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Intracytoplasmic sperm injection [ICSI]

Oocyte is injected during ICSI Intracytoplasmic sperm is an in vitro fertilization procedure in which a single sperm is injected directly into an egg.

Indications
This procedure is most commonly used to overcome male infertility problems, although it may also be used where eggs cannot easily be penetrated by sperm, and occasionally as a method of in vitro fertilization, especially that associated with sperm donation. It can be used in teratozoospermia. Once the egg is fertilized, abnormal sperm morphology does not appear to influence blastocyst development or blastocyst morphology. Even with severe teratozoospermia, microscopy can still detect the few sperm cells that have a "normal" morphology, allowing for optimal success rate.

History
The technique was developed by Gianpiero Palermo around 1991 at the Vrije Universiteit Brussel, in the Center for Reproductive Medicine.

Procedure
The procedure is done under a microscope using multiple micromanipulation devices (micromanipulator, microinjectors and micropipettes). A holding pipette stabilizes the mature oocyte with gentle suction applied by a microinjector. From the opposite side a thin, hollow glass micropipette is used to collect a single sperm, having immobilised it by cutting its tail with the point of the micropipette. The micropipette is pierced through the oolemma and into the inner part of the oocyte (cytoplasm). The sperm is then released into the oocyte. The pictured oocyte has an extruded polar body at about 12 o'clock

indicating its maturity. After the procedure, the oocyte will be placed into cell culture and checked on the following day for signs of fertilization. In natural fertilization sperm compete and when the first sperm penetrates the oolemma, the oolemma hardens to block the entry of any other sperm. Concern has been raised that in ICSI this sperm selection process is bypassed and the sperm is selected by the embryologist without any specific testing. However, in mid 2006 the FDA cleared a device that allows embryologists to select mature sperm for ICSI based on sperm binding to hyaluronan, the main constituent of the gel layer (cumulus oophorus) surrounding the oocyte. The device provides microscopic droplets of hyaluronan hydrogel attached to the culture dish. The embryologist places the prepared sperm on the microdot, selects and captures sperm that bind to the dot. Basic research on the maturation of sperm shows that hyaluronan-binding sperm are more mature and show fewer DNA strand breaks and significantly lower levels of aneuploidy than the sperm population from which they were selected. A brand name for one such sperm selection device is PICSI. 'Washed' or 'unwashed' sperm may be used in the process.

Gamete intrafallopian transfer [GIFT]


Gamete intrafallopian transfer is a tool of assisted reproductive technology against infertility. Eggs are removed from a woman's ovaries, and placed in one of the Fallopian tubes, along with the man's sperm. The technique, which was pioneered by endocrinologist Ricardo Asch, allows fertilization to take place inside the woman's body. With the advances in IVF the GIFT procedure is used less as pregnancy rates in IVF tend to be equal or better and do not require laparoscopy.

Method
It takes, on average, four to six weeks to complete a cycle of GIFT. First, the woman must take a fertility drug to stimulate egg production in the ovaries. The doctor will monitor the growth of the ovarian follicles, and once they are mature, the woman will be injected with Human chorionic gonadotropin (hCG). The eggs will be harvested approximately 36 hours later, mixed with the man's sperm, and placed back into the woman's Fallopian tubes using a laparoscope.

Indications
A woman must have at least one normal fallopian tube in order for GIFT to be suitable. It is used in instances where the fertility problem relates to sperm dysfunction, and where the couple has idiopathic (unknown cause) infertility. Some patients may prefer the procedure to IVF for ethical reasons, since the fertilization takes place inside the body. This is a semi invasive procedure and requires laparoscopy .

Success rate
As with most fertility procedures, success depends on the couple's age and the woman's egg quality. It is estimated that approximately 25-30% of GIFT cycles result in pregnancy, with a third of those being multiple pregnancies. The First GIFT baby in the UK was Todd Holden born on the 23rd October 1986 in the Peterborough District Hospital.(WOW) The First GIFT baby in the US was Kaitlynne Kelley born on the 28th of April in 1987.

