MikeLigalig.com

Blonde..for personal reasons.
LOL

Brown eyes or blue eyes?

yikes! 

Quote from: grazie7y on February 18, 2008, 02:44:53 PM
Isn't her name Angel, Dong?  She is so pretty jud!

Yes her nick name is 'Angel', pero her full name is Lorraine Angeli Lucino.
I should get her to join.

Quote from: jaja on February 18, 2008, 03:05:09 PM
pareha diay me kagamay sa imo sis branbran!

Hahahaha. o really, janjan??

hahaha, she is what my mom likes to call, "Valenciana girl"

'Small but terrible'. She is 4'11 but she can easily take down her Kuya Bran 

Dear ___________________


You may not remember me, but you and I were partners in Organic Chemistry I, and Organic Chemistry II.  You were always rushing to finish mixing reagents, and the Professor would punish us as a group, pero I could not get myself to get angry at you. Your eyes, they were the most greenest things I've ever seen in my life. You may have already forgotten about me, but I still remember you, and the fact that you would always be late to Organic Lab.

You know that Professor Perischini was angry at me one time because you were late to lab? I wanted to express my disappointment in you after lab that day, but the moment you walked in the door, my anger dissipated, as if boiling water evaporating in air.

And I still remember the many times you skipped our study sessions, how you kept me waiting for you. Waiting for your arrival. I lost many lead cartridges that way, clicking my automatic pencil, waiting for you. But you did not come.

Ah, alas, it was not meant to be.

Where are you, o green-eyed beauty,
Your former organic chemistry lab partner,
L. Lucino

TB Mafia?

Please please don't shoot me

This reminds me of 'The Godfather'

Quote from: orChids on February 18, 2008, 02:20:13 PM
Lagi nangawat ko ug oras dalang kuha sa akong baktin sa school.I feel better na Glen,see yah next time.Regards.

Ms. Orchids! Konnichi wa! We missed you as well!

Quote from: Kyle828 on February 18, 2008, 02:10:09 PM
Ya we are chatting on my Y.M. I am doing fine. School is going great! how about yourself?

Hahaha good to know, Kyle. Keep up the studies. I'm doing alright, Kyle. A bit stressed, but its nothing that can't be overcome.

Good luck this week! Make us all proud, kiddo.

These are beautiful beautiful pictures! Mr Ferniz, you have handsome parents! More blessings to you and your family. Many regards from the Lucino Family.

*shrugs*

haha saba dinha!

Yo Kyle? you went phone chatting ha?

How are studies going, kiddo?

very surprised, jan jan. Na mingaw mi nimo diri.

ehem..



ps. missed you

Hahaha, joker jud ka bro.

Scott, come back.

We miss your photographic skills.

And your presence!

Luuy pood intawn ang civilians na namatay ani. Such an unnecessary loss of life.

hahaha hilarious

Musta naman ka Janjan

A PRETTY good way to generate an outcry, as the archbishop of Canterbury learned in Britain recently, is to say that a Western legal system should make room for Shariah, or Islamic law. When the archbishop, spiritual leader of the world’s 80 million Anglicans, commented in a radio interview that such an accommodation was “unavoidable,” critics conjured images of stonings and maimings, overwhelming his more modest point.

The archbishop, the Most Rev. Rowan Williams, did not propose importing Shariah into the criminal law and was referring mostly to divorces in which both sides have agreed to abide by the judgment of a religious tribunal. His proposal was groundbreaking only in extending to Islamic tribunals in Britain a role that Jewish and Christian ones have long played in the judicial systems of secular societies. Courts in the United States have endorsed all three kinds of tribunals.

In 2003, for instance, a Texas appeals court referred a divorce case to a local tribunal called the Texas Islamic Court. In 2005, the federal appeals court in New Orleans affirmed an award in an employment arbitration by the Institute for Christian Conciliation, which uses Biblical teachings to settle disputes. And state courts routinely enforce the decisions made by a Jewish court, known as a bet din, in commercial and family law cases.

The outcry in Britain was apparently something of a visceral reaction to aspects of Islamic law, though the archbishop himself condemned what he called the inhumanity of “extreme punishments” and some Islamic countries’ “attitudes toward women.”

The larger question, legal experts in the United States said, is whether government courts should ever defer to religious ones. The answer may depend on whether the people involved authentically consented to religious adjudication, whether they are allowed to change their minds and whether the decisions of those tribunals are offensive to fundamental conceptions of justice.

All of that, said John Witte Jr., a law professor at Emory University, “is the big frontier question for religious liberty.”

The archbishop speaks in sonorous circumlocutions and he was not a model of clarity when he was interviewed by BBC radio on Feb. 7. Even his followers had a hard time untangling just what he meant.

“I’m an Episcopalian,” said Janice A. Schattman, the lawyer in the Texas case who persuaded the appeals court to defer to the Islamic one. “Rowan Williams, bless his heart, can be quite obscure.”

But the archbishop’s central point seemed to be that people should be able to agree to have family law cases resolved by religious courts if all concerned agree. By Monday the archbishop was backtracking, saying he had spoken clumsily with “a misleading choice of words.”

Azizah Y. al-Hibri, the president of Karamah, an international lawyers’ group based in Washington and made up of Muslim women, said she applauded the archbishop’s initial position.

“Muslims, Christians and Jews should all deal with their own family law issues in their own arbitration councils,” she said. “The government should stay out of the bedroom.”

That notion has met resistance where Islam is involved. After the authorities in Ontario raised the possibility that arbitrators might use Shariah to settle family disputes, formal recognition of all religious arbitrations there, including existing Catholic and Jewish ones, was withdrawn.

“There will be one law for all Ontarians,” Dalton McGuinty, the province’s premier, said in 2005.

Almost no one suggests that criminal law should take into account the defendant’s religion in meting out punishment. At the other extreme, few people object to allowing purely commercial disputes between sophisticated businesspeople to be adjudicated through private arbitrations. The hard questions, as the archbishop learned, arise in the area of family law, where the agreement to arbitrate may be uninformed or obtained by duress. State courts have occasionally refused to enforce separation agreements reached through bet din arbitrations on the ground that the woman involved had been pressured into participating.

Once consent is given, moreover, questions arise about whether and when it may be withdrawn. “People have a right in Western systems to change religions,” said Douglas Laycock, a law professor at the University of Michigan. “Can they opt out after the dispute arises or after the judgment is given?”

Most fundamentally, some judgments from religious tribunals may be at odds with constitutional protections, human rights and basic notions of fairness.

In an article to be published shortly in The Washington and Lee Law Review, Robin Fretwell Wilson, a law professor at Washington and Lee University, wrote that Muslim women who decide to seek a divorce can face harsh financial consequences under Islamic law. “Threatened with the prospect of certain poverty,” she wrote, “some women will surely be forced to stay in an abusive relationship.”

Professor Wilson said in an interview that government courts should refuse to enforce any ruling from a religious tribunal that leaves a woman worse off than she would have been in a conventional divorce.

“Society has a stake in the outcome,” she said. “Some religions are tilted against women.”

In the Texas case, however, it was a woman, Rola Qaddura, who sought arbitration in a dispute over a dowry and the distribution of assets after a divorce. The parties had signed an agreement to arbitrate their case “according to the Islamic rules of law by Texas Islamic Court” in Richardson.

The appeals court said the agreement was valid. Ms. Schattman, who represented Ms. Qaddura, said the appeals court’s ruling was proper and unexceptional. “An agreement to arbitrate is an agreement to arbitrate,” she said.

In the end, though, the parties could not agree on a panel of arbitrators and the effort collapsed, Ms. Schattman added, saying of the Islamic court: “It was kind of a new thing.”

A suicide bomber blew himself up in a large crowd gathered at a dogfighting event just outside this city in southern Afghanistan, killing about 80 people and wounding more than 90 others in the country’s worst single bombing since 2001.

According to witnesses and officials, the bomber killed a local police chief, Abdul Hakim Jan, a number of his guards and scores of villagers attending the event in the Argandab district, just north of the city of Kandahar.

The governor of Kandahar Province, Asadullah Khaled, said 80 people had died and more than 90 had been wounded. A spokesman for the Ministry of Health in Kabul, Dr. Abdullah Fahim, said the Kandahar hospital had received 67 bodies. But some families had taken bodies straight home for burial from the scene of the blast, he said.

“This is the action of the enemies of our country,” Mr. Khaled said. “They do not let Afghans enjoy their lives and have a peaceful life.”

A spokesman for the Taliban, Qari Yousuf Ahmadi, denied that the Taliban had carried out the attack and suggested it was a result of internal fighting within the Afghan government. “We did not carry out this blast in Kandahar â€" we strongly reject that,” he said by telephone.

Kandahar, a former stronghold of the Taliban, has been the scene of some of the country’s worst suicide attacks over the past two years as Taliban insurgents have tried to undermine the government of President Hamid Karzai.

The Argandab district, a rich agricultural valley north of the city, had long kept the Taliban at bay under the leadership of a powerful tribal chief, Mullah Naquibullah, but the district has become a target of the militants in the months since his death. The slain police chief, Mr. Jan, was a prominent anti-Taliban commander who was taking a determined stance against the insurgents.

There has been an exponential increase in casualties caused by suicide bomb attacks in recent months both in Pakistan and Afghanistan, the results of more powerful explosives and more sophisticated and lethal tactics, government officials in both countries have said. In the past two years, suicide bombings in Afghanistan aimed at foreign troops often only killed the bombers and a few bystanders, but in recent attacks, often in crowded public places, local police officers and officials appeared to have been the targets.

The bombers have also used larger amounts of plastic explosives packed in vests on the front and back of their chests and packed ball bearings around the explosives, causing devastating damage to unprotected civilians. A November bombing in Baghlan, in which many were killed by ball bearings, represented a new level of carnage for Afghanistan.

Recent bombings in Pakistan have shown a similar pattern. A double suicide bombing on Benazir Bhutto’s homecoming procession on Oct. 18 in the city of Karachi caused unparalleled casualties, with 140 killed and 500 wounded, according to the government. Many of them were killed by ball bearings, police officials said.

In both countries security officials say the planning and expertise of the bomb attacks can be traced back to figures operating in the tribal areas of Pakistan, in particular North and South Waziristan. Two men cited are the Pakistan Taliban leader Baitullah Mehsud, accused of being responsible for many recent suicide attacks in Pakistan, including the one that killed Ms. Bhutto, and another man, Mullah Abdullah, who was behind an attack on a five-star hotel in Kabul.

