attitude, really, and one ' s attitude in choosing oneself must be sincere, for as one believes, so one is. what counts, says kierkegaard, is the energetic sincerity of choosing the real either / or over the mere either / or, such as choosing to visit the bank or barber. the real choice is ultimately between good / evil, good being the real self. kierkegaard admits that many other worthwhile decisions may be made along the way to finding / receiving one ' s self. we hear the familiar song hand down from ancient times : the treasure is within, in the inner unity where man is reconciled with himself if not his collective self projected as god. kierkegaard criticizes critics for not taking up the inner life, for losing themselves in illusions, professions, callings and other forms of escape. yet his own approach is also an escape. indeed, life is an escapade! are we to deride the philosopher who said, “ as a matter of fact, i am not slightly interested in self - spelunking, given all the objects the universe has to offer. ” " you are capable of spending a whole month reading nothing but fairy tales - kierkegaard writes in his either / or novel - you make a profound study of them, you compare and analyze, and your study is not barren of result - but what do you use it for? to divert your mind ; you let the whole thing fire off in a brilliant display of fireworks. " kierkegaard opines that philosophy mediates the past but cannot mediate what has not occurred yet : it cannot choose the future. and philosophers would have no past to mediate unless there were an absolute either / or. one obviously has to do something besides philosophize to get something done. life would come to a dead stop with philosophy alone. " we have the disgusting sight of young men who are able to mediate christianity and paganism, are able to play with the titanic forces of history, and are unable to tell a plain man what he has to do in life.... " that is true. we must not entirely abandon self - spelunking – and we study others to discover our own nature. nevertheless we find in kierkegaard ’ s philosophy the usual christian activism without specification of particular deeds. kierkegaard ’ s philosophical protagonist offers nothing in particular except marriage and family to the hypothetical young man addressed in the book - long letter constituting the novel either / or


. he is an i or individual, a category of one at dodge city, facing down the multiplicitous world, a world rendered as multiple by the psychological interpretation of diverse inchoate sensations that the mind is delighted to organize and represent as a reflection of its microcosmic god, the individual. v. the russian philosopher of freedom nicolai aleksandrovich berdyaev, the “ russian philosopher of freedom ”, is mistakenly labeled an existentialist because he emphasized anthropocentric subjectivity in his theory of " objectification ". his nods to dostoyevsky, nietzsche, jaspers, and other non - systematic philosophers also served to place him in the existentialist camp. berdyaev, nonetheless, was more of a personalist than an existentialist. according to berdyaev, personalism idealizes the human person. there is no such thing as a national person. that is a self - conceited lie, stupid and ludicrous. the nation is the projection of everything evil as a collective ideal. evil is called good. hate, violence, egoism, pride, will to power are all converted into virtues. do not idolize nation or volk. do not be a slave to " the people. " real personality has an existential center - of conscience. real personalities are individual, and therefore volk will crucify them. egocentricity and personalism are contrary concepts. " egocentricity is the original sin of man... an illusory, distorted universalism... a false perspective... under the power of objectivization, which it seeks to turn into an instrument of self - affirmation... man is the salve of the surrounding external world, because he is the slave of him.... " of course, " the genius lives near to primary reality and to real existence whereas the culture elite is subject to the laws of objectivization and socialization. " " all possession, whether it is possession by base passion or by lofty ideas denotes the loss of the spiritual center in man. " the very notion of the soul as a unity derived from physical processes is false. the unity of the soul - process is the dynamic spiritual principle itself and not arbitrarily conceived points or ' souls. ' the notion that the individual of individualism constitutes resistance to its environment and is thus constituted by isolated exercises of its own will is an illusion. such an individual would have to be exterior


itself and not arbitrarily conceived points or ' souls. ' the notion that the individual of individualism constitutes resistance to its environment and is thus constituted by isolated exercises of its own will is an illusion. such an individual would have to be exterior to the world in order to rebel against it, hence the individual would have to be alienated, impersonal, and self - determined or self - caused, as if it were a god facing a violent enemy. the enmity projected onto a seemingly violent world would define the supposedly self - determining individual who blames the objective world for everything perceived to be in opposition ; thus is he in fact determined by objectivity rather than his presumed subjectivity. as ' subject ' in distinction from ' object ', he thinks he is a free individual, but the emperor has his foot on his neck. since humankind is natural and human beings are a social species, the individual of individualism has dissociated himself from nature and society. for him, nature is dead ; god is dead ; society is dead. since society comprises individuals, the person is dead as well, for the person is a social individual who enjoys not only individuality but the commune as well. ironically, the individualist believes he is free when in fact he is enslaved. today he is in reality the over socialized, bourgeois individual of the militant protestant - capitalist system. he is fully engaged in the war of all against all where each individual crushes other and is crushed in the material arena of economic forces and interests. he has internalized the anti - social rules of engagement that subjugate him. like a prisoner in bentham ' s panopticon - prison, he is constantly under observation and control by unseen wardens. but he is a virtual slave ; a slave not to his circumstances, but to himself. he is, as it were, self - imprisoned, and is like the animal that stays put when the fence is torn down. " the individualist is the slave to himself, he is under the spell of slavery to his own ego, and, therefore, he cannot resist the slavery which comes from the non - ego.... man is always a slave of the non - ego through the ego …. the object world can make a person a martyr but it cannot make him a conformist. " on the other hand, whereas individualists are " wolfish " and vicious, berdyaev ' s person of personalism is virtuously communal and fraternal. for


did “ king ” arthur really exist? the story of king arthur and his knights of the round table is one of western culture ’ s best known. many people assume that they deal with a king from the dark ages of england and that, once the clearly legendary aspects such as the magician merlin are removed, they have some value as history. this is not the consensus opinion of historians, though. many – probably the majority, in fact – are of the opinion that there was no “ king ” arthur, that the stories originate in welsh legend or mythology and that they can tell us nothing of value about the fifth and sixth centuries in britain. on the other hand, a small minority believe that there is a kernel of truth behind the stories and that it is possible to glimpse the activities of a shadowy individual known as arthur behind them. the documentary evidence if there were contemporary documents mentioning arthur, there would be no controversy about his existence ( although there might well be controversy over other aspects of the story ). there are not and it is only by assuming that he lurks under an alias that it is possible to suggest that there might be contemporary evidence. was arthur known as riothamus during his lifetime? if he was, why did later legend change the name? and why is the first source to name him beyond all shadow of doubt dated to the early ninth century? the archaeological evidence if the documents don ’ t tell us about arthur, surely there must be some archaeological evidence? after all, there was the grave found by the monks of glastonbury and there is a sixth - century site at tintagel, where an inscription confirming that people named art … existed in the sixth century was found. and wasn ’ t south cadbury proven to be camelot by leslie alcock ’ s excavation in the 1960s? surely archaeology must be of some assistance …


the bone box of james son of joseph, brother of jesus biblical archaeology seems particularly prone to fraud, especially of fraudulent artefacts that confirm the religious beliefs of the faithful. one of the greatest stumbling - blocks that christians have always experienced is that while they are proud to base their religion on books that they claim to be thoroughly historical in nature, many events of the new testament – especially those of the gospels – have no confirmation outside the bible. roman records are silent about many of the events and personages in stories familiar to all christians. forgeries purporting to confirm some of these details have been around since the first development of the religion and continue to be produced today. the “ discovery ” of the ossuary on 21 october 2002, hershel shanks, editor of the biblical archaeology review, announced at a press conference that an amazing discovery had been made in israel : a contemporary inscription confirming the existence jesus of nazareth and his brother, james the just. the ‘ find ’ was an ossuary, a stone box designed to contain the bones of a deceased individual. jewish practice in the first century ce was to lay out bodies of the wealthy in family tombs until all the flesh had decayed and subsequently to gather up the bones for storage in ossuaries. this was a typical example of the middle for the first century ce, close to the traditional date of james ’ s martyrdom in 62 ce. most ossuaries carry an inscription naming the individual inside together with a patronymic ( name of the father ) ; this example reads יעקוב בר יוםף אחוי ד ישוע ( ya ‘ aquv bar yusef akhui de yeshua ‘, “ james son of joseph brother of jesus ” ). all three names – ya ‘ aquv ( jacob or james ), yusef ( joseph ) and yeshua ( jesus ) were common enough at the time, but what is intriguing is that james is identified not just as the son of joseph, but as the brother of jesus, implying that jesus was somehow more important than joseph within the family for identifying james. this coincidence led those who first studied the ossuary to conclude that it was likely that james was indeed james “ the just ”, brother of jesus of nazareth. the box was brought to hershel shanks ’ s attention by an israeli collector of antiquities, oded golan. he claimed to have bought the box from an arab dealer in jerusalem in the 1960s and to have taken no real interest in


nazareth. the box was brought to hershel shanks ’ s attention by an israeli collector of antiquities, oded golan. he claimed to have bought the box from an arab dealer in jerusalem in the 1960s and to have taken no real interest in it until shortly before the announcement of the inscription. initial studies, including one by the geological survey of israel, concluded that the ossuary was a genuine first - century artefact, made from a limestone local to the jerusalem area and that no evidence for modern forgery could be detected. however, from the outset, there were critical voices. for a start, there are no parallels for the formula used to identify the person contained in the box. a second and more thorough analysis by the israeli antiquities ’ authority found a number of irregularities in the inscription, in the patina covering it and in the circumstances of its discovery. while there is no doubt that the ossuary is genuine, and that the first half of inscription ( ףםוי רב בוקעי – ya ‘ aquv bar yusef ) is also genuine, the epigrapher rochelle altman dismissed the second half as a “ poor forgery ”. in 2003, the israeli police arrested oded golan on suspicion of forging antiquities ; they found his home and a workshop he rented filled with forged objects in various stages of manufacture together with the equipment needed to make the objects. he eventually confessed his guilt during questioning, but denied the charges during his trial in 2004. he was, nevertheless, found guilty. despite this damning evidence against the authenticity of the inscription, hershel shanks continues to defend the ossuary, launching bitter attacks on its critics. the archaeological community, for the most part, does not accept the object as providing contemporary proof for the existence of jesus of nazareth. this story raises important questions. why was the ‘ discovery ’ announced at a press conference before academic study took place? why were the first tests of authenticity conducted by the geological survey of israel rather then the israel antiquities ’ authority? why was so much faith placed in an object without provenance? why has hershel shanks continued to support its authenticity despite the overwhelming evidence that oded golan was an accomplished forger who worked over many years? there is a lesson for us all in this debacle.


