abandonability

Retaining the original color of a hardwood deck extremely difficult

2012-03-15T07:37:00.000-07:00

Q. We recently put in a large mahogany deck (about two monthsago). The builder applied three coats of Thompson's water seal, andit is now starting to gray in places and fade. Please let us know howto restore the nice red color and then what to do for routinemaintenance.

A. Unfortunately, you put wax on your deck, which is just aboutthe worst thing you could do. Technically, once wax is placed on ahardwood, it never leaves completely. You have a very hard wood deck.As you are now aware, all hardwoods gray very rapidly. It isextremely difficult to retain the original color of a hardwood deck.On top of that, many of the imported hardwoods are so dense they donot accept stain …

Using a population-based health information system to study child health

2012-03-15T05:16:00.000-07:00

ABSTRACT

Objective: This paper describes the population-based analyses of measures of child health status used throughout this supplement.

Methods: The articles in this supplement examine health-related data for children 0 to 19 years. Most analyses cover the period from April 1, 1994 to March 31, 1999. Administrative and survey data were used to assess child health and well-being. For regional comparisons, data were broken down by subregions of Manitoba, called Regional Health Authorities (RHAs), and neighbourhoods of Winnipeg, called Winnipeg Community Areas (Winnipeg CAs).

The premature mortality rate (PMR) was used as a proxy of the overall health of the …

Mississippi River spill idles ships near New Orleans

2012-03-15T02:51:00.000-07:00

Dozens of cargo ships, petrochemical tankers and smaller vessels stacked up Thursday near a closed stretch of the Mississippi River, a day after a collision between a barge and tanker spilled more than 400,000 gallons (1.5 million liters) of fuel oil into the nation's busy shipping waterway.

The Coast Guard said reopening nearly 100 miles (160 kilometers) of river to ship traffic could take days, and efforts to clean up the oily sheen left by largest oil spill on the river since 2000 could take weeks. Many of the ships waited at the river's Gulf of Mexico outlet to head upriver to grain and petrochemical terminals above New Orleans, one of the world's busiest …

Adopt Krugman ; The Nobel laureate's trade theory can change India's business paradigm.

2012-03-15T00:27:00.000-07:00

My aim was that i would produce a certain volume of cars andcreate a very low-cost, very low-break-even point plant that a youngentrepreneur could buy. A bunch of entrepreneurs across the countrycould establish such assembly operations and Tata Motors would traintheir people, oversee their quality assurance and they would becomesatellite assembly operations for us. It would be satisfying if thesmall car created 10-15 satellite groups of young engineers whocould get together and do a business. They would never otherwise beable to get into the assembly of cars,Ratan Tata had told anewspaper on January 10, 2008, describing his business plan for theRs 1-lakh Nano.

By design …

Stoichiometry and Substrate Affinity of the Mannitol Transporter, EnzymeII^sup mtl^, from Escherichia coli

2012-03-14T22:07:00.000-07:00

ABSTRACT

Uptake and consecutive phosphorylation of mannitol in Escherichia coli is catalyzed by the mannitol permease Enzymell^sup mtl^. The substrate is bound at an extracellular-oriented binding site, translocated to an inward-facing site, from where it is phosphorylated, and subsequently released into the cell. Previous studies have shown the presence of both a high- and a lowaffinity binding site with K^sub D^-values in the nano- and micromolar range, respectively. However, reported K^sub D^-values in literature are highly variable, which casts doubts about the reliability of the measurements and data analysis. Using an optimized binding measurement system, we investigated the …

Suspicious vehicle closes roads near White House

2012-03-14T19:46:00.000-07:00

WASHINGTON (AP) — A block of Pennsylvania Avenue near the White House in Washington has been closed because of a suspicious vehicle.

Secret Service spokesman Max Milien says there are closures on Pennsylvania …

Harvick wins wild race at Daytona

2012-03-14T17:26:00.000-07:00

Kevin Harvick lined up next to his Richard Childress Racing teammate for a two-lap sprint to the finish at Daytona International Speedway.

He and Clint Bowyer didn't bother to talk strategy.

"It was every man for himself at that point," Harvick said.

The Sprint Cup points leader sailed to his second win of the season, leading a strong RCR contingent in a wild night of racing at Daytona. There were a record 18 leaders, 47 lead changes, a 20-car accident and numerous angry drivers when it was over.

Harvick earned high praise from team owner Childress, who compared his driver to the late Dale Earnhardt, considered one of the best …

Charleston Daily Mail Neediest Cases Appeal: Mother's poor vision makes job difficult; Case No. 13

2012-03-14T15:05:00.000-07:00

This 30-year-old single mother of three desperately needs a pairof glasses.

Just out of beauty school, she landed a good part-time job shehopes will lead to full-time employment so she can make a better lifefor her three children, 4, 8 and 12.

But frequent headaches that often are severe make her jobdifficult.

She is struggling to study for a required state board test, whichshe must pass to get full-time work in her field.

"She can't do that much until she gets her license. They only lether do the basic, little things right now," her caseworker said.

Her caseworker says simple, daily tasks - both at work and at home- prove difficult for this …

Chevron 3Q Profit Plunges 26 Percent

2012-03-14T12:45:00.000-07:00

Chevron Corp. third-quarter profit slid 26 percent, the steepest decline in five years as the energy market's unpredictable pendulum swung against the second-largest U.S. oil company and most of its peers.

The San Ramon-based company said Friday that it made $3.72 billion, or $1.75 per share, in the three months ended in September, down from net income of $5.02 billion, or $2.29 per share, at the same time last year.

Analysts were bracing for a lower profit, but the erosion was far worse than their average earnings estimate of $2.07 per share, based on a survey by Thomson Financial.

Revenue growth also was lackluster during the quarter, edging up …

Oprah Winfrey going to Democratic convention

2012-03-14T10:25:00.000-07:00

Oprah Winfrey will be there when Barack Obama accepts the Democratic nomination for president later this month in Denver.

There are no plans for Winfrey to take the stage at the acceptance speech, longtime friend Gayle King told Entertainment Tonight.

"Oprah has not been asked to introduce him, is not expecting to be asked to …

BOOK BRIEFS

2012-03-14T08:04:00.000-07:00

NOW IN PAPERBACK

Books un the Holocaust - fiction and non-fiction - have recently been made available in paperback.

Prisoners: A Jewish Guard in a Nazi POW Camp. by Burl Zollo (Academy Chicago Publishers. 304 pp., $18.95) is a debut novel by this Chicago writer, originally published in 2003 and now reissued in paperback.

American Jewish soldier Stuart "Sandy" Delman is assigned as a guard at a POW compound housing Germans who know he's Jewish and take every opportunity to curse him.

Delman can't always contain his own responses, and is punished for it. Ironically, circumstances lead the young HoLdier. who had enlisted to fight Germans. to devise a plan to …

Robbie Knievel to jump 24 trucks at Ohio site where his father jumped 14 buses

2012-03-14T05:44:00.000-07:00

Robbie "Kaptain" Knievel plans to jump over 24 delivery trucks at the site of one of his father's most famous stunts.

Knievel says he'll be going 98 mph (158 …

"You Said It"

2012-03-14T03:14:00.000-07:00

The Chicago Defender sends a photographer to some parts of Chicago every week to get your opinion on an issue of the day. This week, the Defender wants to know if the city's gun buy-back initiative is effective in reducing violence.

No. Because they don't show them destroying the guns. They need to show a big picture of them destroying the guns and then that might make a difference.

-Michael

I don't think it's going to decrease violence. You have to do more than... just give a person $100 and think that it's going to be A-okor all-good. They can turn in one gun and still have two or three left. -Tyree

Yes. You have to start somewhere. Why not start somewhere and see where it goes? If it doesn't work, then at least you tried.

-Kevin

No. People have guns everywhere.

-Christophe

No. Because these little brothers out here... the only thing they think about is gang banging. The younger generation... is growing up with no morals, no sense. They only draw on what they see on TV and what these brothers are doing out the streets. -D'Antoine

CUISINE DE BALL PARK: Sampling the fare at Memorial Stadium

2012-03-14T00:22:00.000-07:00

We've succumbed to the "ball park eating syndrome" at the Boise Hawks game. Food that you would immediately run from on the "outside," casts a spell on you as soon as you surrender your ticket at the gate.

None of us has eaten dinner and we have a hot dog craving. Unfortunately, the vendors circulating in the bleachers are only hawking beer and sodas so I'm sent to brave the early inning lines at the concession stand.

My order of hot dogs takes 15 minutes to fill because of a kitchen snafu, but the folks behind the counter are cheerful and keep offering me a chance to change my order to hamburgers, popcorn chix and halibut and fries. But I am on a mission for hot dogs.

The concession stand carries the usual assortment of sunflower seeds, peanuts and Cracker Jacks. I am tempted to order some Cracker Jacks to satisfy childhood memories, but pass after learning they no longer come in boxes--only in bags--some how it isn't the same. The BBQ pit, across from the concession stand, does a bang-up business while I wait. Customers watch their chorizos, Italian sausage and Philly cheese steaks being cooked on an outdoor grill while they banter with the cook.

After finally grabbing our dogs we wander down to the Hawks Den, the restaurant along the left field line, to sit in the plastic green lawn chairs and order some beers. Their selection doesn't disappoint us as they offer us familiar standbys such as Bud and Rolling Rock along with a wide selection of imports and northwest microbrews.

We're watching a disaster unfold for the Hawks--the visiting Eugene Emeralds have scored three runs without taking the bat off their shoulder--as the Boise pitcher comes completely unraveled. Two balks and a passed ball have cleared the bases and our talk turns to the food we've eaten while watching baseball.

Between the three of us, we have over a century of experience gathered from lowly Winston-Salem Warthog and Toledo Mud Hens games to shrines such as Fenway Park and Wrigley Field. We've ingested crab cakes, garlic fries and funnel bread during that time, but we all agree that hot dogs are the signature food for baseball.

"A hot dog is just a hot dog, until you eat one at the ball park," says Joanne.

"Hot dogs always taste better when you get them at a ball game," Sandy chimes in.

They're chomping on the basic "Hawk" dogs while I'm trying to figure out how I'm going to get my mouth around my "German" dog without letting all the accompanying grilled onions, peppers and tomatoes slide onto my lap.

Joanne and Sandy give the Hawk dogs a thumbs up as they are tasty, juicy and hot and the bun is nicely steamed. My German dog isn't very spicy, but what it lacks in quality it makes up in quantity and the accompanying garnishes.

I throw caution to the wind and go back to the BBQ Pit for a Philly cheese steak. Unfortunately, the steak and onions have been sitting in a steamer and aren't fresh off the grill. Besides looking limp they taste as if they have been cooked in Philly and shipped to Boise.

But all isn't lost as there is always something else to try at a ball park. We decide to give the dessert selection a go. I order the ice cream in the plastic helmet from the concession stand while Joanne and Sandy select the Grasshopper Mousse and Skillet Cookie from the Hawks Nest.

The ice cream is real--a true find in ball parks these days--not the softserve you get in most ball parks these days. Joanne's mousse gets the balance of mint and chocolate just right, with neither overpowering the other. Sandy's chocolate chip cookie is the size of a piece of pie and smothered in ice cream and chocolate sauce.

While the Hawks weren't able to send us home happy, the ball park food did.

Illustration (A woman eating a hotdog)

Turner calls hearing on police shootings

2012-03-13T21:30:00.000-07:00

City Councilor Chuck Turner called for a hearing on use of deadly force by the Boston Police Department last week after the sixth death at the hands of a police officer in little more than a year claimed the life of 37-year-old Willie Murray Jr.

"In the last 15 months there have been six fatal shootings by police officers. In five of them, the officers were found to be justified," said Turner eliminating the Murray shooting where the actions of officer Shawn West are currently being investigated by a grand jury.