Hours after the explosion on Sunday, pale and shaken survivors were still at the scene. Abandoned shawls, shoes, caps and bits of human flesh were strewn on the bloodied field. Five vehicles, including police cars, had been badly damaged. The body of a man the police said was the bomber lay mangled.

Muhammad Khan, 25, said he had been knocked over by the blast. “I couldn’t hear or speak or walk,” he said. “My whole body was numb, and I thought I was injured, but my heart is working. Luckily my brother rushed and picked me up and poured water over my head. Thank God, I am fine.”

People had traveled miles to watch the dogfights, a pastime that was banned by the Taliban when they were in power because it entails gambling. But the fights, involving huge Afghan mastiffs, have returned to much of the country, often sponsored by local commanders and landlords. In Argandab, they take place every Sunday during the winter, and huge crowds gather, attracting street vendors who hawk food and drinks.

Noor Muhammad, 32, said he had survived because he was sitting down and a dog in front of him blocked the force of the explosion. The police chief, who was nearby, was killed along with five of his bodyguards, he said.

He said he believed many more people had been killed in the attack. “I counted 30 vehicles carrying the dead bodies; in each car there were seven to eight dead bodies,” he said. “Thousands of people were watching the dogs fighting, including young children and old people. Some people were selling things like oranges and tea and other food from stalls. People had come from different parts of Kandahar Province.”

Hospital officials in Kandahar said that children and teenagers were among the wounded, and that about 15 patients were critically hurt.

The Taliban were ousted from power in 2001 after the United States led an invasion in the wake of the Sept. 11 terrorist attacks, but their insurgent movement has been gathering strength, and suicide bombings attributed to them have been increasing in the past few years.

The November suicide bombing, when the bomber attacked a parliamentary delegation visiting the normally quiet province, killed 72 people, 59 of them schoolchildren gathered to welcome the delegation. After that attack, Mr. Karzai said Taliban insurgents had committed 116 suicide bombings in 2007.

Pakistanis began to vote Monday morning in parliamentary elections that are expected to diminish President Pervez Musharraf further and present Washington with a challenging new political lineup here as it pursues its fight against the Taliban and Al Qaeda in the region.

The vote, which was delayed after the assassination of the opposition leader Benazir Bhutto nearly two months ago, comes amid anxiety of further political turmoil if the government manipulates the results or enters a battle with the opposition parties over forming a government.

At least four candidates, including Ms. Bhutto, and nearly 100 other Pakistanis have been killed during the campaign. Parties have reported kidnappings and arrests of candidates and attempts to intimidate their families, according to Sheila Fruman, director of the National Democratic Institute’s office in Pakistan. The opposition has threatened street protests if the vote is perceived to be unfair, but has also called for a government of national consensus.

American officials and others here hope that the election provides a fresh opportunity for a new civilian government to rally Pakistanis behind the fight against the militants who now threaten the security and stability of the country.

After growing frustration with eight years of military rule, opposition politicians and analysts argue that Mr. Musharraf has lost the support of the people and cannot fight extremism effectively without it.

With the country facing a growing insurgency by the Taliban and Al Qaeda, rising prices and escalating violence, the nationwide vote will now serve as a kind of referendum on Mr. Musharraf, who has grown deeply unpopular.

No matter which party prevails, Mr. Musharraf, who has been Washington’s partner in the campaign against terrorism for the past six years, is almost certain to emerge further reduced in the post-election skirmishing.

He is already much weakened after resigning as army chief in November, and a popularly elected prime minister with the backing of Parliament will emerge as a competitive new force.

The party that has supported Mr. Musharraf for the past five years, the Pakistan Muslim League-Q, is expected to fare considerably worse than it did in the last election.

The party of Ms. Bhutto, the Pakistan Peoples Party, is riding a wave of sympathy after her death and may emerge as the largest party in Parliament, analysts say.

Ms. Bhutto’s party, which is now led by her widower, Asif Ali Zardari, and the party of former Prime Minister Nawaz Sharif, Pakistan Muslim League-N, are moderate parties opposed to terrorism.

They argue that only a popularly elected government can bring the country together to oppose militancy. For his part, Mr. Zardari has also called for a government of national consensus after the election and has not ruled out working with Mr. Musharraf.

The insurgency remains at the top of the Bush administration’s agenda here, and American officials have started to prepare for Mr. Musharraf’s eventual exit. In a series of high-level visits in the past month on how to stem the militants’ efforts to destabilize Pakistan, Washington officials have focused on the new army chief, Gen. Ashfaq Parvez Kayani, an implicit recognition of the shifting balance of power.

Despite the complaints about violence in the campaign and the potential for fraud, the Bush administration has appeared determined to validate the election as a satisfactory exercise in democracy. Richard A. Boucher, the assistant secretary of state for South and Central Asian affairs, recently told a Congressional committee that he was looking for “as fair an election as possible.”

International observers, including several United States senators who arrived Sunday, have already warned of serious flaws in the pre-election process. They have said that at best the election would be deemed “credible,” rather than free and fair.

The biggest question will be to what extent the government apparatus will try to manipulate the results in favor of the pro-Musharraf party. If the elections are skewed too far in its favor, the government risks large protests and violence.

Senator Joseph R. Biden Jr. of Delaware, the Democratic chairman of the Senate Foreign Relations Committee, said after arriving in Lahore on Sunday night as an election observer that he was “mildly optimistic” that the election would be “fairly credible.”

But Mr. Biden said that if the election turned out to be seriously flawed, he would seek to curtail United States military aid to Pakistan. Two other members of the foreign relations committee, Senators Chuck Hagel, Republican of Nebraska, and John Kerry, Democrat of Massachusetts, also arrived to observe the election.

The Birth of A Nation!




The province of Kosovo declared independence from Serbia on Sunday, sending tens of thousands of ethnic Albanians streaming through the streets to celebrate what they hoped was the end of a long and bloody struggle for national self-determination.

Kosovo’s bid to be recognized as Europe’s newest country â€" after a civil war that killed 10,000 people a decade ago and then years of limbo under United Nations rule â€" was the latest episode in the dismemberment of the former Yugoslavia, 17 years after its dissolution began.

It brings to a climax a showdown between the West, which argues that Serbia’s brutal subjugation of Kosovo’s ethnic Albanian majority cost it any right to rule the territory, and the Serbian government and its allies in the Kremlin. They counter that Kosovo’s independence is a reckless breach of international law that will spur other secessionist movements across the world.

As Albanians danced in the streets and fired guns in the air in the capital, Pristina, international reaction was sharply divided, suggesting that the clash between the principles of sovereignty and self-determination was far from resolved.

Britain, France and Germany were expected to be the first recognize the new nation as early as Monday, while other nations, fearing separatist movements within their own borders, have said they will refuse. Russia demanded an emergency meeting of the United Nations Security Council to proclaim the declaration “null and void,” but the meeting produced no resolution.

The United States and additional European Union member states were expected to recognize Kosovo’s independence in the coming days.

President Bush, speaking in Tanzania, said the United States would continue to work to prevent violence in Kosovo, while reaching out to Serbia. He said that resolving the conflict in Kosovo was essential to stability in the Balkans and that “the Serbian people can know that they have a friend in America.”

In declaring independence, Kosovo’s prime minister, Hashim Thaci, a former leader of the guerrilla force that just over 10 years ago began an armed rebellion against Serbian domination, struck a note of reconciliation. Addressing Parliament in both Albanian and Serbian, he pledged to protect the rights of Kosovo’s Serbian minority.

“I feel the heartbeat of our ancestors,” he said. “We, the leaders of our people, democratically elected, through this declaration proclaim Kosovo an independent and sovereign state.”

Kosovo, a desperately poor, predominantly Muslim landlocked territory of two million, has been a United Nations protectorate since 1999, policed by 16,000 NATO troops. Its unemployment rate is about 60 percent and average monthly wage is $250.

Electricity is so undependable that lights go out in the capital several times a day. Corruption is rife and human trafficking threatens to entrench a lawless state on Europe’s doorstep.

Ethnic Albanians from as far away as the United States poured into Pristina over the weekend, braving freezing temperatures and heavy snow to dance in frenzied jubilation. Beating drums, waving Albanian flags and throwing firecrackers, they chanted: “Independence! Independence! We are free at last!”

A 100-foot-long birthday cake was installed on Pristina’s main boulevard.

In an outpouring of adulation for the United States, the architect of NATO’s 1999 bombing campaign against Serbian forces under President Slobodan Milosevic, revelers unfurled giant American flags, carried posters of former President Bill Clinton and chanted, “Thank you, U.S.A.” and “God bless America.”

Hundreds of people, many waving Albanian flags, celebrated in Times Square. Revelers in cars drove in circles around the area, leading chants whenever they passed the crowds gathered on the sidewalks.

That spirit of exaltation contrasted sharply with the despair, anger and disbelief that gripped Serbia and the Serbian enclaves of northern Kosovo. In Belgrade, Serbia’s capital, as many as 2,000 angry Serbs converged on the United States Embassy, hurling stones and smashing windows.

In the Kosovo Serb stronghold of Mitrovica, a grenade was thrown at a United Nations building, the police said. No one was injured.

Vojislav Kostunica, the prime minister of Serbia, which has regarded Kosovo as its heartland since medieval times, vowed that Serbia would never recognize the “false state.”

In an address on national television on Sunday, he said Kosovo was propped up unlawfully by the United States and called the declaration a “humiliation” for the European Union. The Serbian government has ruled out using military force in response, but was expected to downgrade diplomatic ties with any government that recognized Kosovo.

Demonstrations were planned for Monday in Serbian enclaves across Kosovo. Serbs said they were under orders from Belgrade to ignore the independence declaration and remain in Kosovo to keep the northern part of the territory under de facto Serbian control, raising questions about Serbia’s long-term aims.

Useless violence. My condolences to those who died.

Dear______________

Thank you so much for taking me out during Valentines Day, salamat for the 'gift'.
Remember that my arm is always here for you to hold on to. My chest is always here for you to lay your head on.

My affection and my friendship,
Yours Truly,

Cat-fight.