piloncillo is unrefined brown sugar from mexico that comes in a small conical shape. piloncillo is served in drinks such as cafe de olla and champurrado., and the sugar can also be used in moles and sauces. the cone of piloncillo must be shaved or grated before use. piloncillo is the name given in mexico to small blocks or bricks of unrefined solid cane sugar. they are also often seen in the shape of small truncated cones. in central america and south america they are called panela or “ tapa dulce ” ( in costa rica ) because of its bottle - cap shape, and may or may not be cone - shaped. in panama it is also called raspadura, thought to derive from the words “ raspar ” ( to scrape ) and “ duro ” ( hard ), a reference to the way the hard sugar brick is shaven to produce usable shards for cooking. the local dialect often drops the letter “ s ”, resulting in the word we hear as “ ra ’ padura ” or rapadura. the color of piloncillo ranges from light tan to dark brown. for the longest time piloncillo was considered an inferior sweetener. it was used as a cheaper substitute for refined sugar, especially in dishes and desserts that did not require the clear color of refined sugar. despite its inferior reputation, many desserts originating ( and still consumed ) in mexico are made to this day using piloncillo, such as atole, capirotada, sweet potatoes, flan, and more. piloncillo, mexican brown sugar piloncillo is an unrefined sugar from mexico. the name piloncillo refers to the traditional cone shape in which the sugar is produced. it is also know as panela and panocha. there are actually two varieties of piloncillo produced one is lighter ( blanco ) and one darker ( oscuro ). the cone size can vary from as small as 3 / 4 ounce to as much as 9 ounces per cone. the cones shown in the picture above are about 3 ” tall. how to use piloncillo is very hard compared to the brown sugar you purchased in a box at the local grocer. chop the piloncillo with a serrated knife. you can substitute piloncillo in any recipe calling for dark brown sugar. traditional uses for mexican pilonci


the brown sugar you purchased in a box at the local grocer. chop the piloncillo with a serrated knife. you can substitute piloncillo in any recipe calling for dark brown sugar. traditional uses for mexican piloncillo cafe de olla - an earthy mixture of viennese - roast coffee, cinnamon, aniseeds, and piloncillo ( mexican dark brown sugar ). champurrado - a special hot chocolate thickened with masa and flavored with piloncillo and aniseeds. ( oxford english dictionary ) u. s. regional ( chiefly south - west. ). [ < mexican spanish piloncillo ( 1743 ) < spanish pilon pilon n. + - cillo, diminutive suffix. ] a type of coarse brown sugar, often moulded into a hard cone or block, produced esp. in mexico. cf. penuche n. 1. 1844 j. gregg commerce of prairies i. 173 when short of means they often support themselves upon only a real each per day, their usual food consisting of bread and a kind of coarse cake - sugar called piloncillo. 1875 fur, fin & feather 108 you are.. all the days eating piloncillo and learning spanish with the senorites [ sic ]. dictionary of american regional english also pelonce, peloncillo, pilonce, pilonci, pilonsillo [ mexspan ; dimin of pilon a loaf of sugar ] chiefly tx unrefined sugar, usu in the form of a cone or loaf. 1844 gregg commerce 1. 173 nm, when short of means they often support themselves upon only a real each per day, their usual food consisting of bread and a kind of coarse cake - sugar called piloncillo. 1845 green jrl. texian exped. 264, our cook brings us in.. two and a half pounds of brown sugar, ‘ pilonci. ’ 1854 ( 1932 ) bell log tx - ca trail 35. 310, gathered some quinces and stewed them with pelonce or mexican sugar. 1875 fur fin & feather 108 nm, you are all the nights at fandangoes, and all the days eating piloncillo and learning spanish with the senorites [ sic ]. 1892 dn 1. 193 tx, pilon : a loaf of sugar. the usual forms in texas are pilonce amd


york, ny : d. appleton & company the sugar cane grows remarkably well in the bottom lands, and is cultivated in sufficient quantities to supply a small sugar mill. nothing, however, is made but the common pilonce, an article inferior to the most ordinary brown sugar of commerce. the tex - mex cookbook by robb walsh new york, ny : broadway books this mexican raw brown sugar is more flavorful than regular brown sugar. it is sold in a cone and must be grated or dissolved in water before use. piloncillo pecan patties piloncillo is old - fashioned, unrefined raw cane sugar. the cones were once sold in the streets by candy vendors. today you can find it in mexican markets and in the specialty aisles of grocery stores. one 8 - ounce piloncillo sugar cone ( to yield 1 cup grated piloncillo ) 2 - inch - long cinnamon stick 1 / 8 teaspoon salt 1 / 2 tablespoon unsalted butter 1 / 2 teaspoon vanilla extract 8 ounces pecan halves broken in half ( about 1 1 / 2 cups ) aluminum foil, sprayed with nonstick cooking spray (... ) what is piloncillo? posted sep 10th 2006 1 : 03pm by nicole weston piloncillo is an unrefined sugar that is commonly used in mexican cooking. the sugar has been around for at least 500 years, and was being made before the spanish came to mexico around 1500. it is made when sugar canes are crushed, the juice is collected and boiled then poured into molds, where it hardens into blocks. the fact that it comes in block form is one of the reasons why white and brown sugars are more commonly used, even in mexican cooking, than piloncillo once was. to use it, it must be grated or chiseled off the main block - a process which is well worth the resulting flavor boost in food to some, but too time consuming for others to bother. unlike white sugar, which is flat and one - dimensional in its sweet flavor, piloncillo is smoky, caramely and earthy. it has far more flavor than brown sugar, which is generally just white sugar with a small amount of molasses added back to it. it can be uses in moles and other sauces, as well as to simple sweeten coffee or top off buttery toast. texas ( lone star state dictionary ) • ( 0 ) comments


readers who find this page helpful, may also wish to visit these pages. exhibition smartawards that leave no mind behind and reward every participant while educating all of us about art and life. how to install a student show with ordinary masking tape. http : / / www. goshen. edu / ~ marvinpb / arted / tape. html exhibit design and installation seldom have enough space to show things as professionally as it should be. i like to keep exhibit design and presentation very simple and very orderly, even when it too crowded. i want the viewers to notice the artwork — not fancy exhibition design, brightly colored mats, or some distracting exhibition idea. when i see an exhibit with colored mounts, decorative additions, or bizarre arrangements, it makes me feel that there was not much respect for the artwork itself. i avoid colored additions. i have never seen an art museum that used colored mats or mounts for artworks created on paper. i want to see the artwork. i want the exhibition to be so well designed that i do not notice the exhibit design more than the artwork itself. to arrange the artwork, we begin with the largest works and gradually add smaller works to complete the design of the exhibit. sometimes works can be grouped attractively. we often lay it out on the floor to see how a group works before putting who does the work the older students or more advanced classes do the work. when students learn how to design and install the exhibit they are learning art. they are learning very useful life - long choice - making skills and i am ultimately getting lots of help. they have to make choices about color relationships, dominance and subordinance, how to move the viewer ' s attention around, and all the standard compositional questions. i remind students to be particularly careful about the negative spaces between the works. i make the negative spaces ( wall spaces, if any ) equal unless there is a good visual reason not to make them equal. they have to make choices about color relationships, dominance and subordinance, how to move the viewer ' s attention around, and all the standard compositional questions. they are asked to be as fair as possible when they choose where to place their own work and that of see this link for ideas on mounting a display of school artwork where there is no good provision for an exhibit. it illustrates a simple way to use tape to hang temporary exhibits so the work will not fall down, but can still be removed without damaging the wall after hanging a week or so. labels and information london


where there is no good provision for an exhibit. it illustrates a simple way to use tape to hang temporary exhibits so the work will not fall down, but can still be removed without damaging the wall after hanging a week or so. labels and information london, i was fortunate to see a special exhibition of original drawings from the hand of michelangelo, 1475 – 1564. often there was a photograph of the painting or sculpture that michelangelo subsequently created using the ideas he developed with drawing. much of the wall space was given to text that helped me understand the need for each of his drawings. soon after entering, an explanation on the wall said, “ drawing was an essential tool for michelangelo to explore his ideas... his creativity was stimulated by the process of drawing — the flow of ideas quickening as he realized them on paper. ” ( - - from the british museum wall, march, 2006 ) in april, 2006, i viewed an exhibit of the student work of art education students at the college of creative studies in detroit. in addition to titles, artist name, and media, each work or group of artworks was displayed together with the assignment. by sharing the concepts used in the assignment the viewer participates in the learning. our art programs get more of the support from parents, administrators, and other teachers when we help them see the learning benefits of art when we display our student ’ s artwork, students need to see their names with it, and others need to see at least the main goals of the assignment that occasioned the work. why was the work created? what purposes does it fulfill as learning for the student artist and for us the viewers? our students are not assigned the sistine chapel ceiling, the last judgment, or a larger than life marble statue of david, but our assignments shape their work just as michelangelo ’ s commissions shaped his work. as the result of including assignment information, more than one art teacher has heard comments from parents and other teachers saying things such as, “ oh, i didn ' t ’ t know they learned things in art! ” in addition to the student name, it is good to include title, media, and the name of the art class if it is high school work, or year in school if elementary or middle school. if there are several teachers, include the teacher ' s name as well. some situations require a large overall title to the exhibit. some exhibits are divided by categories, and it may be helpful to identify each category. in some schools, it is good to make some way - finding signs


