembri et al., 2000; Watson, 2000). During the freezing process, cells shrink again when cooling rates are slow enough to prevent intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. Though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity. Classical combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson RodrÄ ±Ã‚ ´guez-MartÄ ±Ã‚ ´nez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a boar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to track changes in sperm membrane permeability, has been developed recently by our group (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only after destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al. , 2006), it seems plausible that this is a result of the silencing of a multidrug transporter. This m