Ate Ging, in my personal point of view, the video puts to much negative emphasis, trying to rile up viewers.
Yes they are right about bacteria being present in lemons, but that is the case for everything we eat. The water we drink or the softdrinks that we drink in restaurants are also saturated in water-born bacteria, so too are the meats we order in restaurants, the vegetables etc, are filled with minuscule bacterial presence.  Our very fingers, really, are filled with bacteria, even after we wash them. And under our nails are fecal bacteria, even if washed, still remnants remain.

Not all bacteria are bad, within our bodies there are intestinal bacteria whose job is to digest food; for example we have lactase bacteria, which will digest lactose-based foods; thus allowing us to eat any dairy-based products. Without these bacteria, we would be unable to consume it. On the contrary, people who are lactose-intolerant suffer due to the lack of lactase enzymes and lactose-eating bacteria.  There are also bacteria within our stomachs which inhibit the growth of foreign-based pathogens (bacteria), thus allowing the stomach to remain homeostatic.

In all reality, Ate Ging, the bacteria found in food and water/drinks will be inhibited and destroyed upon entering our salivary tract (which has digestive enzymes in it), and digested in the digestive tract, which produces hydrocholoric acid, that digests food and kills most foreign pathogens, with symbiosis with our internal bacteria.

So long as we remain healthy, Ate Ging, so long as our immune system is not suppressed, the body can fight any bacteria, any virus nature has to offer, to the best of its ability. 

The Portuguese rendition

[mp3tube]01da4dd17eeae916150c6b65433a39f1[/mp3tube]

Quote from: svetlana on February 18, 2008, 11:21:15 AM
I'd prefer more than 1 child, hhhmmm 3 at the most  (nagdamgo, buot2x!) hahaha

Hahaha, I have to beat my dad's record of 4 children. Lima ahong gusto; 3 boys, and 2 girls.
God-willing

Quote from: svetlana on February 18, 2008, 11:21:15 AM

PI wedding or abroad?

Both. I want a church wedding here in the 'states and in the Philippines.


Would you rather have a dog or a cat?

Yea, the video is very informative.

Quote from: svetlana on February 18, 2008, 11:13:26 AM
tagna lagi nako nga you'll find it very very tough 

Natural ra man na, Sheila. A fisherman will catch many fishes in his lifetime, but he may only have one, diba?
It is easy to catch a northern and american fish, but it is very hard and rigorous to find/catch the elusive Boholana fish,

One needs a bigger net, no?

Church wedding, it will be grande, God-willing, like the one of my father and mother, and my grandparents before them.

My wife shall enjoy nothing but the best, that I promise.

To have 1 child or more than one child?

Happy Birthday to both of you two! And may you enjoy blessings upon blessings. Continue to shine your light to your family and friends. Give your best, God will do the rest

Take Care, Much Regards,
Bran Lorenzo

If we want to get free, it's helpful to know how we got in bondage in the first place. Lesson 1 exposes one of the core roots that cause bondage: the fear of man. When we're afraid of people, or they're afraid of us, the chains of bondage begin to tighten around us. Once we recognize the cause of bondage, we can begin to take steps towards true freedom.

"So they put slave masters over them to oppress them with forced labor, and they built Pithom and Rameses as store cities for Pharaoh. But the more they were oppressed, the more they multiplied and spread; so the Egyptians came to dread the Israelites and worked them ruthlessly.    They made their lives bitter with hard labor in brick and mortar and with all kinds of work in the fields; in all their hard labor the Egyptians used them ruthlessly. The king of Egypt said to the Hebrew midwives, whose names were Shiphrah and Puah, "When you help the Hebrew women in childbirth and observe them on the delivery stool, if it is a boy, kill him; but if it is a girl, let her live." The midwives, however, feared God and did not do what the king of Egypt had told them to do; they let the boys live. Then the king of Egypt summoned the midwives and asked them, "Why have you done this? Why have you let the boys live?" The midwives answered Pharaoh, "Hebrew women are not like Egyptian women; they are vigorous and give birth before the midwives arrive."
So God was kind to the midwives and the people increased and became even more numerous.
(Lesson from Exodus 1:1-14)

lol, the two names are the names of my uncles (Roy, Roger)

Steven, Stewie, Salvo

This is a very tough one.
Very.

But I want my children to have the beauty of their ancestors. The Boholano blood must be preserved, so..
Morena.

Would you rather have one grandchild or many?

To give completely to God, Body and Soul
Can you live with only the bare necessities? Yet still worship? We must learn from them~
















He saith to him again the second time, "Simon-Peter, son of Jonas, lovest thou Me?" He saith unto Him, "Yes, Lord, Though knowest that I love Thee." He saith unto him, "Feed My sheep."
(John 21: 15-17)

Motorola

To give power or have power?

Farnesyltransferase-Inhibitors - New drugs for cancer treatment
By: Dr. Bernd Nosse

Abstract:


Posttranslational lipidation is an essential modification for membrane-bound proteins being involved in intracellular signaling pathways. Ras proteins, belonging to the Ras (rat-adeno-sarcoma) superfamily as some of such are known to play a decisive role in the signal transduction cascade resulting in cell differentiation and proliferation. Ras is lacking a transmembrane domain and it can only function when bound near the growth factor receptor at the cell membrane. Therefore it requires farnesylation to increase its hydrophobicity and ability to anchor to the plasma membrane. This posttranslational modification is catalyzed by the enzyme farnesyltransferase (FTase) which transfers a farnesyl (C15) residue to the carboxy-terminal rest of the Ras protein. Scientists nowadays assume that the cause of approximately 30% of all human tumors is mutated Ras, mainly being involved in myeloid malignancies and carcinomas of e.g. the colon, breast, pancreas, lung and thyroid. Point mutations in the genes lead to the inability of Ras to regulate itself and Ras as a molecular switch remains constantly activated. A selective blocking of FTase by farnesyltransferase inhibitors (FTIs) is viewed as a promising target for therapeutic cancer treatment: Preventing the prenylation of Ras hinders it from attachment with the plasma membrane and thus interrupts the oncogenic Ras function.

Signaling pathway:


A novel and hopeful starting point in the development of new therapies to treat malign illnesses is the investigation and modification of processes, which control the proliferation and differentiation of cells. A promising target is the Ras signal transduction pathway: The membrane-bound Ras protein acts as molecular switch, allowing to transfer a signal coming from the outside of the cell (e.g. a growth factor) into the nucleus (see Fig. 1). The first step in this cascade is the dimerisation of the monomeric receptor tyrosine kinase by stimulation with an extracellular ligand. Grb2 as the first adaptor protein recognizes a binding site enabling in turn a second adaptor molecule called Sos ("Son of sevenless") to attach. Sos subsequently stimulates the inactive GDP-carrying Ras to exchange GDP for GTP and thus becomes active. Thereafter the active Ras can forward the signal to other downstream effectors, e.g. Raf which is the first protein kinase in the MAP-kinase signaling cascade [1][2][3], which finally results in the transcription of different genes. [4][5][6]



Ras and cancer:


Approximately 30% of all human tumors arise from a mutated Ras oncogene [7][8][9], particularly point mutations in the corresponding Ras genes are found in over 90% of human pancreatic carcinomas [10] and 50% of human colon cancers [11]. As a biochemical consequence of those mutations the Ras protein loses its GTPase activity (an intrinsic enzyme, hydrolyzing GTP to GDP) and therefore permanently stays in the GTP-bound active state sending constantly signals into the nucleus. This in turn results in uncontrolled cell division being characteristic for cancer diseases.

Posttranslational modifications of Ras:


The crucial point for normal as well as mutated Ras proteins is the necessity to be bound to the cell membrane to ensure signal transduction. The anchorage to the cell membrane is achieved by a series of posttranslational modifications, in which the enzyme farnesyltransferase (FTase) first transfers the lipophilic farnesyl residue of farnesyl pyrophosphate (FPP) to the mercapto function of a cystein. This amino acid is a component of the so called "CaaX-box" located at the C-terminus of Ras, whereat "C" stands for cystein, "a" for an aliphatic amino acid and "X" is methionine, serine, alanine, or glutamine. [12] The mode of binding in the active site of the FTase was investigated with the help of X-ray crystal structures of crystallized complexes from FTase, FPP and a CaaX mimetic [13][14]: A central zinc ion promotes the deprotonation of the cystein to its thiolate which in turn substitutes the pyrophosphate group of FPP generating a C-S linkage and thus connecting the farnesyl residue to the Ras protein. In the next steps the "aaX" part is cleaved off by a specific endoprotease and the free S-farnesylcystein at the C-terminus is finally methylated with the help of a methyltransferase enabling the Ras protein to bind to the cell membrane. [15]

Farnesyltransferase inhibitors (FTIs)


Different classes and strategies to inhibit the enzyme farnesyltransferase have been designed which are listed below.
Inhibitors based on the CaaX motif try to simulate the C-terminal tetrapeptide and compete with Ras to be farnesylated. Their development was initiated by just varying some amino acids (e.g. 1, IC50 = 25 nM in vitro, [16]), which in vivo suffered from membrane permeability and proteolysis. Combinatorial chemistry and high throughput screening eventually led to highly efficient inhibitors, where the rational concept is hardly recognizable (e.g. 2, IC50 = 0.7 nM, [17]).
Farnesyl pyrophosphate analogues (e.g. 3, IC50 = 75 nM, [18]) are in general less attractive due to the possibility to compete with FPP as a substrate also in other enzymatic processes such as squalene synthase.
Bisubstrate inhibitors are based on the theory that a complex consisting of the enzyme and both substrates (FPP and CaaX-box) is formed prior to the catalysis. As a consequence they combine both strategies mentioned above: They incorporate a farnesyl and a CaaX mimetic (see for example 4, MIC = 0.1 mM, [19]) and display enhanced selectivity and activity. Other inhibitors can be found in the pool of natural products, e.g. pepticinnamin E (5, IC50 = 42 mM, [20][21][22]).
Recent studies deal with metal chelating FTIs: The inhibitor design is based on the combination of a Zn-chelating core with a substituent, recognizing the aromatic residues which are situated around the zinc ion in the active site of the FTase (e.g. 6, IC50 = 1.9 mM, [23]).
Beside the aforementioned classes, FTIs from natural product sources (e.g. 7, IC50 = 5 mM, [24]) and from compound libraries (e.g. 8, IC50 = 40 nM, [25]) are also known.