Fertilisation
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A sperm cell fertilising an ovum Fertilisation (also known as conception, fecundation and syngamy), is the fusion of gametes to produce a new organism. In animals, the process involves the fusion of an ovum with a sperm, which eventually leads to the development of an embryo. Depending on the animal species, the process can occur within the body of the female in internal fertilisation, or outside in the case of external fertilisation The entire process of development of new individuals is called procreation, the act of species reproduction.

Fertilisation in animals
The mechanics behind fertilisation has been studied extensively in sea urchins and mice. This research addresses the question of how the sperm and the appropriate egg find each other and the question of how only one sperm gets into the egg and delivers its contents. There are three steps to fertilisation that ensure species-specificity: 1. Chemotaxis 2. Sperm activation/acrosomal reaction 3. Sperm/egg adhesion.

Internal vs. External


Consideration as to whether an animal (more specifically a vertebrate) uses internal or external fertilisation is often dependent on the method of birth. Oviparous animals laying eggs with thick calcium shells, such as chickens, or thick leathery shells generally reproduce via internal fertilisation so that the sperm fertilise the egg without having to pass through the thick, protective, tertiary layer of the egg. Ovoviviparous and euviviparous animals also use internal fertilisation. It is important to note that although

some organisms reproduce via amplexus, they may still use internal fertilisation, as with some salamanders. Advantages to internal fertilisation include: minimal waste of gametes; greater chance of individual egg fertilisation, relatively "longer" time period of egg protection, and selective fertilisation; many females have the ability to store sperm for extended periods of time and can fertilise their eggs at their own desire. Oviparous animals producing eggs with thin tertiary membranes or no membranes at all, on the other hand, use external fertilisation methods. Advantages to external fertilisation include: minimal contact and transmission of bodily fluids; decreasing the risk of disease transmission, and greater genetic variation (especially during broadcast spawning external fertilisation methods).

Sea urchins

Acrosome reaction on a Sea Urchin cell Chemotaxis was discovered as the method by which sperm find the eggs. This chemotaxis is an example of a ligand/receptor interaction. Resact is a 14 amino acid peptide purified from the jelly coat of A. punctulata that attracts the migration of sperm. After finding the egg, the sperm gets through the jelly coat through a process called sperm activation. In another ligand/receptor interaction, an oligosaccharide component of the egg binds and activates a receptor on the sperm and causes the acrosomal reaction. The acrosomal vesicles of the sperm fuse with the plasma membrane and are released. In this process, molecules bound to the acrosomal vesicle membrane, such as bindin, are exposed on the surface of the sperm. These contents digest the jelly coat and eventually the vitelline membrane. In addition to the release of acrosomal vesicles, there is explosive polymerization of actin to form a thin spike at the head of the sperm called the acrosomal process.

The sperm binds to the egg through another ligand reaction between receptors on the vitelline membrane. The sperm surface protein bindin, binds to a receptor on the vitelline membrane identified as ERB1. Fusion of the plasma membranes of the sperm and egg are likely mediated by bindin. At the site of contact, fusion causes the formation of a fertilisation cone.

