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The Human Services certificate/associate degree is a performance-based training program designed to prepare students for employment in the field of human services/social services. Coursework also meets many of the lower division requirements for advanced degrees in related fields. The certificate offers students excellent preparation in the knowledge and skills needed for successful employment. The certificate also provides an overview of local human services agencies including fieldwork in the student's emphasis of choice.
Program Highlights
Employment Outlook
The U.S. Bureau of Labor Statistics reports that employment of social and human services assistants is expected to grow by nearly 34 percent through 2016. Job prospects are expected to be excellent, particularly for applicants with appropriate post secondary education. According to the California Occupational Guides, human service workers with post secondary education should find excellent job opportunities through 2014. In Sonoma County, there is projected to be a 12% increase in employment of substance abuse counselors in the 10-year period ending in 2014.
Typical Starting Salary for Graduates
State government $35,510
Fees/Financial Aid
Santa Rosa Junior College is a public community college. Fees are $26 per unit for California residents. Out-of-state and international students will pay higher fees. The minimum number of units required for a degree is 60 units, and a certificate in Human Services ranges from 43.5 - 46.5 units depending on the certificate emphasis chosen. Books and materials are additional costs. Some students may have their fees waived by the Board of Governors' Grant awarded by the State of California. For information on the Board of Governors' Grant and on the many other forms of aid, visit or contact the Financial Aid Office at (707) 527-4471.
What is the First Step I Need to Take to Enroll in the Program? Take the English reading/writing and mathematics assessments and meet with a counselor for course planning.
Recommended Preparation
English skills are critical for success in the Human Services classes. Students should plan to take the SRJC's English assessment for placement into appropriate English classes. Students entering the Human Services core curriculum would ideally have a score recommending placement in English 100. Good writing and computer skills are necessary for employment within the human services field.
Required Courses
A complete list of required courses are available online (www.santarosa.edu/certificates and www.santarosa.edu/majors), in the class schedule and in the college catalog.
Starting the Program
The Human Services core curriculum begins each fall or spring semester. Students should begin, pending English assessment results, by enrolling in Coun 90, Introduction to Human Services. See recommended sequence of courses.
When the Human Services student has decided on an emphasis, he/she should see the Intern Instructor/Coordinator for intern placement in the final semester of their studies. The 3-unit requirement (6 units if the emphasis is Alcohol and Drugs) requires 12 hours per week for a non-paid (volunteer) position or 15 hours per week for a paid position. Students may develop their own position or utilize the assistance of the placement office. Call (707) 527-4329 for an appointment or stop by Work Experience Department, 3rd floor of the Bertolini Student Center on the Santa Rosa Campus. Plan ahead, because the Intern Instructor's/Coordinator's schedule is usually full at the beginning of each semester. Enrollment in the intern units occurs after the position has been secured and approved. There is an application to complete. Students may enroll until the end of September (for the fall semester) and February (for the spring semester). The intern program is also available during the summer semester.
Placement Services
Assistance with job placement is available through SRJC's Student Employment Office. Students are encouraged to register with the Career Developer to receive guidance with resume development and referral for job opportunities. The phone number is (707) 527-4690 for placement assistance and/or labor market information.
Questions or comments about this page may be sent to firstname.lastname@example.com.
Santa Rosa Junior College is officially accredited by the Western Association of Schools and Colleges


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HARRISBURG >> At a time when the state's economy is struggling to pick up steam, Pennsylvanians see strong connections between public education, state funding and economic development, according to a recent statewide public opinion poll.
"While our state's year-over-year job growth sits among the weakest states and the state's contribution to public education funding is below the national average, Pennsylvanians are making connections that state officials have so far missed," said Jim Buckheit, executive director of the Pennsylvania Association of School Administrators (PASA). "Strong public schools are critical to a strong economy and the investments our state makes should strengthen every community."
Terry Madonna Opinion Research's Spring 2014 Omnibus Survey shows large majorities of registered voters believe public schools have an effect on the economy, the state's investment in public schools should be larger and a school funding formula should be used to ensure fair distribution of state funds to schools in all communities.
"Public education funding must be seen for what it is — an investment in Pennsylvania," said Nathan Mains, executive director of the Pennsylvania School Boards Association. "Without proper funding, it is becoming increasingly challenging for school directors to balance budgets while maintaining the integrity of educational offerings."
Survey findings on education are as follows:
• More than 8 in 10 Pennsylvanians (84 percent) surveyed said they believe public schools have a "very strong" or "some" effect on economic development;
• Nearly three quarters of Pennsylvanians (71 percent) surveyed said they believe the state investment in public schools needs to be "much larger" or "somewhat larger";
• More than two-thirds of Pennsylvanians (67 percent) said schools with greater numbers of students in poverty should "definitely" or "probably" receive more state funding;
• Nearly three quarters of Pennsylvanians (72 percent) said they "strongly favor" or "somewhat favor" using a school funding formula to ensure fair distribution of funding;
• Nearly three quarters of Pennsylvanians (72 percent) said "definitely yes" or "probably yes" to using a school funding formula to ensure fair distribution of state education funding.
"Built upon a patchwork of annually changing supplemental formulas with inconsistent targets for additional support, current basic education funding results in an unpredictable distribution of dollars to school districts across the Commonwealth. This distribution only exacerbates the challenges of budget planning from year to year. We need a funding system that is built on reliable, known and verifiable data that supports long term and consistent budgeting" said Jay Himes, executive director of the Pennsylvania Association of School Business Officials (PASBO).
"Pennsylvania school officials also are holding off on construction projects during the state's moratorium on reimbursements for renovations and new building projects, compounding the operational budget challenges. No projects have been added to the school construction pipeline since October 2012 and Gov. Tom Corbett proposed extending the moratorium through June 30, 2015. Without changes, the existing process threatens to drive up local property taxes, force school boards to cut more from educational programs, and hamper economic growth.
"Some of the oldest school buildings in the state are in rural communities where students are preparing for the future without modern tools for teaching and learning. Schools in every community deserve adequate support for a great education that allows students to stay and rebuild or reinvent their local economies," said Joe Bard, executive director of the Pennsylvania Association of Rural and Small Schools.
Statewide, Pennsylvanians said education funding and adequate state support for all schools should be greater priorities in the state's budget.
"Central and western Pennsylvania deserve the same support and opportunities as all other parts of the state, which depends on changes to ensure adequate funding for all public schools," said Dr. J. Hugh Dwyer, chair of the Central Pa. Education Coalition. "School leaders and parents see the inequities in state support for education and are expressing their beliefs that changes are needed to ensure a better future for their children and communities."
The survey was designed to be representative of the state's registered voter population. It involved telephone interviews with 800 adults between Feb. 10-20, 2014. The telephone poll was conducted using statewide voter registration lists and reflects the opinions of Democrats and Republicans, parents, and residents living in every region of the state. It has a margin of sampling error of plus or minus 3.5 percentage points for all respondents.

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St. Mary's student performs with DYSE
By Molly Moser
The Dubuque Symphony Youth Ensembles (DSYE) opened its 2013 – 2014 concert season at the University of Dubuque's Heritage Center on Sunday, Nov. 17. The DSYE is comprised of three training ensembles: Dubuque Youth String Ensemble, Dubuque Youth Philharmonia, and Dubuque Symphony Youth Orchestra.
Mara Shannon, age eight, was selected to play with the String Ensemble following a live audition. Mara told her violin instructor, Katie Sigwarth, that she'd like to perform with the DYSE. "My mom said, 'Are you sure about it? Are you ready for it? You have to do lots of practicing!' I was like, yes!" Mara told The Press.
The String Ensemble is an introductory honors ensemble for young string players with at least one year of ensemble experience or equivalent ability. The group performs various levels of string orchestra literature from the standard middle school repertoire.
Mara has been taking violin lessons weekly for two years. "My mom showed me the violin on TV, and I wanted to play it," Mara explained. She was drawn to the sound and the finger movements, but says struggled when she first started. "I didn't know how to exactly play it, but then I practiced and got it," she said.
Mara also plays piano, but says she's not as advanced at that as she is at the violin. She started on a quarter-size violin and has moved up to a half-size. "If your violin is the right size, you have to make sure it goes from here, to here," Mara said, tucking her violin under her chin and wrapping her palm around its opposite end. By those standards, the half size instrument fits the third grader-perfectly – but she expects to move up to a full size violin by age 10 or 12.
The String Ensemble, which consists of 24 students playing bass, violin, viola, and cello, is conducted by Ann Duchow. Duchow is associate concertmaster for the Dubuque Symphony Orchestra (DSO) and is the orchestra director at Dubuque Senior High School. In the season's first concert, she led the String Ensemble in three selections that showcased different types of bow strokes.
Mara's own sheet music is full of notes like 'counting,' 'bow position,' and 'brush-offs,' written in a child's scrawl. "I wasn't scared," said Mara when remembering her performance with the DSYE. The best part about it, she says quite simply, was playing music.
She enjoyed practicing for the concert, though she admits it wasn't easy. "At first the songs were hard, and I didn't know what to do," she said. "My teacher Katie Sigwarth helped me with it on Tuesdays." The DSYE rehearses on Sunday evenings throughout the school year at Dubuque Senior High School. Rehearsals began on Sept. 8.
Mara's parents, Dan and Joan Shannon, joined the audience at Heritage Center to cheer for their daughter. "Oh my gosh, it was so good," said Joan. "I was in awe. Those kids are amazing."
Two additional DYSE concerts will be held on March 9 and May 18. To order tickets, call (563) 585-SHOW.


