Putative S-Adenosyl Methionine Dependent Ironâ€Sulfur

Putative S-Adenosyl Methionine Dependent Iron–Sulfur Identification and Characterization of a Putative S-Adenosyl Methionine dependent Iron–Sulfur containing protein from  Methanococcus Jannaschii Qi Liu Research Proposal Dr. MishtuDey Dr. M. Lei Geng Dr. Leonard R. MacGillivray Dr. Amnon Kohen Dr. Daniel Quinn Introduction Biological methane formation is a microbial process that is catalyzed by microbes called methanogens, which belong to the third kingdom of life, the Archaea. Methane is formed at the final catalytic step by methyl-coenzyme M reductase (MCR), in which coenzyme B (CoBSH, N-7-mercaptoheptanoylthreonine) donates two electrons to reduce methyl-coenzyme M. MCR is a 300kDa enzyme, which is composed of three different subunits in an ÃŽ ±2ÃŽ ²2ÃŽ ³2 arrangement and contains 2 mol of the nickel tetrapyrrole coenzyme F430, which are buried deeply within the protein complex[1]. There are five modified amino acids were found out on the ÃŽ ±-subunit and near the active site of MCR from methanothermobacter marburgensis based on the X-ray crystallographic studies. They are 1-N-methylhistitine (HisÃŽ ±400), 5-(S)-methylarginine (ArgÃŽ ±271), 2-(S)-methylglutamine (GluÃŽ ±400), S-methylcysteine (CysÃŽ ±452), where the side chains are methylated and one thioglycine (GlyÃŽ ±445) forming a thiopeptide bo nd (Figure 1). Since the DNA sequence of the encoding MCR gene shows no unsusal condons or unusual codon usages at the positions in which the five modified amino acids were found, the modifications are introduced after translation [1]. According to vivo labeling experiments with L-(methyl-D3)-methionine, people found that the methyl groups from the modified amino acids are introduced biosynthetically from the methyl group of methionine by specific S-adenosylmethionine (SAM) dependent Figure1. Post Translational Modifications in MCR  enzymes. These methyl translational modifications are catalyzed by protein methylases that specifically recognize the amino acid sequences up and downstream of amino acid to be methylated. The genome of methanogens has many open reading frame predicted to be putative methyltransferases, which also agrees with the proposal that there are four different SAM-dependent protein methylases are involved in the post translation modification. A search of six kn own methanogenic genomes led to the identification of conserved open reading frame around the MCR gene cluster. Some methangenic archaea contain two MCR isoenzymes, designated MCR1 and MCR2. This conserved hypothetical protein is found around MCR1 in Methanococcus jannaschii and Methanobacterium thermobacter. The open reading frame from Methanococcus jannaschii, MJ0841 is annotated as a conserved hypothetical protein, which is found to be related to the radical SAM enzyme superfamily. The signature motif of SAM radical enzymes is three cysteine motif â€Å"CX3CX2C† (Figure 2), multiple sequence alignment of the putative gene from methanogens show the conserved CXGFCXXC, which is known to coordinate to [4Fe-4S] cluster. (Figure 3) Figure 2. Multiple Sequence Alignment of MJ0841 Homologues from Different Methanogens Figure 3. [4Fe–4S] cluster coordinated by three-cysteine motif CxxxCxxC. The fourth iron of the cluster interacts with a bound SAM. Specific Aims It is interesting and important to determine the function of this hypothetical protein. We speculate that this hypothetical protein, MJ0841, could be a possible candidate responsible for the post-translation modification of the methylated amino acids, or, is involved in the formation of the thioglycine in MCR. Research Plan, Results and Discussion Expression and Purification of MJ0841 MJ0841, a 1248bp gene, was cloned into pET28a vector. The resulting plasmid was transformed in E.coli BL21(DE3) cells for gene expression. In order to increase the iron content, MJ0841 was also co-expressed