Mammals
Usually mammals rely on internal fertilisation through copulation. After a male ejaculates, a large number of sperm cells move to the upper vagina (via contractions from the vagina) through the cervix and across the length of the uterus toward the ovum. The capacitated spermatozoon and the oocyte meet and interact in the ampulla of the fallopian tube. It is probable that chemotaxis is involved in guiding the sperm to the egg, but the mechanism has yet to be worked out. However, demonstration of formyl peptide receptors (60.000 receptor/cell; higher binding capacity in the tail region) in the surface membrane of human sperms strongly supports, that - besides specific chemoattractant substances i.e. resact - professional chemoattractant ligands like formyl Met-Leu-Phe (fMLF) have also the ability to induce migration of sperm.[6] The zona pellucida of the egg binds with the sperm. In contrast to sea urchins, the sperm binds to the egg before the acrosmal reaction. The zona pellucida is a thick layer of extracellular matrix that surrounds the egg and is similar to the role of the vitelline membrane in sea urchins. A glycoprotein in the zona pellucida, ZP3 was discovered to be responsible for egg/sperm adhesion in mice. The receptor galactosyltransferase (GalT) binds to the N-acetylglucosamine residues on the ZP3 and is important for binding with the sperm and activating the acrosome reaction. ZP3 is sufficient for sperm/egg binding but not necessary. There are two additional sperm receptors: a 250kD protein that binds to an oviduct secreted protein and SED1 which binds independently to the zona. After the acrosome reaction, it is believed that the sperm remains bound to the zona pellucida through exposed ZP2 receptors. These receptors are unknown in mice but have been identified in guinea pigs. In mammals, binding of the spermatozoon to the GalT initiates the acrosome reaction. This process releases the enzyme hyaluronidase, which digests the matrix of hyaluronic acid in the vestments surrounding the oocyte. Fusion between the oocyte plasma membranes and sperm follows, allowing the entry of the sperm nucleus, centriole and flagellum, but not the mitochondria, into the oocyte. The fusion is likely mediated by the protein CD9 in mice (the binding homolog). The egg "activates" itself upon fusing with a single sperm cell, thereby changing its cell membrane to prevent fusion with other sperm. This process ultimately leads to the formation of a diploid cell called a zygote. The zygote begins to divide and form a blastocyst and when it reaches the uterus, it performs implantation in the endometrium. At this point the female's pregnancy has begun. If the embryo implants in any tissue other than the uterine wall, an ectopic pregnancy results, which can be fatal to the mother.

In some animals (e.g. rabbits) the act of coitus induces ovulation by stimulating release of the pituitary hormone gonadotropin. This greatly increases the probability that coitus will result in pregnancy.

Humans
The term conception commonly refers to fertilisation, the successful fusion of gametes to form a new organism. 'Conception' is used by some to refer to implantation and is thus a subject of semantic arguments about the beginning of pregnancy, typically in the context of the abortion debate. Gastrulation, which occurs around 16 days after fertilisation, is the point in development when the implanted blastocyst develops three germ layers, the endoderm, the ectoderm and the mesoderm. It is at this point that the genetic code of the father becomes fully involved in the development of the embryo. Until this point in development, twinning is possible. Additionally, interspecies hybrids survive only until gastrulation, and have no chance of development afterward. However this stance is not entirely accepted as some human developmental biology literature refers to the "conceptus" and such medical literature refers to the "products of conception" as the postimplantation embryo and its surrounding membranes. The term "conception" is not usually used in scientific literature because of its variable definition and connotation.

Fertilisation and genetic recombination


Meiosis results in a random segregation of the genes contributed from each parent. Each parent organism generally has the same genetic make-up, but differs for a fraction of their genes. Therefore, each gamete produced by a person will be genetically different from the others from that person, as well as from the gametes produced by another person. When gametes first fuse at fertilisation, the chromosomes donated by the parents are combined, and, in humans, this means that (2) = 17.6x1012 chromosomally different zygotes are possible for the non-sex chromosomes, even assuming no chromosomal crossover. If crossover occurs once, then on average (4) = 309x1024 genetically different zygotes are possible for every couple, not considering that crossover events can take place at most points along each chromosome. The X and Y chromosomes do not undergo crossover events, so are excluded from the calculation. Note that the mitochondrial DNA is only inherited from the maternal parent.

In vitro fertilisation [IVF]

Oocyte with surrounding granulosa cells

"Naked" Egg

early embryo development following fertilisation

In vitro fertilisation (IVF) is a process by which egg cells are fertilised by sperm outside the womb, in vitro. IVF is a major treatment in infertility when other methods of assisted reproductive technology have failed. The process involves hormonally controlling the ovulatory process, removing ova (eggs) from the woman's ovaries and letting sperm fertilise them in a fluid medium. The fertilised egg (zygote) is then transferred to the patient's uterus with the intent to establish a successful pregnancy. The first successful birth of a "test tube baby", Louise Brown, occurred in 1978. Prior to that, there was a transient biochemical pregnancy reported by Australian Foxton School researchers in 1973 and an ectopic pregnancy reported by Steptoe and Edwards in 1976. The term in vitro, from the Latin root meaning within the glass, is used, because early biological experiments involving cultivation of tissues outside the living organism from which they came, were carried out in glass containers such as beakers, test tubes, or petri dishes. Today, the term in vitro is used to refer to any biological procedure that is performed outside the organism it would normally be occurring in, to distinguish it from an in vivo procedure, where the tissue remains inside the living organism within which it is normally found. A colloquial term for babies conceived as the result of IVF, test tube babies, refers to the tube-shaped containers of glass or plastic resin, called test tubes, that are commonly used in chemistry labs and biology labs. However, in vitro fertilisation is usually performed in the shallower containers called Petri dishes. (Petri dishes may also be made of plastic resins.) However, the IVF method of Autologous Endometrial Coculture is actually performed on organic material, but is yet called in vitro. This is used when parents are having infertility problems or they want to have multiple births.