"The purpose of this hearing is to look at the six deaths and look at the procedures used by the officers."

According to Boston Police Department Superintendent Ann Marie Doherty, the overriding instruction police officers receive when undergoing training on the use of deadly force is "to use the least amount of force necessary" to subdue a suspect.

"But it is impossible to think of each possible scenario that a police officer could come by," said Doherty. "In the end, the individual is the final decision-maker."

According to Turner, of the six shootings in the past 15 months, three of the police officers who caused the death of a suspect were under 25 years of age. Some are concerned that maybe the young age and lack of appropriate training could have led to the deaths of the suspects.

"As I've looked at these cases, I don't think these were cases where issues of racial bias or issues of rogue officers were a factor in determining the deadly outcome," said Turner. "I do believe, however, there are issues about whether protocols are leading to the actions of these young police officers."

The Boston Police Department says none of the police officers were 25 years of age or younger.

"All the officers involved in the shooting were between 26 and 34 years of age," said Mariellen Burns, director of media relations for the Boston Police Department emphasizing that officers attend intensive training for months before making it to the streets.

Boston police officers all carry a firearm. But they also carry batons, mace, and could possibly be carrying a stun gun in the future as well.

"We are currently assessing the use of stun guns," said Doherty, although the weapons are currently illegal in the state of Massachusetts.

Doherty further said that relative to other US cities of equal size to Boston, five fatal shootings is really not that much at all.

"Five deaths in less than two years is a very small number," said Doherty. "Its larger than what we've had in the past but this is not something that goes off the charts."

Still there are those who are concerned.

"This is number six in the last year," said Sadiki Kambone, director of Black Community Information Center. "This shows that the police are still out of control. They are more inclined to reach for a weapon then they ought to use common sense."

As for her reaction to Turner's request for in investigation into police policy and protocol, Doherty says she feels neither threatened nor angry since reviewing police policy is an ongoing process at the department.

Photograph (Chuck Turner)

British government bans radical Islamist group

2012-03-13T18:40:00.000-07:00

LONDON (AP) — The British government on Thursday banned a tiny radical Muslim group that attracted ire by protesting outside the U.S. Embassy on the anniversary of Sept. 11 and burning memorial poppies on Remembrance Day

Home Secretary Theresa May said the group would be outlawed from midnight.

May said she was satisfied the group was "simply another name" for an organization already banned for glorifying terrorism under names including al-Ghurabaa, The Saved Sect, al-Muhajiroun and Islam4UK.

The group's spokesman, Anjem Choudary, accused the government of trying "to cover up the truth."

Muslims Against Crusades posted a message on its website saying it had disbanded and was canceling a "Hate for Heroes" anti-soldiers protest scheduled for Friday — Armistice Day, which marks the end of World War I on Nov. 11, 1918.

The order comes after members of the group barged into a London mosque where Conservative lawmaker Mike Freer was meeting constituents last month, prompting officials to lock him in a room for his own safety.

Before that incident, the group's website referred to the 2010 attack on legislator Stephen Timms, who was stabbed in the stomach by a 21-year-old Muslim woman angered by his support for the Iraq war.

The website said the attack should serve as a "piercing reminder" to politicians that "their presence is no longer welcome in any Muslim area."

Rethinking Jerusalem

2012-03-13T15:45:00.000-07:00

I recall watching footage of the 1994 Rwandan genocide from the comfort of my living room. Images of machete-wielding young people have staying power in the personal video recorder that is my brain. Almost one in seven people perished in just over three horrible months. Most troubling was the sad fact that the vast majority of Rwandans at the time of the genocide would have identified themselves as Christians. How could this be?

A few weeks ago, our church hosted three Rwandan guests. Their ministry to young people and women with HIV/ AIDS is making a difference in the small African country. Facing daunting realities, these servants have acted with vision, rather than wallow or run for sanctuary elsewhere. They took their Jerusalem seriously.

One team member, Luke, had never visited a western nation before. He was like most of us who travel somewhere new, bringing as he did many stereotypes with him. Those assumptions of Canada were shattered as he roamed Vancouver's Lower East Side, Canada's "poorest postal code." He was deeply disturbed. The shock of what he saw on our streets messed with him as much as the jetlag from which he was recovering.

When the group's leader spoke to our church family on Sunday morning, he gave a powerful challenge. Coming to our Jerusalem from the ends of the earth, he spoke of how hope in Rwanda is replacing despair after almost 20 years of reconciliation and repair. He spoke of spiritual renewal and signs of life, and of the persistent need for transformation and healing. He invited our people, so ably fitting Luke's stereotypes, to join in financially supporting their important work. But he also brought things back to our Jerusalem. "Don't come and help us if you won't look after your Jerusalem first," he said with straightforward clarity. That arrow of truth sunk deep.

"But you will receive power when the Holy Spirit has come upon you',' said Jesus in Acts 1:8, "and you will be my witnesses in Jerusalem and in alljudea and Samaria, and to the end of the earth" The world everywhere is a goulash of the beauty and the broken. There is no end of worthy projects to support; of places to send eager servants; and of people who need the wholeness of the gospel that saves sinners, restores dignity and rights wrongs.

All of our churches in some way participate in this immense task in creative and generous ways. Like Luke, some of us will go where our stereotypes will be evaporated and others of us will say our part is to help Luke get there. All this is important, but what are we doing with our Jerusalem? How is our witness of the wholeness of the gospel going there?

So easily do we live with the mess of our own backyard. We can be emotionally moved by stories from far, far away, while the brokenness we pass by in our own Jerusalem is ignored. The familiar is seen with a critical eye. The Sunday morning prophet's challenge sounds deep, like sonar for the soul: We must rethink Jerusalem or our witness to the ends of the earth will ring hollow.

Our Rwandan friends knew firsthand what can happen if the witness in Jerusalem is neglected. These brothers were not outside consultants, but spoke from the credibility gained by enduring the worst and working towards a different future in their Jerusalem. And that was precisely what gave heavenly weight to their message.

[Sidebar]

The shock of what he saw on our streets messed with him as much as thejetlagfrom which he was recovering.

[Author Affiliation]

Phil Wagler (phil_wagler@yahoo.ca) is rethinking life in Surrey, BC as a husband, father, neighbour and pastor.

EUROPE NEWS AT 0700 GMT

2012-03-13T12:50:00.000-07:00

UPCOMING FOR SUNDAY, OCTOBER 3:

ITALY-POPE-SICILY

PALERMO, Sicily — Sicilians hope Benedict XVI's first visit to Sicily as pontiff will help stir consciences on an island where the Mafia long held sway. They are also appealing for the pope to declare an anti-Mafia priest who was slain by the mob in 1993 a martyr, paving the way for possible sainthood. Pope's first appearance at 0800 GMT.

BOSNIA-ELECTIONS

SARAJEVO, Bosnia-Herzegovina — Bosnians vote in a general election likely to further entrench the country's ethnic divisions, threatening closer ties to the West and possible EU entry. Polls close 1700 GMT, first results expected 2100 GMT.

BRITAIN-CONSERVATIVES

LONDON — Britain's Conservative Party holds its annual conference, the first since the British national election. David Cameron speaks on Wednesday.

KOSOVO-SERB CHURCH

PEC, Kosovo — Serbia's President Boris Tadic and other senior officials to attend the enthronement of the head of Serbia's Orthodox Church, Patriarch Irinej in the medieval patriarchy in the western town of Pec, amid security concerns. Pec was among the most hard-hit areas by Serb forces during the 1998-99 Kosovo war.

HUNGARY-ELECTIONS

BUDAPEST, Hungary — Prime Minister Viktor Orban's center-right Fidesz party seeks to repeat its landslide win from April's parliamentary elections in Sunday's nationwide municipal elections. Budapest, guided by retiring liberal mayor Gabor Demszky since 1990, is also expected to go to the Fidesz candidate. Polling stations open from 0400 to 1700 GMT.

GREECE-CHINA

ATHENS, Greece — Chinese Prime Minister Wen Jiabao on second day of official visit to Greece. Delivers address in Greek Parliament at 0730GMT, visits archaeological museum and Acropolis, meets with opposition leader Andonis Samaras.

GERMANY-REUNIFICATION

BERLIN — Germany marks the 20th anniversary of reunification, with events in Bremen and Berlin.

GERMANY-GREECE

BERLIN — Greek Prime Minister George Papandreou to be honored for his "strength of truthfulness" at a ceremony held by a private foundation.

YOUR QUERIES: The Europe & Africa Desk in London can be reached at +44 207 427 4300.

Protein-Water and Protein-Buffer Interactions in the Aqueous Solution of an Intrinsically Unstructured Plant Dehydrin: NMR Intensity and DSC Aspects

2012-03-13T09:49:00.000-07:00

ABSTRACT

Proton NMR intensity and differential scanning calorimetry measurements were carried out on an intrinsically unstructured late embryogenesis abundant protein, ERD10, the globular BSA, and various buffer solutions to characterize water and ion binding of proteins by this novel combination of experimental approaches. By quantifying the number of hydration water molecules, the results demonstrate the interaction between the protein and NaCl and between buffer and NaCl on a microscopic level. The findings overall provide direct evidence that the intrinsically unstructured ERD10 not only has a high hydration capacity but can also bind a large amount of charged solute ions. In accord, the dehydration stress function of this protein probably results from its simultaneous action of retaining water in the drying cells and preventing an adverse increase in ionic strength, thus countering deleterious effects such as protein denaturation.

INTRODUCTION

Protein function in general is manifested via a complex array of interactions between the protein and its molecular environment, most apparent in the case of interactions of the molecule with cofactors, ligands, and substrates, modifying enzymes, allosteric metabolites, targeting proteins, and other mucromolecular binding partners. Underlying all these, however, is an intricate network of interactions with hydration water and solute ions that need to be replaced when the protein folds and interacts with its partners. Quantitative characterization of this latter, thus, is of prime importance for a molecular understanding of protein function.

A special aspect of protein hydration is the case of intrinsically unstructured proteins (IUPs), which do not fold into a well-defined three-dimensional (3D) structure under native, physiologic conditions (1-3). These proteins often realize their functions via molecular recognition, in which structural disorder confers specific advantages, such as specificity without excessive binding strength and many more. Because of their lack of a folded structure and largely exposed interaction surfaces, it is anticipated that the hydration of these proteins is significantly different from that of ordered, globular proteins. In fact, as reported in two recent articles, we used an NMR relaxation technique for characterizing the hydration of IUPs and found that their hydration is significantly higher than that of a globular control protein of similar size (4,5). Our observations provided a direct demonstration that IUP structure is more exposed than globular proteins and able to discriminate between fully and partially disordered elasses of IUPs.

An interesting and thus far ill-characterized class of these proteins is late embryogenesis abundant (LEA) proteins. LEA proteins and one of their subclasses, dehydrins, provide protection to plants and free-living insects against deleterious consequences of water loss and freezing under dehydration stress conditions (6-8). The expression of these proteins is induced by osmotic stresses, such as draft, high salinity, and/or freezing, when they provide protection against hypoosmotic conditions. The mechanism of protection from the loss of water is unclear but could be by acting as hydration buffer, sequestering ions, protecting other proteins, renaturing unfolded proteins, or a combination of any of these (8,9). Probably all these functional aspects are underlined by their highly hydrophilic (10) and intrinsically unstructured nature, demonstrated for a handful of them thus far (11-15). The protein we characterize in this respect is the early responsive to dehydration (ERD) 10, expressed in plants in certain very actively dividing tissues and ubiquitously under drying conditions (16).

