The problem and review of related literature and studies

Introduction
Everyone is naturally pre-determined to have an offspring. This would just exist in a relationship of a man and woman. It is not always true in a relationship if there is a new life that will be formed. Several factors may contribute to the hindrances in the formation of new life especially to man because male (sperm) gamete is the one responsible to initiate fertilization, the union of male (sperm) and female (ovum) gametes.

These may include low sperm counts, poor sperm quality, anatomical problems and hormonal imbalances.

These factors were also described under male infertility which leads to inability to produce offspring. Pre-fabricated drugs to treat this are readily available in the market, but it is associated with some undesirable effects.

Talahib (Saccharum spontaneum) is one of the most abundant plants found in the Philippines especially in open areas. The different parts of talahib are pharmaceutically used as diuretic, poultice, purgative, astringent and emollient. It is also found to be an aphrodisiac that has an effect to arouse sexual desire which may resemble to boost fertility.

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The researchers aim to prove the effectiveness of ethanolic extract of talahib on spermatocyte proliferation in male rabbit (Oryctolagus cuniculus).

The leaves of talahib will be used in this experiment. The researchers will also conduct phytochemical screening for the determination of the different chemical constituents present in the talahib leaves.

Thus, if the ethanolic extract of talahib leaves will be proven effective, it would be beneficial to those couple who desires to have an offspring most especially to male which is the focused of this study on spermatocyte proliferation.

Statement of the Problem
This study deals with the determination of the efficacy of ethanolic extract of talahib leaves on spermatocyte proliferation. Specifically, the study
aims to answer the following:
1. What are the different phytochemical constituents present in the ethanolic extract of talahib leaves? 2. Is there a correlation between testosterone level and sperm count in different treatments of ethanolic extract of talahib leaves? 3. Is there a significant difference on spermatocyte proliferation between the different treatments of ethanolic extract of talahib leaves to that of Proviron? Hypotheses

Ho – There is no significant relationship between testosterone level and sperm count in different treatments of ethanolic extract of talahib leaves. Ho – There is no significant difference between ethanolic extract of talahib leaves and Proviron as commercially used agent for infertility. Significance of the Study

The study has significant bearing information on the spermatocyte proliferation of ethanolic extract of talahib leaves. Thus, the current study is significant to the following: Community. The result of the study will be helpful to the community which can provide an affordable and alternative source for the treatment of infertility problems especially to man. Department of Health (DOH)/ Department of Science and Technology (DOST). The results of the study will give additional information about the use of talahib as a source of treatment for infertility problems to man.

Pharmaceutical Industries. The result of the study would be of great help in the formulation of new agent for treatment of infertility most probably if it would be more potent than the commercially available drug.

Researchers. This study helps the researchers to invest knowledge to the status and have a view on the medicines available in the market that will enhance the production of spermatocytes. Future Researchers. This study will serve as the basis and reference of the future researchers especially if their study focused on spermatocyte proliferation. Setting of the Study The phytochemical screening of the different constituents present in the ethanolic extract of talahib leaves will be conducted at St. Paul University Philippines (SPUP) College-Science Laboratory. The talahib leaves will be collected at Masical, Baggao, Cagayan. Scope and Delimitation

This study deals primarily on the efficacy of ethanolic extract of talahib leaves on the spermatocyte proliferation. The extract will be subjected to phytochemical screening to determine the presence of secondary metabolites such as Alkaloids, Quaternary and/or Amine Oxidase Bases, Flavonoids, Tannins and Polyphenolic Compounds, Carbohydrates, Saponin Glycosides, Proteins, Fixed Oils, Fats, and Volatile Oils and Anthraquinones.

The researchers will administer the extract through oral gavage in the male white rabbit. This study will quantify the testosterone level, number of sperm, morphology of sperm and motility of sperm. This will determine the effect of ethanolic extract of talahib leaves on spermatocyte proliferation. The administration of the ethanolic extract of talahib leaves with different treatments of 0.25 grams, 0.10 grams, 0.05 grams, 0.025 grams and 0.01 grams of one hundred (100) percent concentration will determine the capacity of talahib leaves on spermatocyte proliferation.