Indications
IVF may be used to overcome female infertility in the woman due to problems of the fallopian tube, making fertilisation in vivo difficult. It may also assist in male infertility, where there is defect sperm quality, and in such cases intracytoplasmic sperm injection (ICSI) may be used, where a sperm cell is injected directly into the egg cell. This is used when sperm have difficulty penetrating the egg, and in these cases the partner's or a donor's sperm may be used. ICSI is also used when sperm numbers are very low. ICSI results in success rates equal to those of IVF fertilisation. For IVF to be successful it may be easier to say that it requires healthy ova, sperm that can fertilise, and a uterus that can maintain a pregnancy. Due to the costs of the procedure, IVF is generally attempted only after less expensive options have failed. This also avails for egg donation or surrogacy where the woman providing the egg isn't the same who will carry the pregnancy to term. This means that IVF can be used for females who have already gone through menopause. The donated oocyte can be fertilised in a crucible. If the fertilisation is successful, the zygote will be transferred into the uterus, within which it will develop into an embryo.

IVF can also be combined with preimplantation genetic diagnosis (PGD) to rule out presence of genetic disorders. A similar but more general test has been developed called Preimplantation Genetic Haplotyping (PGH).

Method
Ovarian stimulation
Treatment cycles are typically started on the 3rd day of menstruation and consist of a regimen of fertility medications to stimulate the development of multiple follicles of the ovaries. In most patients injectable gonadotropins (usually FSH analogues) are used under close monitoring. Such monitoring frequently checks the estradiol level and, by means of gynecologic ultrasonography, follicular growth. Typically approximately 10 days of injections will be necessary. Spontaneous ovulation during the cycle is typically prevented by the use of GnRH agonists that are started prior or at the time of stimulation or GnRH antagonists that are used just during the last days of stimulation; both agents block the natural surge of luteinising hormone (LH) and allow the physician to initiate the ovulation process by using medication, usually injectable human chorionic gonadotropins.

Egg retrieval
When follicular maturation is judged to be adequate, human chorionic gonadotropin (hCG) is given. This agent, which acts as an analogue of luteinising hormone, would cause ovulation about 42 hours after injection, but a retrieval procedure takes place just prior to that, in order to recover the egg cells from the ovary. The eggs are retrieved from the patient using a transvaginal technique involving an ultrasound-guided needle piercing the vaginal wall to reach the ovaries. Through this needle follicles can be aspirated, and the follicular fluid is handed to the IVF laboratory to identify ova. It is common to remove between ten and thirty eggs. The retrieval procedure takes about 20 minutes and is usually done under conscious sedation or general anaesthesia.

Fertilisation
In the laboratory, the identified eggs are stripped of surrounding cells and prepared for fertilisation. In the meantime, semen is prepared for fertilisation by removing inactive cells and seminal fluid in a process called sperm washing. If semen is being provided by a sperm donor, it will usually have been prepared for treatment before being frozen and quarantined, and it will be thawed ready for use. The sperm and the egg are incubated together at a ratio of about 75,000:1 in the culture media for about 18 hours. In most cases, the egg will be fertilised by that time and the fertilised egg will show two pronuclei. In certain situations, such as low sperm count or motility, a single sperm may be injected directly into the egg using intracytoplasmic sperm injection (ICSI). The fertilised egg is passed to a special growth medium and left for about 48 hours until the egg consists of six to eight cells.

In gamete intrafallopian transfer, eggs are removed from the woman and placed in one of the fallopian tubes, along with the man's sperm. This allows fertilisation to take place inside the woman's body. Therefore, this variation is actually an in vivo fertilisation, not an in vitro fertilisation.