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Curatorial Projects:
The show is filled with fantastic connections between traditional and contemporary subject matter and styles. The Ethnic Art Foundation is sharing this amazing work throughout the United States for the next two years. Patter Hellstrom Visual Art / Curatorial is assisting with it's management.
2012 FALL California State University/Fresno
2013 WINTER New Delhi, India
2014 FALL Chazen Museum University of Wisconsin
2014 WINTER Syracuse University
MITHILA PAINTING: The Evolution of an Art Form
Early reviewers of the MITHILA PAINTING: THE EVOLUTION OF AN ART FORM :
"How can one generalize about a genre that includes: Baua Devi's lush interpretations of the Krishna stories; Pinki Kumari's de- and re-construction of the classic ritual kohbar motif; Priyansha's extraordinary homage to a Botticelli Nativity; Rani Jha's powerful and disturbing depiction of the abortion clinic; Shalinee Kumari's iconic visions of capitalism and global warming" says John H. Bowles, author of Painted Songs & Stories: The Hybrid Flowerings of Contemporary Pardhan Gond Art.
While Joanna Williams, one of the most respected South Asia Art Historians in the USA and professor of Art History at the University of California, Berkeley stated that, "in 40 paintings on paper this exhibit updates an ancient wall painting tradition ranging from the early deities and marriage images to contemporary narratives and social and feminist themes."
Contact Paul S. Aaronson, Exhibition Manager email@example.com for the details of exhibition booking or more data about the show.
Works on paper sourced for a Luxury condo building on the Hudson River overlooking Manhattan
Patter Hellstrom and Peter Zirnis curated 75 original art works, a large-scale mobile and an enlargement of artist Tomomi Ono's print Seed Fusion into the public spaces of the Westin Hotel Jersey City Newport which opened in 2009.
Exhibitions 2002- 2007
Art Newport - October 2002
New Growth - December 2002
Color Complexities - January 2003
Inside/Out - October 2003
Spirit in Art - March 2004
Encyclopedia of Images - October 2004
Art at Newport - Sculpture Walk - August 2005
Outer Worlds /Inner Thoughts - August 2005
Views from Newport - May - August 2006
Discover Mithila - December 2006 - February 2007
MACK-CALI REALTY Exhibition Series:
- Glimmering Hues February - April 2005
- DIVERGE May - July 2005
- Building Blocks August 2005 - January 2006
PUBLIC ART / Pavonia Arcs
Commissioned Sculpture by Robert Pfitzenmeier, 2004 / project managed by Hellstrom+Zirnis
Sculpture 'surfs' onto waterfront
Bonnie Friedman
Thursday, July 22, 2004
Robert Pfitzenmeier's "Pavonia Arcs," a 16-foot stainless steel and zerconium sculpture in the shape of a sail, was unveiled in front of the Fleet Bank on Pavonia Avenue and Washington Boulevard Tuesday, making it the newest addition to the Newport waterfront. Commissioned by the Queens-based Lefrak Organization, which owns the building, the structure's theme is in keeping with its proximity to water - suggesting the shape and movement of a wind surf.
"I'm a sailor and a windsurfer," said Pfitzenmeier, the 57-year-old Jersey City artist who lives and works just steps away from the statue."And this relates to that dynamic. The wind is such a powerful force here." This is the second outdoor installation for Pfitzenmeier, who said he chose stainless steel and zerconium because of the durable nature of the material. The nine triangular panels inside the sail are brilliant shades of orange, pink, blue and yellow,and flutter with the wind. Jamie LeFrak, a principal in the family-run Lefrak Organization, said he chose this particular design because of how it evokes movement. "I think, aesthetically, it's most pleasing," Lefrak said. "I like the expressive feature of the sail."
During his 30-year career, Pfitzenmeier has created indoor installations around the country, including Alaska, Delaware, Wisconsin, and in New York's Kennedy International Airport. Many of his works have been commissioned for hospitals. "I think hospitals have been responsive because the work has an upbeat quality to it," Pfitzenmeier said. "It is colorful and abstract and has a healing quality to it."
For Pfitzenmeier, the unveiling was especially exciting, given that he lives so close to where the statue stands. "Lots of times, I make a statue and that's it,"he said. "Now I can see it all the time." Pfitzenmeier, who has a studio at 111 First Street, an artist's enclave in the nearby Powerhouse Arts District, was joined by many of his fellow artists at the unveiling.
"We're very proud," said Nancy Wells, a mixed-media artist. "It's so nice to come out and be a part of something happening in the community. " Peter Zirnis, of Hellstrom/Zirnis, the company that headed up the artist search, thanked the Lefrak organization for commissioning the work.


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CHICAGO, IL - NOVEMBER 10: Jay Cutler #6 of the Chicago Bears passes against the Detroit Lions at Soldier Field on November 10, 2013 in Chicago, Illinois. (Photo by Jonathan Daniel/Getty Images) ORG XMIT: 184892800 ORIG FILE ID: 187586754 / Jonathan Daniel Getty Images
View the full article on USATODAY.com.
Copyright 2015 USATODAY.com
Read the original story: Marc Trestman's gamble doesn't work out