the teacher ' s name as well. some situations require a large overall title to the exhibit. some exhibits are divided by categories, and it may be helpful to identify each category. in some schools, it is good to make some way - finding signs to guide the visitors to the room or place in the building where they can see the artwork. end of essay - see related links below smart art awards? giving awards has benefits and pitfalls. this page explains a way to gain motivational and education benefits from awards while avoiding the negative pitfalls ( a teaching invention seen here first ). no good place for an exhibition? how to install an exhibit using tape so it will not fall down. how to build a temporary self - supporting wall in the gym using tackable fiberboard. this space is used as a drawing room in an old gymnasium. also see shop drawings below. the walls are used to tack up drawings during critique sessions. a similar wall can be used for exhibitions. panels are available in different colors and with fabric cover for classroom use. other products are also available at your building suppliers. do not hide exit signs. 4 x 8 foot panels are attached to angle spleens with screws. links to. pdf shop drawings for tackable fiber panels to display artwork. sheet 3 drawing ( 24kb download,. pdf files ) sheet 2 ( 28kb download,. pdf file ) sheet 1 ( 40kb download,. pdf file ) teachers may make one copy of the drawings for school use. commercial or business use restricted without specific permission. caution : find a safe and spacious place to add walls. do not hide exit signs. do not restrict or block school corridors or add combustibles unless your fire marshall and insurance company approves. the author does not make any safety claims for the products illustrated on this page. check the specs of the products before using them in a school setting. panels for classroom use. more links _ of interest to art teachers, administrators, and parents all rights reserved. images, text, and design © marvin bartel 2004. parents and teachers may make one copy of this page for personal study so long as they keep the © information with the copy. permission is required to make copies, publish, or to post on another web site. mention the url or the title of this page in your correspondence with the author. you may make a link to this page from your page author info : as an art teacher, i was also gallery director for the art gallery adjacent to the


henry gray ( 18211865 ). anatomy of the human body. 1918. 6c. 2. the pelvis the pelvis, so called from its resemblance to a basin, is a bony ring, interposed between the movable vertebræ of the vertebral column which it supports, and the lower limbs upon which it rests ; it is stronger and more massively constructed than the wall of the cranial or thoracic cavities, and is composed of four bones : the two hip bones laterally and in front and the sacrum and coccyx behind. the pelvis is divided by an oblique plane passing through the prominence of the sacrum, the arcuate and pectineal lines, and the upper margin of the symphysis pubis, into the greater and the lesser pelvis. the circumference of this plane is termed the linea terminalis or pelvic brim. the greater or false pelvis ( pelvis major ). the greater pelvis is the expanded portion of the cavity situated above and in front of the pelvic brim. it is bounded on either side by the ilium ; in front it is incomplete, presenting a wide interval between the anterior borders of the ilia, which is filled up in the fresh state by the parietes of the abdomen ; behind is a deep notch on either side between the ilium and the base of the sacrum. it supports the intestines, and transmits part of their weight to the anterior wall of the abdomen. the lesser or true pelvis ( pelvis minor ). the lesser pelvis is that part of the pelvic cavity which is situated below and behind the pelvic brim. its bony walls are more complete than those of the greater pelvis. for convenience of description, it is divided into an inlet bounded by the superior circumference, and outlet bounded by the inferior circumference, and a cavity. the superior circumference. the superior circumference forms the brim of the pelvis, the included space being called the superior aperture or inlet ( apertura pelvis [ minoris ] superior ) ( fig. 238 ). it is formed laterally by the pectineal and arcuate lines, in front by the crests of the pubes, and behind by the anterior margin of the base of the sacrum and sacrovertebral angle. the superior aperture is somewhat heart - shaped,


pectineal and arcuate lines, in front by the crests of the pubes, and behind by the anterior margin of the base of the sacrum and sacrovertebral angle. the superior aperture is somewhat heart - shaped, obtusely pointed in front, diverging on either side, and encroached upon behind by the projection forward of the promontory of the sacrum. it has three principal diameters : antero - posterior, transverse, and oblique. the anteroposterior or conjugate diameter extends from the sacrovertebral angle to the symphysis pubis ; its average measurement is about 110 mm. in the female. the transverse diameter extends across the greatest width of the superior aperture, from the middle of the brim on one side to the same point on the opposite ; its average measurement is about 135 mm. in the female. the oblique diameter extends from the iliopectineal eminence of one side to the sacroiliac articulation of the opposite side ; its average measurement is about 125 mm. in the female. the cavity of the lesser pelvis is bounded in front and below by the pubic symphysis and the superior rami of the pubes ; above and behind, by the pelvic surfaces of the sacrum and coccyx, which, curving forward above and below, contract the superior and inferior apertures of the cavity ; laterally, by a broad, smooth, quadrangular area of bone, corresponding to the inner surfaces of the body and superior ramus of the ischium and that part of the ilium which is below the arcuate line. from this description it will be seen that the cavity of the lesser pelvis is a short, curved canal, considerably deeper on its posterior than on its anterior wall. it contains, in the fresh subject, the pelvic colon, rectum, bladder, and some of the organs of generation. the rectum is placed at the back of the pelvis, in the curve of the sacrum and coccyx ; the bladder is in front, behind the pubic symphysis. in the female the uterus and vagina occupy the interval between these viscera. the lower circumference. the lower circumference of the pelvis is very irregular ; the space enclosed by it is named the inferior aperture or outlet ( apertura pelvis [ minoris ] inferior ) ( fig. 239


. the lower circumference. the lower circumference of the pelvis is very irregular ; the space enclosed by it is named the inferior aperture or outlet ( apertura pelvis [ minoris ] inferior ) ( fig. 239 ), and is bounded behind by the point of the coccyx, and laterally by the ischial tuberosities. these eminences are separated by three notches : one in front, the pubic arch, formed by the convergence of the inferior rami of the ischium and pubis on either side. the other notches, one on either side, are formed by the sacrum and coccyx behind, the ischium in front, and the ilium above ; they are called the sciatic notches ; in the natural state they are converted into foramina by the sacrotuberous and sacrospinous ligaments. when the ligaments are in situ, the inferior aperture of the pelvis is lozenge - shaped, bounded, in front, by the pubic arcuate ligament and the inferior rami of the pubes and ischia ; laterally, by the ischial tuberosities ; and behind, by the sacrotuberous ligaments and the tip of the coccyx. the diameters of the outlet of the pelvis are two, antero - posterior and transverse. the antero - posterior diameter extends from the tip of the coccyx to the lower part of the pubic symphysis ; its measurement is from 90 to 115 mm. in the female. it varies with the length of the coccyx, and is capable of increase or diminution, on account of the mobility of that bone. the transverse diameter, measured between the posterior parts of the ischial tuberosities, is about 115 mm. in the female. 58 axes ( fig. 240 ). a line at right angles to the plane of the superior aperture at its center would, if prolonged, pass through the umbilicus above and the middle of the coccyx below ; the axis of the superior aperture is therefore directed downward and backward. the axis of the inferior aperture, produced upward, would touch the base of the sacrum, and is also directed downward, and slightly backward. the axis of the cavityi. e., an axis at right angles to a series of planes between those of the superior and inferior apertures is curved like the cavity itself : this curve corresponds to the con


also directed downward, and slightly backward. the axis of the cavityi. e., an axis at right angles to a series of planes between those of the superior and inferior apertures is curved like the cavity itself : this curve corresponds to the concavity of the sacrum and coccyx, the extremities being indicated by the central points of the superior and inferior apertures. a knowledge of the direction of these axes serves to explain the course of the fetus in its passage through the pelvis during parturition. position of the pelvis ( fig. 240 ). in the erect posture, the pelvis is placed obliquely with regard to the trunk : the plane of the superior aperture forms an angle of from 50° to 60°, and that of the inferior aperture one of about 15° with the horizontal plane. the pelvic surface of the symphysis pubis looks upward and backward, the concavity of the sacrum and coccyx downward and forward. the position of the pelvis in the erect posture may be indicated by holding it so that the anterior superior iliac spines and the front of the top of the symphysis pubis are in the same vertical plane. differences between the male and female pelves. the female pelvis ( fig. 242 ) is distinguished from that of the male ( fig. 241 ) by its bones being more delicate and its depth less. the whole pelvis is less massive, and its muscular impressions are slightly marked. the ilia are less sloped, and the anterior iliac spines more widely separated ; hence the greater lateral prominence of the hips. the preauricular sulcus is more commonly present and better marked. the superior aperture of the lesser pelvis is larger in the female than in the male ; it is more nearly circular, and its obliquity is greater. the cavity is shallower and wider ; the sacrum is shorter wider, and its upper part is less curved ; the obturator foramina are triangular in shape and smaller in size than in the male. the inferior aperture is larger and the coccyx more movable. the sciatic notches are wider and shallower, and the spines of the ischia project less inward. the acetabula are smaller and look more distinctly forward ( derry59 ). the ischial tuberosities and the acetabula are wider apart, and the former are more everted. the pubic symphysis is less deep, and the


new brain scanner helps paralysed people spell words a new brain scanner has been developed to help people who are completely paralysed speak by enabling them to spell words using their thoughts. it uses functional magnetic resonance imaging ( fmri ) to help patients choose between 27 characters - the alphabet and a blank space. each character produces a different pattern of blood flow in the brain, and the device interprets these patterns. the british neurological association called the research " exciting ". the study appears in current biology journal of cell press. fmri is normally used to track brain activity by measuring blood flow. earlier research the new technology is based on earlier applications of the technique, which used free - letter spelling to allow people to answer the equivalent of multiple - choice questions with just a few possible answers. british neuroscientist adrian owen, for instance, used fmri to help a man believed to have been in a vegetative state for five years to answer " yes " and " no " questions by interpreting his brain activity. but the new scanner uses the entire english alphabet and the blank space. end quote elaine snell bna [ the device could be ] a lifeline for patients with persistent vegetitative state ” " this novel spelling device constitutes an alterative approach to motor - independent communication, " bettina sorger of maastricht university in the netherlands, one of the researchers working on the current study, told the bbc. " the work of adrian owen and colleagues led me to wonder whether it might even become possible to use fmri, mental tasks, and appropriate experimental designs to freely encode thoughts, letter - by - letter, and therewith enable back - and - forth communication in the absence of motor behavior. " the team writes in the paper that because the noninvasive device requires " only little effort and pretraining, it is immediately operational and possesses high potential for clinical applications, both in terms of diagnostics and establishing short - term communication with nonresponsive and severely motor - impaired patients ". exciting results elaine snell of the british neuroscience association told the bbc that the technology could become " a lifeline " for patients in a persistent vegetative state, or suffering from other neurological disorders. " this means of communication will make a huge difference to the quality of their life and to that of their families. " this kind of technology can only get better, it ' s very exciting. " dr guy williams from the wolfson brain imaging centre in cambridge agreed. " the technique may need some adaptation