Embryo culture
Typically, embryos are cultured until having reached the 68 cell stage three days after retrieval. In many Canadian, American and Australian programmes[citation needed], however, embryos are placed into an extended culture system with a transfer done at the blastocyst stage at around five days after retrieval, especially if many good-quality embryos are still available on day 3. Blastocyst stage transfers have been shown to result in higher pregnancy rates. In Europe, transfers after 2 days are common. Culture of embryos can either be performed in an artificial culture medium or in an autologous endometrial coculture (on top of a layer of cells from the woman's own uterine lining). With artificial culture medium, there can either be the same culture medium throughout the period, or a sequential system can be used, in which the embryo is sequentially placed in different media. For example, when culturing to the blastocyst stage, one medium may be used for culture to day 3, and a second medium is used for culture thereafter. Single or sequential medium are equally effective for the culture of human embryos to the blastocyst stage. Artificial embryo culture media basically contain glucose, pyruvate, and energy-providing components, but addition of amino acids, nucleotides, vitamins, and cholesterol improve the performance of embryonic growth and development.

Embryo selection
Laboratories have developed grading methods to judge oocyte and embryo quality. In order to optimize pregnancy rates, there is significant evidence that a morphological scoring system is the best strategy for the selection of embryos.[ However, presence of soluble HLA-G might be considered as a second parameter if a choice has to be made between embryos of morphologically equal quality In addition to tests that optimize pregnancy chances, Preimplantation Genetic Diagnosis (PGD) may be performed prior to transfer in order to avoid inheritable diseases.

Embryo transfer
Embryos are graded by the embryologist based on the number of cells, evenness of growth and degree of fragmentation. The number to be transferred depends on the number available, the age of the woman and other health and diagnostic factors. In countries such as Canada, the UK, Australia and New Zealand, a maximum of two embryos are transferred except in unusual circumstances. In the UK and according to HFEA regulations, a woman over 40 may have up to three embryos transferred, whereas in the USA, younger women may have many embryos transferred based on individual fertility diagnosis. Most clinics and country regulatory bodies seek to minimise the risk of

pregnancies carrying multiples. The embryos judged to be the "best" are transferred to the patient's uterus through a thin, plastic catheter, which goes through her vagina and cervix. Several embryos may be passed into the uterus to improve chances of implantation and pregnancy.

Pregnancy rates
Pregnancy rate is the success rate for pregnancy. For IVF, it is the percentage of all attempts that lead to pregnancy, which generally refers to treatment cycles where eggs are retrieved and fertilised in vitro. Statistics referring to "pregnancy" may refer to just a positive pregnancy test, and not necessarily "viable pregnancy" which implies the detection of a fetal heart beat. Pregnancies that are delivered with a viable baby are called live birth rate. Increasingly a distinction is also made between singleton and multiple pregnancies as multiple pregnancies, specifically more than twins, should be avoided because of the associated maternal and fetal risks. With enhanced technology, the pregnancy rates are substantially better today than a couple of years ago. In 2006, Canadian clinics reported an average pregnancy rate of 35%. A French study estimated that 66% of patients starting IVF treatment finally succeed in having a child (40% during the IVF treatment at the center and 26% after IVF discontinuation). Achievement of having a child after IVF discontinuation was mainly due to adoption (46%) or spontaneous pregnancy (42%).

Semen
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Human semen in a Petri dish. Semen is an organic fluid, also known as seminal fluid, that usually contains spermatozoa. It is secreted by the gonads (sexual glands) and other sexual organs of male or hermaphroditic animals and can fertilize female ova. In humans, seminal fluid contains several components besides spermatozoa: proteolytic and other enzymes as well as fructose are elements of seminal fluid which promote the survival of spermatozoa and provide a medium through which they can move or "swim". The process that results in the discharge of semen is called ejaculation.