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Type II NAD(PH) dehydrogenases are located on the inner mitochondrial membrane of plants, fungi, protists and some primitive animals. However, recent observations have been made which identify several Arabidopsis type II dehydrogenases as dual targeted proteins. Targeting either mitochondria and peroxisomes or mitochondria and chloroplasts.
Members of the ND protein family were identified in various plant species. Phylogenetic analyses and subcellular targeting predictions were carried out for all proteins. All ND proteins from three model plant species Arabidopsis, rice and Physcomitrella were cloned as N- and C-terminal GFP fusions and subcellular localisations were determined. Dual targeting of plant type II dehydrogenases was observed to have evolved early in plant evolution and to be widespread throughout different plant species. In all three species tested dual targeting to both mitochondria and peroxisomes was found for at least one NDA and NDB type protein. In addition two NDB type proteins from Physcomitrella were also found to target chloroplasts. The dual targeting of NDC type proteins was found to have evolved later in plant evolution.
The functions of type II dehydrogenases within plant cells will have to be re-evaluated in light of this newly identified subcellular targeting information.
Keywords:Type II NAD(P)H dehydrogenases; Dual targeting; Mitochondria; Peroxisomes; Plastids
Eukaryotic cells are defined by the presence of subcellular compartments termed organelles, the presence of which allows for the partitioning of various biochemical pathways out of the cytosolic environment. Each organelle contains a specific complement of proteins that carry out specialised roles within the cellular landscape. However, while it was thought that specific proteins were only targeted to a single organelle, called location specific proteins, a large number of proteins have now been identified to be targeted to more than one organelle, and as such are termed dual targeted proteins. In plants, over 250 proteins have now been identified as dual targeted (Additional file 1) [1,2], the majority of which are targeted to both mitochondria and plastids. However, in recent years dual targeting of proteins has also been shown to occur between a number of additional organelles including: mitochondria and peroxisomes [1,3-7], plastids and peroxisomes [8-10], plastids and nucleus [11], plastids and endoplasmic reticulum [12], mitochondria and nucleus [3,13,14], cytoskeleton and peroxisomes [15], mitochondria and the cytoplasm [16], and plastids and the cytoplasm [17].
Additional file 1. List of known dual targeted proteins in plants. Indicated for each protein is its known locations, plant species, accession number and functional description. It is also indicated the method used to determine dual targeting. The proteomics column refers to if the protein has been identified in proteomic studies in both of its target organelles.
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Mitochondrial respiration is an essential feature of plant metabolism, resulting in the generation of ATP in the process of oxidative phosphorylation. In addition to the classical electron transport chain coupled to phosphorylation, plant mitochondria contain a non-phosphorylating pathway. A major constituent of this alternative pathway is the type II or rotenone-insensitive NAD(P)H dehydrogenases (ND). Type II ND proteins were first identified in potato [18] and have also been identified in fungi, protists, some bacteria and some primitive animals [19]. The function of these type II ND proteins is that when they are linked to an alternative oxidase (AOX) they constitute a non-phosphorylating respiratory pathway that enables the redox status of the cell to evade adenylate control [20]. Specific studies altering the expression or inactivating the expression of specific type II ND proteins suggest roles in the capacity of NAD(P)H oxidation (nda1) [21]; reactive oxygen species formation and altered phenotypes (ndb4) [19].
The subcellular localisations of plant type II ND proteins have been a topic of debate in recent years. Initial in vivo GFP studies involving the targeting signals fused to GFP from five of the Arabidopsis ND proteins identified all proteins to be located in mitochondria [22]. Independent analyses using in vitro import assays into isolated mitochondria determined that three ND proteins (NDB1, NDB2 and NDB4) are externally located on the inner mitochondrial membrane and three were defined as internal (NDA1, NDA2 and NDC1) [23]. The final protein, NDB3 could not be cloned, and was concluded to be a pseudogene [23], although recent transcriptome analysis during germination suggests that NDB3 is expressed very early in germination [24]. However, subsequent studies have identified that Arabidopsis ND proteins are in fact dual-targeted, NDA1, NDA2 and NDB1 were found to be dual targeted to mitochondria and peroxisomes [25], and NDC1, was determined to be targeted to mitochondria and plastids [25,26]. The different conclusions from these studies can be readily reconciled by the fact that initial studies only used C-terminal GFP tags with the first 50 to 100 N-terminal amino acids of the target proteins [22]. However to ensure that targeting signals are not deleted or blocked, both N- and C-terminal GFP tagging is necessary [9,25]. Also with dual targeted proteins it appears that the mature or passenger protein can influence targeting to mitochondria and chloroplasts [27] so full length coding sequences should be tested to determine full targeting ability [25].
While desirable to use two or more complementing methods to determine subcellular localization [28], this can be difficult with dual targeted proteins in a variety of plant systems, requiring isoform specific antibodies and high purity of organelles. These tools do not exist except for Arabidopsis proteins and organelles. Of the known dual targeted proteins in Arabidopsis only 29 percent have been verified by 2 or more approaches (Additional file 1, 46 out of 165 proteins). This number drops to 15 percent when we look at dual targeted proteins identified in both target organelles by proteomic techniques (Additional file 1, 24 out of 165 proteins). By far the best and most widely used technique to discover dual targeted proteins has been fluorescent tagging (92 percent, Additional file 1, 152 out of 165 proteins). To date, no case of dual targeting using GFP tagging has been shown to be an artifact, in terms that while a false-negative rate may exist, due to the nature of the constructs as outlined above, false-positives have not been reported to date. Furthermore when assessing targeting, prediction and phylogenetic comparisons provides additional avenues to assess targeting, and along with GFP tagging, can be used to determine targeting ability of proteins in a variety of plant species, where direct biochemical approaches with gene families are not feasible.
Given that inactivation of specific ND genes in Arabidopsis has led to altered phenotypes [19], it is important to gain a wider perspective on the location of ND proteins in a variety of plants, as function can only be correctly determined if location is known. In order to gain a better insight into the targeting of plant type II ND proteins a study was undertaken to determine the subcellular localisation(s) of all ND proteins from Oryza sativa (rice) and Physcomitrella patens (Physcomitrella), that spans 500 million years of plant evolution, Thus in order to obtain a phylogenetic view of the dual targeting of ND proteins, subcellular localizations were analysed using GFP fusion proteins.
Phylogenetic analyses and subcellular targeting predictions
Previously, we had determined that a number of ND proteins from Arabidopsis thaliana are dual targeted to several locations including: mitochondria and peroxisomes or mitochondria and plastids [25]. To gain a better understanding of how widespread this dual targeting ability is, we identified all the ND type proteins from a number of plant species, utilised various subcellular prediction programs for each ortholog and identified any peroxisomal type 1 targeting sequences (PTS1) according to several prediction programs [29,30] and the AraPerox database [31] (Figure 1 and Additional file 2). Phylogenetic analysis showed that all ND orthologs are clustered into three distinct clades (Figure 1). Types A, B and C (colored in green, red and blue respectively). It was found that A type ND proteins (in green) are predominantly predicted to be targeted to the mitochondria and is consistent with the previous data from Arabidopsis in which both NDA1 and 2 have cleavable mitochondrial N-terminal targeting signals [23]. However, it was interesting that at least one NDA ortholog from each higher plant species appears to have a PTS1 signal at their C-terminus. None of the prediction programs used predicted peroxisomal targeting for any of the NDA proteins from Physcomitrella (Pp) or Chlamydomonas (Cr).
Figure 1. Phylogenetic and subcellular predictions of plant NAD(P)H dehydrogenases. Phylogenetic tree of NAD(P)H dehydrogenases from Vovox carteri (Vc), Chlamydomonas reinhardtii (Cr), Physcomitrella patens (Pp), Selaginella moellendorffii (Sm), Oryza sativa (Os), Zea mays (Zm), Vitis vinifera (Vv), Arabidopsis thaliana (At), Glycine Max (Gm), Picea glauca (Pg) and Populus trichocarpa (Pt). The sequences of the Saccharomyces cerevisiae (Sc) NAD(P)H dehydrogenases are included as well as NADH dehydrogenases from Synechocystis sp. PCC 6803 (Synechocystis) and Escherichia coli (E. coli). The predicted and experimental localizations of each protein (including this study) are indicated by the coloured boxes. Orange = Mitochondrial, Green = Plastid, Blue = Peroxisomal and Black = no prediction. Shown also are the last 12 amino acids (chosen because the PTS1 predictor uses the last 12 amino acids), putative peroxisomal targeting signals (PTS1) are coloured depending on the strength as determined by the AraPerox database (dark blue - major PTS1, light blue - minor PTS1 and pink - rare PTS1). The known experimental loactions for Arabidopsis thaliana ND proteins were taken from [23,25], for Chlamydomonas reinhardtii NDA1 and NDA2 from [32-34] and for Saccharomyces cerevisiae Ndi1, Nde1 and Nde2 from [35]. The phylogenetic tree was constructed using the neighboring-joining method. Numbers at each node are the percentage bootstrap values of 1000 replicates. The scale bar indicates the number of amino acid substitutions at each site.
Additional file 2. Subcellular localizations of Arabidopsis, rice and Physcomitrella NDC type proteins. Summary of subcellular localization data of plant NAD(P)H dehydrogenases. Indicated is which species and accession number for each protein. The predicted and confirmed subcellular localizations of each protein are indicated. Also shown are the known Peroxisomal type 1 (PTS1) targeting signals from plants which are ordered by the classification from the Araperox database. Blue colour indicates the PTS1 classification. M = mitochondrial, Pl = plastid and Px = peroxisomal.
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Amongst the NDB type proteins, prediction of mitochondrial targeting was less frequent (Figure 1 and Additional file 2). This may be due to the fact that NDB proteins do not contain a cleavable N-terminus mitochondrial targeting signal which is the basis of the prediction programs used [23]. However, the majority (22 out of 29) of NDB proteins, ranging from Physcomitrella to Poplar also contain predicted PTS1 sequences. It is interesting to note that the predicted PTS1 sequences in both NDA and NDB proteins are not conserved in all species. In fact, in different species the PTS1 sequence is completely different in terms of the amino acids making up the last C-terminal tripeptide; however, the properties remain the same. This suggests that the C-terminal regions of NDA and NDB proteins is not crucial for catalytic function. The NDC type proteins, so named for their similarity to a cyanobacterial NADH dehydrogenase, and are mostly predicted to be targeted to plastids [22] (Figure 1). No PTS1 sequences were predicted. This is consistent with the localisation of Arabidopsis NDC1, which is predicted to be plastidial but has been experimentally shown to be targeted to both mitochondria and plastids [25,26].
Subcellular targeting of NDA proteins
It has previously been shown that both NDA proteins from Arabidopsis (AtNDA1 and AtNDA2) are dual targeted to mitochondria and peroxisomes by both GFP-tagging and western blot analysis [25]. To demonstrate the fluorescence patterns expected by a dual targeted protein containing an N-terminal mitochondrial targeting signal and a C-terminal PTS1 sequence we have included the Arabidopsis proteins in our analysis of the rice and Physcomitrella NDA orthologs. When GFP is placed at the C-terminus of both AtNDA1 and 2, the chimeric protein is targeted to mitochondria in both Arabidopsis cell suspension and onion epidermal cells (Figure 2) as visualized by the overlap of the GFP with the RFP signals from the mitochondrial controls. In most instances the GFP signal shows a halo-like shape around the mitochondria. This has been speculated to result when the GFP is not completely pulled into the mitochondria and has previously been observed with AtNDA and AtNDB proteins [25] and is also observed with the GFP tagging of mitochondrial outer membrane proteins [36]. However in this case, as NDA proteins are known to be located in the inner membrane, it is possible that the C-terminus of NDA proteins are located in the intermembrane space (IMS) and may produce a similar result [4]. In contrast to the full-length C-terminal fusions, truncation of both AtNDA1 and AtNDA2 to just the final 10 amino acids fused to the C-terminus of GFP (10 amino acids were chosen to be consistent with previous research on ND targeting in Arabidopsis as well to be consistent with previous work on the targeting of peroxisomal proteins which also used the last 10 amino acids), was found to target solely to the peroxisomes (Figure 2). This co-localization was observed in both tissue types tested.
Figure 2. Subcellular localizations of Arabidopsis and rice NDA type proteins. GFP was fused to the C-terminus of NDA1 and NDA2 from Arabidopsis thaliana (At) and NDA1 and NDA2 from Oryza sativa (Os). Additionally, the last 10 amino acids were fused to the C-terminus of GFP. Subcellular targeting was analyzed in both Arabidopsis cell suspension or onion epidermal tissue along with RFP tagged mitochondrial or peroxisomal controls. The protein accession number, the location of GFP and the number of amino acids used are shown for each construct. Mitochondria (M) and peroxisomes (Px) are indicated. M-RFP - mitochondrial RFP, Px-RFP - peroxisomal RFP. Scale bar indicates 10 μm.
When both NDA proteins from rice (OsNDA1 and 2) were analysed in the same manner, slightly different results were obtained. OsNDA1 was observed to target to mitochondria when GFP was fused to its C-terminus (Figure 2), however when the last 10 amino acids are fused to the C-terminus of GFP, no targeting was observed to any distinct organelle (Figure 2), consistent with the fact that OsNDA1 does not contain a predicted PTS1 signal. When the targeting of OsNDA2 was tested it was seen to target to both mitochondria and peroxisomes in both the tissues tested (Figure 2). Therefore, at least one of the rice NDA proteins displays dual targeting ability. To determine if dual targeting ability is also evident with Physcomitrella NDA1 and NDA2, the same set of experiments were performed. In both onion epidermal cells and Arabidopsis cell suspension both Physcomitrella NDA proteins (PpNDA1 and PpNDA2) displayed dual targeting ability to both mitochondria and peroxisomes (Figure 3) as with AtNDA1, AtNDA2 and OsNDA2. However, based on the bioinfomatic analysis the targeting of PpNDA1 and PpNDA2 to peroxisomes is surprising as they do not contain any PTS1 signal nor are they predicted to be peroxisomal (Figure 1). To test whether this peroxisomal targeting is an artifact, we expressed the Physcomitrella NDA fusions in Physcomitrella tissue, thereby eliminating any incorrect targeting arising from cross-species interference. Again, both PpNDA1 and PpNDA2 were dual targeted to mitochondria and peroxisomes (Figure 3), suggesting that the dual targeting of NDA type proteins arose early in land plant evolution.