Our goal here is to gain information on water molecules in the solution of ERD10 and to characterize its structure by means of the separate study of the hound water fraction in aqueous solution by a combination of two different experimental methods. As far as bound water nomenclature is concerned, the terminology proposed by Cooke and Kuntz (17) is used: we refer to water molecules that are in the vicinity of and interact strongly with macromolecular surfaces and that have properties that are delectably different from those of the medium as "bound water," and the remaining water as "bulk water." By the term "unfrozen water." we denote the actual fraction of water molecules in a mobile state at a given temperature. The unfrozen water term can therefore refer to a phase composed of either hound water molecules only or hound plus hulk water molecules. We combine results from proton NMR intensity and differential scanning calorimetry (DSC) measurements on ERD10, a globular control (BSA), and buffer solution itself as reference to understand the special hydration/solvation properties of this IUP. Our results not only confirm that ERD10 has a high hydration potential hut also provide the unexpected finding that this IUP has a large capacity and broad specificity for ion binding. The contribution of these factors to the function of this dehydration factor are discussed. In addition to these immediate implications, we also pursue this present line of research to develop experimental techniques, which provide quantitative structural and activity-related experimental data that characterize IUPs, as opposed to globular proteins.

EXPERIMENTAL

In order to separate the various water phases present in aqueous solution samples (17). they were frozen. The phases of ice protons, organic protons, anil hound water protons are clearly separated in the FID signal by virtue of large differences in the spin-spin relaxation rate (4). Ice protons yield a signal fraction characteristic of solids with a typical decay rate of the order of 10^sup 5^ s^sup -1^. This signal is completely buried in the dead time of the spectrometer. Organic protons and/or irrotationallv hound water protons (17) also yield a solid-like signal fraction with an order of magnitude smaller, hut still large, decay rale. The proton NMR signal of unfrozen water has a much smaller time-domain decay rate, typically 2000 s^sup -1^. This enables specific recording of the FID signal that belongs to bound water molecules. The zero-time extrapolated peak amplitude of the CPMG train gives the fraction of protons that belong to bound water molecules directly. The effect of freezing on the protein solutions was controlled by the comparison of NMR parameters obtained before and after a freeze-thaw cycle at temperatures above 0�C. We found that the freeze-thaw cycle caused no observablechanges for the studied BSA and ERD10 solutions as far as the measured NMR parameters (FID amplitude, T1 and T2 relaxation times) are concerned.

The temperature was controlled by an open-cycle Oxford cryostat with an uncertainty better than � 1 K. ^sup 1^H NMR measurements and data acquisition were accomplished by a Bruker SXP 4-100 NMR pulse spectrometer at frequencies of 44.1 and 82.6 MHz with a stability of belter than �10^sup -6^. The data points in the figures are based on spectra recorded by averaging signals to reach a signal/noise ratio >50. We varied the numher of averaged NMR signals to achieve the desired signal quantity for each sample and for unfrozen water quantities. We controlled the sensitivity of the NMR spectroscope by measuring the length of the π/2-pulse during measurements (18) to obtain reliable M^sub 0^ values. The extrapolation to zero time was done by lining a stretched exponential.

In a multifraction system such as our samples below 0�C, one can write similar lerms for every fraction, namely for ice, for bound water, and for hulk or free water. The enthalpy change ΔH values (see Fig. d) have an error of �5%.

The temperature is an intensive parameter of the system, so the relation between temperature and heat supplied to the sample describes its thermodynamic stale. To obtain the true temperature of the sample in the calorimelric experiment, particularly for the ease of phase transitions, the analysis of thermal paths among the sample, the temperature sensor, and the heat source is necessary. The DSC results presented here were temperature corrected using a procedure worked out during experiments with aqueous NaCI solutions. The details of temperature correction and enthalpy calculations are given elsewhere (19).

The DSC measurements were earned out at heating rate of q = 2 K min^sup -1^ on a TA Instruments heat-flux DSC. Before the experiment, the samples were cooled down with the same rate of q = 2 K min^sup -1^. Cooling at an uncontrolled rate when the temperature changed exponentially from 15 K min^sup -1^ to a rate of less than 2 K min^sup -1^ at low temperatures had no observable influence on thermal behavior at heating. The NMR measurements were done at equilibrium state.

Sample

Double-distilled water was measured as a starting material to obtain calibration data and parameters for the temperature correction: 20 �l of liquid was used in DSC measurements and 100 �l in NMR. NaCl solution, 150 mM, was prepared by dissolving the appropriate quantity of NaCl (alt.) in distilled water. The Tris solution contained 50 mM Tris (Sigma, St. Louis, MO) and 1 mM EDTA (Sigma). pH 7.0. The buffer solution contained 150 mM NaCl, 50 inM Tris, and 1 mM EDTA, pH 7.0.

The aqueous protein solutions were prepared by dissolving the proper amounts of bovine serum albumin ((BSA); Sigma) or early responsive to dehydration 10((ERD10), prepared as described previously (4)) in the above buffer solution. For determining the amount of water bound per unit protein, and having noted that the concentration measurement of IUPs is error-prone because of their unusual reactions with colorimetric dyes, we directly measured the amount of protein dissolved by determining the mass of samples lyophilized from distilled water. This measure provided the absolute concentration of the protein, which could he directly used for calculating the absolute average concentration of its constituent amino acids. Given that NMR enabled the direct measurement of the concentration of bound water molecules, this allowed the calculation of the hydration of proteins normalized to amino-acid units, i.e., n^sub water^,/n^sub amino acid^.

For each solution composition, we curried out the NMR and USC measurements on three to live samples prepared independently. The obtained data were reproducible within the given statistical errors.

RESULTS AND DISCUSSION

We measured the thermal curves of pure water, NaCl solution, and buffer solution by DSC (Fig. 1). The heat capacity of ice determines the heat How of water below 0�C. In this range, the specific heat of ice changes slightly from 1.703 J K^sup -1^ g^sup -1^ at -55�C to 2.07 J K^sup -1^ g^sup -1^ at +5.4�C (20). At 0�C. water exhibits a rapid increase of the heat flow that reflects the melting. At this point, the scale of the specific heat does not represent a real value. It is known that heat capacity at melting is extremely high. The heat flow returns exponentially to the level determined by the heat capacity of the liquid when melting is completed. This is a transient effect, which indicates how the sample temperature increases from the melting temperature to the stationary stale of any transition. The heat capacity of the liquid (4.185 J K^sup -1^ g^sup -1^ at 22�C (21)) then determines the level of the heat flow on the curve of 150 mM NaCl solution. There are two marked differences, a small endothermic peak (SEP) at about -22�C and the continuous increase of heat flow up to 0�C. The Tris solution has a SEP at -13.6�C of much smaller amplitude and wider wings. The buffer solution shows a similar behavior with a small shift of the SEP toward lower temperatures than the NaCl solution of the same NaCl concentration.

We used double-distilled water as a control of NMR intensity measurements (Fig. 2 a). The unfrozen fraction changes from one to zero in the near vicinity of 0�C as expected. Traces of impurity cause a small deviation compared to a discrete jump from one to zero at exactly 0�C. The unfrozen water fraction in the NaCl solution (Fig. 2 a) shows characteristics absent in pure water because of the solute ions: a wide thermal hysteresis between -33�C and -22�C and then a smooth rise up to 0�C. The step in the unfrozen water fraction occurs at the same temperature in the heating direction as the SEP on the heal flow curve (also detected during healing). The smooth rise of the unfrozen water fraction above the hysteresis loop means that the melting starts at low temperatures and proceeds continuously up to 0�C. The Tris solution has a smaller unfrozen water traction (Fig. 2 b) than the NaCl solution. The hysteresis loop extends to a wider temperature range between 40�C and -13�C and is quite shallow compared to that of NaCl. The curve of the buffer solution (Fig. 2 c) resembles more closely that of the NaCl solution, although its hysteresis loop gets wider and shifts to lower temperatures between -41�C and -25�C. In the case of both Tris and the buffer solution, the upper border of the hysteresis loop coincides with the corresponding SEP of the heating-mode DSC curve.

In the cointerpretation of these results, one can start from the NMR intensity data in pure water (Fig. 2 a). In the ice phase, there are no mobile water molecules at all: the zero-intensity range of the curve represents this state. We suppose that the small knee below 0�C is a consequence of the impurity hydration. In the NaCI, the Tris, and the buffer solution samples, the nonzero NMR intensity indicates the existence of unfrozen mobile water molecules. We calculated the number of unfrozen water molecules per solute units (Fig. 2). In the NaCl solution (Fig. 2 a), the result of n^sub H^sub 2^O^n^sub NaCl^ = 9.2�0.5 at - 22�C (upper side of the hysteresis loop) is in a quite reasonable agreement with 10 water molecules being in the first hyclration shell of a watermediated Na+-Cl- ion pair in liquid water as obtained by molecular dynamics simulations (22). And the value of n^sub H^sub 2^O^n^sub NaCl^ = 7.4�0.5at - 33�C (lower side of the hysteresis loop) agrees excellently with the quantity of seven water molecules in the first hyclration shell of a close Na+-CP ion pair (without a water molecule between the two ions) in liquid water (22). Other authors calculated coordination numbers ranging from 10.68 to 18.33 for a NaCl unit (23). The n^sub H^sub 2^O^n^sub NaCl^ values obtained from our NMR intensity data fall within this interval in the temperature range determined by the freezing point in the cooling direction and the eutectic melting point in the healing direction. The comparison of the n^sub water^^n^sub solute^ curves made between the buffer solution (Fig. 2 c) and its constituents (Fig. 2. a and b) reveals qualitative differences among the three solutions. The n^sub water^^n^sub solute^ curve of the buffer solution cannot be reproduced as a weighted sum of the curves of its constituents.

An important inference from these observations is that buffer constituents (Tris and NaCl) are not independent in terms of hydration but interact with each other and partially replace water molecules that hydrate them. Further, the relatively large difference between the two sides of the hysteresis curves points to a significant change, and thus freedom of movement of hydration water around these solute ions. Our results demonstrate a novel experimental approach for the direct characterization of solute entities by measuring the number of water molecules that hydrate them at any temperature. The SEP and the sudden step in the unfrozen water fraction (NMR intensity) on heating are both attributed to the change in the motional state of hydration H2O molecules, most presumably in the first hydration shell. The great endothermic peak is the DSC response to the solid-liquid first-order phase transformation. The low-temperature shift and the widening of the hysteresis loop in the buffer solution are consequences of the interaction of solute molecules Tris, EDTA, and NaCl, as the NaCl concentrations are the same in the NaCl solution and in the buffer solution.

The DSC and unfrozen water fraction (NMR intensity) data obtained for aqueous solutions of the proteins BSA (c^sub protein^ = 50 mg/cm^sup 3^ Fig. 3) and ERDH) (c^sub proteins^ = 25 mg/cm^sup 3^, Fig. 4) are in striking contrast to the results for the NaCl, the Tris or the buffer solutions (Figs. 1 and 2). The quantity of bound (unfrozen) mobile water and the heat How change continuously below 0�C as expected. The real surprise is the absence of hysteresis in the mobile water fraction and the lack of the SEP in the protein solutions! The bound water fraction for the BSA solution has a gentle slope below -25�C with values of 0.040 � 0.002 at -25.5�C and 0.020 � 0.002 at -53.5�C. The water content of BSA solution freezes completely between -53.5 and -55.6�C. In contrast to the BSA solution, the hound water fraction for the ERD10 solution changes significantly even below -25�C. It is diminishing steadily from the value of 0.055 � 0.002 at -25.5�C to the value of 0.024 � 0.002 at -46.7�C. The remaining mobile water fraction freezes completely at around -48�C, at a much higher temperature than in the case of the BSA solution.