a pouch that forms in the wall of the large intestine is called a diverticulum. when this pouch becomes infected or swollen, it is called diverticulitis. copyright © nucleus medical media, inc. it is not clear why the pouches form. it may be due to a constant build up of pressure when food moves too slowly through the bowel. this pressure increases and pushes along the sidewalls of the bowel creating pouches. digested food or stool can become trapped in one of the pouches. this leads to swelling and infection. the following may contribute to diverticulitis : - low - fiber diet — fiber softens stools and makes them pass through the bowel more easily - increased pressure in the bowel from straining to pass a hard stool - defects in the colon wall the doctor will ask about your symptoms and medical history. a physical and rectal exam will be done. finding the disease early is important. the pouch can break, releasing stool into the abdomen. this is a medical emergency that requires surgery. tests may include : - your bodily fluids and waste products may be tested. this can be done with : - stool sample analysis - blood tests - images may need to be taken of your bodily structures. this can be done with : after the inflammation subsides, other tests may be performed to examine the colon, including : the goals of treatment is to : - resolve the infection and inflammation - rest the bowel - prevent complications antibiotics and other drugs are given to fight the infection. pain medications and drugs are given to decrease the abdominal pain. you may also be given medication to help control vomiting. for mild swelling, you can drink clear liquids for the first two to three days. for a more severe case, you will be admitted to the hospital, where fluids are given by iv. antibiotics will also be given to you through iv. changes in your diet can help prevent future attacks. - increase the amount of fiber you eat by eating more fruits, vegetables, and whole grains - supplement your diet with a fiber product, as advised by your doctor - avoid laxatives and enemas - avoid narcotic medications — they can slow down bowel movement and can cause constipation to remove the section of the bowel with pouches may be recommended if : - you have had multiple attacks during a two - year period - a pouch breaks and the contents spread into the abdominal cavity, which requires that the cavity be cleaned out surgery is also used to treat complications of divert


with pouches may be recommended if : - you have had multiple attacks during a two - year period - a pouch breaks and the contents spread into the abdominal cavity, which requires that the cavity be cleaned out surgery is also used to treat complications of diverticulitis, such as : - abscess — occurs if the infected pouch fills with pus - blocked bowel — scar tissue that forms and blocks movement of stool through the intestine - fistula — occurs if the infection spreads and colon tissue attaches to another organ, such as the bladder or the uterus / vagina when surgery is done on an elective basis, the surgeon will remove the part of the bowel that is diseased and connect the normal parts of the bowel back together. when surgery is done on an emergency basis, the diseased part of the bowel will be removed. the healthy parts of the bowel will not be connected right away. your bowel will need time to rest and heal. the upper part of the bowel will be attached to the abdominal wall. a port will allow waste to pass from the intestine to a bag outside of your body. if possible, the healthy bowel will be reconnected after 6 - 12 weeks. if you are diagnosed with diverticulitis, follow your doctor ' s


carnitine is a substance used by the body uses to turn fat into energy. it is not normally considered an essential nutrient because the body can manufacture all it needs. however, supplemental carnitine could in theory improve the ability of certain tissues to produce energy. this has led to the use of carnitine for various muscle diseases as well as heart conditions. carnitine is primarily used for heart - related conditions. some evidence suggests that it can be used along with conventional treatment for to improve symptoms and reduce medication needs. when combined with conventional therapy, it may or may not help prevent medical complications or sudden cardiac death in the months following a lesser evidence suggests that it may be helpful for a condition called ( pain in the legs after walking due to narrowing of the arteries ), as well as congestive heart failure in addition, a few studies suggest that carnitine may be useful for carnitine may also be helpful for improving exercise tolerance in people with chronic pulmonary obstruction disease ( copd ), more commonly known as emphysema. you should not attempt to self - treat any of these serious medical conditions, nor should you use carnitine as a substitute for standard drugs. growing, if not entirely consistent, evidence suggests that l - carnitine or acetyl - l - carnitine, or their combination, may be helpful for improving sperm function and thereby provide benefits in male infertility. carnitine has also shown promise for improving male sexual function81, 82, 93 and improving mental and physical two studies found evidence that carnitine is helpful for, a condition affecting the penis. some studies have found evidence that one particular form of carnitine, acetyl - l - carnitine, might be helpful in but the two most recent and largest studies found no benefit. one review evaluated published and unpublished trials, and concluded that acetyl - l - carnitine may only be helpful for very mild alzheimer ' s disease. in preliminary trials, acetyl - l - carnitine has shown some promise for treatment of or dysthymia ( a milder condition related to depression ). some evidence suggests that carnitine may be useful for improving blood sugar control in people with type 2 ( adult - onset ) better evidence suggests benefit with acetyl - l - carnitine for a major complication of diabetes, diabetic peripheral neuropathy ( injury to nerves of the extremities caused by diabetes ). acetyl - l - car


evidence suggests benefit with acetyl - l - carnitine for a major complication of diabetes, diabetic peripheral neuropathy ( injury to nerves of the extremities caused by diabetes ). acetyl - l - carnitine might help prevent diabetic cardiac autonomic neuropathy ( injury to the nerves of the heart caused by diabetes ). however, one study found that carnitine supplements had an adverse effect on in people with diabetes. much weaker evidence suggests possible benefits for neuropathy caused by the chemotherapy drugs weak evidence hints that carnitine might help reduce liver and heart toxicity caused by the chemotherapy drug some evidence suggests that carnitine may be able to improve one small study demonstrated a beneficial effect of l - carnitine on anemia and high cholesterol in patients on hemodialysis for chronic renal failure. a genetic condition called fragile x syndrome can cause behavioral disturbances such as hyperactivity, along with intellectual disability, autism, and alterations in appearance. a preliminary study of 17 boys found that acetyl - l - carnitine might help to reduce hyperactive behavior associated with this condition. evidence for the effectiveness of l - carnitine in attention deficit and hyperactivity disorder ( adhd ) has been mixed. celiac disease is an autoimmune disease affecting the digestive tract. fatigue is a common symptom of the disease. one small double - blind trial found evidence that use of l - carnitine at a dose of 2 g daily might help alleviate this symptom. weak evidence hints that carnitine may help people with degeneration of the cerebellum ( the structure of the brain responsible for voluntary muscular movement ). one very small study suggests carnitine may be helpful for reducing symptoms of chronic fatigue syndrome another study suggests that carnitine may be of value for treating and for severe liver disease. a substantial study marred by poor design ( specifically, far too many primary endpoints ) found equivocal evidence that l - carnitine, taken at dose of 500 mg three times daily, might be more effective than placebo for the treatment of other weak evidence suggests that carnitine may be helpful for decreasing the muscle toxicity of azt ( a drug used to treat other weak evidence hints that the acetyl - l - carnitine might reduce nerve - related side effects caused by hiv drugs in general. one study failed to find carnitine effective for promoting although another found that carnitine


used to treat other weak evidence hints that the acetyl - l - carnitine might reduce nerve - related side effects caused by hiv drugs in general. one study failed to find carnitine effective for promoting although another found that carnitine might lead to improvements in body composition ( fat - muscle ratio ). carnitine is widely touted as a physical sports performance enhancer, but there is no real evidence that it is effective and some research indicates that it is not. little to no evidence supports other claimed benefits such as treating irregular heartbeat, down ' s syndrome, muscular dystrophy, and alcoholic fatty liver disease. however, in a randomized trial involving 25 patients with and early brain dysfunction ( hepatic encephalopathy ) associated with severe forms of this condition, carnitine appeared to significantly improve the function of both the liver and the brain after 3 months of treatment. what is the scientific evidence for carnitine? carnitine might be a good addition to standard therapy for. in one controlled study, 200 individuals with angina ( the exercise - induced variety ) took either 2 g daily of l - carnitine or were left untreated. all the study participants continued to take their usual medication for angina. those taking carnitine showed improvement in several measures of heart function, including a significantly greater ability to exercise without chest pain. they were also able to reduce the dosages of some of their heart medications ( under medical supervision ) as their symptoms decreased. unfortunately, the results of this study can ' t be fully trusted because researchers didn ' t use a double - blind protocol. ( for more information on why double - blinding is so important, see why does this database rely on double - blind studies? ) another trial did use a double - blind, placebo - controlled, design tested l - carnitine in 52 people with angina and found evidence of benefit. in addition, several small studies ( some of them double - blind ) tested propionyl - l - carnitine for the treatment of angina, and also found evidence of benefit. people with advanced hardening of the arteries, or, often have difficulty walking due to lack of blood flow to the legs, a condition called. pain may develop after walking less than half a block. although carnitine does not increase blood flow, it appears to improve the muscle ' s ability to function under difficult circumstances. a 12 - month, double - blind, placebo - controlled trial of 485 patients with


less than half a block. although carnitine does not increase blood flow, it appears to improve the muscle ' s ability to function under difficult circumstances. a 12 - month, double - blind, placebo - controlled trial of 485 patients with intermittent claudication evaluated the potential benefits of propionyl - l - carnitine. participants with relatively severe disease showed a 44 % improvement in walking distance as compared to placebo. however, no improvement was seen in those with mild disease. another double - blind study followed 245 people and also found benefit. similar results have been seen in most but not all other studies of l - carnitine or propionyl - l - carnitine. propionyl - l - carnitine may be more effective for intermittent claudication than plain carnitine. congestive heart failure several small studies have found that carnitine, often in the form of propionyl - l - carnitine, can improve symptoms of congestive heart failure in one trial, benefits were maintained for 60 days after treatment with carnitine was stopped. after a heart attack l - carnitine has shown inconsistent promise for use after a a double - blind, placebo - controlled study that followed 101 people for 1 month after a heart attack found that use of l - carnitine, in addition to standard care, reduced the size of the infarct ( dead heart tissue ). in the months following a severe heart attack, the left ventricle of the heart often enlarges, and the pumping action of the heart becomes less efficient. some evidence suggests that l - carnitine can help prevent heart enlargement, but that it does not improve heart function. in a 12 - month, double - blind, placebo - controlled study of 472 individuals who had just undergone a heart attack, use of carnitine at a dose of 6 g / day significantly decreased the rate of heart enlargement. however, heart function was not significantly altered. a 3 - month, double - blind, placebo - controlled study of 60 individuals who had just undergone a heart attack also failed to find improvements in heart function. ( heart enlargement was not studied. ) results consistent with those of the studies above were seen in a 6 - month double blind, placebo - controlled study of 2, 330 people who had just had a heart attack. carnitine failed to produce significant reductions in mortality or heart failure ( serious decline in heart function ) over the 6