Figure 3. Subcellular localizations of Physcomitrella NDA type proteins. GFP was fused to the C-terminus of NDA1 and NDA2 from Physcomitrella patens (Pp). Additionally, the last 10 amino acids were fused at the C-terminus of GFP. Subcellular targeting was analyzed in both Arabidopsis cell suspension, onion epidermal tissue and Physcomitrella protonemal tissue along with RFP tagged mitochondrial or peroxisomal controls. The protein accession number, the location of GFP and the number of amino acids used are shown for each construct. Mitochondria (M) and peroxisomes (Px) are indicated. M-RFP - mitochondrial RFP, Px-RFP - peroxisomal RFP. Scale bar indicates 10 μm.
Subcellular targeting of NDB proteins
The three Arabidopsis NDB proteins (AtNDB1, AtNDB2 and AtNDB4) have previously been determined to be mitochondrial [22,25], which was confirmed in this study (Figure 4). However, the last 10 amino acids of AtNDB1 fused to the C-terminus of GFP targeted to peroxisomes, whilst the last 10 amino acids of AtNDB2 and AtNDB4 did not target GFP to any distinct organelle and appeared cytosolic, indicative of no targeting (Figure 4). This confirms that AtNDB1 is dual targeted to both mitochondria and peroxisomes and that AtNDB2 and AtNDB4 can only be targeted to mitochondria. To determine if dual targeting of NDB proteins also occurs in rice, the two rice orthologs (OsNDB1 and OsNDB2) were also analysed. Both OsNDB1 and OsNDB2 were observed to target GFP to mitochondria and the last 10 amino acids can target GFP to peroxisomes in all the tissues tested (Figure 4). This suggests that the dual targeting of NDB proteins is conserved in rice and Arabidopsis.
Figure 4. Subcellular localizations of Arabidopsis and rice NDB type proteins. GFP was fused to the C-terminus of NDB1, NDB2 and NDB4 from Arabidopsis thaliana (At) and NDB1 and NDB2 from Oryza sativa (Os). Additionally, the last 10 amino acids were fused at the C-terminus of GFP. Subcellular targeting was analyzed in both Arabidopsis cell suspension or onion epidermal tissue along with RFP tagged mitochondrial or peroxisomal controls. The protein accession number, the location of GFP and the number of amino acids used are shown for each construct. Mitochondria (M) and peroxisomes (Px) are indicated. M-RFP - mitochondrial RFP, Px-RFP - peroxisomal RFP. Scale bar indicates 10 μm.
Unlike rice and Arabidopsis, Physcomitrella contains three NDB homologs (PpNDB1, PpNDB2 and PpNDB3) and when GFP was fused to their C-termini, GFP was targeted to the mitochondria with a distinctive halo-like fluorescence in Arabidopsis cell suspensions (Figure 5). However, when these same constructs were expressed in onion epidermal cells a very different picture emerged for PpNDB1 and PpNDB2. Not only were PpNDB1 and PpNDB2 targeted to mitochondria but they could also target GFP to plastids (Figure 5). For PpNDB3, only mitochondrial targeting was observed (Figure 5). When the last 10 amino acids of PpNDB1, PpNDB2 and PpNDB3 were fused to the C-terminus of GFP, both PpNDB1 and PpNDB2 could target GFP to peroxisomes and PpNDB3 showed targeting of GFP to the cytosol (Figure 5). The GFP assays were repeated with Physcomitrella tissue (Figure 6) and as seen in the onion epidermal cells and Arabidopsis cell suspensions both PpNDB1 and PpNDB2 were able to target GFP to mitochondria, plastids and peroxisomes (Figure 6). PpNDB3 was only observed to target GFP to mitochondria, even in a homologous system (Figure 6). The lack of peroxisomal targeting by the C-termini of PpNDB3 is not surprising, as no PTS1 sequence is predicted (Figure 1). However, the targeting of PpNDB1 and PpNDB2 to plastids was unexpected. However, they are closely related to the Chlamydomonas NDA2 protein (Figure 1), which has been demonstrated to be a solely plastidic protein [32,33]. Notably, Physcomitrella NDB proteins are also considerably longer at the N-terminus compared to Arabidopsis (Additional file 3), suggesting additional targeting information and as the ambiguous targeting predictor (ATP) predicts both PpNDB1 and PpNDB2 to be dual targeted (Figure 1 and Additional file 2).
Figure 5. Subcellular localizations of Physcomitrella NDB type proteins. GFP was fused to the C-terminus of NDB1, NDB2 and NDB4 from Physcomitrella patens (Pp). Additionally, the last 10 amino acids were fused at the C-terminus of GFP. Subcellular targeting was analyzed in both Arabidopsis cell suspension or onion epidermal tissue along with RFP tagged mitochondrial, peroxisomal or plastid controls. The protein accession number, the location of GFP and the number of amino acids used are shown for each construct. Mitochondria (M), Plastids (Pl) and peroxisomes (Px) are indicated. M-RFP - mitochondrial RFP, Px-RFP - peroxisomal RFP, Pl-RFP - platid RFP. Scale bar indicates 10 μm.
Figure 6. Subcellular localizations of Physcomitrella NDB type proteins in Physcomitrella tissue. GFP was fused to the C-terminus of NDB1, NDB2 and NDB4 from Physcomitrella patens (Pp). Additionally, the last 10 amino acids were fused at the C-terminus of GFP. Subcellular targeting was analyzed in Physcomitrella protonemal tissue along with RFP tagged mitochondrial, peroxisomal or plastid controls. The protein accession number, the location of GFP and the number of amino acids used are shown for each construct. Mitochondria (M), Plastids (Pl) and peroxisomes (Px) are indicated. M-RFP - mitochondrial RFP, Px-RFP - peroxisomal RFP, Pl-RFP - platid RFP. Scale bar indicates 10 μm.
Additional file 3. Multiple sequence alignment of the N-terminal regions of NDB proteins. Shown is a multiple sequence alignment of the N-terminal regions for the Chlamydomonas reinhardtii (Cr), Physcomitrella patens (Pp), Oryza sativa (Os) and Arabidopsis thaliana (At) NDB protein sequences. Highlighted by the red boxes are the N-terminal extensions found in Chlamydomonas and Physcomitrella sequences.
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Subcellular targeting of NDC proteins
Phylogenetic analysis showed that among the three clades made up of NDA, NDB and NDC proteins, NDC is the most ancestral group and branched out early in evolution. With regard to the targeting ability of NDC proteins, only one ortholog was found in Arabidopsis, rice and Physcomitrella. When Arabidopsis NDC1 (AtNDC1) was fused to GFP and expressed, two different patterns emerged. In Arabidopsis cell suspensions a clear plastidic localization could be observed, however, in onion epidermal cells clear targeting to both mitochondria and plastids was observed (Figure 7). As AtNDC1 was previously shown to be imported into isolated Arabidopsis mitochondria [23,25,26] it can be concluded that AtNDC1 is dual targeted to mitochondria and plastids [25,26].
Figure 7. Subcellular localizations of Physcomitrella NDC type proteins. GFP was fused to the C terminus of the NDC type proteins from Arabidopsis thaliana (At), Oryza sativa (Os) and Physcomitrella patens (Pt). Subcellular targeting was analyzed in Arabidopsis cell suspension, onion epidermal tissues and for the Physcomitrella NDC1 also in Physcomitrella protenemal tissue along with RFP tagged mitochondrial or plastid controls The protein accession number, the position of GFP and the number of amino acids used are shown for each construct. Mitochondria (M) and plastids (Pl) are indicated. M-RFP - mitochondrial RFP, Pl-RFP - plastid RFP. Scale bar indicates 10 μm.
To determine when dual targeting of NDC1 arose we analysed the targeting ability of rice (OsNDC1) and Physcomitrella (PpNDC1). For OsNDC1 we see a similar pattern as with AtNDC1 in that plastid targeting was observed in Arabidopsis cell suspensions and dual targeted to mitochondria and plastids in onion epidermal cells (Figure 7). For PpNDC1 only plastid targeting was observed in both Arabidopsis suspension and onion epidermal cells (Figure 7). GFP tagging of PpNDC1 in Physcomitrella tissue resulted in only plastid targeting (Figure 7). These results suggest that in lower plants such as Physcomitrella, NDC1 proteins are only targeted to plastids, and in higher plants such as Arabidopsis and rice NDC1 has additionally acquired targeting to mitochondria during plant evolution.
This aim of this study was to determine the targeting ability of plant ND proteins in land plants. Aquatic green algae and land plants diverged over a billion years ago [37]. Examination of genes encoding ND proteins in Chlamydomonas revealed five genes. The closest NDA ortholog between Chlamydomonas and the higher plant NDA proteins is CrNDA1 (Figure 1). CrNDA1 has been identified as an inner mitochondrial protein, with roles in the oxidation of the mitochondrial matrix NADH in the absence of complex I [34]. In this study, it was observed that both PpNDA1 and PpNDA2 are dual targeted to mitochondria and peroxisomes (Figure 3), indicating that the dual targeting of NDA arose early in land plant evolution. Although the tripeptide SRF in PpNDA1 and PpNDA2 is not predicted to direct peroxisomal import (Additional file 2), it has been experimentally validated as a plant PTS1 tripeptide [30,38]. The contradiction of targeting between predictive and experimental data may be due to the fact that for non-canonical PTS1 tripeptides targeting ability depends largely on the upstream sequence [39]. The reason of prediction inaccuracy for non-canonical PTS1 proteins is that the proteins containing canonical PTS1 tripeptides dominate the dataset, most of which lack upstream enhancing sequences for targeting [30,39]. In Angiosperm plants such as Arabidopsis and rice, dual targeting of NDA proteins is not only conserved, but peroxisomal targeting is "strengthened" by the acquisition of canonical PTS1 targeing signals. In AtNDA1, AtNDA2 and OsNDA2, all contain the PTS1 tripeptide SRI; a feature of many proteins known to target to the peroxisomes [38]. In summation, while the acquisition of dual targeting ability in land plants cannot be dated precisely, it arose early and is conserved.
The closest NDB ortholog between Chlamydomonas and the higher plant NDB proteins is CrNDA2 (we have kept the name from previous publications however it clusters with NDB family (Figure 1)). It has been demonstrated that CrNDA2 was located within the thylakoid membranes of plastids and is crucial for nonphotochemical plastoquinone reduction and associated processes in Chlamydomonas [32,33]. The plastid localisation is interesting, as Physcomitrella NDB proteins (PpNDB1 and 2) are also targeted to plastids (Figures 5 and 6), in addition to mitochondria and peroxisomes. It is speculated that the Chlamydomonas and Physcomitrella proteins are able to target plastids because they contain an N-terminal extension, in contrast to the Arabidopsis and rice NDB proteins (Additional file 3). This extension ranges from 26 amino acids in Chlamydomonas and up to 60 amino acids in PpNDB1 and PpNDB2; which may carry extra targeting signals for plastids. In regards to the peroxisomal targeting of plant NDB proteins, it can also be hypothesized that this targeting ability arose very early in plant evolution. Analysis of the C-terminal end of CrNDA2 while not predicted to be peroxisomal, it does contain the tripeptide SRV which has been demonstrated to target peroxisomes previously [38]. However previous studies only used western blots on isolated plastids and mitochondria, and the testing of peroxisomes was not performed [32,33]. It is possible that even in Chlamydomonas, NDB type proteins contained peroxisomal targeting ability. In terms of plastid targeting, this ability has been lost in the higher plants of Arabidopsis and rice, possibly due to the loss of the N-terminal extension found in the Chlamydomonas and Physcomitrella sequences. A possible reason for this loss of plastid targeting is that in higher plants, plastids contain a type I ND which functions in non-photochemical plastoquinone reduction, however, such genes are absent from the genomes of many green algae including Chlamydomonas [40]. The situation in Physcomitrella would be the intermediate form, as Physcomitrella contain both the type I and type II ND proteins. This is consistent with plants starting out with only type II ND proteins in plastids followed by the gain of type I ND proteins which would facilitate the loss of the type II ND proteins. Therefore the role of NDB like proteins in the plastids of higher plants has become redundant.
The evolutionary history for NDC type proteins appears simpler as each plant genome encodes only a single copy of the gene. However the targeting ability of these proteins across the different model plants is not the same. Currently, there is no existing data on the targeting of Chlamydomonas CrNDA5, the closest homolog to NDC proteins in higher plants. Recently, the role of AtNDC1 in plastids has been analysed. AtNDC1 was found to be involved in the reduction of a plastoquinone analog in vitro and affect the overall redox state of the total plastoquinone pool in vivo[26]. AtNDC1 was also demonstrated to be required for normal platochromal-8 accumulation and is essential for viatmin K(1) production [26]. AtNDC1 has also been found to associate with the plastoglobules within plastids [41,42]. It is tempting to speculate that NDC1 proteins perform these same functions in all plant species but as yet no functional data for the role of NDC1 protein outside of Arabidopsis exists. However the fact that a NDC1 protein is found in all species analyzed (Figure 1) suggests that it does have a required function in plants. It appears that the targeting ability of NDC1 proteins and functional data for AtNDC1 support the hypothesis that the mitochondrial targeting of NDC1 proteins is a newly acquired phenomenon. However, the exact role for NDC1 proteins within the mitochondrial inner membrane has not yet been defined.
The dual targeting of plant ND proteins to mitochondria (and chloroplasts) and peroxisomes appears to have arisen early within the evolution of plants. We cannot at this time date this accurately, as we cannot rule out the possibility of the NDA and NDB type proteins from Chlamydomonas reinhardtii being targeted to peroxisomes. Based on the results presented on the targeting of ND proteins to peroxisome from Physcomitrella, the lack of a predicted PTS1 sequence is not reliable enough to say that ND proteins are not targeted to peroxisomes. Thus ND proteins from Chlamydomonas reinhardtii may also be targeted to peroxisomes.
Various studies have been undertaken to try and determine the cellular roles of type II ND proteins in higher plants. By utilising over expression and inactivation of different ND proteins it has been demonstrated that ND proteins play roles in the capacity of NAD(P)H oxidation (nda1) [21], modulating the total leaf NADPH/NADP + ratios (NDB1 over expressor) [43], reative oxygen species formation and salinity tolerance (ndb4) [19]. The functions of ND proteins have been proposed presuming an exclusive mitochondrial localization. However, as shown in this study, and other studies [1,44,45], the targeting ability of a protein depends on the test tissue and the constructs used, and cell specificity cannot be ruled out, i.e. dual targeting may occur in specific types of cells. Previous studies analyzing the location of NDB1 in potato tubers concluded that there was no NDB1 in peroxisomes [46,47]. This difference to this study may be due to several reasons, firstly, there is a difference between targeting and accumulation, and the specialized storage tissue of potato tubers may not be an ideal tissue to study the location of an enzyme, especially if stem/shoot phenotypes are being examined. Secondly there are a number of technical problems with the methods used. The method used to purify the peroxisomes from the mitochondria [48], is claimed to produce pure organelles with only 2% or less cross- contamination. However in both cases only catalase activity is used to verify organelle purity [46,47]. Subsequent proteomic studies in yeast, mammalian and plant systems show that mitochondrial contamination is the most abundant contaminant of peroxisomes and vice versa [49]. Also Catalase is present in even highly purified mitochondria [50]. This suggests that the immunological approaches used in previous studies were not quantitative or sensitive enough [46,47].