with PDB1282. Overnight cell culture grown at 37oC in Luria-Bertani (LB) medium containing both kanamycin (50ug/ml) and ampicillin (100ug/ml) was inoculated, in a 100-fold dilution, into Terrific Broth (TB) media aerobically at 37oC. FeCl3 was also added to a final concentration of 100uM to be as the iron content for iron-sulfur cluster during the growth when OD600 was around 0.3. Protein expression was then induced at OD600 of 0.6 to 0.7 with addition of Isopropyl ÃŽ ²-D-1-thiogalactopyranoside (IPTG) to final concentration of 0.5mM. After overnight incubation at 37oC around 18hours, the cells were harvested by centrifugation at 5000rpm for 30mins at 4oC, and stored at -80oC. The following procedures were all carried out in oxygen free environment at 20oC. Purification was conducted anaerobically at oxygen level always below 2.0ppm in anaerobic chamber. 23g Cell were moved into anaerobic chamber and resuspended in 120mL lysis buffer (50mM tris-HCl, 300mM NaCl, 5% glycerol pH 7.5), and Phenylmethanesulfonyl fluoride (PMSF) 1mM final concentration, 3 tablets of protease inhibitor, 2-mercaptomethanol 10mM) for 15mins. The cells were lysed by sonication for 15mins followed by centrifugation at 30,000 rpm for 40mins at 4oC to remove the cell debris. The supernatant was applied to a packed 15mL Ni-NTA resin column equilibrated with lysis buffer. The column was then washed with 5 column volume of wash buffer (50mM tris-HCl, 300mM NaCl, 10mM IMD, 5% glycerol, pH 7.5). The brownish protein was eluted by gradient elution with 5 column volume of wash buffer and 5 column volume elution buffer (50mM tris-HCl, 300mM NaCl, 200mM IMD, 5% glycerol, pH 7.5). SDS-PAGE was applied to analyze the desired clean protein fractions, which were then combined an d set for overnight dialysis with dialysis buffer (50mM tris-HCl, pH 7.5, 5% glycerol) with slow stirring. The pooled fractions were concentrated using an Amicon centrifugal filter with a 30kDa molecular weight cut off (MWCO). The collected protein was further purified with 20mL packed Q-sepharose column equilibrated with lysis buffer (50mM tris-HCl, 5% glycerol, pH 7.5). The column was then washed with 5 column volume of wash buffer (50mM tris-HCl, 200mM NaCl, 5% glycerol, pH 7.5). The brownish protein was eluted by gradient elution with 5 column volume of wash buffer and 5 column volume elution buffer (50mM tris-HCl, 700mM NaCl, 5% glycerol, pH 7.5). SDS-PAGE was applied to analyze the desired clean protein fractions, which were then combined for reconstitution (Figure 4). Figure 4. SDS-PAGE gel electrophoresis analysis of MJ0841 purification fractions Reconstitution of the [4Fe-4S] Cluster of MJ0841 in Vitro The above apo-protein (16uM, 40ml) was incubated with final concentration of 5mM DTT for 1h with slow stirring at room temperature. Then, cystein was added into the above solution by dropwise to reach 10 molar folds excess of protein. After 30mins incubation, 10 molar excess of Fe(NH4)2(SO4)2 was added slowly to provide enough iron content for iron sulfur cluster and incubated for 30mins. The resulting solution was incubated with 10 molar excess of Na2S finally, and the brownish protein solution changed to dark brown after adding Na2S. The above final protein solution was kept in 4oC overnight around 14 hours for building up enough [4Fe-4S] clusters. In order to remove the unbounded iron and sulfur, the overnight reconstituted protein was concentrated to 2.5 – 3ml of final volume and loaded onto a 5mL PT10 column equilibrated with lysis buffer (50mM tris-HCl, 5% glycerol, pH 7.5), and the final pure protein was combined. Reduction of [4Fe-4S] Cluster UV-vis spectroscopy was applied here for detecting the reduction of [4Fe-4S] cluster. A characteristic peak for [4Fe-4S] cluster was shown up near 412nm before reducing. Sodium Dithionite was used as the reducing agent and the stock solution was prepared freshly right before adding into the protein. 