Conclusion:


Ras as one of the most frequent oncogenes found in human cancers emerged as a valuable target for anti-cancer therapy. One approach to decrease the activity of Ras-proteins is to avert its anchorage to the plasma membrane with the help of farnesyltransferase inhibitors (FTIs). Those substances prevent the enzyme farnesyltransferase (FTase) from adhering a lipophilic farnesyl residue to the C-terminus of Ras, and hence to impede Ras to bind to the cytosolic side of the membrane. Apart from a number of lead structures, showing potent inhibitory activities, have been investigated, also at least three FTIs are currently undergoing clinical evaluation [26][27][28] (Zarnestra® (9) [29][30], Sarasar® (10) [31] and BMS-214662 (2) [32]) which demonstrates in a striking way the potential of this strategy to treat malign illnesses.

By: Dr. R. Bandichhor, M.D. 


Abstract

HIV epidemic continues to involve researchers and clinicians to unearth new facts about the virus and develop new treatment regimens for patients. The status of anti-HIV treatments is never static but constantly changing due to complicated genetic diversity of the virus and the inability of the body to clear the virus. The development of combination therapy is viewed as an essential step to increase the life span of the HIV positive individuals. Tremendous strides have occurred in the HIV/AIDS arena, yet daunting challenges remain. This article presents a glimpse of recent advances in combination therapy for AIDS.

Introduction

Since the AIDS epidemic began, the FDA has approved 47 drugs for the treatment of HIV/AIDS, including 19 antiretroviral drugs for use singly or in combination therapy.[1] Highly Active Anti-Retroviral Therapy (HAART) regimens utilizing protease inhibitors (PIs) are a major factor in reducing the number of AIDS deaths from the mid-1990s to the present. It has been identified a pipeline of 52 drugs for HIV infection and AIDS, including extended release formulations of existing antiretroviral therapies, immune modulators, fusion inhibitors, vaccines, topical microbicides and drugs targeting dementia [2], lipodystrophy [3] and other complications of AIDS.
HAART regimens

The most common initial regimens consist of two nucleoside analogs, combined with either a PI, an NNRTI (Non Nucleoside Reverse Transcriptase Inhibitors) or a third nucleoside analog. The combination of two nukes and a PI is supported by data from randomized studies with clinical endpoints.[4] These regimens, however, often involve a considerable pill burden and relatively frequent side effects, which makes compliance difficulties. They are possibly quite robust with regard to immunological efficacy [5] which has yet to be demonstrated for NNRTIs or nukes. NNRTs with two nucleoside analogs regimen is found to have advantages as they are effective with low pill burden and good tolerability. However, the significantly emerging disadvantage is rapid development of cross resistance. Most explored nucleoside analogs combination is Trizivir [ AZT ( Azidothymidine), 3TC ( Epivir) and abacavir] but studies have shown that in patients with a high viral load, efficacy was inferior to PI combinations.[6]

More recently Bristol-Myers Squibb [7] has submitted an NDA (New Drug Application) for Zerit, [8] an extended-release preparation of stavudine, which is a NRTI (Nucleoside Reverse Rranscriptase Inhibitor). The one-capsule, once daily formulation should improve quality of life and compliance relative to the FDA-approved immediate-release formulation of Zerit, with similar virologic activity. In a recent phase III multinational, randomized, doubleblind, placebo controlled trial, BMS 099   (a type of studies performed by Bristol-Myers Squibb) combining Zerit with efavirenz and lamivudine, 80% of the 392 patients in the treatment arm containing Zerit extended release achieved viral load suppression below 400 copies/mL after 48 weeks of treatment, compared with 75% of the 391 patients in the arm containing Zerit immediate release. More than half of the patients in each treatment arm reduced viral load to fewer than 50 copies/mL, and adverse events were similar in both groups.[9]

Coviracil (emtricitabine) is another NRTI for which Triangle Pharmaceuticals [10],[11] has submitted an NDA. This drug has potent selective activity against both (HBV) hepatitis B virus  and HIV. In a 48-week, double-blind, placebo-controlled phase III trial [FTC-301, The FTC 301 study is a randomised, double-blind, double dummy comparative study of FTC (Emtricitabine) vs d4T with a backbone of didanosine and efavirenz in 571 treatment naive individuals] comparing once-a-day Coviracil to immediate-release Zerit given twice daily, each combined with efavirenz and Videx EC (didanosine), interim analysis of safety and efficacy led to unblinding the trial and offering Coviracil to all 571 subjects. Patients with Coviracil in the arm had significant improvements in immunologic function, and 87% had persistent virologic response through six months.[12],[13]

To tackle the emerging problem of antiretroviral resistance, Pfizer [14] is in phase III testing of capravirine, [15] a second-generation non-nucleoside reverse transcriptase inhibitor (NNRTI) which has been shown in several studies to retain activity against HIV that has developed some NNRTI-resistant mutations, including K103N.[16] After the unexpected development of vasculitis in animal models, the FDA placed human clinical trials of capravirine on partial clinical hold in January, 2001.[17] Agouron Pharmaceuticals, [18] a Pfizer company slated to market capravirine, believes that it may offer unique benefits to HIV infected patients and therefore remains committed to its clinical development and to additional toxicology studies.

One of the problems with available protease inhibitors (PI) is their tendency to increase blood lipid concentrations. In hopes of circumventing this problem, Bristol-Myers Squibb has begun phase III testing of atazanavir, an experimental PI formerly called BMS-232632. [19] A randomized trial in 467 patients at 51 sites suggests that concentrations of total cholesterol, LDL cholesterol and triglycerides do not increase significantly in treatment naive patients taking a HAART regimen including atazanavir, and that the drug is effective, safe and well tolerated compared to nelfinavir. Other advantages include once daily dosing and fewer gastrointestinal effects, but the drug is more likely to induce jaundice than nelfinavir.[20]

Unlike currently approved antiretroviral drugs, which target viral enzymes involved in replication, the fusion inhibitors are designed to block HIV from fusing with a host cell before the virus begins replication within the cell. Trimeris [21]has submitted an NDA for fusion inhibitor Fuzeon (enfuvirtide; formerly T-20) [22] and has received fast track designation from the FDA. Physician enrollment for the U.S. Early Access Program for Fuzeon began August 19, 2002, for a limited number of patients with advanced HIV disease. Data from two large, international phase III trials suggest that patients on combination therapy with Fuzeon are twice as likely to achieve undetectable blood levels of HIV and a significant increase in immune cells at 24 weeks compared with patients who received combination therapy without Fuzeon.[23]

PRO 542, known as CD4-IgG2, is a novel fusion protein that incorporates the HIV-binding region of the human cell surface receptor for HIV into a human antibody molecule, [24] from Progenics Pharmaceuticals [25] is a novel fusion protein shown to safely reduce viral load by 60% to 80% in selected patients. Multi-dose phase II studies are under way of PRO 542 as salvage therapy for HIV-infected patients refractory to currently available antiretroviral medications.

Rather than targeting the HIV virus directly, immune system modulators boost the ability of the immune system to pause viral spread. These include Celgene’s drug Thalomid (thalidomide) [26]; Chiron’s product IL-2 Proleukin (aldesleukin), [27] a recombinant form of interleukin-2; Alferon N Injection (interferon a-n3), a human leukocyte derived natural interferon treatment, and WF10 (an immuno modulator) designed by Dimethaid Research [28] to enhance macrophage function and to thereby eliminate reservoirs of HIV-infected CD4 cells and macrophages. All these drugs are in phase III development. WF10 [29], intended for use as adjunctive treatment, is a chemically stabilized chloride matrix called tetrachlorodecaoxygen (TDCO). Studies to date suggest that WF10, administered intra-venously, is safe in patients with HIV and that it may be especially useful in moderate to advanced AIDS. [30] By promoting phagocytosis and reducing macrophage production of TNF-a (Tumor Necrosis Factor-a), WF10 may slow AIDS progression and reduce the likelihood of opportunistic infections.

Based on phase II trials by Immune Response Corporation, Remune (HIV-1 immunogen) [31] is a promising adjunctive therapy using inactivated virus to restore HIV specific immune responses, with positive effects on viral load and on counts of CD4 helper T lymphocytes.

Vaccines designed to prevent AIDS rather than to slow its progression are AIDSVAX B/B and AIDSVAX B/E, both in phase III development by VaxGen.[32],[33]

An alternate approach to prevention is the spermicidal microbicide BufferGel (polyacrylic acid), in phase III development by ReProtect. [34] By increasing the acidity of semen, this product tends to kill sperm and to inactivate pathogens including HIV, HPV, herpes, syphilis and gonorrhea. [35] Data on the anti-HIV activity of BufferGel are expected in two years.
Post treatment issues

Complications of AIDS include lipodystrophy syndrome associated with HAART, for which Ark Therapeutics [36] is in phase II testing of EG005, a drug that reduces cellular levels of Angiotensin II and thereby increases mitochondrial efficiency.

Drugs targeting AIDS-related dementia for which phase II testing is completed include CPI-1189 (4-acetylamino-N-tert-butyl-benzamide) from Centaur Pharmaceuticals [37] and memantine (3,5-dimethyl-adamantan-1-ylamine) from Forest Laboratories.[38]

CPI-1189, prevents apoptosis and reduces glial fibrillary acidic protein immunostaining in a TNF-a infusion model for AIDS dementia complex, [39]  is an oral drug that potentially inhibits neuroinflammation, has safely produced statistically and clinically significant cognitive improvements in AIDS dementia in double-blind, placebo- controlled clinical studies lasting five months.

Memantine, which modulates N-methyl-D-aspartate (NMDA) receptor activity, has been used to treat other types of dementia, with rapid and lasting improvement in cognitive, psychological, social and motor impairments. Animal models including transgenic mice suggest the applicability of this drug to AIDS dementia, for which clinical trials are ongoing.[40]
Conclusion

Drugs identified in the pipeline may continue to improve the outlook for AIDS patients however significant progress in HAART has reduced the number of AIDS deaths in recent years. New antiretroviral drugs and their combination regimens may improve tolerability and compliance. Emergence of fusion proteins, which prevent HIV entry into the cell may prove as one of the major tools in the management of the disease. Additionally immune modulators work synergistically in order to reduce the viral load as well as immunogens are supposed to improve immunity.