seen in a 6 - month double blind, placebo - controlled study of 2, 330 people who had just had a heart attack. carnitine failed to produce significant reductions in mortality or heart failure ( serious decline in heart function ) over the 6 - month period. however, the study did find reductions in early death. ( unfortunately, for statistical reasons, the meaningfulness of this last finding is questionable. reduction in early death was a secondary endpoint rather than a primary one. ) carnitine is used along with conventional treatment, not as a substitute for it. high levels of blood sugar can damage the nerves leading to the extremities, causing pain and numbness. this condition is called diabetic peripheral neuropathy nerve damage may also develop in the heart, a condition called cardiac autonomic neuropathy. acetyl - l - carnitine has shown considerable promise for diabetic peripheral neuropathy and some promise for cardiac autonomic neuropathy. two 52 - week double - blind, placebo - controlled studies, involving a total of 1, 257 people with diabetic peripheral neuropathy, evaluated the potential benefits of alc taken at 500 mg or 1000 mg daily. the results showed that use of alc, especially at the higher dose, improved sensory perception and decreased pain levels. in addition, the supplement appeared to promote nerve fiber regeneration. a small study found some potential benefits for cardiac autonomic neuropathy. for more information, including full dosage and safety issues, see the full male sexual function carnitine has shown promise for improving male sexual function. one double - blind, placebo - controlled study of 120 subjects compared a combination of propionyl - l - carnitine ( 2 g per day ) and acetyl - l - carnitine ( 2 g per day ) against testosterone for the treatment of male aging symptoms ( sexual dysfunction, depression, and fatigue ). the results indicated that both testosterone and carnitine improved erectile function, mood, and fatigue, as compared to placebo. however, no improvements were seen in the placebo group. this is an unusual occurrence in studies of erectile dysfunction, so it casts some doubt on the study results. a double - blind study of 40 men evaluated propionyl - l - carnitine ( 2 g per day ) in diabetic men with erectile dysfunction who had not responded well to viagra. the results indicated that carnitine significantly enhanced the effectiveness of via


of 40 men evaluated propionyl - l - carnitine ( 2 g per day ) in diabetic men with erectile dysfunction who had not responded well to viagra. the results indicated that carnitine significantly enhanced the effectiveness of viagra. in another double - blind study, a combination of the propionyl and acetyl forms of carnitine enhanced the effectiveness of viagra in men who suffered from erectile dysfunction caused by prostate surgery. growing evidence suggests that l - carnitine or acetyl - l - carnitine or their combination may be helpful for improving sperm quality and function, thereby benefiting for example, in one double - blind, placebo - controlled study of 60 men, use of combined l - carnitine ( 2 g per day ) and acetyl - l - carnitine ( also at 2 g per day ) significantly improved sperm quality. chronic obstructive pulmonary disease ( copd ) evidence from three double - blind, placebo - controlled studies enrolling a total of 49 people suggests that l - carnitine can improve exercise tolerance in, presumably by improving muscular efficiency in the lungs and other muscles. numerous double - or single - blind studies involving a total of more than 1, 400 people have evaluated the potential benefits of acetyl - l - carnitine in the treatment of and other forms of dementia. however, while early studies found evidence of modest benefit, two large and well - designed studies failed to find acetyl - l - carnitine effective at all. the first of these was a double - blind, placebo - controlled trial that enrolled 431 participants for 1 year. overall, acetyl - l - carnitine proved no better than placebo. however, because a close look at the data indicated that the supplement might help people who develop alzheimer ' s disease at an unusually young age, researchers performed a follow - up trial. this 1 - year, double - blind, placebo - controlled trial evaluated acetyl - l - carnitine in 229 patients with early onset alzheimer ' s. unfortunately, no benefits were seen here either. one review of the literature concluded that acetyl - l - carnitine may be helpful for mild cases of alzheimer ’ s disease, but not more severe cases. a double - blind study of 60 seniors with dysthymia ( a mild form of ) found that treatment with 3 g of carnitine daily over a 2 - month period significantly improved symptoms as compared


s disease, but not more severe cases. a double - blind study of 60 seniors with dysthymia ( a mild form of ) found that treatment with 3 g of carnitine daily over a 2 - month period significantly improved symptoms as compared to placebo. positive results were seen in two other studies as well, one of depression and one of dysthymia. enlargement of the thyroid ( goiter ) can be due to many causes, including cancer and iodine deficiency. in some cases, thyroid enlargement occurs without any known cause, so - called benign goiter. treatment of benign goiter generally consists of taking thyroid hormone pills. this causes the thyroid gland to become less active, and the goiter shrinks. however, there may be undesirable effects as well. symptoms of ( too much thyroid hormone ) can develop, including heart palpitations, nervousness, weight loss, and bone breakdown. a double - blind, placebo - controlled trial found evidence that use of l - carnitine could alleviate many of these symptoms. this 6 - month study evaluated the effects of l - carnitine in 50 women who were taking thyroid hormone for benign goiter. the results showed that a dose of 2 g or 4 g of carnitine daily protected participants ' bones and reduced other symptoms of hyperthyroidism. carnitine is thought to affect thyroid hormone by blocking its action in cells. this suggests a potential concern — carnitine might be harmful for people who have low or borderline thyroid levels to begin with. this possibility has not been well explored as yet. is an inflammatory condition of the penis that develops in stages. in the first stage, penile pain occurs with erection ; next, the penis becomes curved ; finally, erectile dysfunction may occur. many medications have been tried for peyronie ’ s disease, with some success. one such drug is tamoxifen, which is better known as a treatment to prevent breast cancer recurrence. a 3 - month, double - blind study compared the effectiveness of acetyl - l - carnitine to the drug tamoxifen in 48 men with peyronie ' s disease. acetyl - l - carnitine ( at a dose of 1 g daily ) reduced penile curvature while tamoxifen did not ; in addition, the supplement reduced pain and slowed disease progression to a greater extent than tamoxifen.


| evolutionary conserved micrornas are ubiquitously expressed compared to tick - specific mirnas in the cattle tick rhipicephalus ( boophilus ) microplus. | | jump to full text | | pmid : 21699734 owner : nlm status : medline | | background : micrornas ( mirnas ) are small non - coding rnas that act as regulators of gene expression in eukaryotes modulating a large diversity of biological processes. the discovery of mirnas has provided new opportunities to understand the biology of a number of species. the cattle tick, rhipicephalus ( boophilus ) microplus, causes significant economic losses in cattle production worldwide and this drives us to further understand their biology so that effective control measures can be developed. to be able to provide new insights into the biology of cattle ticks and to expand the repertoire of tick mirnas we utilized illumina technology to sequence the small rna transcriptomes derived from various life stages and selected organs of r. microplus. results : to discover and profile cattle tick mirnas we employed two complementary approaches, one aiming to find evolutionary conserved mirnas and another focused on the discovery of novel cattle - tick specific mirnas. we found 51 evolutionary conserved r. microplus mirna loci, with 36 of these previously found in the tick ixodes scapularis. the majority of the r. microplus mirnas are perfectly conserved throughout evolution with 11, 5 and 15 of these conserved since the nephrozoan ( 640 mya ), protostomian ( 620mya ) and arthropoda ( 540 mya ) ancestor, respectively. we then employed a de novo computational screening for novel tick mirnas using the draft genome of i. scapularis and genomic contigs of r. microplus as templates. this identified 36 novel r. microplus mirna loci of which 12 were conserved in i. scapularis. overall we found 87 r. microplus mirna loci, of these 15 showed the expression of both mirna and mirna * sequences. r. microplus mirnas showed a variety of expression profiles, with the evolutionary - conserved mirnas mainly expressed in all life stages at various levels, while the expression of novel tick - specific mirnas was mostly limited to particular life stages and / or tick organs. conclusions : anciently acquired mirnas in the r. microplus lineage not only tend to accumulate


all life stages at various levels, while the expression of novel tick - specific mirnas was mostly limited to particular life stages and / or tick organs. conclusions : anciently acquired mirnas in the r. microplus lineage not only tend to accumulate the least amount of nucleotide substitutions as compared to those recently acquired mirnas, but also show ubiquitous expression profiles through out tick life stages and organs contrasting with the restricted expression profiles of novel tick - specific mirnas. | roberto a barrero ; gabriel keeble - gagnere ; bing zhang ; paula moolhuijzen ; kazuho ikeo ; yoshio tateno ; takashi gojobori ; felix d guerrero ; ala lew - tabor ; matthew bellgard | related documents : | 23618514 - molecular determination of tritrichomonas spp. in aborted bovine foetuses in eastern an... 22139924 - an in silico model for identification of small rnas in whole bacterial genomes : charact... 20238054 - a hypoplastic retinal lamination in the purpurin knock down embryo in zebrafish. 2691504 - the primary structure of porcine liver acylamino acid - releasing enzyme deduced from cdn... 20381534 - sybr green based real - time rt - pcr assay for detection and genotype prediction of bovine... 17846794 - molecular and phylogenetic analysis of mads - box genes of mikc type and chromosome locat... | type : journal article ; research support, non - u. s. gov ' t date : 2011 - 06 - 24 | | title : bmc genomics volume : 12 issn : 1471 - 2164 iso abbreviation : bmc genomics publication date : 2011 | | created date : 2011 - 07 - 22 completed date : 2011 - 11 - 15 revised date : 2013 - 05 - 24 | medline journal info : | nlm unique id : 100965258 medline ta : bmc genomics country : england | | languages : eng pagination : 328 citation subset : im | | centre for comparative genomics, murdoch university, wa, australia. email @ example. com | | apa / mla format download endnote download bibtex | drosophila / genetics gene expression profiling gene expression regulation * larva / genetics micrornas / genetics, metabolism * ovu