Conceptual Framework

Figure 1: Paradigm of the Study

Theoretical Framework
The ethanolic extract of talahib leaves will be used in this study to determine its effect on spermatocyte proliferation. The researchers will be using sexually mature male white rabbits as the test animal from which the seminal fluid and the blood will be withdrawn to be used for semen analysis and testosterone level analysis respectively. The test animal will be given equal amounts of food and water and placed in the same environment for twenty eight (28) days then initial analysis of the semen and blood of rabbits will be conducted to eradicate physiologic factors that may affect the results of the study.

The seminal fluid will be collected through artificial vagina (AV) for semen analysis and the blood will be collected through the marginal ear veins for testosterone level analysis. After the initial analysis, different treatments will be prepared for administration for another twenty eight (28) days. Each treatment consists of three (3) replicates given in five doses of ethanolic extract of talahib leaves, the positive control (Proviron) and negative control for twenty eight (28) days.

Review of Related Literature and Studies Wild sugarcane (Saccharum spontaneum) is a coarse, erect, perennial, usually more or less tufted or gregarious grass, attaining a height of 1 to 3.5 meters, and rising from stout underground rootstock. The leaves are harsh, linear, 0.5 to 1 meter long, 6 to 15 millimeters wide. The panicles are white, erect, 15 to 30 centimeters long, with the branches slender, whorled, fragile, and the joints covered with long, soft, white hairs.

The spikelets are about 3.5 millimeters long, very much shorter than the copious, long, white hairs at the base. It is locally known as talahib (Tag.), sikal (Ibn), lidda (Ilk).

Talahib is found in open areas at low and medium altitudes, ascending to 1,500 meters, and often gregarious, occupying almost exclusively large areas throughout the Philippines. A decoction of the roots is used in the Philippines as a diuretic. According to Nadkarni, the roots are used as a galactagogue and diuretic (an agent that promotes renal excretion of electrolytes and water).

Sulit reports that the warm pulp of the stem is used as a poultice and applied to painful parts of the legs, and in cases of suffering from beriberi. Talahib is not just an ordinary plant, but a useful plant for it possesses many medicinal properties which it could really help people especially those who are suffering from financial problems because talahib can be found anywhere and can be prepared easily by themselves. Researchers from the Natural News consider environmental pollution, plastic chemicals, pharmaceutical drugs, pesticides, unhealthy diets; technologies such as cell phones can contribute to an epidemic decline in sperm counts among modern men.

Researchers from the Cleveland Clinic Lerner College of Medicine at Case Western Reserve University in Ohio found that regular cell phone use leads to a significant decline in sperm count. According to reports, an estimated eight million US couples now have fertility problems, and about half of these cases involve men with poor sperm quality. Last year, a five-year study found that bisphenol-A (BPA), a chemical widely used in many plastic products, is responsible for destroying sperm.

Another study presented at the 2006 conference of the American Society for Reproductive Medicine found that common selective serotonin reuptake inhibitor (SSRI) antidepressant drugs can literally bring a man's sperm count to zero. And a recent study out of the University Hospital San Cecilio, Granda, Spain, found that common pesticides used on food crops lead to poor quality and lowered sperm counts. Obesity, eating junk food and drinking sodas, using a laptop on your lap, and even taking too many hot baths can also destroy sperm.

Male infertility isn't something you hear much about on the news, you may be surprised to know that male infertility is almost as likely as female infertility to be involved in a couple's inability to achieve pregnancy. The good news is that most cases of male infertility can be resolved either by treating the problem or using fertility treatments. When this is not the case, a couple facing male infertility may turn to a sperm donor or adoption to help build their family.

About ten to fifteen percent (10-15%) of couples will not be able to achieve pregnancy after one year of unprotected intercourse. Out of this group, the following statistics on the cause of infertility generally apply: Thirty to forty percent (30-40%) of couples will discover fertility problems in both partners. About twenty percent (20%) of couples will discover fertility problems in only the man. About thirty to forty percent (30-40%) will discover fertility problems in only the woman. About ten percent (10%) of couples will not find a reason for their infertility (also known as "unexplained infertility”.