100 equivalents of sodium dithionite were mixed with concentrated protein, and the peak at 412nm was reduced (Figure 5). According to the results from UV-vis, the 4Fe-4S cluster was built up by reconstitution, also the [4Fe-4S]2+ was reduced to [4Fe-4S]+ by the reduction of sodium dithionite. Figure 5. UV-vis spectra of purified reconsituted MJ0841 Figure 6. EPR spectrum of as-isolated MJ0841 (blue trace) and purified reconstituted MJ0841 originating from a [3Fe-4S]+ cluster. reduced with 100 equiv sodium dithionite (red trace) EPR samples preparation and spectral collection EPR spectrums of as-isolated, oxidized and reduced form of MJ0841 are detected at 10K. All samples are prepared anaerobically. As-isolated protein was prepared with the protein without reconstitution, and exhibits a strong isotropic EPR signal, which is centered at g=2.01 same as the g=2.01 signal of the [3Fe-4S]+ cluster form (Figure 6). The oxidized form protein containing [4Fe-4S]2+ was prepared by injecting 200ul concentrated protein purified through PD10 column into EPR tube, and it normally shows silent EPR signal (Figure 7). Reduced form of as-isolated protein was performed by mixing with 100 equivalents of sodium dithionite with 200ul concentrated protein, which gives the reduction form of [4Fe-4S]+ cluster and shows the characteristic EPR signal with g factors of g=2.03 and g=1.92 (Figure 8). Figure 7. EPR spectrum of oxidized form of MJ0841. Figure 8. EPR spectrum of reduced form of MJ0841. SAM cleavage activity of MJ0841 The characteristic reaction for detecting radical SAM enzymes is reductive cleavage of SAM into S-adenosylhomocysteine (SAH) and 5’-deoxyadenosyl radical (5’dAdo). Assays were conducted under strict anaerobic conditions. The reaction assay contains the following: 50mM Tris-HCl, 5mM DTT, 5Mm sodium dithionite, 0.5mM SAM. Reactions were initiated by addition of SAM and carried out at 20oC for 20 hours. The control reaction was run under the same conditions as the above assay, but without presence of protein. Trifluoroacetic acid (TFA) (final concentration 5% v/v) was added to quench the reactions, which then were identified by HPLC analysis. HPLC analysis of SAM cleavage assay products After quenching by TFA, the reaction mixture was centrifuged and the supernatant was applied into HPLC analysis. 10ul of assay mixture was injected into C18 column, which had been pre-equilibrated with equilibrium buffer (40mM ammonium acetate, pH=6.2). Then the column was washed with a linear gradient from 0-50% acetonitrile for 30mins at room temperature to detect SAH and 5’dAdo. The UV-detector was set at wavelength 258nm, and the standard samples, SAM, SAH, and 5’dAdo, were run with the same condition as SAM cleavage reaction products. According to the retention time comparison between standard samples and products, formation of SAH and 5’dAdo were all detected via HPLC analysis when enzyme was present. In the absence of enzyme MJ0841, SAM was not consumed at all and there were no any products peak formed, which confirmed the SAM was cleaved by enzyme. The dark blue, red, light blue traces show the relative intensities of 5’dAdo, SAH, and SAM standards. The green trace shows the assay with the use of reconstituted MJ0841, and the SAH and 5’dAdo were both observed. The purple trace shows the control assay without MJ0841, and there was not any of 5’dAdo formed (Figure 9). Figure 9. HPLC analysis of the SAM cleavage assays Conclusions Future Work Initial results seem to show the [4Fe-4S] cluster and the enzyme activity. Since SAM was cleaved enzymatically, the products will be detected by mass spectroscopy to confirm the formation SAH and 5’dAdo. Furthermore, probable substrates of MJ0841 will be prepared, which should contain the amino acids that would be modified. The activity assays with substrates will be examined to detect the desired methylation reaction on substrates.