, australia. email @ example. com | | apa / mla format download endnote download bibtex | drosophila / genetics gene expression profiling gene expression regulation * larva / genetics micrornas / genetics, metabolism * ovum / metabolism rna precursors / genetics rhipicephalus / genetics *, growth & development | 0 / micrornas ; 0 / rna precursors | journal id ( nlm - ta ) : bmc genomics publisher : biomed central copyright ©2011 barrero et al ; licensee biomed central ltd. received day : 15 month : 4 year : 2011 accepted day : 24 month : 6 year : 2011 collection publication date : year : 2011 electronic publication date : day : 24 month : 6 year : 2011 volume : 12first page : 328 last page : 328 publisher id : 1471 - 2164 - 12 - 328 pubmed id : 21699734 | evolutionary conserved micrornas are ubiquitously expressed compared to tick - specific mirnas in the cattle tick rhipicephalus ( boophilus ) microplus | | roberto a barrero1 | | email : firstname. lastname @ example. org | | gabriel keeble - gagnere1 | | email : email @ example. com | | bing zhang23 | | email : bing. zhang @ deedi. qld. gov. au | | paula moolhuijzen13 | | email : firstname. lastname @ example. org | | kazuho ikeo4 | | email : email @ example. com | | yoshio tateno4 | | email : firstname. lastname @ example. org | | takashi gojobori4 | | email : email @ example. com | | felix d guerrero5 | | email : felix. guerrero @ ars. usda. gov | | ala lew - tabor1236 | | email : a. lewtabor @ uq. edu. au | | matthew bellgard13 | | email : firstname. lastname @ example. org | 1centre for comparative genomics, murdoch university, wa 6150, australia 2department of employment, economic development and innovation ( deedi ) biotechnology laboratories, the university of queensland, st lucia, qld 4067, australia 3crc for beef genetic technologies, university of new england, armidale, nsw 2351, australia 4center for information biology and dna databank of japan


laboratories, the university of queensland, st lucia, qld 4067, australia 3crc for beef genetic technologies, university of new england, armidale, nsw 2351, australia 4center for information biology and dna databank of japan, national institute of genetics, yata 1111, mishima, shizuoka 411 - 8540, japan 5us department of agriculture, agricultural research service, 2700 fredericksburg, rd., kerrville, tx 78028, usa 6queensland alliance for agriculture and food innovation institute, the university of queensland, c / o deedi, st lucia, qld 4067, australia the arthropods are a diverse group of organisms including chelicerata ( ticks, spiders ), myriapoda ( centipedes, millipedes ), crustacea ( crabs, shrimps ), and insecta ( flies, beetles ). molecular estimates indicate that ticks emerged 300 ± 27 mya, while the prostriate and metastriate hard tick lineages diverged 241 ± 28 mya. rhipicephalus ( boophilus ) microplus is considered to be the most economically important tick parasite in the world. r. microplus is a hard tick associated with cattle infestations but can also occasionally be found on other hosts including horses, goats, sheep, pigs and some wild animals living in subtropical and tropical regions worldwide [ 2, 3 ]. r. microplus is a member of the metastriate lineage of ticks that includes numerous genera and species of medical and veterinary importance. in comparison, i. scapularis is a member of the prostriate lineage that comprises the single genus ixodes. the prostriate and metastriate lineages differ markedly in many aspects of their biology such as type of developmental cycle ( i. e., three - versus one - host ticks ), host range and vector competence. comparative analyses between prostriate and metastriate gene sets including mirnas may reveal the genetic basis for fundamental differences in the biology of these tick lineages. r. microplus is generally a single host tick spending all parasitic life cycle stages on cattle. the eggs hatch in the environment and the larvae crawl up grass or other plants to find a host. in the summer, r. microplus can survive for as long as 3 to 4 months without feeding. in cooler temperatures, they may live without food for up to six


environment and the larvae crawl up grass or other plants to find a host. in the summer, r. microplus can survive for as long as 3 to 4 months without feeding. in cooler temperatures, they may live without food for up to six months. newly attached seed ticks ( larvae ) are usually found on the softer skin inside the thigh, flanks, and forelegs. after feeding, the larvae molt twice, to become nymphs and male or female adults. each developmental stage ( larval, nymph and adult ) feeds only once, but the feeding takes places over several days. adult male ticks become sexually mature after feeding, and mate with feeding females. an adult female tick that has fed and mated detaches from the host and deposits a single batch of many eggs in the environment. typically, these eggs are placed in crevices or debris, or under stones. the female tick dies after ovipositing. ticks in the subgenus boophilus have a life cycle that can be completed in 3 to 4 weeks ; this characteristic can result in a heavy tick burden particularly on tick susceptible cattle in tropical areas [ 2, 3 ]. micrornas ( mirnas ) are small 19 - - 25 nucleotide regulatory rnas that act as post - transcriptional modulators of gene expression in animals and plants. they are estimated to represent 1 % of the transcriptome in higher eukaryotes and predicted to control the expression of up to 30 % of messenger rnas [ 6, 7 ]. most mirnas are encoded in intergenic regions and are transcribed by rna polymerase ii as long primary nuclear mirnas ( pri - mirnas ), which range from hundreds to thousands of nucleotides in length. one pri - mirna typically contains a single or several mirna precursors ( pre - mirnas ) as stem - loop, hairpin structures flanked by unstructured, single stranded rna sequences. pre - mirnas are cleaved near their loops by the cytoplasmic rnase iii enzyme dicer to generate a heteroduplex of two ~ 23 - nt rnas that are then packed into the risc complex. mature mirna sequences are encoded either in the 5 ' - arm or the 3 ' - arm of pre - mirnas. the decision as to which sequence is incorporated into the silencing complex is influenced by the difference in pairing stabilities between the two ends of the mirna : mirna star ( mirna *


3 ' - arm of pre - mirnas. the decision as to which sequence is incorporated into the silencing complex is influenced by the difference in pairing stabilities between the two ends of the mirna : mirna star ( mirna * ) duplex, with preferential incorporation of the strand whose 5 ' end is less stably paired [ 11, 12 ]. in some cases both strands of the mirna : mirna * duplex were found expressed at similar levels. about half of the mirna genes in drosophila melanogaster are clustered and transcribed from a single polycystronic pri - mirna. recently, next generation sequencing technologies have been utilized to profile and discover mirnas genome - wide. to assist in this process it is normally required to have a reference genome sequence. currently, there is no reference genome sequence for r. microplus, with the ixodes scapularis genome draft ( iscaw1. 1 ) the closest reference genome in which 37 mirnas have been identified. some mirnas are highly conserved throughout evolution including let - 7, present in metazoan lineages such as arthropods and vertebrates that diverged 641 - 686 mya. thus, the use of the d. melanogaster reference genome for which 152 mirnas are currently annotated may allow not only the identification of highly conserved tick mirnas, but also to discover arthropod - specific mirnas. previous studies have shown that mirnas are continuously being added to metazoan genomes through time, and once these are integrated into gene regulatory networks, show only rare nucleotide substitutions within the mature mirna sequence at predictable positions and are only rarely secondarily lost [ 17 - 19 ]. this is likely related to the strong purifying selection against changes in secondary structure of pre - mirnas. wheeler and colleagues documented evolutionary stable shifts to the determination of position 1 of the mature sequence that can be displaced towards either the 5 ' or 3 ' end, a phenomenon called seed shifting, as well as the ability to post - transcriptionally edit the 5 ' end of the mature read, changing the identity of the seed sequence and possibly the repertoire of downstream targets. currently there are no known micrornas reported for r. microplus and we aimed at the identification and discovery of evolutionary conserved as well as novel tick - specific mirnas in r. microplus by using a combination of next generation high throughput sequencing, comparative gen


micrornas reported for r. microplus and we aimed at the identification and discovery of evolutionary conserved as well as novel tick - specific mirnas in r. microplus by using a combination of next generation high throughput sequencing, comparative genomics and de novo computational screening. we constructed eight small transcriptome libraries derived from various cattle tick life stages and from selected organs including gut, salivary glands and ovaries. we aim to characterize changes in gene set and expression levels of mirnas at various tick life stages as well as selected organs. we also conducted an evolutionary analysis to identify subsets of r. microplus mirnas and mirna * sequences that are perfectly conserved since either the nephrozoa ( 641 - 686 mya ), protostomia ( 618 - 653 mya ), arthropoda ( ~ 540 mya ) or ixodidae ( ~ 241mya ) ancestor [ 1, 16, 21 ]. we also provide evidence of seed shifts and gene duplications unique to the r. microplus lineage. currently there are 37 known mirnas in ixodes scapularis, a species belonging to prostriate hard tick lineage [ 15, 21 ], but there are no known mirnas identified for r. microplus or other metastriate hard tick species. in order to identify r. microplus mirnas and expand the repertoire of cattle tick mirnas and to obtain insights into changes in mirna expression throughout the cattle tick life stages and in selected adult female tick organs, a high throughput sequencing approach was conducted. this approach generated more than 35 million short reads derived from the tick small rna transcriptome from eggs, unfed larvae, larvae exposed for six hours to the host without being allowed to feed ( frustrated larvae ), and adult ticks as well as selected adult female tick organs ( table 1 ). to identify tick mirnas we anticipated that a fraction of the known arthopoda mirnas should be conserved in both the chelicerata ( cattle ticks ) and insecta lineages regardless of their estimated divergence time of more than 500 mya [ 16, 21 ]. under this assumption, sequenced short reads from tick samples could be mapped onto the d. melanogaster genome in order to identify identical or nearly identical conserved tick mirnas. to be able to distinguish true sequence polymorphisms from non - specific mapping artefacts the performance of several short read aligners were initially evaluated to define the tool