To make a quantitative comparison between the two proteins, the number of mobile water molecules per amino acid unit (n^sub water^/n^sub amino acid^) was calculated for the concentration-normalized number of amino acids (see Experimental), which provides a direct and comparable measure of the hydration capacity of the two proteins. The results are plotted against temperature in Fig. 5. We decided on the norm to be the amino acid unit to obtain a parameter independent of the actual length or amino acid composition of the particular protein and of the protein concentration of the solution. The hytlration (bound unfrozen water fraction) differs markedly for the globular protein BSA and the IUP ERD10. The n^sub water^/n^sub amino acid^ parameter has a value of 4.9 � 0.2 at -25.5�C for the BSA solution, and it reaches its lowest value of 2.5 � 0.2 at -53.5�C. We measured n^sub water^/n^sub amino acid^ = 5.9 � 0.5 for the ERD10 solution just before complete freezing, which is 2.4 times greater. The value 13.5 � 0.5 for the ERD10 solution at -25.5�C is 2.8 times higher than it is for the BSA solution. These quite large differences between the IUP and the globular protein relied a much larger binding capacity for water of the IUP molecule. In principle, such a difference is expected from previous two-dimensional (2D) electrophoresis studies (24). which have placed ERD10 among IUPs. The novel 2D technique, however, only provides a qualitative assessment of the gross structural status of the proteins, whereas the approach presented here enables a quantitative estimation of the exposure of the polypeptide chain manifest in its hydration. In effect, this higher hydration is quantitative evidence for the unstructured nature of this protein. Our previous comparison of two IUPs (calpastatin and MAP2c) (5) has shown that quantification of hydration permits a deeper insight into the structure of IUPs. which segregate into structural subclasses of different levels of disorder, such as random coil, molten globule, and premolten globule (25). Taken at face value, the data calculated here suggest that ERD10 is practically fully disordered, best approximated by the random coil state.

In theory, these measured values could also be interpreted in terms of the average solvent accessible surface area (SASA) of ERDK). This would provide information on what portion of SASA is actually accessible to water, or how effectively residues in contact with water modify water structure and result in hydration resistant to freezing alongside bulk water molecules. Because of the inherent structural heterogeneity of an IUP, however, there is no independent estimate of SASA other than measuring its hydration directly, as carried out here. Thus, at present one cannot tell if this or any other IUP hinds more or less water than expected for a globular protein of similar SASA. As discussed below, the hydration of the IUP and the globular BSA do show qualitative differences in terms of heterogeneity, which can be seen from the slope of unfrozen water fraction versus temperature curves and the ultimate freezing point, which may point to their principally different mechanisms of water binding. Further studies, however, will he needed to elucidate these points.

Although these differences among proteins of different structural status were expected in light of prior data (4.5). the contrasting behavior of the protein solution and its buffer is very surprising. A reasonable question is what quantity of protein molecules are necessary to cancel the thermal hysteresis of the unfrozen water fraction and the SEP. To address this issue, we studied a scries of protein solutions by changing the protein concentration only. The integral areas of SEPs plotted in Fig. 6 are proportional to the enthalpy change of the eutectic phase separation (Eq. 3). The integral area of the SEP decreases and the hysteresis fades away as the protein concentration is increased for both types of proteins. The most striking difference between the two protein types lies in the minimal protein concentration necessary to suppress the SEP or the hysteresis: it is between S and 12.5 mg/cm^sup 3^ for ERD10, whereas there is a big SEP even at a BSA concentration of 25 mg/cm^sup 3^. These results prove the more pronounced water-binding activity of the IUP ERD10 than of the globular protein BSA.

These results, however, also point to an unexpected aspect of protein action in that the absence of the hysteresis/SEP from the curves suggests that independent hydration of solute ions is very effectively abolished by ERD10. This suggests that the hydration layer of Tris/NaCl is replaced by the protein, which "solvates" the ions. This finding lits nicely with prior suggestions on the molecular mechanisms of how LEA proteins counter the damage caused by dehydration/ freezing conditions. These proteins may offer various ways for maintaining homeostasis under such conditions, such as membrane protection, chaperone action, water retention and ion sequestration (8.9). None of these putative mechanisms has received unequivocal and general experimental verification thus far. Our results presented here, and in a previous paper (4). demonstrate the high hydration capacity of this and probably other LEA proteins. Further, hydration of this protein and that of BSA show significant differences, in the sense that hydration of the IUP ERDK) is rather heterogeneous, with an array of distinct binding sites. The steady and steep increase in the amount of unfrozen water on heating can be interpreted as melting of ice phases at different temperatures. The melting of different ice phases can be interpreted as a change in the motional state (dynamics) of water molecules bound at different binding sites. BSA, on the other hand, behaves very differently in that it contains fewer and more homogeneous binding sites, the hydration water of which freezes at a temperature 7�C lower. The lowest mobile water concentration of 2.5 � 0.2 water molecules per amino acid of the BSA solution equals the threshold level of hydration (0.40 g of water per gram of protein ) required to fully activate the dynamics and functionality of globular proteins (26-28).

Large-scale binding of solute ions to the protein is a novel finding that deserves further consideration. Shrinking of cytosol volume as a result of water loss is detrimental to cells for several reasons, one being protein denaturation and salting-out, which occur because of an increase in ionic strength. For this reason, the cell's defense strategy is to produce small organic osmolytes (sugars) for maintaining osmotic equilibrium and lowering ion concentration by sequestering ions. This latter may be yet another function of largely disordered LEA proteins, underscored by the evidence presented here that ERD10 binds a large number of ions. A similar but more specific capacity has been shown for its homolog ERD14 (29) and has been suggested as a rather general function of IUPs acting as "metal sponges" (1). This effect, in combination with the high hydration capacity of IUPs and their noted chaperone activity (30), may explain the general observation that high-level expression of "hydrophilins" (10) is a general and evolutionarily conserved response to a variety of dehydration stress conditions, such as draft, salinity, and freezing.

Conclusions and outlook

The main results of our work can be summarized as follows:

We propose a novel method for quantitative measurement of water molecules in the hydration shell directly based on NMR intensity data for NaCl and protein-butter aqueous solutions.

The thermal hysteresis in the NMR intensity curve of the NaCl-water solution is connected with changes in the motional state of protons belonging to hydration-shell water molecules, probably the start of rotational motion of these water molecules.

The DSC-versus-temperature curve consists of contributions from changes in the motional state of water molecules, heat capacities of the constituent phases, and the heat of melting, where appropriate.

A small endothermic peak was detected in the DSC curves at the same temperature range where the thermal hysteresis exists in the NMR intensity curves. The ordinary explanation of bordering steps of the NMR-intensity hysteresis loop is to assume activation and deactivation of rotational molecular motion of hydration water molecules at the appropriate temperatures. The rotational motion is probably the initial step of the eutectic phase separation (the literary interpretation of thermal properties of salt solutions (brines) (31,32)).

Tris and EDTA additives to NaCl solution cause shifts in NMR-intensity hysteresis and temperature of the small endothennic peak. These qualitative changes refer to interaction of NaCl with the small molecular constituents.

Both the thermal hysteresis in the NMR intensity curve and the SEP disappeared in the protein-buffer solution at a given protein concentration. The limits and character of disappearance are markedly different for the selected globular protein and IUP.

The thermal behavior and the value of NMR intensities themselves are also characteristically different for the two types of proteins, which reveals the differences in the quantity and the kinetic behaviors of hydration shell water molecules.

Open questions are the following:

Do the same effects exist in the cases of other proteins? How far can the above-summarized results be generalized? It is of immediate note that cooling and freezing applied in the present experiments may affect the protein's native conformation. Reversibility does hold, as repeated NMR experiments done before and after the freeze-thaw cycle leave the NMR parameters unaffected. A major conformational change can also be ruled out, as the reported parameters (FID intensity and DSC response) do not depend on the speed of cooling. Further experiments with extremely fast cooling will be required to control and rule out liner structural effects.

What is the role of the other members of the buffer in the processes that result in the disappearance of the NMRintensity hysteresis and the SEP?

NMR measurements on ^sup 23^Na and ^sup 35^Cl nuclei can give direct proof of bound water molecules in the immediate neighborhood of ions or of interacting ion-protein units.

We intend to look tor other physical methods offering similar responses to these questions.

This work was supported by the International Senior Research Fellowship GR067595 from the Wellcome Trust. Peter Tompa acknowledges the suppoit of the Kolyai J�nos Scholarship.

[Reference]

REFERENCES

1. Dunker, A. K., C. J. Brown, J. D. Lawson, L. M. Iakoucheva, and Z. Ohradovic. 2002. Intrinsic disorder and protein function. Biochemistry. 41:6573-65X2.

2. Tompa. P. 2002. Intrinsically unstructured proteins. Trends Biochem. Sci. 27:527-533.

3. Dyson, H. J., and P. E. Wright. 2005. Intrinsically unstructured proteins and their functions. Nat. Rev. Mol. Cell Biol 6:197-208.

4. Bokor, M., V. Csizmok, D. Kovacs, P. Banki, P. Friedrich, P. Tompa. and K. Tompa. 2005. NMR relaxation studies on the hydrate layer of intrinsically unstructured proteins. Biophys. J. 88:2030-2037.

5. Csizmok, V., M. Bokor, P. Banki, �. Klement, K. F. Medzihradszky, P. Friedrich. K. Tompa. and P. Tompa. 2005. Primary contact sites in intrinsically unstructured proteins: the case of calpastatin and microtuhule-associaled protein 2. Biochemistry. 44:3955-3964.

6. Ingram, J., and D. Bartels. 1996. The molecular basis of dehydration tolerance in plants. Annu. Rev. Plum Physiol. Plant Mol. Biol. 47:377-403.

7. Thomashow, M. F. 1999. Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu. Rev. Plant Physiol. Plant Mol. Biol 50:571-599.

8. Wise, M. J., and A. Tunnacliffe. 2004. POPP the question: what do LEA proteins do? Trends Plum Sci. 9:13-17.

9. Bray, E. A. 1993. Molecular responses to water deficit. Plant Physiol. 103:1035-1040.

10. Garay-Arroyo, A., J. M. Colmenero-Flores. A. Garciarrubio. and A. A. Covarrubias. 2000. Highly hydrophilic proteins in prokaryotes and eukaryoles are common during conditions of water deficit. J. Biol. Chem. 275:5668-5674.

11. Eom, J., W. R. Baker, A. Kintanar, and E. S. Wurtele. 1996. The embryo-specific EMB-1 protein of Daucus carota is flexible and unstructured in solution. Plain Sci. 115:17-24.

12. Lisse, T., D. Bands, H. R. Kalbitzer, and R. Jaenicke. 1996. The recombinanl dehydrin-like desiccation stress protein from the resurrection plant Craterostigma plantagineum displays no defined three-dimensional structure in its native state. Biol. Chem. 377:555-561.

13. Soulages, J. L., K. Kim, C. Walters, and J. C. Cushman. 2002. Temperature-induced extended helix/random coil transitions in a group 1 late emhryogenesis-abundant protein from soybean. Plant Physiol. 128:822-832.

14. Goyal, K., L. Tisi, A. Basran, J. Browne, A. Burnell, J. Zunlo, and A. Tunnacliffe. 2003. Transition from natively unfolded to folded state induced by desiccation in an anhydrobiotic nematode protein. J. Biol. Chem. 278:12977-12984.

15. Koag, M. C., R. D. Fenton, S. Wilkens, and T. J. Close. 2003. The binding of maize DHN1 to lipid vesicles. Gain of structure and lipid specificity. Plant Physiol. 131:309-316.

16. Kiyosue, T., K. Yamaguchi-Shinozaki, and K. Shinozaki. 1994. Characterization of two cDNAs (ERD10 and ERD14) corresponding to genes that respond rapidly to dehydration stress in Arabbidopsis thaliana. Plant Cell Physiol. 35:225-231.