Compare contrast essay Essay

Novelist Paul Sheldon has plans to make the difficult transition from writing historical romances featuring heroine Misery Chastain to publishing literary fiction. Annie Wilkes, Sheldon’s number one fan, rescues the author from the scene of a car accident. The former nurse takes care of him in her remote house, but becomes irate when she discovers that the author has killed Misery off in his latest book. Annie keeps Sheldon prisoner while forcing him to write a book that brings Misery back to life. Compare & Contrast Essay â€Å"A Rose for Emily† by: William Faulkner & â€Å"Misery† by: Stephen King I’ve chosen to compare & contrast â€Å"A Rose for Emily† By: William Faulkner with â€Å"Misery† By: Stephen King, Obsessive women, broken ankles, trapped men & Suspenseful endings both of these stories have a lot in common. In Stephen King’s â€Å"Misery† A Famous Novelist named Paul Sheldon is transitioning his writing from Romance to Fiction. While on his way to Colorado from his winter hide away he is in a horrific car accident brought on by the harsh weather conditions. Thankfully the writer’s number one fan and former nurse Annie Wilkes lived nearby and saved Paul from freezing to  death. Annie Wilkes just like Miss Emily Grierson lived isolated and away from the world, though Miss Emily lived in the town & Annie Wilkes didn’t Miss Emily Isolated herself socially by not leaving her home nor opening the door when people of the town came by. Annie However, actually lived in an isolated cabin in the mountains. In â€Å"A Rose for Emily† people of the town feel sorry for her because after losing her father (page: 207 she was sick for a long time), But not long after they began seeing her with a gentle man named Homer Barran. A construction worker who had come into town to pave the sidewalks. After a year of being with Homer the towns folk began saying they would marry even though Homer claimed to like men (page: 209, He was not a marrying man). One evening after  ending their relationship Homer went to see Miss Emily using rat poison she killed him and let everyone believe he left and went on with his life while she hid his remains. Annie Wilkes didn’t kill Paul Sheldon, but took advantage of the fact that he had been put in critical condition due to the car accident she removed him from. For weeks she held him captive until he gave her favorite novel the proper ending it deserved. After noticing his several, sneaky attempts to escape Annie straps Paul to the bed and cripple s him by breaking both his ankles with a sledge hammer. Both of these women felt the need to keep these men hidden, trapped, and injured so they may stay with them forever. Miss Emily trapped Homer in death to stay with her since he had no interest in marrying her. In â€Å"Misery† Prior to having his ankles broken Paul finds newspaper clippings of previous victims of Annie where she was never convicted, He then realizes he may be next on her list. Soon after crippling Paul an officer drops by to see if Annie was aware that Novelist Paul Sheldon was missing and they were searching for him. Paul hears the officer and yells for help when Annie hears his screams she kills the officer and Paul is left once more to endure her gruesome punishment. Neither one of the men could predict what was going to happen to them, but Homer didn’t endure being tortured nor was he held captive for weeks. In â€Å"A Rose for Emily† Miss Emily was never caught nor suspected for the  disappearance of Homer. After the death of Miss Emily and after her services the town folk were curious to see   the inside of her home. (page 211, there was one room that no one had seen in forty years, and which would have to be forced). After entering the room they noticed mens toiletries, suits, ties and a pair of shoes even more so they noticed the man in her bed, His well over rotten body lay there in a nightshirt and next to him they could see where Miss Emily would lay beside him every night after his death. Unlike Miss Emily Annie didn’t get away with what she did to Paul, after killing the officer Paul tricks her into buying Champagne and cigars to celebrate the return of Misery, while setting up for their night of celebration Paul lights the re-written novel on fire and throws it. Annie begins to put out the flame when Paul slams his typewriter on the top of her head and shoves pieces of the burned novel pages into her mouth, she then dies of a fractured skull. Sometime after Paul is rescued by police. Both of these stories where great and though they are in different settings the characters shared similar physiological problems and depression. I think if both of these women met and could ever be in the same story there would be gory, suspenseful, torturing of men. I would like to read a book where Stephen King and William Faulkner write a story sharing both their ideas, I think it would be very interesting, thrilling and great to read.

Dota Thesis Essay