Physiological aspects
Internal and external fertilization
Depending on the species, spermatozoa can fertilize ova externally or internally. In external fertilization, the spermatozoa fertilize the ova directly, outside of the female's sexual organs. Female fish, for example, spawn ova into their aquatic environment, where they are fertilized by the semen of the male fish. During internal fertilization, however, fertilization occurs inside the female's sexual organs. Internal fertilization takes place after insemination of a female by a male through copulation. In low vertebrates (amphibians, reptiles, birds and monotreme mammals), copulation is achieved through the physical mating of the cloaca of the male and female. In marsupial and placental mammals, copulation occurs through the vagina.

Composition of human semen


During the process of ejaculation, sperm passes through the ejaculatory ducts and mixes with fluids from the seminal vesicles, the prostate, and the bulbourethral glands to form the semen. The seminal vesicles produce a yellowish viscous fluid rich in fructose and

other substances that makes up about 70% of human semen. The prostatic secretion, influenced by dihydrotestosterone, is a whitish (sometimes clear), thin fluid containing proteolytic enzymes, citric acid, acid phosphatase and lipids. The bulbourethral glands secrete a clear secretion into the lumen of the urethra to lubricate it. Sertoli cells, which nurture and support developing spermatocytes, secrete a fluid into seminiferous tubules that helps transport sperm to the genital ducts. The ductuli efferentes possess cuboidal cells with microvilli and lysosomal granules that modify the semen by reabsorbing some fluid. Once the semen enters the ductus epididymis the principle cells, which contain pinocytotic vessels indicating fluid reabsorption, secrete glycerophosphocholine which most likely inhibits premature capacitation. The accessory genital ducts, the seminal vesicle, prostate glands, and the bulbourethral glands, produce most of the seminal fluid. Seminal plasma of humans contains a complex range of organic and inorganic constituents. The seminal plasma provides a nutritive and protective medium for the spermatozoa during their journey through the female reproductive tract. The normal environment of the vagina is a hostile one for sperm cells, as it is very acidic (from the native microflora producing lactic acid), viscous, and patrolled by immune cells. The components in the seminal plasma attempt to compensate for this hostile environment. Basic amines such as putrescine, spermine, spermidine and cadaverine are responsible for the smell and flavor of semen. These alkaline bases counteract the acidic environment of the vaginal canal, and protect DNA inside the sperm from acidic denaturation. The components and contributions of semen are as follows: Gland Approximate % Description Approximately 200- to 500-million spermatozoa (also testes 2-5% called sperm or spermatozoans), produced in the testes, are released per ejaculation. amino acids, citrate, enzymes, flavins, fructose (the main energy source of sperm cells, which rely entirely on seminal sugars from the seminal plasma for energy), 65-75% vesicle phosphorylcholine, prostaglandins (involved in suppressing an immune response by the female against the foreign semen), proteins, vitamin C acid phosphatase, citric acid, fibrinolysin, prostate specific antigen, proteolytic enzymes, zinc (serves to help to stabilize the DNA-containing chromatin in the prostate 25-30% sperm cells. A zinc deficiency may result in lowered fertility because of increased sperm fragility. Zinc deficiency can also adversely affect spermatogenesis.) bulbourethral < 1% galactose, mucus (serve to increase the mobility of sperm glands cells in the vagina and cervix by creating a less viscous

channel for the sperm cells to swim through, and preventing their diffusion out of the semen. Contributes to the cohesive jelly-like texture of semen.), preejaculate, sialic acid A 1992 World Health Organization report described normal human semen as having a volume of 2 ml or greater, pH of 7.2 to 8.0, sperm concentration of 20106 spermatozoa/ml or more, sperm count of 40106 spermatozoa per ejaculate or more, and motility of 50% or more with forward progression (categories a and b) of 25% or more with rapid progression (category a) within 60 minutes of ejaculation.

Appearance and consistency of human semen

Human semen Most semen is white, but grey or even yellowish semen can be normal as well. Blood in the semen can cause a pink or reddish colour, known as hematospermia, and may indicate a medical problem which should be evaluated by a doctor if it does not readily disappear. After ejaculation, semen first goes through a clotting process and then becomes more liquid. It is postulated that the initial clotting helps keep the semen in the vagina, but liquefaction frees the sperm to make their long journey to the ova. Immediately after ejaculation semen is typically a sticky, jelly-like liquid often forming globules. Within 5 to 40 minutes it will become more watery and liquid before finally drying.[

Semen quality
Semen quality is a measure of the ability of semen to accomplish fertilization. Thus, it is a measure of fertility in a man. It is the sperm in the semen that is the fertile component, and therefore semen quality involves both sperm quantity and sperm quality.