Potential causes of male infertility includes complete absence of sperm (azoospermia), low sperm count (oligospermia), abnormal sperm shape (teratozoospermia), problems with sperm movement (asthenozoospermia), sperm that is completely immobile (necrozoospermia); the sperm may be alive and not moving, or they may be dead, problems with sperm delivery, due to sexual dysfunction, an obstruction, previous vasectomy, or retrograde ejaculation, and problems with erections or other sexual problems.

There are a variety of conditions that may lead to male infertility. The most common cause of male infertility is varicoceles. A varicocele is a varicose vein found in the scrotum. The extra heat caused by the vein can lead to low sperm count and impaired sperm movement. Several causes of male infertility came from what was taken in and doings in everyday life. One of the causes of male infertility is the low production of sperm cells which is considered to be the focus of this study.

These would help the researchers and readers for it provides information for the prevention of such problems especially on the part of the male. (http://infertility.about.com/od/causesofinfertility/a/male_infertility.htm) In men, the terms “erectile dysfunction, sterility and infertility” are often confused and mistakenly inter-exchanged with each other. However, in reality, these denote three different conditions.

It is, therefore, very important for every adult man who is concerned about his sexual and overall health to know and understand the basic difference among these three disorders that could have a profound psychosocial impact on each of any man. Sterility is often taken as a synonym to infertility, though they both mean disability to bear a child yet they have a very basic difference. Sterility means that it is impossible for a couple to conceive a child. A diagnosis of sterility is given after a thorough examination concludes that there is no sperm production and ovulation cannot occur.

In simpler words, sterility means that the person is unable to reproduce sexually; this mostly is because of problem in the reproductive system. Sterility is very restricted to the reproductive system like damage in ovaries or veins in the testicles might cause sterility. Infertility refers to inability to conceive or not being able to get pregnant after one year of trying. It also means that a couple is not sterile but for some reason has not been able to conceive a child. Infertility might exist because of certain abnormalities that might not be related to the reproductive system.

At times infertility exists because of stress, emotional shocks or side effect of any treatment. There are three conditions that need to be met for conception to be possible: sperm must be present, the fallopian tubes must be open, and ovulation must be able to occur. If one or more of these conditions is not met, the couple suffers from “true infertility.” If all three conditions are met but the couple has failed to conceive, the diagnosis is “sub-fertility.” Infertility in men is not an uncommon condition. About one-third of infertility cases are caused by women’s problems.

Another one-third of fertility problems are due to the man. The other cases are caused by a mixture of male and female problems or by unknown problems. Infertility in men is most often caused by a problem called varicocele. This happens when the veins on a man’s testicle(s) are too large. This heats the testicles. The heat can affect the number or shape of the sperm. Other factors that cause a man to make too few sperm or none at all includes the movement of the sperm which may be caused by the shape of the sperm. Sometimes injuries or other damage to the reproductive system block the sperm. Sometimes a man is born with the problems that affect his sperm.

A man's sperm can be changed by his overall health and lifestyle. Some things that may reduce the health or number of sperm include: heavy alcohol use, drugs, smoking cigarettes, age, environmental toxins, including pesticides and lead, health problems such as mumps, serious conditions like kidney disease, or hormone problems, medicines, and radiation treatment and chemo therapy for cancer. Erectile dysfunction or impotence is the persistent or recurrent inability to attain or maintain an erection sufficient to complete sexual intercourse or another chosen sexual activity.

It is very common and affects at least one in every ten men. This means that there are an estimated 2.3 million men in the UK suffering from erectile dysfunction. Regrettably, only about 10% of sufferers actually receive treatment. The number of men suffering erectile dysfunction increases with age. Most men have an occasional failure to get or keep an erection. It usually results from stress, tiredness, anxiety, depression, or excessive alcohol consumption.

The statements above represent the differences between the commonly mistaken or confused terms, erectile dysfunction and infertility. It would help the researchers and the readers to distinguish these terms from each other. Erectile dysfunction is defined as the inability of a male to maintain an erection sufficient to complete sexual intercourse or activity while for infertility, it is referred the inability to conceive due to low amount or absence of sperm cell production. (http://smallpenises.blogspot.com/2011/01/what-are-core differences-between.html) During spermatogenesis, meiosis, or reduction division occurs.