Here in the Philippines, we believe in the saying of our national hero Dr. Jose P. Rizal that â€Å"Youth is the hope of our Mother land†. In their hand lies the future generation to follow. But as we see from the situation nowadays, what we are expecting from the youth is the contrary if the saying. Teenagers are prone to pregnancy. Almost every year there is a rapid increase on the number of pregnant youths. One of the major causes of this problem is that most of the youths nowadays grew up in broken homes. Parents either the father or mother of the youths chose to leave their children behind because of the fact that they cannot handle anymore their obligation as parents. In relation to this problem, our government is now working on the Reproductive Health Bill (RH Law) as one of the most effective solution not only to teenage pregnancy but also to the increasing population of our country. The literature review was conducted to identify risk factors associated with teenage pregnancy and parenthood, including patterns of teenage sexual behaviour, as well as current knowledge about effective strategies in teenage pregnancy prevention. The impact of teenage parenthood on parents and children and ways of supporting pregnant and parenting young women was also explored. The review relied predominantly on Australian literature in order to reflect local issues and perspectives. However, the need to consider more recent or extensive research in some areas required the use of overseas literature, mainly from the US and the UK.Teenage pregnancies in the Philippines surged by 70 percent over one decade, a ranking official of the United Nation Population Fund Agency (UNFPA). Ugochi Daniels, UNFPA country representative, said the rising number of teenage pregnancies in the country is an area of concern that the agency is giving serious attention to, exposing as it does adolescent girls (10 to 19 years old) to high risk. Government statistics on female adolescent pregnancies indicate that the total number of annual births changed little over the preceding 10 years, but the number of teenage pregnancies rose 70 percent, from 114,205 in 1999 to 195,662 in 2009. Based on data compiled from birth certificates, of the 1.75 million live births in 2009, the latest review year, over 11 percent involved teenage mothers. Teenage pregnancy is widespread in the Philippines, especially amongst the poor. It is astory that may not be too pleasant, but one that is repeated all too often.Teen pregnancy is anobvious rising issue in our society. During the 21 st century, the statistics about this issue continues to rise even though there are a lot of disadvantages that young mothers and fathers can obviously get in the said issue. Fox (2008) defined teenage pregnancy as: Teenage pregnancy is one of the most important and alarming issues in today’s world. A teenage pregnancy, also referred to as teen pregnancy, comes to be when a girl attains motherhood before she becomes a major or adult, as laid down by the law of the land Some 16.5 million Filipinos belong to the 15-24 year old age group. We are forced with a glaring truth that at a very young age, a lot of young people today have children of their own. 30% of all births belong to this age group; and by the age of 20, 25% of the youth are already mot hers. This shocking truth about teenage pregnancy in the Philippines should wake the government and parents themselves to take action (Flores, 2008). According to the 2002 Young Adult Fertility and Sexuality Study by the University of the Philippines Population Institute (Uppi) and the Demographic Research and Development Foundation, twenty six(26%) of our Filipino youth nationwide from ages 15 to 25 admitted to having a premarital sex experience. What ¶s worse is that 38 percent of our youth are already in alive-in arrangement. Another is the 1998 National Demographic and Health Survey (NDHS)reveals that 3.6 million of our teenagers (that’s a whopping 5.2 percent of our population!) got pregnant. In 92 percent of these teens, the pregnancy was unplanned, and the majority, 78 percent, did not even use contraceptives the first time they had sex(Singsong, 2008).Most teenagers don’t plan to get pregnant, but many do. Teen pregnancies carry extra health risks to the mother and the baby. Risks for the baby include premature birth and a low birth weight. To control this, we must help each other by following the suggested solutions. I. Cause Causes of teenage pregnancy are occurring due to following reasons according to Fox (2008): Consequence of raging hormones. Most teenagers experience sudden and hitherto unknown emotions and feelings during their early puberty. All this, coupled with a sudden sense of new-found freedom and sexuality, results in many of them giving vent to their feelings through sexual expressions and experiences. Of course, most countries stress on sex education in schools, but yet, some teenagers involve themselves in unsafe sex, which could lead to unwanted pregnancies. Peer pressure. A lot of teenagers indulge in early sexual behaviour due to peer pressure. This is due to the fact that they feel the great need to be ‘hip’ and ‘accepted’ by their circle of friends. The only way they could probably achieve that would be by having a boyfriend or girlfriend or at least by dating and indulging in sexual acts often. Contraceptive Failure. Though there are many types and brands of contraceptives and emergency birth control pills easily available in the market, these are never a guarantee for complete safety. Only Mother Nature knows when she might want to get into action, so no form of birth control is going to be completely foolproof! (Fox, July 18, 2008).Other causes of teenage pregnancy are: 1. Lack of Parental Guidance. Most people evade their children from talking about sex. In some cases, they provide false information regarding sex and discourage their children to participate in any informative discussion about sex. In some cases, teenage mothers are not well educated about sex before getting pregnant and thus this leads to lack of communication between the parents and the children. 2. Inadequate Knowledge about Safe Sex. Most adolescents are unaware of safe sex. They probably have no access to the traditional methods of preventing pregnancy. And the main reason behind is that they are either too embarrassed or fear to seek information about it. It was found that more than 80% of teenage pregnancies are unintended. 