Before You Read, take note: God Is Brilliant, in all of his creations. Disregard not that, and heed it.



The genes sense time? A clock ticks inside us!
-S. Valeri


Until I peeled apart my eye lids, didn't realize that it is nothing but the cosmic rays emanating from sun only has interrupted my cozy early morning dreams. Every morning is the same, there is a literal "soft-war" inside the brain, arguing for and against to get up and start the daily routines. Though there always remains a residual desire to sleep more in the warmth of the plumage, the irresistible pull from within never lets us to lay down beyond a limit. Assumably most of us might have been experiencing it in every morning. Have you ever wondered about what really exerts from within, and demands to wake up out of the bed every morning? If it is said that nothing but the clock ticks inside hauls us from within, it won't be an exaggeration at all. Yes, there really exists a clock inside our skull! Now, a counter question may pop up - could the springs and cogs of the whole clock machinery identifiable inside the network of myriad of neurons in the brain? It should be a perplexing task to convince for a moment, but daring to answer 'yes' would be appropriate in case it can be agreeable to imagine that the networking of proteins is analogous to the 'springs' of a clock and the genes, which are responsible for the expression of these proteins, function as the main cogs of the clock mechanism. In this era of proteomics, the chronomics (the science of time) has gained much impetus than ever since. In this brief account, a cross-section of the important developments in the chronobiology and its implications on human health and welfare are being presented.

The only known life hosting planet, earth, revolves on its own axis. This planetary movement has endowed a rhythmically oscillating level of light on earth. Consequently all the organisms on earth experience the rhythmic changes in day and night, which is an easily perceivable signal for most organisms on earth and not to miss at all. This rhythmic change in day and night has constrained the organisms on earth to organize their daily activities into a defined temporal pattern, which repeats in cycles roughly close to a period of 24 h. Therefore, this rhythmic activities of an organism is termed circadian rhythm (from Latin 'circa' = about; dies = day). For e.g. the daily rest-activity rhythm in Drosophila have a period of ~ 24.3 h (Fig.1).


figure 1 depicts the daily rest-activity pattern of a fruit fly - Drosophila. Top black and white bars represent 12 h night and 12 h day respectively. The multiple small horizontal black bars filled in the figure represent the event records. The fly was kept in a small tube that contain food in one end and was capped at the other end with a cotton plug. When the fly moves along the tube, it crosses a laser beam, that recorded as an event. The contiunous horizontal black bar represents the constant darkness. The yellow shaded area indicates further the time which is in dark period. The fly exhibits a typical bimodal activity pattern, when light comes 'on' and 'off'. It continues through also in constant darkness


In classical terms, circadian rhythm is defined as oscillations in the biochemical, physiological, and behavioral functions of an organism with a periodicity of ~ 24 h. Circadian rhythms have three basic features. First, the rhythm is an innate property of the organism and is maintained under constant conditions. Second, the period length is temperature compensated, thus it is ensured to maintain the rhythm at a constant rate throughout the physiological range of external temperature. Third, it is synchronizable with the outside world by external cues. Light plays a major role as a circadian zeit geber (German:zeit =time, geber = giving), may be because light is an essential factor to maintain the life on earth. The circadian rhythms are exhibited by organisms from every level of the 'life-tree', the blue green algae, cyanobacteria, to the advanced level of life, the mammals, and indeed humans too. The conservation of biological clocks throughout evolution has suggested that it confers a selective advantage to the organism. It is important that the organism must be well fit to the environmental settings to survive through the pressures of life challenges. Therefore, the organism should have a highly functional biological clock, which can sense and respond quickly to the environmental fluctuations. The activities of different species of organisms on earth have been temporally discrete, in such a way that it should benefit the organism maximum. For e.g. the bees go in search for nectar only after the dawn, because the blossom is in correlation to the sun light. In a different perspective, some lower animals are avoided being preyed by higher predatory animals by simply scheduling their foraging out of phase with the predator, thus compromising for the potential risk of predation and feeding. It might be noticed that humans are normally active during the day time (diurnal), with a period of 25.1 h, whereas some lower animals like mice are active during the night (nocturnal), with a period of 23.7 h.

History:

The concept of time has always perplexed and fascinated people. The research on biological rhythms has drawn the attention of scientists since 1729, when Jean Jacques d'Ortous de Mairan (a French astronomer) has started to look at the changes in plants in relation to day and night (1) . Thereon the quest was continuing relentlessly through the centuries. Though it was widely accepted only ~ 60 years ago that the life forms have an internal timekeeping device, a genetic underpinning for circadian clock in plants was first proposed by Buenning in 1935 (2). It took another 30 years for Pittendrigh (1967) to infer that the animals (insects) also have an internal clock that too geared by a genetic gadgetry (3).

The 1971 have witnessed a discovery that ever since connected a behavioral trait to a gene, thereby gave birth to a novel branch of biology- the behavioral neurogenetics. Konopka R J (Caltech, USA) was the first to report a 'clock gene' from Drosophila melanogaster (the common fruit-fly)(4). His discovery of period mutant (a defective gene) has marked the real beginning of molecular chronobiology. Flies carrying mutation in this gene exhibited an altered rhythm in their daily rest-activity pattern. However, as recently as 1994 only the clock gene period in Drosophila was molecularly characterized (5). In last decade there was an explosion in this field which was aptly dubbed by some clock researchers as "Clockwork Explosion". As a culmination of the efforts of many researchers, presently there is a wealth of knowledge available to the scientific community. A family of genes called clock controlled genes (ccg) exists today. The ccg exceed more than seven by now and every day breaks into a new surprise as well. Most of the discoveries about circadian clock have been achieved in the model organism Drosophila, because of the vast repertoire of genetic and molecular tools available for this simple fruit fly. Its genome was completely sequenced and many genes have already been identified and much more are annotated. For better correlation of the findings in fruit fly to higher animals researchers have embarked onto mammals, like rodents, in the recent past, nevertheless the progress made is amazing than predicted. The uncanable achievements in the field of bioinformatics and molecular biology have fuelled the pace of rhythm research exponentially.

This realm of research has come to light when its paramount importance has been emphasized by recognizing 'biological clock' as the first runner-up in the list of break-through of the year in 1998 by the American Association for the Advancement of Science.


Modeling the circadian clock:

The simple clock model accommodates three components, an input, an oscillator and an output. The most potent input in the environment is the light signals. However, temperature and even the food could work as an input to set the clock. The input feeds the internal oscillator, and informs about the external environment, where the organism lives. The oscillator 'reads' and 'translates' the information about the external environment and generates an output in compliance with the external settings. The ability to read correctly the external world is very important for an organism to reorganize its physiology and behavior to have a better fitness. Fitness is the key to survival of an organism according to the classic theory of Darwin. The well organized will have better survival in an ever-changing world, where it can swiftly adapt to the challenges.

Photoreceptors like rhodopsin and cryptochrome function as the input into the central oscillator. Mutants for various photoreceptor molecules or genes down stream to them (in a functional cascade) have proven that light is an important signal for setting the internal clock. The eyes play a major role in circadian photoreception, besides its primary function, vision.

The central oscillator is composed of multiple genes and their proteins. In Drosophila the ccg genes express in clock neurons in the brain and other peripheral tissues. These genes show a rhythmic expression pattern, which is the basis of circadian functions. Drosophila has two clusters of important clock neurons called lateral and dorsal neurons. The clock proteins peak in the clock neurons in midnight (Fig. 2).


Figure 2 shows a full brain of Drosophila. The brain was stained with anti-PER antibody, which is having a fluorescent label. The brain was dissected out at midnight to check for the expression level of PERIOD protein. The arrows indicate the various clock neurons in the brain, which express the clock protein PERIOD. sLNvs means small lateral ventral neurons; lLNvs, large lateral ventral neurons; LNds, lateral neurons dorsal; DNs, dorsal neurons. The sLNvs are believed to be the pacemaker neurons in Drosophila

and trough in the daytime. This could be visualized by staining these proteins using fluorescent-labeled antibodies against the proteins.

The output is the general behavioural or physiological function of an organism. Drosophila researchers routinely study two main outputs. First, the daily rest-activity rhythm, which have a period of ~ 24.3 h; it continues in constant darkness as well. Second, the eclosion rhythm (emergence of a fly from a puparium).

A clock-mutant (having defect in a clock gene) fails to show an overt rhythm in rest-activity pattern or even the eclosion rhythm. That's how really in the beginning researchers have 'spotted' the clock mutants, and later on identified and characterized the 'clock genes' involved in the clock mechanisms.



Molecular mechanisms:

The basic mechanisms behind the orchestration of circadian timekeeping are achieved by the unique wiring of proteins. Presently it has been envisaged that the clock machinery works based on a transcriptional-translational feed back loop (6 - recent review; references therein). That literally means, the transcription of a gene - the copying of DNA sequences, is negatively influenced by the translation of the same gene- the process of production of a protein from a gene. There is a temporal separation of approximately 6 h between these two processes. Besides, at least one of the clock protein is sensitive to light signals. The interplay of light and the clock proteins eventually accomplishes a circadian pattern of clock gene expression.

Presently, there are at least seven clock controlled genes known in Drosophila. The concerted action of these genes achieve the 'jerk and torque' of the clock machinery. They are namely, period (per), timeless (tim), Clock (Clk), cycle (cyc), vrille (vri), double-time (dbt) and shaggy (sgg). Most of them have counterparts in higher animals including man. For e.g. there are three period genes in humans, hper1, hper2 and hper3.