The spermatogonia are stem cells containing 46 chromosomes that divide by mitosis to produce two daughter cells. One remains a stem cell. The other becomes a progenitor spermatogonium, destined to develop into sperm. This spermatogonium divides by mitosis to form two primary spermatocytes, each with 46 chromosomes. The primary spermatocytes then complete the first meiotic division, producing two secondary spermatocytes, eachof which contains 23 chromosomes (1N). The secondary spermatocytes subsequently undergo the second meiotic division to produce four spermatids, each with 23 chromosomes, 22 autosomes, and one sex chromosomes (X or Y).

Spermatogenesis and male fertility are dependent upon the presence of testosterone in the testis. In the absence of testosterone or the androgen receptor, spermatogenesis does not proceed beyond the meiosis stage. The major cellular target and translator of testosterone signals to developing germ cells is the Sertoli cell. (http://www.landesbioscience.com/journals/47/article/16956) Some hormones are involved in the sperm production and proliferation of sperm cells.

Gonadotropic hormones secreted by the hypothalamus stimulate the Leutinizing hormone (LH) and the Follicle Stimulating Hormone (FSH). Leutenizing hormone (LH) goes to their target cells in the testes called the Leydig cells causing the production of testosterone that can influence spermatogenesis. Follicle Stimulating Hormone (FSH) goes to seminiferous tubules where the sperm cell is produced causing spermatocyte to develop into the sperm.

Increase in the level of follicle stimulating hormone results to adequate amount of testosterone level and the development of spermatogenesis proceeds. Testosterone is a hormone that is secreted in the testes influencing the production of sperm and producing secondary sex hormones. In the seminiferous are also sertoli cell which are designed to nourish, mature and develop the sperm.

Increase production of sperm stimulates the release of inhibin from the sertoli cells. Inhibin is a hormone that feeds back to pituitary gland, modulating FSH production, without affecting LH production. (http://www.sglearnonline.com/reproductive-system.html ; Essential of anatomy and physiology 6th edition by Seeley, Stephen, Tate Testosterone is a hormone found in men. It is produced in the testis in men and in the adrenal gland in relatively smaller amounts.

It is not directly responsible for the production of sperm cells, but it is however still a part of the process: when testosterone levels drop so does the production of sperm. It is also an integral part of men’s sexual activity, tied in with his libido and sex drive.

Hormone levels that are affected by the testosterone in the body can cause problems with ejaculation and erections. This is the primary cause of erectile dysfunction. There are many reasons for the decline of testosterone levels: age, stress, fatigue, and medical conditions. This is especially problematic if you’re trying to conceive a baby with your wife. Testosterone isn't the only fuel for a man's sex drive and performance. But low testosterone can reduce your ability to have satisfying sexual intercourse. Lack of sex drive and erectile dysfunction are sexual problems that can result from low testosterone.

In a large study of men in Massachusetts, about 11% overall said they had a lack of sex drive. The researchers then tested all the men's testosterone levels. About 28% of men with low testosterone had low libido. These men were relatively young, with an average age of 47; older men might have worse sexual symptoms. (http://www.enduranz.ph/blog/the-link-between-testosterone-and-sperm-production/)

Testosterone can reduce the ability to have a satisfying intercourse due to lack of sex drive and erectile dysfunction which results from the low level of this hormone. It is also considered as a part of the process for sperm production wherein a low level of this hormone, may lead to a reduced amount of sperm cell. This information would help the researchers since the parameters include testosterone level in determining the amount of sperm produced.

According to the Fertility Institutes, Laboratories performing sperm "counts", in general, vary in the details that they provide the physician requesting the "count". A general sperm count as part of a fertility evaluation should include the total density or count (20 million per ml or above), and the motile density (8 million per ml or higher). The motile density is perhaps the most important part of the semen analysis, as it reports the total number of sperm thought capable of progressing from the site of sperm deposition to the site of fertilization.

This allows determining if the male have normal semen value. Sperm morphology is the evaluation of sperm size, shape and appearance. It should be assessed by carefully observing a stained sperm sample under the microscope. The addition of colored “dyes” (stains) to the sperm allows the observer to distinguish important normal characteristics as well as abnormal findings. Several methods of staining sperm are used, and the method employed should be one with which the examiner is comfortable. WHAT ARE THE METHODS? Several shapes or forms of human sperm have been identified and characterized. These