. melanogaster genome in order to identify identical or nearly identical conserved tick mirnas. to be able to distinguish true sequence polymorphisms from non - specific mapping artefacts the performance of several short read aligners were initially evaluated to define the tool that mapped the largest amount of true positives and introduced a limited amount of false positive aligned short reads. we created simulated 36 - bp short reads using mutation rates from 0. 1 % to up to 16 % ( additional file 1 ) containing both snps and / or insertion / deletions ( indels ) and evaluated the ability of each tool to correctly align simulated short reads. our results indicated that novoalign produced the best overall short read mapping performance ( additional file 2 ). thus, we used this tool to align the generated tick short reads onto the drosophila genome. to ensure reliable short read alignments base quality scores were taken into account to conduct an interative alignment approach aiming to identify the best mapping position for each read. novoalign generates a mapping quality score for each aligned read so that ambiguously mapped short reads can be removed from downstream analyses. out of 35 million short reads derived from various cattle tick life cycle stages and three key organs, 3. 5 million reads were aligned onto known mirna loci on the drosophila genome with at least a quality alignment score of q = 1 ( table 1 ). we manually inspected the aligned reads onto mirna loci and removed non - specific alignments. to identify possible duplicated r. microplus mirnas all reads mapped onto a single d. melanogaster mirna locus were inspected and if mirna isoforms were observed these were required to be cloned in at least two distinct libraries to be validated. this approach identified 46 r. microplus mirnas including four duplicated copies for rmi - let - 7 ( a, f, m and n ), and two duplicated copies for rmi - mir - 219 and rmi - mir - 285. in addition we also detected the expression of 7 mirna * sequences, of these four have counterparts in d. melanogaster ( additional file 3 ). it has been recently reported that there has been gain and loss of mirna families in arthropod lineages. thus some known mirnas may have been lost in the d. melanogaster genome but still be present in tick genomes. to evaluate this possibility and also to identify novel tick - specific mirnas we conducted a computational screening using mir


understanding of changes in gene regulatory networks during r. microplus life stages and in vital organs. in the above analysis, we also identified 36 novel r. microplus mirnas and of these 12 were conserved in the i. scapularis genome ( additional file 3b ). examples of pre - mirnas of novel tick - specific mirnas showing typical drosha - processed features including localization of mature mirna within few nucleotides of the loop are shown in figure 1. interestingly we detected the expression of mirna * sequences in four novel tick pre - mirnas common to both r. microplus and i. scapularis ( additional file 3b and figure 1 ). to determine if the 36 identified novel r. microplus mirnas are restricted to cattle tick, we aligned mature mirna and pre - mirnas sequences onto the genomes of anopheles gambiae, apis mellifera, aedes aegypti, tribolium castaneum, nasonia vitripennis, pediculus humanus, culex quinquefasciatus and nine drosophila genomes ( additional file 4 ). the mapping coordinates of aligned reads were then used to retrieve genomic segments to evaluate for typical pre - mirna structures. none of the 36 novel r. microplus mirnas were found conserved in the 16 tested genomes suggesting that these may represent tick - specific mirnas. the genome size of the cattle tick r. microplus is three times larger than that of i. scapularis, thus we anticipate that other novel r. microplus mirnas are likely to be identified once the complete genome becomes available. overall, we identified 87 r. microplus mirnas with 72 of these expressed during various cattle tick life stages and 67 in selected adult female tick organs ( figure 2a and additional file 3 ). we found 52 mature mirnas expressed in both the life stage and tick organ samples with the majority of these ( 44 ; 84. 6 % ) corresponding to evolutionary conserved mirnas ( figure 2a ). in contrast, the majority of the 20 life stage - and 15 organ - specific mirnas corresponded to novel cattle tick - specific mirnas suggesting that these mirnas play unique roles at specific life stages and key tick organs. a recent study suggested that mirna invention is closely related to the evolution of tissue identities in bilaterian species. further investigation is required to determine if novel tick - specific mir


these mirnas play unique roles at specific life stages and key tick organs. a recent study suggested that mirna invention is closely related to the evolution of tissue identities in bilaterian species. further investigation is required to determine if novel tick - specific mirnas contribute to the implementation of biological features unique to tick species. similar to what has been observed in other species we found mirnas that show expression from both arms of the pre - mirna. in cattle ticks we observed co - expression of 15 mature and complementary star sequences including rmi - mir - 10, rmi - mir - 71, rmi - mir - 993 and rmi - mir - 5308 that are expressed in all cattle tick life stages evaluated in this study ( table 2 ). co - expression of rmi - mir - 153 and rmi - mir - 5314 mature and star sequences was restricted to larval stages while rmi - mir - iab - 5p was only detected in adult ticks. these mirna * sequences were highly conserved between r. microplus and i. scapularis despite their estimated divergence of 241 ± 28 mya indicating that these sequences in these species are under selective pressure to avoid nucleotide changes. interestingly rmi - mir - 79 *, rmi - mir - 281 * and rmi - mir - 993 * are expressed at significantly higher levels than their mature counterparts suggesting that some of the currently annotated mature mirnas in reference databases may correspond to mirna * sequences or in some lineages like r. microplus mirna * sequences become the primary transcript from the mirna / mirna * duplex. to evaluate global changes in mirna expression during r. microplus life stages, short read counts overlapping the 87 mirnas and 15 mirna * sequences were normalized as reads per million and compared against the overall expression found in eggs. our results indicate that in the transition from egg to larval stages there is nearly a 90 - fold increase in mirna transcripts ( figure 2b ). interestingly, larvae exposed to the host for six hours showed a significant reduction in the accumulation of mirna transcripts as compared to unexposed larvae. the largest accumulation of mirna transcripts was found in female adults, showing 2. 7 - fold higher level than males. we also compared changes in expression between evolutionary conserved and tick - specific ( unique ) mirnas. evolutionary conserved r. microplus mirnas are significantly more highly expressed as compared to the identified


, showing 2. 7 - fold higher level than males. we also compared changes in expression between evolutionary conserved and tick - specific ( unique ) mirnas. evolutionary conserved r. microplus mirnas are significantly more highly expressed as compared to the identified novel tick - specific mirnas ( figure 2c ), only during larval stages there was an increment in the expression of tick - specific mirnas. overall in the life stage samples, we identified 72 r. microplus mirnas, of these 37, 63 and 61 mirnas were expressed in egg, larval and adult stages, respectively ( figure 2d ). interestingly, 32 ( 44. 5 % ) of the identified mirnas were expressed in all cattle tick life stages with 31 of these evolutionary conserved in a range of species and only one rmi - mir - 5308 unique to tick species. in contrast most of the stage - specific mirnas corresponded to novel tick - specific mirnas. these findings suggest that evolutionary conserved mirnas play a ubiquitous role through out cattle tick life stages, while most novel tick - unique mirnas are restricted to specific life stages. we next evaluated the percentage fraction of expression of each mirna in each sample. in eggs there are five abundantly expressed mirnas including rmi - mir - 1, rmi - mir - 310 / mir - 92, rmi - mir - 279, rmi - mir - 275 and rmi - mir - 71 that accounted for 80. 6 % of the total amount of mirna transcripts ( figure 3a and additional file 3 ), while in all other life stages mir - 1 was the most abundantly expressed mirna ( figure 3 ). among the top ten most abundant mirnas in eggs we found two novel cattle tick - specific mirnas, rmi - mir - 5316b and rmi - mir - 5331, that accounted for 4. 3 % and 3. 1 % of mirna transcripts, respectively ( figure 3a ). these novel mirnas were only expressed at statistically significant levels ( p < 9. 62e - 06 ) during the egg stage ( additional file 5 ) suggesting they play critical roles during this stage. although rmi - mir - 1 is the most abundantly expressed mirna in cattle tick larvae ( figure 3b ), we found eight other mirnas, including rmi - mir - 133, rmi - mir - 87, rmi - mir - 10, and rmi - mir - 252, that had a higher fold - change


( figure 3b ), we found eight other mirnas, including rmi - mir - 133, rmi - mir - 87, rmi - mir - 10, and rmi - mir - 252, that had a higher fold - change ratio in mirna transcript levels as compared to that in eggs suggesting that these mirnas may play a role during larval development ( table 3 and additional file 6 ). we also found two novel tick - specific mirnas, rmi - mir - 3931 and rmi - mir - 5308, among the top ten expressed larval mirnas representing 2. 7 % and 1. 2 % of the mirna transcripts ( figure 3b ). interestingly, in drosophila mir - 10 is encoded within the hox cluster downstream of its target sex combs reduced ( scr ), which is a gene required for proper embryo and adult development in flies [ 27, 28 ]. it has also been reported that the mir - 10 binding site in the src 3 ' utr is conserved across a large number of arthropod species, some with an estimated divergence time of over hundreds of millions years. similar to the observation in larval stages, rmi - mir - 1 was vastly abundant in adult ticks accounting for 87. 4 % and 86. 2 % of mirna transcripts in females and males, respectively ( figure 3c and 3d ). the second most abundant mirna in adult ticks is rmi - let - 7a, a mirna known to be involved in the transition from late larval to adult stage in worms. it remains to be elucidated if rmi - let - 7a is also expressed during tick nymph stage associated with the transition to adult fate. in contrast to egg and larval stages, no novel tick - specific mirnas were observed among the top ten most abundant mirnas in adult ticks. these findings suggest that novel tick - specific mirnas may play key roles during early embryo and larval development stages. the relative changes in expression of 27 mirnas and 3 mirna * expressed in all life stages tested in r. microplus are shown in figure 3e. interestingly let - 7, mir - 100 and mir - 125 are known to be clustered in the same genomic location in the d. melanogaster and a. gambiae genomes within 1 kb and 4. 5 kb, respectively. in general it is accepted that clustered mirnas are likely to share


are known to be clustered in the same genomic location in the d. melanogaster and a. gambiae genomes within 1 kb and 4. 5 kb, respectively. in general it is accepted that clustered mirnas are likely to share highly correlated expression profiles if these are within 50 kb of each other. our results show that these three mirnas present similar expression trends ( figure 3e ) suggesting that rmi - let - 7a, rmi - mir - 100 and rmi - mir - 125 may also collocate to the same genomic region in the r. microplus genome. to validate the observed mirna expression profiles we conducted real time pcr amplification of five randomly selected mirnas that showed perfect sequence conservation between ticks and flies. these included rmi - let - 7a, rmi - mir - 1, rmi - mir - 7, rmi - mir - 12, and rmi - mir - 124. as controls we selected u14 and snorna - 442 non - coding rnas, which are reference genes used in drosophila studies ( ambion, applied biosystems ), but none of these were either expressed or conserved in cattle ticks. we then normalized the relative expression of all mirnas in eggs, frustrated larvae and female ticks against the expression of rmi - let - 7 in female ticks ( additional file 7 ). our results validated the abundant expression of rmi - mir - 1 in eggs, frustrated larvae and female samples. we also verified that rmi - let - 7 is highly expressed in female ticks as compared to eggs or frustrated larvae samples ( additional file 5 ). overall we observed a good agreement between the trends observed in real time pcr quantification and high throughput small rna sequencing. r. microplus normally parasitizes a single host and it is believed this host specificity depends on the specific detection of bovine phenolic compounds. to evaluate if host odour recognition by ticks may trigger changes in mirna expression we exposed tick larvae to its host for six hours without allowing the larvae to feed ( frustrated larvae ), and then we collected small rna samples for high throughput sequencing. we found 57 mirna expressed in frustrated larvae, of these 54 were also expressed in unexposed larvae ( additional file 3 ). there were six mirnas expressed in unexposed larvae that were not detected in frustrated larvae, particularly rmi - mir - 5315 that showed