17. Cooke, R., and I. D. Kuntz. 1974. The properties of water in biological systems. Annu. Rev. Biophys. Bioeng. 3:95-126.

18. Tompa, K., P. B�nki. M. Bokor. G. Lasanda, and J. Vas�ros. Diffusible and residual hydrogen in amorphous Ni(Cu)-Zr-H alloys. 2003. Journal of Alloys and Compounds. 350:52-55.

19. Kamasa, P., M. Bokor, M. Pyda, and K. Tompa. 2006. Temperature correction for heat capacity and phase transformation studies of water solutions in heat-flux-type calorimeters. Thermohimica Acta. In press.

20. Giauque, W. F., and J. W. Stout. 1936. The entropy of water and the third law of thermodynamics. The heat capacity of ice from 15 to 273 K. J. Am. Chem. Soc. 58:1144-1150.

21. Jessel, A. R. C. S. 1934. The effect of dissolved air on the specific heal of water. Proc Phys Soc: 46:747-763.

22. Khalack, J. M., and A. P. Lyuhartsev. 2004. Car-Parinello molecular dynamics simulations of Na+-Cl- ion pair in liquid water. Condens. Matter Phys. 7:683-698.

23. Zhou, J., X. Lu, Y. Wang, and J. Shi. 2002. Molecular dynamics study on ionic hydration. Fluid Phase Equilib. 194-197:257-270.

24. Csizmok, V., E. Szollosi, P. Friedrich, and P. Tompa. 2006. A novel two-dimensional electrophoresis technique for the identification of intrinsically unstructured proteins. Mol. Cell. Proteomics. 5:265-273.

25. Uversky, V. N. 2002. Natively unfolded proteins: a point where biology waits for physics. Protein Sci. 11:739-756.

26. Kuntz, I. D., Jr., and W. Kauzmann. 1974. Hydration of proteins and polypeptides. Adv. Protein Chem. 28:239-345.

27. Rupley, J. A., and G. Careri. 1991. Protein hydration and function. Adv. Protein Chem. 41:37-172.

28. Poole, P. L., and J. L. Finney. 1983. Sequential hydration of a dry globular protein. Biopolymers. 22:255-260.

29. Alsheikh, M. K., B. J. Heyen, and S. K. Randall. 2003. Ion binding properties of the dehydrin ERD14 are dependent upon phosphorylation. J. Biol. Chem. 278:40X82-40889.

30. Tompa, P., and P. Csermely. 2004. The role of structural disorder in the function of RNA and protein chapcrones. FASEB J. 18:1169-1175.

31. Koop. T., A. Kapilashrami. L. T. Molina, and M. J. Molina. 2002. Phase transitions of sca-salt/water mixtures at low temperatures: implications for ozone chemistry in the polar marine boundary layer. J. Geophys. Res. 105:26393-26402.

32. Cho, H., P. B. Shepson, L. A. Barrie. J. P. Cowin, and R. Zaveri. 2002. NMR investigation of the quasi-brine layer in ice/brine mixtures. J. Phys. Chem. B. 106:11226-11232.

[Author Affiliation]

P. Tompa,* P. B�nki,[dagger] M. Bokor,[dagger] P. Kamasa,[dagger] D. Kov�cs,* G. Lasanda,[dagger] and K. Tompa[dagger]

*Institute of Enzymology, Biological Research Center, and [dagger] Research Institute for Solid State Physics and Optics, Hungarian Academy of Sciences, Budapest, Hungary

[Author Affiliation]

Submitted March 8, 2006, and accepted for publication June 1, 2006.

Address reprint requests to P. Tompa, Institute of Enzymoldgy, Biological Research Center, Hungarian Academy of Sciences, H-1518 Budapest, PO Box 7, Hungary. Tel.: 361-279-3143: Fax: 361-466-5465: E-mail: tompo@enzim.hu.

Jury Indicts Jefferson in Bribery Probe

2012-03-13T06:17:00.000-07:00

WASHINGTON - Rep. William Jefferson, D-La., was indicted Monday on federal charges of racketeering, soliciting bribes and money-laundering in a long-running bribery investigation into business deals he tried to broker in Africa.

The indictment handed up in federal court in Alexandria., Va., Monday is more than an inch thick and list 16 alleged violations of federal law that could keep Jefferson in prison for up to 200 years.

This is a breaking news update. Check back soon for further information. AP's earlier story is below.

WASHINGTON (AP) - An indictment charging Rep. William Jefferson, D-La., in a long-running bribery investigation is being announced Monday, federal officials said.

The indictment is being handed up in U.S. District Court in Alexandria. A press conference was being organized for late Monday in Washington to discuss the case.

A Justice Department official familiar with the case said the indictment outlining the evidence against Jefferson is more than an inch thick and charges the congressman with crimes that could keep him in prison for up to 200 years. The official spoke on condition of anonymity because he was not authorized to discuss the case.

Almost two years ago, in August 2005, investigators raided Jefferson's home in Louisiana and found $90,000 in cash stuffed into a box in his freezer.

Jefferson, 63, whose Louisiana district includes New Orleans, has said little about the case publicly but has maintained his innocence. He was re-elected last year despite the looming investigation.

Jefferson, in Louisiana on Monday, could not immediately be reached for comment.

Two of Jefferson's associates have already struck plea bargains with prosecutors and have been sentenced.

Brett Pfeffer, a former congressional aide, admitted soliciting bribes on Jefferson's behalf and was sentenced to eight years in prison.

Another Jefferson associate, Louisville, Ky., telecommunications executive Vernon Jackson, pleaded guilty to paying between $400,000 and $1 million in bribes to Jefferson in exchange for his assistance securing business deals in Nigeria and other African nations. Jackson was sentenced to more than seven years in prison.

Both Pfeffer and Jackson agreed to cooperate in the case against Jefferson in exchanges for their pleas.

The impact of the case has stretched across continents and even roiled presidential politics in Nigeria. According to court records, Jefferson told associates that he needed cash to pay bribes to the country's vice president, Atiku Abubakar.

Abubakar denied the allegations, which figured prominently in that country's presidential elections in April. Abubakar ran for the presidency and finished third.

Court records indicate that Jefferson was videotape taking a $100,000 cash bribe from an FBI informant. Most of that money later turned up in a freezer in Jefferson's home.

In May 2006, the FBI raided Jefferson's congressional office, the first such raid on a sitting congressman's Capitol office. That move sparked a constitutional debate over whether the executive branch stepped over its boundary.

The legality of the raid is still being argued on appeal. House leaders objected to the search saying it was an unconstitutional intrusion on the lawmaking process. The FBI said the raid was necessary because Jefferson and his legal team had failed to respond to requests for documents.

Some but not all the documents seized in the raid have been turned over Justice Department prosecutors.

Associated Press writer Cain Burdeau in New Orleans contributed to this report.

Edwards Admires Parish's Longevity

2012-03-13T02:35:00.000-07:00

CHARLOTTE, N.C. When you've played in the NBA for 19 years andyour number of games is in four digits, there aren't many nights whenyou face an older player who has played in more games.

But that was the case of veteran center James Edwards on Fridaynight as the Bulls played the Charlotte Hornets.

Edwards, who was activated from the injured list earlier in theday, played in his 1,161st game.But that paled in comparison to Charlotte's Robert Parish, whotied Kareem Abdul-Jabbar's NBA record by playing in his 1,560th game.Parish, in his 20th season, will break the record tonight when theHornets visit Cleveland.Is catching Parish, 42, a goal for Edwards, 40?"No, I'm not going to catch him," Edwards said. "I was justtrying to stay around longer than him, but he just keeps going."It's a great accomplishment. I don't think anybody's going toget that."With nine games left in the regular season, Edwards has a littlemore than two weeks to show he is the team's best option for a thirdcenter in the playoffs. In a sense, this is his last push to getready."I should hope so," Edwards said. "I haven't heard one way oranother, but I hope so."Roster MoveTo make room for Edwards, the Bulls placed John Salley on theinjured list with a strained groin muscle. He must sit out a minimumof five games.TributesThroughout the game, videotaped tributes to Parish were playedon the Charlotte Coliseum scoreboard. The first was from NBAcommissioner David Stern, and others were from Parish's formerteammates - most of whom have retired."You really have no idea," Edwards said of having a long career."You have to be lucky and you have to be injury-free. You have totake care of your body and you have to be lucky to last this long."At halftime, the Hornets gave Parish a grandfather clock.For StartersToni Kukoc again was in the Bulls' starting lineup at powerforward and again played splendidly, scoring 16 points in the firsthalf on 5-for-5 shooting and finishing with 20.Bulls coach Phil Jackson isn't saying what he's planning to dowith his lineup against Orlando on Sunday."It's worked both ways, so I think that's another option thatPhil can rely upon," Michael Jordan said. "I think that's somethinghe's going to experiment with. As a coach, you like to have alldifferent kinds of options."Cat CallsOne of the biggest ovations Jordan has received lately cameagainst Miami on Thursday, when he was forced to leave the gamebecause he had blood on his shorts. He changed his shorts on thebench because he always wears navy blue University of North Carolinashorts under his oversized uniform shorts.Still, that didn't prevent a host of whistles."There are a lot of nosy people out there," he said. "But I wasprepared for it with my shorts. They didn't see anything other thanthe UNC shorts."

Rice presses US Congress to OK India nuclear deal

2012-03-12T23:35:00.000-07:00

U.S. Secretary of State Condoleezza pressed Congress to approve a nuclear deal with India after the agreement won the endorsement Saturday from nations that supply nuclear material and technology.

"The main thing is that the international work is now done. I certainly hope we can get it through. It'd be a huge step for the U.S.-India relationship," the top U.S. diplomat told reporters as she traveled from Tunisia to Algeria during a trip to North Africa.

"It would be good to get everything finalized. I hope we can get it through Congress," Rice said.

But time is drawing near for U.S. lawmakers to consider the deal. Congress has only a few weeks of work in September before it is scheduled to break for the rest of the year to campaign for November elections.

The Nuclear Suppliers Group, which governs the legal world trade in nuclear components and know-how, approved the deal after contentious talks and concessions to countries that feared the pact could set a dangerous precedent. Some opposing countries, including Austria, Ireland, New Zealand, Norway and Switzerland, had expressed fears that a reversal of more than three decades of U.S. policy toward India could set a dangerous precedent in the struggle to discourage other nations from pursuing weapons of mass destruction.

"This is an important step forward," Rice said after hearing of the approval by representatives meeting in Vienna, Austria. "This is really a very good step forward for the nonproliferation framework. The India deal is landmark. It's no secret that India has been outside the nonproliferation regime for the entire history of its program."

The deal would reverse three decades of U.S. policy by shipping atomic fuel to India in return for international inspections of India's civilian reactors.

This past week, a leading Democratic lawmaker made public a Bush administration document that says the U.S. had the right to immediately stop nuclear trade with India if India conducted an atomic test. The U.S. position appeared at odds with Indian officials' insistence that the accord would not ban Indian nuclear tests.

Supporters of the civilian nuclear deal say atomic cooperation with India would provide crucial energy to a democratic, economically vibrant country. Critics say it would ruin global efforts to stop the spread of atomic weapons and boost India's nuclear arsenal.

Rice said she had made "a lot of calls" to help win approval. Among the holdouts were Austria, Ireland and New Zealand _ each of which heard from Rice.

She also spoke with China's foreign minister on Saturday, which a senior State Department official said was important in reaching the compromise.