While various roles for ND protein in mitochondria and plastids have been ascribed as outlined above, no function(s) has been described in peroxsiomes. Type II ND proteins are able to oxidize both NADH and NADPH and both are produced in peroxisomes via Isocitrate dehydrogenase, Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. Malate dehydrogenase and Hydroxyacyl-CoA dehydrogenase can also produce NADH in plant peroxisomes [51]. While other enzymes can oxidise NADH and NADPH in peroxisomes, this is also true for mitochondria and plastids, that have several systems to oxidise NADH or NAD(P)H [52,53].
The nature of the acquisition of dual targeting to peroxisomes and mitochondria of plant ND proteins is unknown, but as it only requires the modification of the terminal 3 amino acids of a protein, it can be readily envisaged that it can occur. It can also be seen that not all the PTS1 sequences are the same in the different ND proteins from different species (Figure 1). This indicates that the last three amino acids are not conserved due to functional constraints. This is supported by the crystal structure of the yeast type II ND protein Ndi1, which illustrated that the C-terminal region is not required for enzymatic activity [54,55]. Rather, the C-terminal region is required for attaching Ndi1 to the inner mitochondrial membrane. This may mean that the C-terminal region of type II NDs may be able to readily undergo mutations which does not affect the catalytic activity of the protein. But may change the function as the protein is now targeted to an additional location, where it can carry out a new function.
The dual targeting of ND proteins in plants to mitochondria and peroxisomes is interesting because it involves two distinct targeting signals at opposite ends of the proteins. Examples of this have been previously reported to occur naturally, in Arabidopsis acyl-activating enzymes which display an identical arrangement of targeting signals except the mitochondrial signal is replaced with a chloroplast signal [56], and an artificially created protein containing the chloroplast targeting signal of Small subunit unit of rubisco (SSU) attached to both RFP and GFP followed by a PTS1 signal, could in fact target to both chloroplasts and peroxisomes in vivo[57]. This suggests that both signals can be recognized in the cell environment, even though the N-terminal signal would emerge from the ribosome before the C-terminal this differs slightly from previous work performed on the targeting signals of dual targeted proteins [58,59]. However these studies were focused on dual targeting between mitochondria and chloroplasts, where the targeting signals are both located at the N-terminus. Thus regulation of import or binding factors may occur, to distribute dual targeted proteins.
In conclusion this study shows that ND proteins are targeted to multiple organelles in land plants (Figure 8). We assessed targeting from three major groups of plants, Dicots = Arabidopsis, a Moss = Physcomitrella, and a Monocot = rice, where we used Allium (onion). Thus targeting was assesses for all three species that belonged to that major plant group. This multiple targeting ability arose early in the evolution of land plants. It is important that the understanding of the location of protein(s) be taken from a wide variety of sources as possible. While models like Arabidopsis have provided much information, it should not be presumed that all plants are the same with respects to the location of orthologous proteins. Targeting signals of proteins are less constrained by specific sequence requirements than active sites of proteins, and therefore may be acquired and/or altered over time. While it is not feasible to interrogate the subcellular proteomes of a variety of plants species to the same degree as has been obtained in Arabidopsis, discounting evolutionary differences would mean that all plants should be the same, and by definition that is not the case.
Figure 8. Summary of the subcellular localizations of type II ND proteins in plants. Combining the results from this study and other previous studies of the subcellular locations in four model plant species (Chlamydomonas reinhardtii, Physcomitrella patens, Arabidopsis thaliana and Oryza sativa) of the Type II ND proteins. Question marks denote predicted possible localizations. Chloroplasts - green, Mitochondria - red and Peroxisomes - blue.
Bioinformatic analyses
Protein sequences for all Arabidopsis type II NAD(P)H dehydrogenase proteins were obtained from the TAIR website (http://www.arabidopsis.org webcite) (NDA1; At1g07180, NDA2; At2g29990. NDB1; At4g28220, NDB2; At4g05020, NDB3; At4g21490, NDB4; At2g20800, NDC1; At5g08740) and used in BlastP [60] searches against Volvox carteri, Chlamydomonas reinhardtii (Chlamydomonas), Physcomitrella patens, Selaginella moellendorffii, Oryza sativa, Zea mays, Vitis vinifera, Glycine Max, and Populus trichocarpa protein databases using the Phytozome website (http://www.phytozome.net webcite). Blastx searches were used in order to identify proteins from Picea glauca on the NCBI database (http://www.ncbi.nlm.nih.gov webcite) [60]. The known sequences for the Escherichia coli, Saccharomyces cerevisiae, and Synechocystis sp. PCC 6803 NADH dehydrogenases were downloaded from the NCBI website (http://www.ncbi.nlm.nih.gov webcite). Predictions of subcellular localizations were carried out using Predotar [61], TargetP [62] and Wolfpsort [63]. The ATP predictor [64] was also utilized to predict the probability of a given protein to be dual targeted to mitochondria and plastids. Peroxisomal targeting was predicted using the PTS1 predictor [29], PredPlantPTS1 [30] and by comparing the last 3 C-terminal amino acids with the known PTS1 sequences from the AraPerox database [31]. Phylogenetic analysis was carried out by first creating a multiple sequence alignment of all proteins using MAFFT [65]. The phylogenetic tree was calculated using MEGA 5 [66] using statistical method of neighboring-joining with the number of bootstrap replications of 1000.
Cloning and constructing of vectors
RNA extraction from Arabidopsis thaliana, Oryza sativa and Physcomitrella patens was carried out using the RNeasy kit (Qiagen, Melbourne) according to manufacturer's instructions. Reverse transcription was carried out using SuperScript™ III First-strand synthesis system (Invitrogen, Sydney). For some genes, cDNA clones were obtained from the Knowledge-based Oryza Molecular biological Encyclopedia (KOME) [67] or the RIKEN Bioresource Center [68]. cDNA or cDNA clones were used as templates to amplify full-length cDNAs or the last C-terminal 10 amino acids prior to the stop codon for cloning into N and C-terminal GFP vectors [3,25] using recombinant Gateway technology (Invitrogen, Sydney). The targeting signals of the Pisum sativum small subunit of 1,5 – ribulose bisphosphate carboxylase/oxygenase (SSU) and Cucubita sp. malate synthase were fused to RFP and used as a plastid and peroxisomal markers respectively [3,69]. For mitochondrial controls three different constructs were utilised. For Arabidopsis suspension cells the targeting signal of the Glycine max alternative oxidase (AOX) was used [3]. For onion cells the cytochrome c oxidase IV targeting signal from Saccharomyces cerevisiae fused to mCherry was utilised [70]. Finally the full length coding sequence from the Physcomitrella alternative oxidase (Pp1s183_11V6) fused to mCherry was used in Physcomitrella tissue [45].
Subcellullar localizations
Biolistic co-transformation of GFP and RFP/mCherry fusion vectors was performed on Arabidopsis cell suspensions and onion epidermal cells as previously described [3]. Briefly, 5 μg of each GFP and RFP plasmid were co-precipitated onto gold particles and bombarded onto 4-day-old Arabidopsis cell suspensions and freshly peeled onion epidermal cells using the PDS-1000/He biolistic transformation system (Bio-Rad, Sydney). For putative Physcomitrella proteins, transformation was also performed on 7-day-old protonemal tissues. Following incubation for 12–24 h at 22°C (25°C for Physcomitrella) in the dark, GFP and RFP expression was visualized at 100X magnification using a BX61 Olympus microscope (Olympus, Melbourne) with the excitation wavelengths of 460/480 nm (GFP) and 535/555 nm (RFP), and emission wavelengths of 495–540 nm (GFP) and 570–625 nm (RFP). Images were captured using CellR imaging software (Olympus, Melbourne) as previously described [3].
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
LX performed the experimental work with the assistance of SRL and MM. CC and JW oversaw the analysis, design and implementation of the study. CC and JW drafted the manuscript. All authors read and approved the final manuscript.
This work was supported by a WA state government Postgraduate scholarship to LX and the Australian Research Council Discovery Grant DP0664692 to JW. CC is supported by a Humboldt Research Fellowship for Postdoctoral Researchers from the Alexander von Humboldt Foundation.
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Mol Biol Evol 2007, 24(4):956-968. PubMed Abstract | Publisher Full Text
Ytterberg AJ, Peltier JB, van Wijk KJ: Protein profiling of plastoglobules in chloroplasts and chromoplasts, A surprising site for differential accumulation of metabolic enzymes.
Plant Physiol 2006, 140(3):984-997. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Lundquist PK, Poliakov A, Bhuiyan NH, Zybailov B, Sun Q, van Wijk KJ: The functional network of the Arabidopsis plastoglobule proteome based on quantitative proteomics and genome-wide coexpression analysis.
Plant Physiol 2012, 158(3):1172-1192. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Liu YJ, Norberg FE, Szilagyi A, De Paepe R, Akerlund HE, Rasmusson AG: The mitochondrial external NADPH dehydrogenase modulates the leaf NADPH/NADP + ratio in transgenic Nicotiana sylvestris.
Plant Cell Physiol 2008, 49(2):251-263. PubMed Abstract | Publisher Full Text
Van Aken O, Zhang B, Carrie C, Uggalla V, Paynter E, Giraud E, Whelan J: Defining the mitochondrial stress response in Arabidopsis thaliana.
Mol Plant 2009, 2(6):1310-1324. PubMed Abstract | Publisher Full Text
Xu L, Carrie C, Law SR, Murcha MW, Whelan J: Acquisition, conservation, and loss of dual-targeted proteins in land plants.
Plant Physiol 2013, 161(2):644-662. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Liu YJ, Nunes-Nesi A, Wallstrom SV, Lager I, Michalecka AM, Norberg FE, Widell S, Fredlund KM, Fernie AR, Rasmusson AG: A redox-mediated modulation of stem bolting in transgenic Nicotiana sylvestris differentially expressing the external mitochondrial NADPH dehydrogenase.
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Physiol Plant 1993, 88(1):19-28. Publisher Full Text
Eubel H, Meyer EH, Taylor NL, Bussell JD, O'Toole N, Heazlewood JL, Castleden I, Small ID, Smith SM, Millar AH: Novel proteins, putative membrane transporters, and an integrated metabolic network are revealed by quantitative proteomic analysis of Arabidopsis cell culture peroxisomes.
Plant Physiol 2008, 148(4):1809-1829. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Eubel H, Lee CP, Kuo J, Meyer EH, Taylor NL, Millar AH: Free-flow electrophoresis for purification of plant mitochondria by surface charge.
Plant J 2007, 52(3):583-594. PubMed Abstract | Publisher Full Text
Igamberdiev AU, Lea PJ: The role of peroxisomes in the integration of metabolism and evolutionary diversity of photosynthetic organisms.
Phytochemistry 2002, 60(7):651-674. PubMed Abstract | Publisher Full Text
Rasmusson AG, Geisler DA, Moller IM: The multiplicity of dehydrogenases in the electron transport chain of plant mitochondria.
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Livingston AK, Cruz JA, Kohzuma K, Dhingra A, Kramer DM: An Arabidopsis mutant with high cyclic electron flow around photosystem I (hcef) involving the NADPH dehydrogenase complex.
Plant Cell 2010, 22(1):221-233. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Feng Y, Li W, Li J, Wang J, Ge J, Xu D, Liu Y, Wu K, Zeng Q, Wu JW, et al.: Structural insight into the type-II mitochondrial NADH dehydrogenases.
Nature 2012, 491(7424):478-482. PubMed Abstract | Publisher Full Text
Iwata M, Lee Y, Yamashita T, Yagi T, Iwata S, Cameron AD, Maher MJ: The structure of the yeast NADH dehydrogenase (Ndi1) reveals overlapping binding sites for water- and lipid-soluble substrates.
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Hooks KB, Turner JE, Graham IA, Runions J, Hooks MA: GFP-tagging of Arabidopsis acyl-activating enzymes raises the issue of peroxisome-chloroplast import competition versus dual localization.
J Plant Physiol 2012, 169(16):1631-1638. PubMed Abstract | Publisher Full Text
Sommer M, Rudolf M, Tillmann B, Tripp J, Sommer MS, Schleiff E: Toc33 and Toc64-III cooperate in precursor protein import into the chloroplasts of Arabidopsis thaliana.
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J Mol Biol 2009, 393(4):803-814. PubMed Abstract | Publisher Full Text
Chew O, Rudhe C, Glaser E, Whelan J: Characterization of the targeting signal of dual-targeted pea glutathione reductase.
Plant Mol Biol 2003, 53(3):341-356. PubMed Abstract
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ: Basic local alignment search tool.
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Small I, Peeters N, Legeai F, Lurin C: Predotar: A tool for rapidly screening proteomes for N-terminal targeting sequences.
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Emanuelsson O, Nielsen H, Brunak S, von Heijne G: Predicting subcellular localization of proteins based on their N-terminal amino acid sequence.
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Years ago, the East Tokyo United soccer team was a force to be reckoned with, captained by the talented and charismatic Takeshi Tatsumi. But after leaving suddenly to join an amateur team in England, ETU's record and morale quickly plummeted, their meager status kept afloat only by the efforts of new captain Murakoshi. Now, several years have passed and Takeshi is in town once more, this time to bring the team back to victory. But with fans resentful of the man's former departure, clashing personalities on the team and Takeshi's unorthodox and lackadaisical coaching style, ETU's rise to fame won't be easy...
4 votes
3 votes
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Giant Killing was a show that I had high hopes for that in the end just couldn't deliver. The premise of the show managed to be both worn out and new at the same time. You have a lack luster sports team that is struggling to stay afloat and a new coach comes with lofty goals and a plan to turn things around through strategy. I've seen this played out in western media a lot but this is the... read more
I'd like to say its a typical sports anime but it is in some ways and it isn't...The general plotline is typical. A weak soccer team meets an eccentric but talented coach and soon, they begin winning games and fulfilling their potential. Nothing crazy and unexpected here.The animation wasn't the typical type. The art style was different, which is, dare I say, refreshing. Nothing special... read more
|Giant Killing
|Original Manga
Character Design
Original Manga Creator