in frustrated larvae, of these 54 were also expressed in unexposed larvae ( additional file 3 ). there were six mirnas expressed in unexposed larvae that were not detected in frustrated larvae, particularly rmi - mir - 5315 that showed a significant lost in expression ( table 4 and additional file 8 ). only rmi - mir - 31a, rmi - mir - 285a and rmi - mir - 5329 were specifically detected in frustrated larvae, but none of these had significant expression levels ( p < 9. 62e - 05 ) ( additional file 8 ). the major difference in mirna expression found in frustrated larvae, as compared to un - exposed larvae, is the down - and up - regulation of 26 and 23 mirnas, respectively ( p < 9. 62e - 05 ) ( additional file 8 ). among the most down regulated mirnas in frustrated larvae are rmi - mir - 317, rmi - mir - 315, rmi - mir - 33, rmi - mir - 87 and five tick - specific novel mirnas including rmi - mir - 5306, rmi - mir - 5309, rmi - mir - 5310, rmi - mir - 5312 and rmi - mir - 5314 ( table 4 and additional file 8 ). other significantly up - regulated mirnas in frustrated larvae compared to unexposed larvae are rmi - mir - 279, rmi - mir - 190, rmi - mir - 79 *, rmi - mir - 96, rmi - mir - 5307 and rmi - mir - 5308 ( p < 9. 62e - 05 ) ( table 4 and additional file 8 ). the identification of novel tick - specific mirnas showing up and down - regulation upon exposure to the host make these molecules attractive candidates for further host - recognition functional studies. there are significant morphological and behavioural differences between female and male ticks. to evaluate whether there are gender differences in the expression of mirnas we inspected changes in the mirna transcriptome between female and male adult ticks. we found 55 mirnas expressed in adult cattle ticks, of these 46 were expressed in both females and males. we found two mirnas, rmi - mir - 5334 and rmi - mir - 5336, specifically expressed in male ticks, while another seven mirnas were detected in female ticks with four of these also expressed in larval stages ( figure 2d ). the three mirnas uniquely expressed in females were rm


being rather pro - apoptotic. interestingly, mir - 1 level was shown to significantly increase in response to oxidative stress. in ticks blood digestion has been suggested to be a source of oxidative stress [ 40, 41 ]. this is particularly relevant for female ticks that ingest large volumes of blood ( about 100 - fold their own weight ) in preparation for the laying of about 2, 000 to 3, 000 eggs, while males rarely feed on blood. interestingly, apoptosis of salivary glands prior to oviposition in female ticks was reported for dermacentor variabilis and r. microplus. surprisingly r. microplus female ticks presented apoptosis just after 24 hours of host detachment contrasting with the 5 days required for d. variabilis female ticks. these observations suggest that in r. microplus molecular changes are likely to occur prior to female detachment from the host that predispose them for a rapid onset of apoptosis and consequently facilitating egg laying. this notion correlates with the observed significant up regulation of rmi - mir - 1 in semi - engorged females as compared to males ( table 5 ). it remains to be elucidated if blood - mediated oxidative stress in r. microplus contributes to the increased expression level of rmi - mir - 1 in females and if this mirna exercise pro - apoptotic activities similar to its x. laevis mir - 1 counterpart that would ultimately facilitate egg laying. primary target organs for cattle tick control are the gut, salivary glands and ovaries of adult female ticks. nearly 13. 3 million short reads were generated for these three organs, of these 790, 821 and 5, 759 short reads were mapped onto evolutionary conserved and novel tick - specific mirna loci, respectively ( table 1b ). to compare global mirna expression levels between these samples we normalized all mapped mirna transcripts in each sample as reads per million. the tick gut sample showed 2 - fold and 5 - fold higher relative amount of mirna transcripts as compared to salivary glands and ovaries samples, respectively ( additional file 10a ). these differences were due to the larger amount of transcripts mapped onto evolutionary conserved mirna loci in the gut sample as compared to salivary gland and ovary samples ( additional file 10a ). we found 68 mirnas expressed in all tested tick organs with 50 ( 6


the larger amount of transcripts mapped onto evolutionary conserved mirna loci in the gut sample as compared to salivary gland and ovary samples ( additional file 10a ). we found 68 mirnas expressed in all tested tick organs with 50 ( 6 novel ), 50 ( 9 novel ) and 49 ( 13 novel ) expressed in gut, salivary glands and ovaries, respectively ( additional file 10b ). a comparison of r. microplus mirnas found in female tick organs with those found in whole semi engorged adult females revealed that 36 mirnas were commonly expressed in all samples ( additional file 10b ). interestingly the majority of the commonly expressed r. microplus mirnas ( 94. 6 % ) in tick organs corresponded to known mirnas in other species, while 18 of the 20 organ - specifically expressed mirnas found in this study correspond to novel tick - specific mirnas ( additional file 10c ). most of the novel tick specific mirnas were expressed at low levels except for rmi - mir - 5307 found in all three organs and rmi - mir - 3931 present in adult female tick ovaries ( additional file 11 ). similar to what we observed for life stage samples, rmi - mir - 1 was the most abundantly expressed mirna in all sampled organs accounting for ~ 81 - 83 % of the identified r. microplus mirna transcripts in ovaries and gut, and for 43. 1 % of the mirna transcripts in salivary glands ( additional file 11 ). other highly expressed mirnas in salivary glands are rmi - let - 7a ( 15. 1 % ), rmi - mir - 275 ( 10. 5 % ), rmi - mir - 263a ( 5. 0 % ) and rmi - mir - 71 ( 4. 2 % ) ( additional file 11 ). recently, two studies have reported the secretion of mirnas into the saliva [ 46, 47 ]. interestingly, among the most abundantly expressed r. microplus mirnas in salivary glands, only let - 7 orthologs were found expressed in human salivary glands [ 46, 47 ]. the expression of let - 7 in the salivary glands of ticks and vertebrates suggests a functional role for this mirna throughout the evolution of this organ. to evaluate similarities in the global tick mirna expression pattern among all samples we conducted a hierarchical clustering as described by eisen et al. and found


the ancestral sequences using a maximum likelihood approach [ 49, 50 ] under the junkes - cantor model using a tree topology similar to that shown in figure 5a ( modified from ). we found that 73 % of the r. microplus mirnas were perfectly conserved since at least 241 mya with 11, 5, 15 and 6 mirnas conserved since the nephrozoan, protostomian, arthropoda and ixodidae ancestor, respectively ( figure 5b ). only 27 % of the r. microplus mirnas presented nucleotide substitutions unique to the r. microplus lineage. these unique mirnas can be further divided into three groups : 1 ) unique mirna variants in r. microplus for which a protostomian ancestral sequence is available ( rmi - i ), 2 ) duplicated copies of r. microplus mirnas ( rmi - ii ), and 3 ) unique r. microplus mirnas for which an ancestral sequence is not available and only ortholog sequences in the insecta lineage are known ( additional file 13 ). we next examined the number of nucleotide substitutions found in each of the above subsets of r. microplus mirnas by comparing against an ancestral sequence if it can be unambiguously determined ( i. e. a r. microplus mirna perfectly conserved since the arthropoda ancestor was compared against the protostomian ancestral mirna sequence or an earlier ancestral sequence if available ). in cases where no ancestral sequences are available owing to the gain of that mirna in a more recent ancestor the number of base substitutions was recorded as zero ( i. e. a mirna that was gained in the arthropoda ancestor and since then have not accumulated nucleotide substitutions in r. microplus ). the average number of nucleotide substitutions was then calculated for each subset. we determined 0, 0. 25, 0. 8 and 1. 0 average nucleotide changes for r. microplus mirnas conserved since the nephrozoan ( rmi - na ), protostomian ( rmi - pa ), arthropoda ( rmi - aa ) and ixodiade ( rmi - ia ) ancestor, respectively ( figure 5c ). furthermore we found 2. 2 and 1. 3 average nucleotide changes in r. microplus mirna unique variants ( rmi - i ) and duplicate


r. microplus mirnas showing one - nucleotide seed shifting with two of these ( rmi - mir - 79 and rmi - mir - 137 ) and a third mirna ( rmi - mir - iab - 4 - 5p ) displacing the seed towards the 3 ' and 5 ', respectively. interestingly seed shifting in mir - 79 was not observed in the sister taxon i. scapularis nor the crustacean daphnia pulex with which rmi - mir - 79 shares perfect sequence identity. seed shifting of mir - 79 was also found in capitella teleta ( annelida ) and insecta species, but not in their sister taxon, namely, mollusca and crustacea, respectively. these findings suggest that mir - 79 underwent three independent seed shifting events of one - base displacement towards the 3 ' end in r. microplus, c. teleta and insecta ancestor lineages. we also found two independent seed shifting events for mir - 137 one in r. microplus and another among insecta species ( a. mellifera and t. castaneum ) but not in their sister taxon daphnia pulex ( additional file 13 ). the observed consistency in the seed shifting direction in independent events suggests that this phenomenon may be under selective pressure. in our data only rmi - mir - iab - 4 - 5p represents an example of seed shifting unique to the r. microplus lineage, but this mirna is an exceptional example as both mature and star sequences have also accumulated a large number of base substitutions ( additional file 13 ). despite the observed seed shifting and nucleotide substitutions changes the expression level of the mature rmi - mir - iab - 4 - 5p is 5. 2 - fold higher than that of rmi - mir - iab - 4 - 3p, which is consistent with the 5. 4 - fold higher expression level of dme - mir - iab - 4 - 5p as compared to dme - mir - 4 - 3p. these findings suggest that despite the various molecular changes that can take place in a mirna - mirna * duplex, the expression ratio at which these molecules are found expressed in distinct taxa remains unchanged reflecting that other mechanisms may regulate the dosage at which each molecule is present to properly control downstream targets. this study has identified 87 r. microplus mirnas being 51 of these known in other species including 36