The principles of molecular timekeeping in a nutshell: A dimer formed by the mutual binding of CYCLE (CYC) and CLOCK (CLK) proteins reach and attach to the promoter (a specific DNA sequence that drives a gene for expression) of per and tim genes. In turn, the transcription of the per and tim switched on. This event happens in the nucleus. Then the mRNAs of per and tim are transported for protein synthesis into the cytoplasm, where PERIOD (PER) and TIMELESS (TIM) proteins are generated. They get accumulated from early evening through the night in the cytoplasm. As their levels shoot up in the cytoplasm, some 'modifier' proteins like DOUBLE-TIME (DBT) and SHAGGY (SGG) change the physical properties of the PER and TIM by adding a phosphate group (phosphorylation) onto them, later on the PER and TIM form a heterodimer complex. The phosphorylation of PER-TIM initiated the translocation of the PER-TIM heterodimer into the nucleus. As long as the PER-TIM dimer is shuttled to the nucleus, their production switch will be kept on. However, once the concentration of PER-TIM exceeds a limit in the nucleus (by mid night), it interferes with CLK-CYC dimer molecules present in the nucleus, which have been driving the per and tim. The binding of PER-TIM to CLK-CYC distorts the latter's functioning, and results in the switching off the per and tim transcription. Consequently, the further production of PER and TIM is suspended. This state remains as long as the levels of PER and TIM are high in nucleus. That is one side of the story, on the other side, when the light irradiates in the dawn, the TIM in the nucleus (already phosphorylated) gets degraded through a light mediated action. The cryptochrome (a photoreceptor molecule) transduces the light signals to the TIM. Once the TIM is degraded, the PER loses its existence as such. In conclusion, the light rays eliminate the protein complex, PER-TIM, from the nucleus and in turn releases their 'disturbing' action on CLK-CYC. Once the CLK-CYC relieved from the interference of PER-TIM, the former resumes its functioning once again, thus paves the way for another bout of the same expression of per and tim in the day light. (Fig. 3).


Figure 3 is the schematic representation of the clock mechanism in Drosophila. At night, the CLK-CYC dimer binds to the per and tim genes' promoter and start their transcription - produces the respective mRNAs. The mRNAs are translated into proteins. The proteins get phosphorylated and form a dimer:PER-TIM. This dimer translocated into the nucleus, where it binds with the CLK-CYC and disturbs the latters action and stops the production of mRNAs. At day, the dimer PER-TIM is degraded by the light, through a CRY mediated mechanism. This relieves the negative impact on CLK-CYC dimer, thus resumes the transcription of per and tim again.

Medical implications of circadian clock:
The circadian clock has far reaching implications on human health and welfare. There are many emerging issues, which have greater concern in the present day scenario due to the advanced life styles adopted by many populations around the world. In this competitive world, to increase the production rate people have been put through in constant shift work routines; and to promote more business the personals have been travelling across the meridians quite frequently. These activities have bestowed few serious health risks like sleep disturbances, jetlag etc. to the subjects involved in. It is debatable that whether our internal clock machinery has evolved that quickly as we have technically advanced. Few major issues of the human health connected to the circadian clock have been discussed briefly.

The melantonin, which displays a clear circadian pattern, peaks during the night, has been routinely tested for most circadian rhythm related investigations (7). The recent report that circadian clock genes hPer1, hPer2, and hPer3 are expressed in a circadian manner in human peripheral blood mononuclear cells (PBMCs), with the peak level occurring during the habitual time of activity (, offers few more candidates for the studies on human circadian rhythms and their associated disorders.

Reproductive fecundity: The incidence of infertility cases are rapidly increasing among the human population. But, the exact reason for this phenomenon has not yet been identified. It was always believed that there is a relation between the fitness of partners and the success of a pregnancy. In this context, it is well recognized today that circadian clock influences the fitness of an organism (9). The reproductive fecundity is the best output to assess the fitness of an organism. The elegantly designed experiments in Drosophila have proved that defect in a clock gene(s) afflicted the sperm release from the testes to seminal vesicles (these tissues displayed rhythmic and autonomous expression of clock genes), which in effect reduced the successful fertilization frequencies. However, the influence of circadian clock on human reproductive function has not yet been investigated.

FASPS: The human sleep-wake cycle remains as a mystery black box to the entire medical world. Though most of us have a regular pattern of sleep-onset at the fall of night and awakening by about 1-2 h after sunrise, there do exist peculiar marginal cases. In a particular case, called familial advanced sleep phase syndrome (FASPS), the subject goes into bed an hour after sunset and is wide-awake by 4.00 in the morning. Recently, FASPS has been linked to a mutation in human clock gene, hper2, the homolog to Drosophila per (10). The mutation in hper2 blocks a phosphorylation site, which is required for the nuclear translocation of the protein.

DSPS: The opposite case of FASPS is the delayed sleep phase syndrome (DSPS), where the patients go to bed around 4:00 in the morning and wake up around the noon. Though the genetic mechanism underlying DSPS is yet to be elucidated, it is conceivable that there is a 'misinterpretation' about the day by the internal clock mechanism. Either the input or the oscillator may have been corrupted, because the bright light therapy to this patients has found to be quite efficacious.

Fragile X syndrome: The absence of a protein, called fragile X mental retardation protein (FMRP), predisposed the new born baby to become mentally retarded. As the name indicates it is a X chromosome linked inheritable disease. It has been hypothesized that FMRP is involved in the processing and/or translation of mRNAs. To better understand the molecular mechanisms behind fragile X syndrome, a Drosophila homolog of hFMR1, the dfmr1 has been mutated and studied in depth. It is well documented that the fragile X syndrome patients show a variety of sleep disorders. The studies on Drosophila has revealed that the mutation in dfmr1 affects an output function of the circadian clock (11), and the Drosophila become arrhythmic in constant darkness (normally rhythmic). Moreover, the Drosophila model for fragile X syndrome provides new insights into the sleep-wake cycles of animals, because it revealed additional molecular mechanisms connected to this particular genes.


SAD: The seasonal affective disorder (SAD)- winter depression- is a syndrome characterized by recurrent depressions, mainly in countries where there is a severe winter, and relatively less sun light. This syndrome harbors hypersomnia, hyperphagia and carbohydrate craving. SAD prevalence in the United States is estimated to be ~6%, about 11 million people. The females are more affected than males (12). Bright light therapy has been suggested to be beneficial for these patients to recover from their depressive mood, at least to certain extent. Assumably, some circadian clock functioning is impaired in these patients. The sleep and melatonin release is under the control of a central clock in the brain; the supra chiasmatic nuclei (SCN) is believed to receive some light signals from the retina, and that triggers the internal circadian clock function.


Jetlag: Approximately 78% of the subjects have reportedly been suffered sleep disturbances during the first night after a transmeridian flight (13). An accompanying gastrointestinal disturbance has also been reported by ~60% of these subjects. It has been demonstrated that the transmeridian flying is an acute stress for female aircrews, they have been suffering a delayed ovulation and associated menstrual disturbances, at in 30-35% of them. Traveling across meridians imposes a new light-dark phase on the traveler. In this newer day-night phase, the traveler's internal clock is set to the earlier time zone, and he/she needs more time to re-synchronize to local environment. As a consequence, the individual suffers from insomnia, fatigue and irritability. At this moment, the social demands in a new time zone and the person's ability to coping with it are in conflict, because the internal clock is out of phase. The individual takes at least a day or more to adjust to the local environment. In mammals including humans, the pineal hormone melatonin level peak during the night, and its release could be suppressed by exposure to light pulses. The SCN expresses high-affinity melatonin receptors; and to circumvent the effects of jet-lag melatonin has been widely prescribed in clinics, though its usefulness in treatment of jetlag and other ailments is controversial. Studies on pilots and cabin crews have exhibited cognitive deficits, possibly in working memory that became apparent after several years of chronic disruption of circadian rhythms due to the constant flights across the transmeridian (14).

For airline pilots, sleep deprivation and circadian desynchronization due to rapidly rotating shift schedules, rapid time zone changes, and unorganized sleep/wake cycles can result in severe performance decrements. Twelve airline accident cases in the past suggest that pilot scheduling might have been a factor for these accidents; and the investigation committees have came up with suggestions for the chronohygiene of the pilot scheduling (15). In conclusion, the rapid time-zone transitions impaired performances and consequently the safety of both aircrews and the frequent flyer. Therefore, more prudent practices should be implemented in the air traffic systems to avoid further incidence of airline crashes.

Rotating Shiftwork: Impaired mental and physical performance is always a spin-off of the sleep deprival, which is inevitable in a shiftwork routine. One of the compounded effect of sleep disturbance is fatigue. The individual's ability to perform a task optimally as dictated by the circadian clock is often skewed from the demand of a shiftwork regime. In realistic terms, the person fails to pay attention to the minute and often important details of his task. Many dreadful disasters occurred in the past were regarded as errors committed by workers who had not been synchronized to work in the night shift. For example the disasters at Three Mile Island, Chernobyl, and Bhopal were occurred between midnight and dawn (16). The ailments arising out of a shiftwork routine could be ameliorated to a greater extent by simply exposing the subject to high-intensity light during the night, to shift the phase of the circadian clock (17).

Breast Cancer: The incidence of breast cancer has steadily increased during this century in industrialized countries. The projected reason is that the individuals are exposed to light for longer periods than that afforded by the natural daily light-dark cycle. The melatonin hypothesis proposes that the suppression of melatonin secretion at night by artificial light increase the chances for breast cancer by increasing the exposure to estrogen (18). The lifetime exposure to estrogen is a major risk factor for breast cancer. To further support this hypothesis, it has been reported that severely blind women had about half the incidence of breast cancer than women with normal vision (19). Further research is warranted to arrive at a precise answer to the many open questions.


Learning and memory: per mutation in Drosophila has been linked to robust changes in certain drug sensitivities and to defects in the patterns of expression of some genes that are tightly connected to the control of learning and memory (20). It is well known that the mental acuity is not the same throughout the day. Therefore, it is advisable to find out that at what time a person can learn the best and according to that he/she should pursue his/her learning. Understanding more about the role of different genes on learning and memory would open novel avenues for pharmacological interventions to manipulate the learning process in the future.
Issues related to diagnosis and treatment: The success of any treatment depends on how accurately the physician can diagnose the cause of the problem. The art of diagnosis relies basically on observing the physiological and behavioural variables. These variables change in a rhythmic manner over the course of a day. The relevance of considering the circadian rhythmic variance, while performing diagnosis is not yet clear. But, physicians keeping in mind this fact would certainly give an advantage. The efficacy of cancer treatment drugs has been found to be largely depend on the time of delivery (21). Thus, it is possible to improve the therapeutic potential and minimize toxic side effects by optimizing schedules for administering drugs. As a matter of fact, it is reminded that in Ayurveda practices the time of administration of a drug is strigently followed.