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(l-r); Camden residents Mangaliso Davis (left) and Eulisis Delgado protest signs to express their displeasure of the appointment State Superintendent Paymon Rouhanifard, prior to the Camden Board of Education meeting Tuesday held at the Preston O.R. Toliver II Broadcast Center in Camden.
- Filed Under
The new head of Camden's school district was a no-show at a city board of education meeting Tuesday.
The absence of State Superintendent Paymon Rouhanifard upset some in the crowd of about 80 residents and teachers who attended the meeting to express displeasure at his appointment by the state board of education Monday.
Isabelle Ross of Camden called the hiring a "slap in the face." She wanted Camden residents to have a say in the decision to hire Rouhanifard. ...


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Gainful Employment Disclosures
The US Department of Education requires colleges to disclose information for financial aid eligible programs that prepare students for gainful employment in a recognized occupation. At the University of Alaska Anchorage (UAA), these programs include undergraduate certificates, occupational endorsement certificates, graduate and post-baccalaureate certificates. UAA is pleased to provide information including related employment occupations, program costs, graduation rates, and where applicable, median loan debt and job placement rates.
Occupational Endorsement Certificate
- GE PWSCC OEC Direct Services Specialist (261.9 KiB)
Undergraduate Certificates
- GE PWSCC UC Computer Information Office Systems (262.4 KiB)
- GE PWSCC UC Industrial Technology (261.1 KiB)