What is manga?: The influence of pop culture in adolescent art

2012-03-12T20:40:00.000-07:00

One of the major problems facing art education is the loss of children's interest in art as the get older. This loss of interest may be due to internal struggles of adolescence or to external stresses, oftentimes in the art classroom itself. Artistic developmental theories explain this loss of interest in art as a universal, cross-cultural tendency. However, recent research (Toku 1998, 2000; Wilson, 1997, 1999, 2000) has suggested that Japanese children may be an exception to this tendency. Throughout adolescence Japanese children tend to continue to acquire skills to express visual narratives in the form of comics, or manga (pronounced "mahngah"). This brings up an important question for art educators: How can this interest in comic books be utilized as a teaching tool in Japan or even extended to countries like the United States?

Internal and external disruptions in children's artistic development

Piagetians, including Lowenfeld and Brittain (1970) describe children's artistic development as a hierarchical linear progression called the stage theory of cognitive development. However, there is an argument that children's artistic development does not always show a linear progression and artistic ability often stops during the transition period from child to adolescent art. For example, Read (1958) explains that most children's artistic abilities decline around 11 to 14 years old due to the loss of interest in and motivation to create art. He called it the "period of oppression." Gardner (1980, 1990) and Davis (1997) also explain the tendency of decline during middle childhood in the pattern of a U-curve of cognitive development. Why do children lose their interest in art at a certain age?

There is no simple answer, rather complex internal and external factors. Internal disruptions relate to issues of self-awareness; children start to realize their own limitations in producing realistic art. They also start to compete with their peers and to judge the relative value of their artwork. External disruptions relate to the social environment. Art is often undervalued as an academic subject. Children may be sensitive to criticism from art teachers. Art curricula are often developed without regard for students' interests.

But do all children really tend to lose their interest in art due to these internal and external disruptions? Is there no hope for teachers to support artistic development through this difficult period? Cross-cultural analysis of children's artistic development in the United States and Japan may suggest such a hope.

The influence of manga in children's artistic development in Japan

Wilson and I have found that there is an impressive movement of young amateur Japanese comic book, or manga artists, whose sheer numbers suggest that Japanese children may be less vulnerable to the "period of oppression." Many of these artists participate in the phenomenon of the Japanese comic markets that were developed to provide young people with an opportunity to exchange their ideas by creating and selling their own original manga magazines. By reflecting Japanese young people's desire to depict their own stories in manga and communicate with peers through these original manga, the comic market has rapidly expanded since 1975 (Schodt, 1996). During 3 days of summer 2000 in Tokyo, more than 300,000 young people from all over Japan participated in the market, with more than 20,000 booths selling original manga magazines. The large number of young people involved in these amateur publishing ventures is one example of their continued interest in graphic narrative. It appears that while attending and after graduating school, Japanese children continue to acquire skills to express visual narratives through this model of pop culture, rather than through art education in schools. Instead of ceasing to express themselves through art, they develop their problem-solving skills and learn visual techniques to replicate their thoughts in the visual narratives of manga (Wilson, 1999). From the early elementary school period through secondary school, it is easy to find the influences of manga, which strongly appear in their graphic narratives (Toku, 1998, Wilson, 1988).

Many questions emerge concerning the particular phenomenon of manga. Why are Japanese children attracted to manga? What are the characteristics of manga? Are they different from comic books in the United States? What are the advantages and disadvantages of manga's influence? If there are advantages to the influence of manga, is it possible to adapt them for art educational curricula? In the following pages I would like to discuss some answers to these questions and the possibility of creating an attractive art curriculum to encourage children's motivation for art. By understanding the mechanism of influence of this element of pop culture in Japanese children's artistic development, it may be possible to predict trends in U.S. children's artistic development in the near future. Nowadays, it is impossible to ignore the influence of pop culture in children's artistic development regardless of the culture; the visual attractions of mass media including TV, movies, and computer games are omnipresent.

This tremendous influx of visual information could be a great resource for creating art educational curricula in the United States and could be more attractive to students and possibly more effective.

The characteristics of manga as the original style of Japanese comic books

Manga-literally meaning "humorous picture"-originally started as a simple caricature, just as in other countries such as the United States. The origin of manga possibly goes back to the 12th century "Chojugiga (the Animal Scrolls) "-literally, "humorous pictures of birds and animals"-depicted by an artist-priest Kakuyu, or Toba (1053-1140) (Akiyama, 1990, Schodt, 1983). Manga developed as a graphic narrative through the work of Hokusai Katsushika (1760-1849), who depicted the Ukiyoe or "floating world." With the expansion of manga's readers' ages from children to adolescents and adults, manga gradually developed into an original style of Japanese comic books reflecting the needs of depicting the complexity of human dramas in graphic narrative. The trend was especially notable after World War II with the influence of U.S. comic books and Disney animation and reached the peak development of the original style of Japanese manga from the 1980s to the 1990s. (Yonezawa, 1997).

In comic books of other countries (such as U.S. comic books), there is a composition of the visual story on the two-dimensional surface. Manga also has what is called the elements of manga or the grammar of manga: 1) picture (depicting objects and figures), 2) word (including onomatopoeia), 3) balloon (indicating words), and 4) frame (surrounding pictures) (Natsume, 1997). The function of each element of manga is a little different from those of the U.S. comic book since manga developed from a simple caricature or good vs. evil story into a complicated story that contains diverse themes including politics, religious, historical, social, cultural issues, and more.

In manga, each element has an important function to explain the meta-levels of space and mind in response to the complicated story. The picture is the content of manga's expression, which is basically constructed of lines. It is generally divided into positive shapes (figures) and negative shapes (background). The word, including onomatopoeia, is generally divided into the outside voice (sound), which comes directly from subjects, and the inside voice, which appears only in the mind. Word also appears independently outside of the frame with or without balloons as a directional connector between frames. The balloon ("fukidashi") used to be the container of the inside voice of the mind or an outside voice from subject/object differentiated from the narration. However, it developed to support the expression of manga on the meta-level. This means both the inside and the outside voice or thought of the subject or object could be shown simultaneously in the same frame, differentiated by the shape of the balloons. The frame ("koma") has a role as a container, which includes the picture as the content, and the word, which is namely "format." It also has a function to integrate time and space. Frame used to be simple square or rectangle shapes displayed in alignment on a page. Later it developed into diverse shapes aligned dynamically to express different psychological situations (Natsume, 1995, 1997, Yomota, 1994).

Thus, the roles of picture, word, and frame have created the unique characteristics of Japanese manga. Put simply, the complexity and drama of the story is the reason young people are strongly attracted to manga. As a result of reading manga and discussing it with their peers, young people's literacy is growing (Nakazawa and Nakazawa, 1993).

The development of more complex stories in manga is related to the expansion of the age range of readers. Regardless of the country, comic books are generally assumed to be for children who eventually progress beyond childish habits, generally after high school. Usually, children stopped buying and reading when they graduated elementary school. However, Japanese children did not stop even after high school because manga was more attractive than other media, such as TV and movies. A new manga generation, which did not stop reading manga when they became adults, emerged during the1960s. The new generation exposed to manga started to have high expectations of the story as they got older. With the reader's expectations, the story of manga developed to express more human drama than a caricature or a simple strip. As a result, manga started to produce many different types of stories, fiction and non-fiction to please readers: science fiction, sports, love stories, history, and so on (Yonezawa, 1997). The manga market started to publish monthly manga in the mid-1950s and weekly manga by the end of the 1950s. For example, the first boys' weekly magazine was published (The Shonen Magazine) in 1959.With the development of the economy in Japan in the1970s and1980s, the manga market rapidly developed, and manga itself became a popular culture in Japan. In 1994, the circulation of one of the boys' manga, The Shonen Jump, with about 20 serial stories and 500 pages in each issue, finally reached 6 million (Nakano, 1997).

For Japanese children, manga was attractive as a visual means to live their dreams and be anything and anyone they wanted to be in the virtual reality (Wilson, 1988, 1997). For adults, manga were visual textbooks that satisfied their curiosity about the world (Toku, 1998, 2000).

Implementing manga in art educational curricula

How can this children's interest in comic books and the advantage of visual literacy through manga be utilized as a teaching tool in Japan or even extended to countries like the United States? In response to this pop cultural phenomenon, the Ministry of Education in Japan decided to adapt the benefit of pop culture to the national art educational curricula for 8th and 9th grades in compulsory education in 1998.

In a course of study of the national art educational curriculum for 8th and 9th grades that will be implemented in April, 2002, the Ministry of Education proposes, "adapting pop culture (manga, illustration, photo, video, computers) to express students' thoughts/ ideas of what they think and what they want to be" (Spring, 1998). In Japan, the Ministry of Education has changed six times (1947, 1951, 1958, 1968, 1976, and 1989) in theory and practice in the field of art education since World War II (Okazaki, 1984; 1995). Through more than 50 years of educational history, this seventh reform (2002) is the first art educational reform that considers children's preferences for the subject matter in the curriculum.

In response to this proposal for the course of study in art education, art educators have started to look for programs with manga's techniques and contexts (see Aida, 2000; Izumiya, 2000). For example, Izumiya (2000) proposes a visual comprehension test as the first step of implementing manga's technique in lower secondary art education. The test involves drawing objects by memory in order to communicate a common subject or situation to a second person. The purpose of this practice is to make students realize that they tend not to pay attention to such common subjects and that drawing simple subjects is not so easy. As the second step, Izumiya also recommends drawing a common situation in ordinary life by using manga's techniques of composition such as closeups and distant views in diverse alignments of frames on a single page. Izumiya recommends utilizing manga's visual grammar as a visual communicative tool to encourage children's visual literacy in art education.

On one hand, the techniques of manga give art teachers an opportunity to support the development of students' visual thinking skills such as observation, articulation, and critical thinking skills. By drawing a scene of ordinary life in sequential frames, students will pay attention to life and nature around them. These manga activities make art more meaningful to students and will give them a chance to find their identities by depicting themselves in a narrative story. On the other hand, it is also true that adapting manga in art educational curricula brings controversial issues from mangaka (cartoonists) and art teachers themselves. Mangaka and editors fear that young people may lose their interest for manga if manga is taught through art education in schools. Some think that manga is so popular among young people because manga has contextual and expressive freedom in narrative apart from the constraint of art education in schools. By implementing the techniques in art education as a communicative tool to express their thoughts, manga ironically may lose the attraction and the quality. This argument may never be resolved (this is a destiny of adapting any new methodology into a school system). Nevertheless, the most important thing for art education is that this is the first attempt to consider children's preferences in art educational curricula, and teachers are beginning to consider the value of pop culture and the advantages and disadvantages of manga. Art teachers in Japan have begun to reevaluate the role of art education to encourage children's interest and motivation for art.

Conclusion

Internal and external disruptions cause the loss of interest and motivation in creating art, especially during the middle of their artistic development in the adolescent period. Although art education curricula are frequently developed in the belief that they help students with issues of identity and fulfillment, it appears that this objective is not being met.

The role of the art teacher is not just how to teach either the techniques of art making or the absolute value of art by making or looking at art. Rather, it is how to draw students' interest and motivation to create art to find themselves in their own ways. While thinking of what the artwork is through the process of art making and critique of artworks, they will find how art is meaningful for themselves in their lives. To do so, we as teachers have to introduce diverse ways to create art as self-expression by including diverse values from different cultures.

This may be easier said than done, since teachers themselves often have difficulty implementing the proper art program to encourage students to make it meaningful for them. Especially in secondary education, many teachers struggle to face the imbalance between students' physical growth and mental growth. Their physical abilities reach almost the level of adults, but their mentality is still in the process of developing. As a result, we tend to bring our own values to art without thinking of students' own preference and aesthetics for art. It's time to open our eyes to look at what's going on in this world to know what are the most attractive artistic themes for students, although they may not necessarily be the same for us. If there is a pop cultural phenomenon that begins to attract children, we should learn the advantages and the disadvantages of the particular pop culture to look at the possibility of implementing it in art education.