Health effects

In addition to its central role in reproduction, various scientific findings indicate that semen has certain beneficial effects on human health, both proven benefits and possible benefits:

Antidepressant: One study suggested that vaginal absorption of semen could act as an antidepressant in women; the study compared two groups of women, one of which used condoms and the other did not. Cancer prevention: Studies suggest that seminal plasma both prevents and fights cancer, particularly breast cancer, reducing risk by "not less than 50 percent." This effect is attributed to its glycoprotein and selenium content, with apoptosis being induced by TGF-Beta. A related urban legend parodied these findings and claimed that performing fellatio at least three times a week reduced the risk of breast cancer. Preeclampsia prevention: It has been hypothesized that substances in semen condition a mother's immune system to accept the "foreign" proteins found in sperm as well as the resulting fetus and placenta, keeping blood pressure low and thereby reducing the risk of preeclampsia. A study shows that oral sex and swallowing sperm may help make a woman's pregnancy safer and more successful, because she is absorbing her partner's antigens.

Semen and transmission of disease


Semen can be the vehicle for many sexually transmitted diseases, including HIV, the virus that causes AIDS. Further research, such as that by Mathur and Goust, demonstrated that non-preexisting antibodies were produced in humans in response to the sperm. These antibodies mistakenly recognized native T lymphocytes as foreign antigens, and consequently T lymphocytes would fall under attack by the body's B lymphocytes. Semen contains proteins with potent bactericidal activity but these proteins are not active against Neisseria gonorrhoeae, a common cause of sexually transmitted disease.

Blood in the semen (hematospermia)


The presence of blood in semen or hematospermia may be undetectable (it only can be seen microscopically) or visible in the fluid. Its cause could be the result of inflammation, infection, blockage, or injury of the male reproductive tract or a problem within the urethra, testicles, epididymis or prostate. It usually clears up without treatment, or with antibiotics, but if persistent further semen analysis and other urogenital system tests might be needed to find out the cause.

Semen allergy
In rare cases, people have been known to experience allergic reactions to seminal fluids, known as human seminal plasma hypersensitivity. Symptoms can be either localized or systemic, and may include vaginal itching, redness, swelling, or blisters within 30 minutes of contact. They may also include generalized itching, hives, and even difficulty breathing. One way to test for human seminal plasma sensitivity is to use a condom during intercourse. If symptoms dissipate with the use of a condom, it is possible that a sensitivity to semen is present. Mild cases of semen allergy can often be overcome by repeated exposure to seminal fluid. In more severe cases, it is important to seek the advice of a physician, particularly in the event that a couple is trying to conceive, in which case, artificial insemination may be indicated.

Nutritional value
Semen is primarily water, but contains trace amounts of almost every nutrient the human body uses. It has somewhat higher amounts of commonly deficient minerals, such as potassium, magnesium, and selenium.[35] One typical ejaculation contains 150 mg of protein, 11 mg of carbohydrates, 6 mg fat, 3 mg cholesterol, 7% US RDA potassium and 3% US RDA copper and zinc. When metabolized, protein yields 4 kcal/g, carbohydrate also yields 4 kcal/g, and fat yields 9 kcal/g. Hence the food energy in the typical ejaculation is 0.7 kcal (2.9 kJ).

Quantity
The volume of semen ejaculate varies. A review of 30 studies concluded that the average was around 3.4 milliliters (ml), with some studies finding amounts as high as 4.99 ml or as low as 2.3 ml.