Conclusion:



Since the discovery of period gene, the circadian clock research has undergone a quantum leap. Presently our understanding about the circadian clock both in lower animals and mammals is quite appreciable. The components of the circadian clock, once seemed to be quite elusive and mysterious, were brought to light by ingenious experiments. The clock picture is more clear now with a transcriptional-translational feedback loop functioning in the center. A family of clock genes and their functions are widely known today. However, the clock puzzle is not yet finished absolutely. The bridging of knowledge on molecular mechanisms of the circadian clock in Drosophila and human illnesses like fragile X syndrome is a commendable achievement. It points toward the feasibility to extract valuable information from lower model systems, that can find applications in medical science. Rational drug design, aimed at many disorders connected to the circadian clock, like insomnia and FASPS could be further rectified by unraveling the molecular mechanisms under laying these disorders. Eventually, it is presumable that further development in this filed may be improving the general well being of the modern man, who abodes in a sophisticated environment or rather in a controlled environment where programmed machines set all the boundaries. The future generations may be in a position to program, as per wish, their internal clock. We can anxiously await to watch what time holds in its store!


Sources:
1.DeMairan, J. 1729. Observation botanique. Historie de L'Academie Royale des Sciences. 35-36.
2.Buenning E. 1935. Zur Kenntnis der erblichen Tagesperiodizitaet bei den Primaerblaettern von Phaseolus multiflorus. Jb. wiss Bot 81:411-418.
3.Pittendrigh CS. 1967. Circadian systems. 1. The driving oscillation and its assay in Drosophila pseudoobscura. Proc. Natl. Acad. Sci. USA 58:1762 - 1767.
4.Konopka RJ, Benzer S. 1971. Clock mutants of Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 68:2112 - 2116.
5.Konopka RJ, Hamblen-Coyle MJ, Jamison CF, Hall JC. 1994. An ultrashort clock mutation at the period locus of Drosophila melanogaster that reveals some new features of the fly's circadian system. J. Biol. Rhythms. 9:189-216.
6.Ralf Stanewsky: Genetic analysis of the circadian system in Drosophila melanogaster and mammals. J. Neurobiology. 54:111-147,2003.
7.Nakahara D, Nakamura M, Iigo M, Okamura H. 2003. Bimodal circadian secretion of melatonin from the pineal gland in a living CBA mouse. Proc. Natl. Acad. Sci. USA 100:9584-9589.
8.Boivin DB, James FO, Wu AF, Cho-Park PF, Xiong H, Sun ZS. Circadian clock genes oscillate in human peripheral blood mononuclear cells. 2003. Blood Jul 31.
9. Beaver LM, Gvakharia BO, Vollintine TS, Hege DM, Stanewsky R, Giebultowicz JM. 2002. Loss of circadian clock function decreases reproductive fitness in males of Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 99:2134-2139.
10. Toh LK, Jones CR, He Y, Eide EJ, Hinz WA, Virshup DM, Ptacek LJ, Fu YH. 2001. An hPer2 phsophorylation site mutation in familial advanced sleep phase syndrome. Science :291:9 Feb. 1040-1043.
11. Inoue SB, Shimoda M, Nishinokubi I, Siomi MC, Okamura M, Makamura A, Kobayashi S, Ishida N, Siomi H. 2002. A role for the Drosophila Fragile X-related gene in circadian output. Current Biology, vol. 12 . August 6. 1331-1335.
12. Rosethal NE, Sack DA, Gillin JC, Lewy AJ, Goodwin FK, Davenport Y, Mueller PS, Newsome DA, Wehr TA . 1984. Seasonal affective disorder. A description of the symptoms and preliminary findings with light therapy. Arch. Gen. Psychiatry. 41: 72-80.
13. Winget CM, DeRosha CW, Markley CL, Holley DC. 1984. A review of human physiological and performance changes associated with desynchronosis of biological rhythms. Aviat. Space Environ. Med. 55: 1086-1096.
14. Cho K, Ennaceur A, Cole JC, Suh CK. 2000. Chronic jet lag produces cognitive deficits. J. Neurosci. March. 15,20(6): RC 66.
15. Price WJ, Holley DC. 1990. Shiftwork and safety in aviation. Occup. Med. Apr- June;5(2): 343-77.
16.Folkard S. 1990. Philos. Trans. R. Soc. London Ser. B 327: 543-53.
17. Dawson D, Campbell SS. 1991. Sleep 14: 511-16.
18. Stevens RG, Davis S, Thomas DR, Anderson LE, Wilson BW. 1992. FASEB J. 6:853-60.
19. Hahn RA. 1991. Epidemiology 2: 208-13.
20. Abarca C, Albrecht U, Spanagel R. 2002. Cocaine sensitization and reward are under the influence of circadian genes and rhythm. Proc. Natl. Acad. Sci. USA Jun. 25: 99 (13): 9026-30.
21. Levi F. 1999. Cancer chronotherapy. J. Pharm Pharmacol 51: 891-898.
22. For general interest:Michael W. Young and Steve A. Kay : Time Zones: A comparative genetics of circadian clocks. Nature Reviews Genetics. Vol. 2,702-715, Sept. 2001.

For the first time in biology, a team of researchers at The Scripps Research Institute (TSRI), California is reporting that the human body makes ozone. Led by Professor Paul Wentworth, Jr, Ph.D., the team has been slowly gathering evidence over the last few years that the human body produces the reactive gasâ€"most famous as the ultraviolet ray-absorbing component of the ozone layerâ€"as part of a mechanism to protect it from bacteria and fungi.

This was a completely unexpected development, since for the last 100 years, immunologists believed that antibodiesâ€"proteins secreted into the blood by the immune systemâ€"acted only to recognize foreign pathogens and attract lethal "effector" immune cells to the site of infection. Two years ago, Lerner & Wentworth demonstrated that antibodies are capable top produce ozone and other chemical oxidants when they are fed with singlet oxygen (a reactive form of O2). Last year, Babior, Lerner & Wentworth evidenced that oxidants produced by antibodies can destroy bacteria by poking holes in their cell walls. Now, Babior & Wentworth found where the singlet oxygen comes from ~ it’s the Neutrophils, one of the effector immune cells which are little cellular factories that produce singlet oxygen and other oxidants. Their research work got recently published in one of the most prestigious science journal, Proceedings of the National Academy of Sciences (PNAS).

Ozone is a reactive form of oxygen that exists naturally as a trace gas in the atmosphere. It is perhaps best known for its crucial role absorbing ultraviolet radiation in the stratosphere, where it is concentrated in a so-called ozone layer, protecting life on earth from solar radiation. Ozone is also a familiar component of air in industrial and urban settings where the gas is a hazardous component of smog. However, ozone has never before been detected in biology.

Ozone is also reported to be a selective & rapid oxidizer of viruses (HIV, Herpes simplex and zoster, cytomegalovirus, Epstein-Barr, myxoviruses & retroviruses), bacteria (coliform & staphylococcus) and virally infected cell membranes. Healthy self cells may be spared because they have the built in protection of the oxidation buffers, tocopherol, ascorbic acid, superoxide dismutase (SOD), uric acid and glutathione peroxidase. It has been documented that cancer cells have little or no expression of oxidation buffer enzymes, catalase, SOD & peroxidase. Hence ozone is particularly damaging to cancerous cells.

The question still remained, however, as to how the antibodies were making the ozone. The TSRI team knew that in order to make the ozone and other highly reactive oxidants, the antibodies had to use a starting material known as singlet oxygen, a rare, excited form of oxygen. During an immune response, the neutrophils, where this singlet oxygen is generated, engulf and destroy bacteria and fungi by blasting them with these oxidants.

The work of the TSRI scientists suggests that the antibacterial effect of neutrophils is enhanced by antibodies. In addition to killing the bacteria themselves, the neutrophils feed singlet oxygen to the antibodies, which convert it into ozone, adding weapons to the assault.

The research paper brings out something that is really new, but there are still a million questions that follow. What does the ozone do to the body's proteins and nucleic acids? Can neutrophils make ozone without the antibodies? How long does ozone last in the body? And, most importantly, how will these discoveries help to cure disease? The research team continues to investigate.

When life gives you a lemon, then take a pinch of salt, sprinkle it on the lemon and suck the juice out.

Make due with what you can, with the situation you are in.

In any case, lemon, though bitter as it is, is a superb source of vitamin C.

Find the goods in any situation

I believe in Evolution, and its divine inspiration.

Science is witness to the unassailable brilliance of God.

The Radiance of Allegheny~

Japanese Hibachi @ Aoyama,
With the buds,

Celebrating post-Midterm Examinations   






Udon, anyone?


Takeshi, the chef; creative dude




'Valentines day heart' lol




Lisa and my sister

im leaving in 30 minutes for a trip to Erie, PA. Gonna get some hibachi tonight then maybe..movies?

Peace out folks!

So? Tubag! 

Four jobs I have had in my life:
    1. Security Operator
    2. College Tutor
    3. College Teacher's Assistant
    4. Nursing Home Activities Supervisor

Four Places I have lived:
    1. Bohol
    2. New York City,
    3. New Jersey
    4. Pennsylvania

Four places I have been:
    1. Philippines
    2. China
    3. Canada
    4. Ukraine

Four of my favorite foods:
    1. Monggo   
    2. Pancit Bihon
    3. Cari Cari
    4. Chicken Adobo

Four places I'd rather be:
    1. Medical School
    2. In my dorm
    3. Bohol
    4. Home

Four friends or relatives I think will respond first:
    1. Hazel
    2. Ate Belle
    3. Ate Ging
    4. Happy

Now thats a romantic site, 'te. Enjoy the night

Owowowow! The city of Love!

Ayaya

sakit!

hahaha, c'mon haps. Whats your dream destination?

Quote from: PrincessSaDagat on February 16, 2008, 06:49:03 AM
Lorenzo, Don't lick to much you'll end up with dry, crack lips.

hahahaha, ill keep that in mind, Princess.

2008 Porsche Carrera GT




I can't stop licking my lips

hmm, pupils dilating? interesting.

Quote from: BlueAngel on January 26, 2008, 06:23:10 AM


Dong, kani lang akoa Cabrio kana tiptonic or automatik
unya na kon makadaug ug Lotto hahahahaha (wishful thinkin)

W. O. W

That took my breath away

Ma'am Bambina,

The singer is: Marvin M. Ang
it is the Filipino rendition of the Japanese version by Ted Ito.