text:
[Edited by crsojo, 7/29/2010 9:56:14 PM]
Read the trainer instructions, this is what the Numpad 3 option does.
"Numpad 3: Troops - PLAYER ONLY - press key to have unlimited Troops."
i thought thats what he was referring too, guess old age is getting to him, that or he hasnt slept since the game was released
Thanks in advance.
Solution ? anyone?


text:
Wine »
This section is for wine and champagne, the symbol of elegance among the drinking world. Here you will find articles for wine reviews and recommendations along with advice, news, vineyard visits and more! Anything and everything to do with wine! Grab the RSS Feed Here
A Taste of Colorado's Wine Country
Perfectly straight row after row of greenery follows you around every bend in the road, an orchard on the left, a vineyard on the right, the occasional farm house in between. If you travelRead More »
Wine Enthusiasts Top 100 Wine Restaurants of 2013
Wine Enthusiasts has sent out experts and sommeliers to rate and review the best wine and wine restaurants from around the country. For their release of the Top 100 Wine Restaurants of 2013, theyRead More »
Q and A with Magnifico Giornata CEO and founder Bob Manfredonia
Italian for "magnificent day", Magnifico Giornata is a line of flavor-infused sparkling champagnes. Created by Bob Manfredonia and Ralph Giannella, Magnifico Giornata was the result of a mutual desire to create something completely unique.Read More »
Club W – Great Wines Delivered Direct
Zane Lamprey has joined Club W as a special guest curator for their wine club program. Zane fans that head on over to http://clubw.com/zane can receive a FREE bottle of wine with their newRead More »
How to Enjoy a Chianti (Without the Hannibal Lecter References)
Thirty or forty years ago, Chianti was famous only for being a classic Italian red from the central region of Tuscany. With a newly-classified D.O.C.G. Appellation, Chianti began to reinvent itself in the 1980sRead More »
Wine and Cheese Pairings for National Wine Day
According to Maxine Borcherding, Certified Sommelier and Wine Educator at Oregon Culinary Institute, a principle of cheese and wine pairing is that wine can either complement cheese flavors or go in the other directionRead More »
Wooden Wine Holders Shaped Like Animals
Displaying your wine can almost be as good as drinking it. Well, not really but still, showing off your wine is part of the experience and it is important to use something that matchesRead More »
Don't Break The Bottle Puzzle – Wine Gift
Now why in the world would you want to make it harder to get to your wine? Perhaps if you are a cruel friend who delights in torturing your friends, then you might enjoyRead More »
Unspillable Wine Glass
Sometimes it seems that wine glasses were designed to tip over. Like some elaborate scheme to stain your rug or clothes. Well, these wine drinkers have had enough. The Saturn Wine Glass by Christopher YamaneRead More »
Win a Trip to A Top Wine Destination
Wine Enthusiast is holding a contest to send a few lucky individuals to some of the top wine destinations in the world. Learn more about the contest below or visit their website for moreRead More »


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Keep Morristown Hamblen Beautiful will be hosting the Annual Sanitation Workers Appreciation Breakfast on April 19 at 7 a.m., at the Hamblen County Highway Department.
Sanitation employees in the city and county work long, hard hours to ensure trash is collected, brush and miscellaneous items are kept off the curbs and roads are safe for travel. They also do little "extras" that sometimes go unnoticed.
KMHB would like to recognize sanitation workers who have gone over and above their job responsibilities and requirements in the community. Anyone wishing to submit the name of a city or county sanitation worker, contact KMHB at 423-586-6422. Submissions can be emailed to email@example.com or faxed to 423-586-6576. Deadline is April 5. Include a brief summary of why this person should be nominated.


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100g/3½oz caster sugar
50ml/2fl oz water
3 nectarines, peeled, stones removed and chopped
½ a ready-made Madeira cake, sliced
75ml/3fl oz sweet dessert wine
100g/3½oz mascarpone cheese
60g/2oz icing sugar, sieved
225ml/8fl oz double cream, whipped until stiff peaks form when the whisk is removed
1 nectarine, skin left on, stone removed, chopped
Heat the caster sugar and water in a pan for 4-5 minutes, until the sugar has dissolved and the liquid forms a syrup.
Transfer to a food processor and allow to cool slightly.
Add the chopped peeled nectarines, then blend to a puree. Allow to cool.
Place 1-2 slices of Madeira cake into the bottom of each of four serving dishes or bowls and drizzle some of the wine over each. You should use about half of the wine.
Place the mascarpone, icing sugar, the remaining wine and whipped cream into a clean bowl and fold together until smooth.
Add the nectarine puree and gently stir through the mascarpone mixture to create a marbled effect.
Add two thirds of the pieces of unpeeled nectarine and fold through the mixture.
To serve, spoon the mixture over the wine-soaked sponge in each bowl and top with a few pieces of the remaining nectarine.
Type the ingredients you want to use, then click Go. For better results you can use quotation marks around phrases (e.g. "chicken breast"). Alternatively you can search by chef, programme, cuisine, diet, or dish (e.g. Lasagne).


text: US Air Force developing 'smarter' drones Technology 26 July 10 by Spencer Ackerman Bryan William Jones Unmanned aircraft, for all their utility, are fairly simple beasts. They're good at taking direction, but they're not so good at processing information on their own. Now the Air Force figures it's time for drones to get a lot smarter, especially as they take off or land.As anyone who's ever flown knows, the runway is a crowded place. Planes on the runway queue up to get airborne. Planes in the air have to coordinate with Air Traffic Control for the order in which they can safely land, taking precautions not to get in anyone's way until it's their turn. There's a fair amount of information to rapidly process in order to avoid collisions and other accidents. Pilots can handle that information load. Drones can't. Yet. It's one of the big reasons why the Federal Aviation Administration has been so reluctant to allow unmanned aircraft to fly over the US. Even robotic flights over relatively unpopulated areas along the southern border have been cancelled when there's the most routine technical hiccups.On Tuesday, the Air Force Research Laboratory at Ohio's Wright-Patterson Air Force Base said it'll soon solicit engineers to design an algorithm to allow drones to "integrate seamlessly" with piloted planes for takeoff and landing. In the algorithm-driven future that the labs want to build, drones will be equipped with a database of terminal procedures; link up with Air Traffic Control; and "recognise the intent of other aircraft."For instance: aircraft landing on parallel runways can appear to be on a collision course before they turn and land. Right now, a drone would simply perceive that a plane's trajectory is going to remain unchanged, making it a threat for collision. But a capable algorithm would let the drone process Air Traffic Control information like basic airfield maps to know that there's no actual danger from the oncoming piloted plane."The developed algorithm(s), optimally, would require no more a priori information than a human pilot," the labs instruct. "Intent analysis should be accurate, reliable and real-time, enabling quick and appropriate decisions that are necessary in this time critical environment."There's a clear commercial application here. As we mentioned on last week, FedEx is starting to think about an airfleet of linked-up drones that can fly in formation at the direction of a piloted aircraft. Building algorithms that can let drones process complex information in congested airspace sounds like a useful step toward that futuristic cargo fleet. Edited by Olivia Solon Latest on wired.co.uk