Embryo Cloning
Embryo cloning is an area of stem-cell research that focuses on embryos and is the process of harvesting human stem cells for scientific study. Embryonic stemcell research is sometimes also referred to as research cloning and therapeutic cloning. Cloning is a general term that scientists use to describe various processes for duplicating biological organisms.. Introduction A common factor between different cloning technologies is that all cloning techniques produce genetic twins of living organisms. The three basic types of cloning include recombinant DNA technology or DNA cloning, reproductive cloning, and therapeutic cloning. DNA cloning (also called molecular and gene cloning) refers to the process of transferring DNA fragments from one organism to another, according to the U.S. Department of Energy Office of Science. Reproductive cloning is the process of generating an animal with same nuclear DNA as another, or previously existing animal (the technique used to create the sheep clone, Dolly). And therapeutic cloning (also referred to as embryo cloning) is the production of human embryos for scientific research. Dolly Dolly was the first mammal to ever be cloned. She was cloned using the technique of reproductive cloning (not embryo cloning) at the Roslin Institute in Scotland in 1997. This significant scientific breakthrough generated much scientific and ethical debate. Dolly was cloned from adult DNA and lived to the age of 6. Towards the end of her life, she suffered from lung cancer and arthritis. Other than that, she lived quite a normal life for a sheep, with few physical problems, according to the U.S. Department of Energy Office of Science. Embyro Cloning Embryo cloning involves collecting eggs from ovaries and removing genetic material from the eggs. After genetic material is removed, a skin cell is inserted into the enucleated egg in order to serve as the egg's new nucleus. When the chemical ionomycin is applied, the egg begins to divide. Stem cells are extracted from the egg five days after division. During this stage of development, the egg is a blastocyst, and the extraction of stem cells destroys the embryo. The end goal of embryo cloning is not to clone a human being, but rather to harvest stem cells for medical research.

Function In November 2001, Advanced Cell Technologies was the first company to clone a human embryo for the purpose of embryo-cloning research. Since then there have been certain breakthroughs in therapeutic cloning research, such as 2003's production of therapeutic nerve cells that cure Parkinson's like disease in mice. Furthermore, in 2006, Children's Hospital Boston announced that it begun experiments with somatic cell nuclear transfer in order to clone diseases such as sickle-cell anemia and diabetes. Benefits Stems cells are important new area of biomedical research because they have the ability to grow into virtually any type of specialized cell. As a result, scientists hope that they can be used to treat a variety of diseases and disorders. Embryo cloning may some day be used to produce healthy cells to replace damaged cells and to create entire organs from a single stem cell. A lot of research still needs to be done, however, embryo cloning has the potential to be a cure for many degenerative disease including cancer and Alzheimer's.

Human embryo cloning


Identification
Human embryo cloning, also called "therapeutic cloning," is different that the conventional understanding of cloning. Scientists use human embryo cloning not to create another human, but to grow an embryo to harvest the cells for research. The stem cells that are removed from the embryo can be used to study any type of human cellular disease, as stem cells have the potential to become any cell in the human body. Scientists can also experiment with cellular therapies to cure a variety of diseases.

Process
Embryo cloning does not require a fertilized embryo as traditional cloning does. Instead, scientists get a sample of cells from the individual they wish to clone, usually by scraping the inside of the cheek for DNA, and insert it into a donated ovum that has been stripped of its genetic material. A slight electric shock is then sent through the ovum, which sometimes results in cell division. A pre-embryo then forms, which can produce a group of stem cells. The embryo can be replanted into a woman's womb, resulting in a pregnancy and a traditional clone, but this step is omitted in embryo cloning. After the pre-embryo has divided, scientists use a needle that is less than 1/5,000 of an inch wide to remove the stem cells. This process kills the embryo. These stem cells can then

be stimulated to grow into any human tissue, which may then be transplanted onto the patient.

Therapeutic Uses
An organ grown from stem cells can be transplanted into a patient with no fear of rejection, since the genetic material of the organ matches that of the host. Patients would also no longer need to wait on the organ transplant list before a donor becomes available, as each organ can be grown whenever it is needed. The organ would also be brand new, meaning doctors would not have to worry about decreased function or any viral or bacterial diseases from the donor being transplanted with the organ.

Controversy
Despite its potential therapeutic benefits, embryo cloning is still controversial. This is primarily because the embryo that results from the cloning process is killed when its stem cells are harvested. Because the pro-life position is that human life starts at conception, the destruction of an embryo is viewed as murder. While stem cell research has gained traction in countries with a very small pro-life movement, like China, countries with a stronger tendency toward pro-life, like the United States, have largely rejected stem cell therapy.