The Japanese critic of pop culture Natsume says, "when we learn something, we need a motive power, which should be to like something, to be interested in it, and for it to be fun" (1999, p.8).

For students who are in such a period of losing interest or motivation for art especially during secondary grades, we should find something to provide the motivational power of learning and to develop students' critical thinking skills. By using the mechanism of manga, it might be possible to help students find their own identities through narrative art. It's time to reevaluate pop culture to give students motivation to create their own values and identities through making and criticizing art.

[Sidebar]

The new generation exposed to manga started to have high expectations of the story as they got older. With the reader's expectations, the story of manga developed to express more human drama than a caricature or a simple strip.

[Reference]

REFERENCES

[Reference]

Aida, T. (2000). A study on the standing of comics in art education. The Journal for the Society ofArt Education. 21. 1-11. Tokyo: Tokyo Gakugei University.

Akiyama, T. (1990).Japanese painting. New York: Rizzoli International Publications. Davis, J. (1997). The "U" and the wheel of "C":

Development and devaluation of graphic symbolization and the cognitive approach at Harvard Project Zero. InA. Kindler (Ed). Child Development in Art. (pp.45-58). Reston, VA: the National Art Education Association.

Gardner, H. (1980). Artful scribbles: The significance of children's drawings. New York: Basic Books.

Gardner H. (1990). Art education and human development. Santa Monica, CA: The Getty Center for Education in the Arts.

Izumiya T. (2000). Bijyutsushitsu de no manga no tsukiaikata [How to work with manga in art room]. In Art in Education. 61 (1). Tokyo: The Society for Art in Education.

Kikuyama, S. (1993). Cartoon as "formative vocabulary." Bulletin of Society ofArt Education in University. A separate volume of 25. 287-294. Tokyo: Society of Art Education.

Lowenfeld, V. & Brittain, W.L (1970). Creative and mental growth. (5th ed.). New York: Macmillan.

Ministry of Education. (1998). Course of Study of Lower Secondary School. 176. Tokyo: Ministry of Education, Science, Sports and Culture. December 14,1998.

Nakano, H. (1997). Comic zasshi market no koubou [Rise and Fall of Comic Market]. In Comic gaku no mikata [How to look at Comic]. Asahi Shinbun Extra Report & Analysis Special. 1997.24,103-109.

Nakazawa, J. & Nakazawa, S. (1993). How children understand comics?: Analysis of comic reading comprehension. In Annual Research Early Childhood. 15. (pp. 35-39).

Natsume, F. (1995). Manga no Yomikata [How to read manga]. Tokyo: Takarajimasha. Natsume, F. (1997). Manga no Bunpo

[Grammar of Mangal. In Comicgaku no mikata [How to look at Comic]. Asahi Shimbun Extra Report & Analysis Special. 1997. 24, 86 - 95. Tokyo: Asahi shinbun

Natsume, F (1997). Why manga are fascinating?: Their visual idioms and grammar [Manga wa naze omoshiroi]. Tokyo: NHK library.

[Reference]

Natsume, F. (1999). Mango ga suki [I like mango]. In Magazine for Art Education. 49 (8). 7-12. Tokyo: Biiku Bunka

Okazaki, A (1984, Fall). An overview of the influence of American art education literature on the development of Japanese art education.Journal of Multi-cultural and Cross-cultural Research in Art Education, 2 (1),82-95.

Okazaki, A (1995). American contemporary art in Japanese school art: Play activity in the 1980s. In H. Kauppinen & IR Diket, (Eds.), Trends in Art Education From Diverse Cultures. (pp. 175 - 180). Reston, VA National Art Education Association.

Read, H. (1958). Education through art. (3rd Ed.). New York: Pantheon.

Schodt, F.L (1983). The world of Japanese comics. New York: Kodansha America, Inc. Schodt, F.L (1996). Dreamland Japan: Writings

on modern manga. Berkeley, CA. Stone Bridge Press.

Toku, M (1998). Why do Japanese children draw in their own ways? Doctoral dissertation. University of Illinois at Urbana-Champaign.

Toku, M. (2000). Cross-cultural Analysis of Artistic Development: Drawings by Japanese and U.S. children. Paper accepted Visual Arts Research, 26 (1) or (2).

Tsukakoshi, Y. (1997). What is art?: Manga and art education. Thejournal of Master's Thesis. (pp, 21-28). Kariya, Japan: Aichi University of Education.

Wilson, B. (1988). The artistic tower of Babel: inextricable links between culture and graphic development In G.W. Hardman & T. Zernich, (Eds.), Discerning art: concepts and issues. (pp. 488 - 506). Champaign, IL Stipes Publishing Company.

Wilson, B. (1997). Child art, multiple interpretations, and conflicts of interest. In AM. Kindler (Ed.). Child development in art. (pp. 81-94). Reston, VA National Art Education Association.

Wilson, B (1999). Becoming Japanese: Manga, children's drawings, and the construction of national character. In Visual Arts Research 25 (2). (pp. 48-60).

Yomota, I. (1994). Mangagenron [The principles of manga]. Tokyo: Chikumasyobo. YonezawaY. (1994), Shojo manga no sekai [The

world of girls' mango]. Tokyo: Heibonsha. Yonezawa Y. (1997). Shonen manga no sekai [The world of boys' mango]. Tokyo: Heibonsha.

[Author Affiliation]

Masami Toku is Assistant Professor, Department ofArt and Art History, at California State University, Chico. E-mail: mtoku@csuchico. edu. She is a first-time contributor to Art Education.

STATE REGULATION OF ON-FARM ANAEROBIC DIGESTION

2012-03-12T16:51:00.000-07:00

CALIFORNIA AND NEW YORK

Introduction of off-farm waste streams to agricultural manure digesters is leading to some hybrid approaches in state permitting and rule development.

A NUMBER of states have granted operating permits to on-farm anaerobic digester facilities that process farm manure and other organics for biogas production and subsequent on-site electrical generation. Some of those facilities are also permitted by the agencies to codigest nonagricultural feedstocks such as fats, oil, grease and food processing residuals, which are usually managed as municipal solid waste. When such materials are added, the farm may generate tipping fees and can produce significantly more methane per ton than by using manure alone, and thus can produce more electrical power.

Regulation of such AD systems has required states to come up with a hybrid of permitting requirements involving regulations applicable to CAFO (confined animal feeding operations), AFOs (animal feeding operations), municipal solid waste, wastewater treatment, air emissions, and nutrient management plans. On top of such state regulations are local zoning and building code requirements. Unlike the U.S. Environmental Protection Agency's Part 503 regulations that set minimum standards for reuse of biosolids, there are no comparable federal standards for operation of anaerobic digesters (AD) for processing manures and food waste. Therefore, regulatory agencies are cutting their teeth on permit applications submitted for such AD projects. Another article in this month's BioCycle Energy section about the Mead, Nebraska facility that processes only on-site generated material, illustrates the importance of the permitting process in development of AD facilities, particularly its impact on capital costs.

As the first in a series of articles on regulations and permitting for anaerobic digesters, BioCycle reviewed requirements in California and New York. From a regulatory perspective, AD may be considered a variation of composting because it is a biological process which maximizes anaerobic bacteria instead of aerobic bacteria and fungi. However, many AD systems operate at around 100�F rather than 131�F as required by the USEPA Part 503 regulations and some state composting regulations. The AD process extracts methane and produces a solid material (digestate) that can be processed, usually through composting, to achieve further pathogen reduction.

CALIFORNIA REGULATORY STRUCTURE

Scott Anders at the University of San Diego's Energy Policy Initiatives Center recently released a paper titled Biogas Production and Use on California's Dairy Farms, A Survey of Regulatory Challenges (August 2007). The paper identifies regulatory challenges in the areas of water, air, electricity, natural gas, and solid waste management. Anders notes that California has 22 anaerobic digesters located on dairy farms, and he identifies the following California state agencies that have some jurisdiction over major portions of biogas production and consumption:

* California Integrated Waste Management Board (CIWMB) has jurisdiction over solid waste disposal, recycling, and composting.

* California Air Resources Board and 35 local air pollution control districts (APCDs) regulate air emissions from mobile and stationary sources.

* State Water Resources Control Board and nine regional water quality control boards are charged with protecting and preserving the water resources of the state.

* California Public Utilities Commission regulates privately-owned electric and natural gas companies.

The CIWMB does not have a statutory or regulatory definition for anaerobic digestion, something that would help to resolve jurisdictional questions for project developers. Anders points out ambiguity over the extent to which the CIWMB has jurisdiction over AD systems.

The only APCD that is currently regulating ADs at dairies in California is the San Joaquin Valley APCD. This district is located in the southern portion of the Central Valley which has significant air pollution issues related to various sources. The district has identified ADs as a good control method for air emissions from manure storage facilities, but does not mandate their use due to the high costs relative to the expected air quality benefits.

With respect to regulatory programs for protection of water quality, Anders notes that historically in California, animal feeding operations such as dairies, feedlots, and poultry facilities operated under a waiver of permit requirements. However, as described below, that situation has changed.

John Menke is a Senior Environmental Scientist with the California Water Resources Control Board and is the Board's technical expert on AFOs and ADs. He notes that California regulations prohibit dairies from discharging manure to surface waters, even during a 25-year, 24-hour storm. Although most AFOs in the state voluntarily comply with the discharge prohibition, some dairy operators did not construct adequately-sized lagoons or storm water containment facilities and as a result had discharges, primarily during storm events. To address such discharges, the state formed an Attorney Task Force to initiate enforcement action. According to Menke, there have been over 100 enforcement actions against dairies in California over the past eight years. Fines are generally in the tens of thousands of dollars for discharges but have been over a million dollars for repeat offenders.

Since few dairies in California discharge manure to surface water, few AFOs are required to obtain NPDES permits. Instead, most dairies are now regulated through Waste Discharge Requirement (WDR) Orders - essentially state permits issued by the regional water quality control boards. Menke notes that there is great regional diversity between dairies within California, where North Coast dairies typically milk 250 to 300 cows and are on pasture, and many dairies in the Central Valley milk more than 1,000 cows and have lagoons that can exceed 40 million gallons. "Given that diversity, it makes sense for each region to develop its own regulations," he explains.

An exception to the use of WDRs is the approximately 170 dairies in the Santa Ana Region in southern California. That region includes the Chino Basin where, due to urban expansion, little cropland remains for manure application. As a result of several manure discharges into the Santa Ana River, all dairies and feedlots in that region are under a general NPDES permit.

Menke notes that WDR Orders have an advantage over NPDES permits in that the WDRs are more appropriate for protection of groundwater. "We're very concerned about salts and nitrate in groundwater in California," he says. "We try to maintain water quality at its ambient concentrations, and we work to ensure that a discharge activity does not make the water unusable." Menke explains that there is an upper limit of 1,000 ppm salt, above which the water is not suitable for use as drinking water. "However, for some beneficial uses the acceptable upper salt limit is lower, and for some specific salts or ions the upper limit may be a low value." Using a WDR Order means that all AFOs, not just concentrated animal feeding operations (CAFOs), can be required to utilize nutrient management plans.

"We recognize that ADs have great benefits for reducing emissions of greenhouse gases and improving air quality," adds Menke. "But the digester does not decrease the salts and nutrients in the manure, and we want to ensure proper management of the organic residuals and effluent that are discharged." He notes that they are particularly concerned about codigestion because the imported feedstocks may add salts and nutrients. A few of the approximately 20 ADs at dairies in California currently use codigestion.