3. Exploitation by Older Men. This is another major factor that contributes to pregnancyamong the teenagers. Those girls who date older men are more likely to become pregnant before they attain womanhood. Rape, sexual exploitations etc. also takes place th at leadsto unwanted pregnancy among teenage girls. 4. Socio Economic Factors. Teenage girls who belong to the poor families are more likelyto become pregnant. Researchers have found that even in the developed countries teenage pregnancy occurs most commonly among the deprived sections. 5. Glamorization of Pregnancy. The movie industry and the media contribute to teenage pregnancy by glamorizing teen pregnancy in news stories and movies. Movies that depict teen pregnancy as something to be desired encourage teens to engage in reckless sexual activity, according to ABC’s â€Å"Good Morning America.† During adolescence, teens become more focused on their appearance and how their peers perceive them. They want to be seen as part of the group, so if teen pregnancy is viewed as acceptable in their school or amongst their friends, they may seek to become pregnant as a way to gain social acceptance. 6. Teenage Drinking. Teen drinking can cause an unexpected pregnancy, according to the website Love to Know. Many teens experiment with drugs and alcohol. Drinking lowers a teen’s ability to control her impulses, contributing to 75 percent of pregnancies that occur between the ages 14 and 21. Approximately 91 percent of pregnant teens reported that although they were drinking at the time, they did not originally plan to have sex when they conceived. 7. Sexual Abuse or Rape. Teens can become pregnant as a result of sexual abuse or rape. The Guttmacher Institute states that between 43 and 62 percent of teens acknowledge that they were impregnated by an adult male, and two-thirds report that their babies’ fathers are as old as 27. Approximately 5 percent of all teen births are the result of a rape. 8. Inconsistent use of birth control Even though the use of contraceptives has increased among teenagers, its use remains spotty. Teens may use birth control to help prevent teen pregnancy, but most do not use contraceptives consistently. Erratic contraceptive use, however, is better than none. A sexually active teen that does not use any birth control has a 90 percent chance of becoming pregnant within a year. Some of the reasons that birth control is not used include the following: uneducated about birth control, uncomfortable using birth control methods and unable to access reliable birth control. 9. Abstinence-only sex education. Despite recent government efforts to fund abstinence-only sex education, the vast majority of teenage boys and girls (62 percent and 70 percent) have initiated vaginal sex by the age of 18. Because of their nature, abstinence-only programs leave out educational information on birth control. These programs have never been shown to actually reduce the incidence of teen pregnancy. However, they are the programs that receive the most funding from the government. There are state governments that forbid any sort of teen sex education in schools at all. This means that teens that experiment may not have information on birth control and are more likely to engage in activity that results in teen pregnancy. 10. Psychological factors. The immature and irresponsible behaviour arising duet complex teenage psychology is another important cause of teenage pregnancies. Teenagers often go through a number of emotions because of their own transition from childhood and peer pressure. The causes above are also similar to the causes here in the Philippines. II. Solution a) Existing Solutions – since abortion is immoral and legally wrong here in the Philippines, they consider adoption for the baby or acceptance for the parents. If not yet born, then they suggest that the baby not be killed. Currently, the government proposes sex education but it is still debated whether it would be taught or not. One way to prevent pregnancy is by practicing safe sex. In addition to it, birth control tools such as condoms, pills, intra-uterine devices or supremacies can regulate the fertility of a woman’s eggs. Even though it decreases the chances of not being pregnant, still it is not 100%sure and effective to avoid pregnancy. Some are even desperate that they will come to a point that they have to abort their babies without knowing the effects it may cause to the mother and it will kill the conscience of who will ever do this abortion thing. With the help of Science, Medicine has developed a new pill contraceptive namedRU486, the morning after solution to a sex encounter. Teenagers nowadays are sexually active without their parents knowing their sexual activities. Often times, media promotes sexuality to the young which may cause them to be mislead with wrong information about sex and they might be misguided, that’s why parents play an important role to the lives of their children. For all we know, sex is for those couples who already undergo to the Sacrament of Marriage, because doing sex needs a lot of sense of maturity not only physical but mental and spiritual basis. Once a pregnancy occurs from premarital sex, the couple is bound to live together for the rest of their lives with or without marriage because they have now the obligation to raise the child. The best way to avoid teen pregnancy is by avoiding or abstaining from sex before marriage. One thing is for sure, abstain from sex will keep you 100% safe from diseases, undesired relationship, and of course teenage PREGNANCY (Life Challenges -AllAboutLifeChallenges.org, 2011). Other possible Solutions are: 1. Abstinence from sex until you are old enough to do so. 2. Usage of contraceptives and other forms of birth controls. 