The original is called "Saigo No Iiwake"

http://youtube.com/watch?v=ZXO3KCUt7Io

Lyrics:

Neta furi ga konna ni tsurai koto da to wa
Ima ochita shizuku wa namida da ne
Soshite kimi ga dete iku yoake wo matte
Kurayami ga kowai kimi no koto dakara

Ichiban daiji na mono ga ichiban tooku e iku yo
Konna ni oboeta kimi no subete ga omoide ni naru

Dare kara mo kimi nara sukareru to omou
Shinpai wa iranai sabishii yo
Muri ni boku no tame da to sayonara no wake
Omotteru kimi dakara setsunakute

part2:
Ichiban chikaku ni itemo ichiban wakariaenai
Konna ni ai shita boku no subete ga iiwake ni naru

Ichiban daiji na mono ga ichiban tooku e iku yo
Konna ni oboeta kimi no subete ga omoide ni naru

Subete ga omoide ni

Ichiban chikaku ni itemo ichiban wakariaenai
Konna ni ai shita boku no subete ga iiwake ni naru

Ichiban daiji na mono ga ichiban tooku e iku yo
Ichiban chikaku ni itemo ichiban wakariaenai
Ichiban daiji na mono ga ichiban tooku e iku yo

Same age nako pre, 22 siya. Sige pre, good luck on your hunting dinha ha! May your nets catch the biggest fish in the sea


More power to you, my friend.

Take a look at this map, circa during the 17th century.

It clearly includes not only Philippines-proper, but Sabah (northeastern Borneo).

Sabah was part of the Philippines then. And must be part of it now, and in the future!

Quote from: ligiah on February 15, 2008, 03:58:47 PM

hahaha! unya Bran, diha ka'y nakagakus gahapon?

haha, why do you want to know? LOL

seriously oo i got a hug and a kiss on the cheeks. Na red baya.

Ikaw, Gai?

I don't think she'll get out of this one. The United States will have her extradited if she ever flees the country to the Philippines. She's looking at about 30+ years of cold hard jail time.

Its not worth some 100k. Money can't buy life.

Unfortunately for her, she didn't realize that until its too late.

The motto is: practice discretion.

haha ug gusto ka pre, i pa introduce ka nako niya.


di ko gusto i-pa heighten ang relationship kai i know her very well. the relationship will not work jud.
Amigo-amiga ra

'te Blue. did you deliver without analgesics or did you have it?

Tinuud jud, there is something about a pure-blooded boholana..very very very attractive jud.

The morena skin is just so.

Quote from: bol-anon_ko on February 16, 2008, 05:17:48 AM
aw naa ra man diay na.. she's a cathc man kaha wla diay nmu gisalo pre.. ug dli ka kasalo pre atua nangPUKOTON kaila ka pukot pre? kanang NET btaw..kana jdu FINE NETS kana gamit sa muro ami.. syaro ug dli mapukotan..

hahaha dili posible, pre. She calls me 'Kuya'

LOL

Luud pood if i pa pursue nako.

Here's a pure Boholana.





Sexy kaayo..

hahaha, o syiempre smart gud na siya kai Pilipina gud. Hahahaha Isug pa kai boholana gud na.

Ug ma suko na, ma suko jud. Bisaya gud.

LOL

Quote from: bol-anon_ko on February 16, 2008, 05:08:55 AM
pre bumper kaayu c Megan pre.. perteng trapika ana pre..

Hahaha, mo singit na siya dali pre kai talawan na. Sa una galing, ning adto mi sa bar. Wa pa man na siya ning turn 21--gi carded siya sa pulis--na hilak hilak jud.hahaha. Luuy pood kai na hadlok siya ipa prisonan for underage drinking. Gi taga-an lang siya ug warning.

Bitaw pre, she is a catch. She likes filipino guys or Chinese guys.

mestiza iyang looks, pero pinay jud sa puso.

Quote from: bol-anon_ko on February 16, 2008, 05:07:12 AM
nya syaro pre ug wala kay nagustuhan bsag isa ana.. puros man mga malami-an nang chicks gi POST nmu dri.. or napul-an na baka ani nila pre? 

player jud ka pre dah... saludo ko nmu

di uy, amigo ra mi. LOL

Pinay cowboy 





Vamanos!

Quote from: Happy on February 16, 2008, 04:31:38 AM
Pure boholana

Dong, daghan man lagi nimo chicks oi hehe

in the words of Mike, blood smells blood

'te Megan looks foreign but she speaks Bisaya perfectly. Her father is Boholano, her mother is Italian.


New years celebration

You see God

hahaha suka sa banyo ha? LOL

did he have fajitas the night before?

ROFLMAO

George

Megan, half Italian, half boholana

Nikky


isug ning bajiha 

Some of the Boholanas that I know, sizzlin... 


Leilani to the right (half spanish)




smokin'     

coffee. It is my drug of choice. LOL

a.k.a "Dopamine in a cup"

would you rather marry a boholano or a foreigner?

hahaha, pre, pero some mixed breeds are very attractive.

Specially Spanish mestizas. Aguy~

This song is so romantic.
Beautiful~

http://youtube.com/watch?v=yMeZbcXyTec

Lyrics:

Nang mawalay ka sa aking puso
Kung bakit hanap-hanap ka pa
Ang 'yong mukha'y lagi
Lagi na lamang sa isipan ko
Bakit di magawa nitong damdamin
Ang paglimot sa mga nagdaan
Sadya nga bang ganyan pag nagmahal
Ay di matatakasan
CHORUS:
Nais ko'y makapiling kang muli
Nais ko'y mayakap kahit sandali
Kung pangarap ay tatanggapin ko
Ikaw pa rin ang iibigin ko
Repeat 1st Verse and Chorus (2x)

kenny rogers rocks!

Here is a pure Boholana in the Singkil

Yes she's definitely mestiza, but dayang! She's one guapang mestiza

Mike, is she part Spanish?

Beautiful

Remark: The first Philippine coins came into existance around 1830. At that time many South American Spanish colonies became independent republics. However, they continued to trade with their former trade partners -- the Philippines being one of them -- but using their new coinage. The Spanish authorities were wary that these new coins, which did not bear the profile of the Spanish monarch but carried words like "LIBERTAD" (liberty), would incite a rebellion in the Philippines. To make them "Spanish" issues, they added the seal of the king or queen of Spain at the time: King Ferdinand VII and Queen Isabella II (hence, F7 and YII). These counterstamps were used to legitimize the coins that arrived (as Spain did not recognize those country's independence until several years after). Growing from the Philippines' practise of punching holes in coins (so as to attach them to money belts), coins which bear counterstamps on the holes are extremely rare and desireable.




Remark: Silver coins bearing Isabella's image were struck from 1864 to 1868, and then again from a few years after using the 1868 dies (hence the reason 1868 is the most common year for this series). Coins in this issue are very similar to their Spanish counterparts. The only difference was the denomination they had: centimos de peso instead of pesetas.



Remark: Silver coins bearing the image of King Alfonso XII (who succeeded Isabella) were struck from 1880 to 1885 and are similar to their earlier Isabella counterparts. Here too, minting continued for a few years using the 1885 die. Coins of this issue differ from their Spanish counterparts in the denomination and the fact that King Alfonso faces in the other direction.



Remark:A very small number of gold coins bearing Alfonso's image were minted. Many mintage years are reported, but only 1882 and 1885 are confirmed. All of these issues from any year are rare.



Remark: A new one peso coin was struck in 1897 with the profile of the boy King Alfonso XIII. This coin was only minted for 1 year, and is, at the same time, the first silver coin to bear the name of the Philippines (Islas Filipinas) and the very last coin issued by Spain in the Philippines.

The thanks goes to Joseph Noriek, Mike. He is the one that took them, and his story is so different from many of ours, he doesn't know Tagalog (only some phrases), nor does he know family members in the Philippines. He was adopted by an American family, and is completely Americanized. But even with this--despite the fact that he was raised in middle America, exposed to all American things, culture, friends, family, love, etc.

In his heart, deep inside him, knew that he had to go back to visit where he came from. To see, with his own eyes, what his people look like.

I admire him, Mike, because he is very faithful. A religious guy, very intelligent and a good friend. And I believe that the Lord guided him back to the Philippines to see where he was from, to see the nuns and priests that took care of him when he was only a kid.

It brings me to tears, jud Mike. He didn't have to go to Philippines, he could have forgotten it as he has no living memory of it (besides that he was born to Filipino parents). But he went back.

It humbles me, Mike. So very much.

The Orphanage














Some friends over there in Manila




I will try my best to get Joe to join Tubag Bohol. So he can be part of our Tubag Bohol family.

The following pictures were taken by my good friend Joseph Noriek, he is also Filipino and a senior in college like me. Joe was adopted as a baby and has never been to the Philippines or seen his biological parents. His story is that of a heart warming one, and one that brings me tears just talking to him about it. He left for the Philippines (Manila) last summer 2007 and I was able to help him a bit in learning basic Tagalog.

I just wanted to share with you guys, his story, and his deep longing for the Philippines. Truly, when I say this, that our roots yearn to find the mother soil. Despite where we are in the present.

Joseph's Journey Back to the Philippines:


Joe in New York, with friends




Joe bringing baby materials for the orphanage that used to house him. This brought me to tears


Leaving Wisconsin


Destination: Manila, The Republic of the Philippines


In his own words, "..and some more of God's beautiful creation"


Landing in the 'Homeland'





Joe and his friends; in Manila


The orphanage, 


The Philippines




The children

What is your dream Honey-Moon destination spot?

My first kiss was with a girl named Pam P, a girl i went to high school with.

Well, it wasn't much of a first kiss, she stole it from me. But i stole one back afterwards
She caught me off guard.

haha, daku man gud ang ulo 'tong bataa to'.
My head was too big to go through kono. LOL

'Hard headed' jud
Pariho sa Papa

LOL

What did you do, and who was it with?

bitaw you're right, now that i remember it, i have friends who say tampons give them more freedom during the menstrual cycle.

so who as it ?

C'mon we won't tell. 

Quote from: Ginger on February 15, 2008, 07:59:30 PM
Dong, girls can still go swimming even on red tide days. hehehe

Thanks to tampax tampons!

haha, i didnt think about that. pero dili ba na uncomfortable?

haha, ill do that, pre. promise.

studying pre. im just taking a break--ning TB na pood lagi. haha

good night, my friend!

haha thats kinky, yo

ah no problem, natural ra man nang mo itch. just put some lotion ma wa ra man na