text:
25 U.S. Code § 4237 - Remedies for noncompliance
prev | next
(a) Actions by Secretary affecting grant amounts
(1) In general
Except as provided in subsection (b) of this section, if the Secretary finds after reasonable notice and opportunity for a hearing that the Department of Hawaiian Home Lands has failed to comply substantially with any provision of this subchapter, the Secretary shall—
(B) reduce payments under this subchapter to the Department by an amount equal to the amount of such payments that were not expended in accordance with this subchapter; or
(2) Actions
If the Secretary takes an action under subparagraph (A), (B), or (C) of paragraph (1), the Secretary shall continue that action until the Secretary determines that the failure by the Department to comply with the provision has been remedied by the Department and the Department is in compliance with that provision.
(b) Noncompliance because of a technical incapacity
The Secretary may provide technical assistance for the Department, either directly or indirectly, that is designed to increase the capability and capacity of the Director of the Department to administer assistance provided under this subchapter in compliance with the requirements under this subchapter if the Secretary makes a finding under subsection (a) of this section, but determines that the failure of the Department to comply substantially with the provisions of this subchapter—
(c) Referral for civil action
(1) Authority
In lieu of, or in addition to, any action that the Secretary may take under subsection (a) of this section, if the Secretary has reason to believe that the Department of Hawaiian Home Lands has failed to comply substantially with any provision of this subchapter, the Secretary may refer the matter to the Attorney General of the United States with a recommendation that an appropriate civil action be instituted.
(2) Civil action
Upon receiving a referral under paragraph (1), the Attorney General may bring a civil action in any United States district court of appropriate jurisdiction for such relief as may be appropriate, including an action—
(A) to recover the amount of the assistance furnished under this subchapter that was not expended in accordance with this subchapter; or
(d) Review
(1) In general
If the Director receives notice under subsection (a) of this section of the termination, reduction, or limitation of payments under this chapter, the Director—
(A) may, not later than 60 days after receiving such notice, file with the United States Court of Appeals for the Ninth Circuit, or in the United States Court of Appeals for the District of Columbia, a petition for review of the action of the Secretary; and
(2) Procedure
(A) In general
(3) Disposition
(A) Court proceedings
(i) Jurisdiction of court The court shall have jurisdiction to affirm or modify the action of the Secretary or to set the action aside in whole or in part.
(ii) Findings of fact If supported by substantial evidence on the record considered as a whole, the findings of fact by the Secretary shall be conclusive.
(B) Secretary
(i) In general The Secretary, by reason of the additional evidence referred to in subparagraph (A) and filed with the court—
(4) Finality
(A) In general
Except as provided in subparagraph (B), upon the filing of the record under this subsection with the court—
Source(Pub. L. 104–330, title VIII, § 818, as added Pub. L. 106–568, title II, § 203,Dec. 27, 2000, 114 Stat. 2891, and Pub. L. 106–569, title V, § 513,Dec. 27, 2000, 114 Stat. 2985.)
References in Text
This chapter, referred to in subsec. (d)(1), was in the original "this Act", meaning Pub. L. 104–330, Oct. 26, 1996, 110 Stat. 4016, known as the Native American Housing Assistance and Self-Determination Act of 1996. For complete classification of this Act to the Code, see Short Title note set out under section 4101 of this title and Tables.
Pub. L. 106–568, § 203, andPub. L. 106–569, § 513, enacted identical sections 818 ofPub. L. 104–330. This section is based on the text of section 818 ofPub. L. 104–330, as added by Pub. L. 106–569, § 513.
LII has no control over and does not endorse any external Internet site that contains links to or references LII.


text:
Stefan Evans wrote:what "same" code do you write in scriptlet?
If its not giving the same result, it can't be the same :-)
Bear Bibeault wrote:The EL for <%= userBean.getName()%> would be ${userBeanString.name}.
The EL and JSTL have been around for about 12 years. Is it not time to start using them?


text:
Latest News and Events
29 November 2013
Grosjean and Maldonado to drive for Lotus F1 Team in 2014
Lotus F1 Team is pleased to announce that Venezuelan driver Pastor Maldonado [28] will join the Enstone squad alongside Frenchman Romain Grosjean [27] for the 2014 Formula 1 seasonREAD MORE
28 November 2013
Lotus Enthusiast Racers Take to Track in Support of the 60th Anniversary Macau Grand Prix
30 Lotus Racers competed on Macau's Guia Street Circuit, where for one lucky family, it was a double podium winREAD MORE
27 November 2013
Forging Forces for Vital Norwich Foodbank
Lotus Cars and Waitrose Supermarkets collaborate across the local region in support of the Norwich FoodbankREAD MORE
25 November 2013
Multi-Million Pound Boost for Lotus
Lotus has secured a £10 million government grant which will see the firm create more than 300 new jobs.READ MORE
24 November 2013
Out with a bang in Brazil
Bad luck in São Paulo sees Romain Grosjean retire after only 4 laps, whilst guest driver Heikki Kovalainen struggled for pace, finishing in P14. It was a disappointing end to the 2013 F1® season.READ MORE
21 November 2013
Lotus Gets Supercharged for Guangzhou Motorshow
Lotus showcases the jewels from its product line-up at the 2013 Guangzhou International Automobile Exhibition, further asserting its presence in the Chinese market as its popularity grows amongst China's fashion-conscious elite.READ MORE
20 November 2013
Lotus Showcases Road, Racing and Engineering Wares at Los Angeles International Auto Show
Fresh off a very strong performance by the Lotus F1 Team at the United States Grand Prix in Austin, Texas last weekend, Lotus races into Los Angeles to demonstrate the full breadth of its automotive engineering competencies.READ MORE
19 November 2013
Brazilian Grand Prix 2013 - Preview
Heikki & Romain share their thoughts on the Interlagos CircuitREAD MORE
18 November 2013
YeeHahh! The Lotus F1® Team displays top flight form and settles the beef in Texas
Romain Grosjean delivers a hard-nosed drive and celebrates with a P2 finish and his 5th consecutive top spot result whilst Heikki Kovalainen battles and takes the flag in P15READ MORE
14 November 2013
Heikki Kovalainen to drive for Lotus F1 Team
Finnish driver Heikki Kovalainen [32] will join Lotus F1 Team for the final two races of the 2013 season as a stand-in for regular race driver Kimi Räikkönen, who is currently undergoing back surgery.READ MORE
12 November 2013
Lucas Auer will race with Lotus Praga LMP2 at the 6 Hours of Bahrain
The 19-year-old Austrian finished 4th overall in the 2013 FIA F3 European Championship.READ MORE
11 November 2013
Looking ahead to the United States GP 2013
Grosjean, Boullier & Permane consider the team's prospects at the Circuit of the AmericasREAD MORE
11 November 2013
Räikkönen to miss final two races of 2013 for back surgery
Lotus F1 Team have confirmed that Kimi Räikkönen will miss the final two rounds of 2013 in the USA and Brazil after electing to undergo back surgery.READ MORE
11 November 2013
Hancock and Claessens club together for Lotus 6 Hour win
Sam Hancock and Michael Claessens' Track Club 3 team won the Lotus 6 Hour race at Brands Hatch on Saturday 9 November, having led the majority of the distance from pole position.READ MORE
7 November 2013
Lotus Motorsport announces latest addition to its racing portfolio; The Lotus Elise S Cup R
Following the success of the Exige V6 Cup and its hardcore sibling the Exige V6 Cup R, the Lotus Elise S is adorned with the Lotus Motorsport magic to present the track-only Elise S Cup RREAD MORE
3 November 2013
Abu Dhabi Drama
Kimi is forced to retire in Lap 1, and Romain narrowly misses a podium finish. Its bad luck all round in the Middle East.READ MORE
30 October 2013
Lotus Cars at Auto Zürich 2013
Demonstrating Lotus' continued commitment to the exclusive Swiss market, highlights of the stunning Lotus range will proudly be on display between 31st October and 3rd November in hall 2, stand B05 of the Messe Zürich for Auto Zürich 2013READ MORE
30 October 2013
Kimi & Romain look forward to the Abu Dhabi Grand Prix 2013
After a challenging Indian Grand Prix, our Finn heads to the scene of his first race victory for Lotus F1 Team fired up for a strong result, while Romain Grosjean heads to Abu Dhabi with four podiums in-a-row very much the target.READ MORE
29 October 2013
Eric Boullier: "We'll give some headaches to the other teams"
It was another strong result for the team in India – albeit with a little drama along the way – leaving Team Principal Eric Boullier buoyed by the positives heading to Abu DhabiREAD MORE
28 October 2013
Deacon and Napier are crowned Lotus Cup UK Champions as Kershaw wins in the dark
Simon Deacon and Andy Napier claimed the SuperSport and Production championships respectively in an incident-packed final round of the 2013 Lotus Cup UK season at Snetterton yesterday (Sunday 27 October)READ MORE
27 October 2013
Colourful and Chaotic, but a Trophy none-the-less
Lotus F1® Team driver Romain Grosjean delivers yet another ace-wielding drive to celebrate a P3 finish, whilst Kimi's hopes of a top spot triumph are cut short due to tyre degradation in the final laps of an enthralling 2013 Indian GPREAD MORE


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Free Commit to B-Fit Health Fair at Binghamton University
Maybe you're like me and you told yourself that this was the year you were going to get into shape, but then life happened and things didn't go exactly as planned, right? Well guess what? The year is still young and there's still time to get your health where it needs to be.