3. Sex Education. 4. Affection and Guidance of parents. 5. Counselling. 6. Discouraging early dating What Ever the solutions may be, we must all keep in mind that we must be pro-life. b) How it is done in other countries. Existing solutions do not only exist on the Philippines but also on other parts of the world. For Example, abortion in the United States has been legal in every state since the United States Supreme Court decision in Roe V. Wade, on January 22, 1973. Prior to â€Å"Roe†, there were exceptions to the abortion ban in at least 10 states; â€Å"Roe† established that a woman has a right to self-determination (often referred to as a â€Å"right to privacy†) covering the decision whether or not to carry a pregnancy to term, but that this right must be balanced against a state’s interest in preserving fetal life (Wikipedia, 2011). In Egypt, they teach knowledge about male and female reproductive systems, sexual organs, contraception and STDs in public schools at the second and third years of the middle-preparatory phrase (when students are aged 12-14)(Wikipedia, 2011). In France, They promote sex education and it has been part of school curricular since 1973. Schools are expected to provide 30 to 40 hours of sex education, and pass out condoms, to students in grades eight and nine (Wikipedia, 2011). In Britain, which has one of the highest teenage pregnancy rates in Europe ,sex education is a heated issue in government and media reports In Sweden, sex education has been a mandatory part of school education since 1956. The subject is usually started between ages 7 and 10, and continues up through the grades, incorporated into different subjects such as biology and history In some countries, such as America, Falcon punch is done by the teens. The Falcon Punch is most often used to end the untimely pregnancy of a loved one in a humane manner, especially if the loved one happens to be under the age of sixteen. (EncyclopediaDramatica,2011). Active euthanasia, commonly referred simply as euthanasia, is only legal in the Netherlands and Belgium. (Wiki Answers, 2011) Euthanasia is the intentional killing by act or omission of a dependent human being for his or her alleged benefit, and over the years it has been one of the favourite tools of those who think society is better off when we take active steps to kill the poor, the disabled, the elderly, or the  ³unwanted ´. Today the debate over euthanasia most commonly revolves around the issue of physician-assisted suicide. There are many forms of euthanasia but the assisted suicide is its most common from. This method has also been done to prevent teenage pregnancy. (Wikipedia,2011) There are many possible solutions that can be done, some countries perform different methods that are immoral for Filipinos. One example is abortion. But amidst all this, we must learn to give importance to life because it is God’s wonderful gift to all of us STATEMENT OF THE PROBLEM: This study answers the following questions: 1. What is your reaction on teen pregnancy? 2. Do you know anything about RH Law? 3. Did you ever think of aborting a baby? 4. Do you still continue to pursue your education? 5. Do you know of anywhere in your local area where free condoms are available to younger couples? 6. What is the best solution or prevention on Early Pregnancy? 7. How the other country preventing early pregnancy. Locale of the Study The Study was done in STI College Laguna, The IT students give an survey about their reaction about Early Pregnancy and what they say about passing the RH Law. Importance: The fact is that the RH Bill is now a law of the Republic, and the President, barring judicial intervention, has the obligation of seeing to its effective, just and balanced implementation. Indeed, it would be tragic if, after all the acrimony, we end up with a law that is not implemented like many good legislation in this country. According to Dean Tony La Vina, in her view, the protracted debate, even if it was too long (lasting 13 years), actually improved the bill and, thanks in part to the opponents of the bill, we have a much more balanced and a better law than originally proposed. Among others, it is clear that abortifacient methods are prohibited (although the definition of abortion as preventing implantation of a fertilized ovum is controversial), freedom of conscience is respected, and there is neither a mandate to reduce our population nor a preference for smaller families. Below is a summary of what I believe are the most important provisions under the RH Law: * It affirms in a balanced way various state policies related to the right of health, including reproductive health, the rights of women, couples and families, and the protection of the life of the mother and the unborn. The law provides for spousal and parental consent in specific instances although safeguards to protect the individual rights of mothers or those availing of a service or procedure are also included. * In my view, the RH Law’s most important provision is the guarantee by the State to provide â€Å"universal access to medically-safe, non-abortifacient, effective, legal, affordable, and quality reproductive health care services, methods, devices, supplies which do not prevent the implantation of a fertilized ovum as determined by the Food and Drug Administration (FDA) and relevant information and education thereon according to the priority needs of women, children and other underprivileged sectors.