CODIGESTION CONSIDERATIONS

For codigestion of manure and off-farm wastes in the Central Valley, Menke says there is a case-by-case review and development of an individual WDR Order. "The big problem is a lack of characterization of the waste stream. For example, if a farmer wants to add 10 percent tomato processing waste to a manure anaerobic digester, they have to provide the expected concentrations of salts and other pollutants in the proposed waste and demonstrate that water quality is protected. We have asked how salts are handled in Europe where they have more anaerobic codigestion systems, and have found that salts do not seem to be much of a concern in Europe. Since California has a greater dependence on groundwater for domestic consumption and irrigated agriculture, we need to ensure that down-gradient water users are not impacted by salts from both natural occurrence in the soil, as well as from upstream farm fields."

In Southern California, the Inland Empire Utility District operates a regional digester that utilizes manure from about eight dairies. Some of the energy produced is used to power a reverse osmosis system that treats drinking water for the region. Effluent from the AD is discharged to a "brine line" that goes to the regional wastewater treatment plant, which discharges its treated effluent into the Pacific Ocean. No such brine line exists for dairies in the Central Valley.

About 1,600 of the state's 2,000 dairies are located in the Central Valley Region, and the Central Valley Water Regional Board has identified the following water quality concerns related to anaerobic digestion: (1) Use of off-site waste in codigestion, which can increase the salt and nitrate content of digestate; (2) Ability of lagoons to safely contain digestate, in particular from codigestion; and (3) Potential groundwater quality impacts from the effluent stored in lagoons.

In May 2007, the Central Valley Water Board adopted a General WDR Order with requirements that apply to the construction of holding ponds, the operation of ADs, and the discharge of manure to cropland. Any dairy operator in the Central Valley wanting to cover an existing storage pond to capture methane gas or to construct a new AD must file a Report of Waste Discharge (ROWD) that describes the digester and how digestate and effluent from the digester will be managed to protect water quality. Board staff will review the ROWD to ensure adequate protection of water quality. For a new lagoon system to be approved, an engineering study must prove through modeling that there will be no unacceptable degradation of groundwater quality.

Recognizing regional water board's regulations, Ander's report recommends that "California should evaluate the possibility of developing a permit process for complex and cross-cutting projects like biogas production... Because biogas production and use requires approval from so many different state and local agencies, centralizing and standardizing the permit process could help to ease the regulatory burden for participants and help California attain its greenhouse gases reduction goals."

Echoing Ander's conclusions is Ruth MacDougall of the Sacramento (CA) Municipal Utility District (SMUD), an electric utility with a commitment to greenhouse gas reduction. SMUD's official policy is to "pursue renewable energy supplies and other non-greenhouse gas emitting energy resources," which currently includes the purchase of 5 MW of power from the Sacramento County Regional Sanitation District's biosolids digester. "We want to get farms to put in anaerobic digestion systems and we have sponsored some research which shows farms can be a good source of renewable energy for our electrical district," says MacDougall. "We have one farm putting in an AD system, but a big problem is that the regulations are not well defined, and we believe it would facilitate more AD projects if we had consistent standards."

NEW YORK STATE'S COOPERATIVE AGREEMENT

New York State has several anaerobic digestion biogas systems operating on farms, including CAFOs. Some of the facilities have been permitted in accordance with the state's solid waste facility regulations, and some under the CAFO regulations, and at least one under both. To avoid such "double regulation" in the future, the two divisions of the State Department of Environmental Conservation (DEC) that regulate solid waste and CAFOs recently drafted a cooperative agreement that allows farms with a CAFO permit to process off-site generated food waste, with no solid waste facility permit required.

Sally Rowland, DEC Environmental Engineer charged with overseeing the state's Part 360 rules for organics recycling projects, explains that the Part 360 regulations are in the process of being amended to reflect food processing in accordance with CAFO permits. "Under a farm's CAFO permit we are confident that the permit addresses all of the issues of concern," she says. "However, for farms that do not have a CAFO permit, a Part 360 solid waste permit will be required to handle food materials." Part 360 permits address the design and operation of a facility, its capacity, and potential environmental impacts of noise, odor, and run-off.

A Part 360 solid waste permit was issued to the Cayuga County Soil and Water Conservation District in 2006 to construct and operate a facility in the Town of Auburn that will anaerobically digest 40,000 gallons/day of dairy manure and straw bedding, 4,200 gallons/day of potato processing wash water, 2 tons/day of potato clippings, and 6 tons/day of grease/cooking fats. The District anticipates using a German mesophilic hydraulic mix digester technology (GBU) that agitates the manure by downward flow of the liquid, minimizing moving parts. Once completed, the methane will be burned in a Jenbacher gas engine to produce electricity and hot water to heat an adjacent nursing home, jail, office building, and maintenance shop, with excess power sold to the electrical grid. Digestate will be dewatered and the solids composted on a 2-acre pad in accordance with pathogen reduction requirements of a Class A compost for use on sites other than farmland. The liquid will be transferred to local farms for land application as fertilizer, in accordance with the farm's nutrient management plan or a land application permit under Part 360.

New York State also requires the Cayuga County facility to have a storm water discharge permit for construction activities, as well as implementation of a storm water pollution prevention plan. Although the facility is not a CAFO, the Part 360 solid waste permit requires any CAFO permitted facility "importing or exporting waste to or from this digester facility to update their CAFO required Comprehensive Nutrient Management Plan."

Due to the gas engine, a DEC Part 201 air facility registration certificate for the exhaust emissions is also required. The air registration also covers a safety flare that will be operated when the generator system is not operating, or when the biogas storage system cannot accept biogas.

Jim Hotaling, who manages the Cayuga County Soil and Water Conservation District's community digester project, explains that several other local permits were required, including state historical commission review, a wetland permit from the U.S. Army Corps of Engineers, and county permits for the building and the access road. Illustrating the project specific nature of such innovative projects, the host Town of Auburn raised concerns about the potential for explosions associated with the new facility. hotaling says that this issue was eventually resolved. "The Town's main concern about the project was liability from explosions, so a couple of months were required to have the attorneys for both parties negotiate an agreement where the County assumes liability for such an accident."

Also located in the Town of Auburn, the Patterson Farm received a Part 360 approval in 2005 for a "research, development and demonstration permit" for a manure and 18,000 gallon/day food waste anaerobic digester. The digested food waste/manure mixture is separated into solids, which are used for bedding, sold as compost, or land applied per the farm's CAFO Manure Management Plan. The Part 360 permit states that "only food processing waste approved under the farm's CAFO permit can be processed at this facility. Land application of digester residue must be in accordance with the CAFO permit, and other treatment and disposal options must be in accordance with Part 360."

The DEC, through its Division of Water, has issued a General State Pollutant Discharge Elimination System (SPDES) Permit for CAFOs. The permit includes minimum standards for protection of water resources that are applicable to all CAFOs, as well as specifications for developing Comprehensive Nutrient Management Plans in accordance with the Natural Resource Conservation Service's Conservation Practice Standard No. NY312. For a CAFO to be in compliance with the CAFO permit, "there shall be no discharge of process wastewater pollutants to the surface water of the State, except for releases in excess of the 25-year, 24-hour storm event. Process water is defined as any wastewater, leachate or precipitation that comes into contact with manure, bedding, grain, or other organic materials."

Therefore, each CAFO farm must have lagoons and treatment systems to comply with those requirements, even if no food waste is handled. In addition, any farm with a CAFO permit must amend its nutrient management plan if any significant changes are made in the facility, such as adding food waste.

In addition to providing legal protection for its manure management practices, as well as savings in fertilizer costs associated with nutrient management plans, the DEC recognizes that another potential benefit to CAFO permitees is renewable energy production. Notes DEC's website: "If green power, such as electric generation from an anaerobic digester, or CO2 trading becomes popular, the comprehensive nutrient management plans needed for the CAFO permit may be required to certify the farm is green."

Information on how other states and Canadian provinces are regulating onfarm digesters and processing of off-farm feedstocks in those systems will be addressed in future articles. If New York's approach of regulating on-farm digesters through the CAFO process is deemed successful, it is likely other states will follow suit. Other states may follow California's approach to empower regional boards to determine permit conditions that are based on regional geographic differences such as size of farms, annual precipitation, water quality, soil conditions, and agricultural practices.

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Permitting anaerobic digesters is sometimes done in accordance with an existing CAFO permit.

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Among the Central Valley Water Regional Board's water quality concerns is possible increase in salt and nitrate content of digestate if off-site waste is codigested with manure.

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Patterson Farm received approval for a "research, development and demonstration permit" for a manure and food waste anaerobic codigester.

Lamont Marvels at Thomas' Skills as Batter

2012-03-12T13:09:00.000-07:00

Frank Thomas' dramatic two-run home run Friday night won acrucial game for the White Sox and continued his blistering RBI pace,tying him with Detroit's Cecil Fielder for the league lead at 96.

Thomas leads the American League in RBI ratio (RBI to at-bats)and is fourth in home run ratio. He became only the 12th Sox to hit30 home runs in a season.

Manager Gene Lamont, who coached Barry Bonds, said he hasn'tseen a player who combines Thomas' power and patience at the plate.

"I don't know if I've ever seen anyone who came into the gamewith his kind of power and his patience at the plate, and hisrelative inexperience - he's only in his fourth season," Lamont said."It's almost an odd combination."

Lamont is as pleased with Thomas' improvement defensively atfirst base.

BACK IN ACTION: Ozzie Guillen started at shortstop Saturdayafter a day of tests Friday on his left ankle, which was injuredWednesday in a play at second.

Tests showed inflammation but no serious injury.

Guillen played defense in the ninth inning Friday but had hisankle taped tightly Saturday and was told by trainer Herm Schneiderto wear high-top spikes to provide extra support.

BELL BACK: George Bell was due back Saturday from Florida wherehe was going through his initial rehabilitation from arthroscopicknee surgery.

Bell will continue his rehabbing with the team.

BO AVAILABLE: Bo Jackson took batting practice again and wasavailable to hit if needed despite a bandaged right hand cut whilerepairing his motorcycle.

Jackson is scheduled to have the stitches removed Wednesday.

SECOND CHANCE: Steve Sax hadn't taken a ground ball at secondbase all year when Lamont inserted him there for defense in theeighth inning Friday.

But Lamont had told Sax he might be playing there before thegame.

"I knew the situation might come up because I was giving Frank(Thomas) a day off at first," he said. "I told the coaches I knewwhat I could run into, but when Ozzie said before the game he couldplay if I needed him, it made it easier."

As it turned out, Sax - who batted in the seventh as the DH -came out in the ninth, and had the Sox had to bat in the bottom ofthe ninth, pitcher Roberto Hernandez would have been at-bat since theSox DH had been pulled.

Lamont said he probably would have pinch hit for Hernandez hadthe situation occurred.

POSTSEASON TICKETS: The Sox, hoping to cash in on the pennantdrive, have announced a sales plan to purchase 1994 full seasontickets with the guarantee of seats to any 1993 postseason games atComiskey Park.

Split or bleacher season ticket-holders also can upgrade theircurrent plans to full 1994 tickets for guaranteed 1993 playoff seats.Information is available at 312-924-1000.

ACE VS. ACE: Jack McDowell will face the Red Sox' Roger ClemensTuesday in the first of the two-game series at Boston. Lamontdecided to keep to his pitching rotation despite Monday's off daywhen Wilson Alvarez would have been scheduled.

The plan keeps pitchers on their work schedule. The Sox haveindicated Alvarez will be recalled from Class AAA Nashville in timefor his next scheduled start, but Lamont could use Kirk McCaskill orJose DeLeon as spot starters if necessary.

FLOAT ON: Bo Jackson, Tim Raines, Lance Johnson and Ellis Burksrode in the Sox float in Saturday's Bud Billiken Parade.