† * In implementing the above policy, the State shall promote and provide information and access, without bias, to all methods of family planning, including effective natural and modern methods which have been proven medically safe, legal, non-abortifacient, and effective in acco rdance with scientific and evidence-based medical research standards. If this â€Å"no bias† principle is actually implemented, it will go a long way in reducing acrimony in the law’s implementation. * The RH law also provides that the State shall also provide funding support to promote modern natural methods of family planning, especially the Billings Ovulation Method, consistent with the needs of acceptors and their religious convictions. * The RH Law does not set demographic or population targets, and in fact, states that the mitigation, promotion and/or stabilization of the population growth rate is incidental to the advancement of reproductive health. Further, each family has the right to determine its ideal family size: although the State is mandated to equip each parent with the necessary information on all aspects of family life, including reproductive health and responsible parenthood, in order to make that determination. * Religious freedom is actually respected in the RH Law. Hospitals owned and operated by a religious group do not have to provide services contrary to its beliefs although these hospitals shall immediately refer the person seeking such care and services to another health facility, which is conveniently accessible. The conscientious objection of a health care service provider based on his/her ethical or religious beliefs are also respected, accompanied by an obligation for referral. These exceptions do not, however, apply in emergency cases. * The RH Law provides that no person shall be denied information and access to family planning services, whether natural or artificial. Minors however will not be allowed access to modern methods of family planning without written consent from their parents or guardian/s except when the minor is already a parent or has had a miscarriage. * Finally, age- and development-appropriate reproductive health education to adolescents (10-19 years) is mandated for public schools, which program shall be based on consultations with parents-teachers-community associations, school officials and other interest groups. Private schools may adopt the program at their own option, but consistent with religious freedom, are not obliged to do so. Scope and Limitations: This investigation is conducted to determine the rate of STI students that is reacted on Early or Teen Pregnancy. This way, we could determine the status and profile of the teenager Reaction in teenage pregnancy. The aspects looked into were the quantitative information of number of STI IT students Reaction in teenage pregnancy and the data behind this. This investigation focuses on number of IT students Reacted in teenage pregnancy in STI College Laguna portrayed by our group as respondents during February 2013. Definitions of Terms Teenage pregnancy – refers to any pregnancy on women that took place between the ages 13 – 19 years old. Teen aged Students – refers to students that ages a number that end in â€Å"teen† as the last syllable such as13, 14, 15, 16, 17, 18, and 19. Rate – refers to a quantity from a recent quantity STIS – refers to Sexually Transmitted Infections STDS – refers to Transmitted Diseases Sex – the sum of the structural, functional, and behavioural characteristics of organisms that are involved in reproduction marked by the union of gametes and that distinguish males and females RH – Reproductive Health Contraceptives – A device or drug serving to prevent pregnancy. Reference: http://www.dcsi.sa.gov.au/pub/ http://www.scribd.com/doc/51595306/Teenage-Pregnancies-in-the-Philippines-2, http://www.rappler.com/thought-leaders/18972-reconciliation-post-rh-law

The Effects Of Air Pollution On Human Health And / Or The...

Air pollution refers to a â€Å"broad range of contaminates† (Brimblecombe Atkins, 2013, p.20) found in the atmosphere that are harmful to human health and/or the environment. While air pollution can, at its most simplistic, be defined as â€Å"any input of chemical, aerosol, or particulate† (Blauvelt Gates, 2011, p. 17) found to be deleterious and airborne, the term can be subdivided in many ways by source, components, and mode of formation. (Blauvelt Gates, 2011, p. 17) Typically, an essential distinction is made between primary and secondary sources of emission. Primary air pollution is released directly from its source (e.g. carbon monoxide–CO– from a car s exhaust pipe) while secondary emissions are formed in the atmosphere itself (for example, ozone–O3–is almost entirely created in the atmosphere), almost always through photochemical (i.e. involving light, in this case sunlight) processes (WHO, 2006). In addition to source, air pollution is further classified by basic composition: either in a gaseous or particulate form. Gases (and vapors) are analyzed at the molecular level and because of their small size they â€Å"are readily taken into the human respiratory system† (WHO, 2006, p.10) which presents a health risk. Common gaseous pollutants include carbon monoxide, carbon dioxide, ozone, methane, sulfur dioxide, and nitrogen dioxide. Particulate matter (PM) is defined as material in solid or liquid phase suspended in the atmosphere (WHO, 2006, p. 11) and is divided byShow MoreRelatedThe Effects Of Air Pollution On Human Health850 Words   |  4 Pagesincreasing, so does pollution. The debate associated with this contamination on the environment has to do with how much should be done to stop pollution, and whether or not this is a serious enough issue to be concerned with. Well, this issue should disturb people. 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