text:
A spat over growth and density in 1966 led to new neighbors, a childhood friendship and a legacy
"How do you look for the good? Because there will be good."
Neighbors shared what they knew about a developer's plans and issued a call to action filled with exclamation points, eloquent capitalization and trepidation.
It could have been yet another salvo in any number of current neighborhood debates in our fast-changing city. For instance, the furor that greeted plans to put restaurants, shops, offices and hundreds of homes — 150 of them designated affordable — where the Colorado Department of Transportation once had its headquarters in Virginia Village. The City Council approved the proposal for the CDOT plot after a tense meeting at which one area resident said the project would mean more crime. He added, referring to the expected newcomers: "We're gonna have to live with them and we're all gonna resent them forever."
A nearly lifelong Virginia Village resident who attended that meeting was no fan of the development plan. But she was troubled by the rancor. Florence Sebern found herself thinking about change in Virginia Village when she was young and messages were tapped out on a typewriter, copied and delivered door-to-door instead of posted on Nextdoor. Back in 1966, a spat over growth and density led to a new neighbor who was an internationally renowned literary figure; a childhood friendship; and a bond for Sebern's mother that, while brief, left a legacy.
"How do you look for the good?" Sebern wanted to tell her present-day neighbors. "Because there will be good."
"We're always better when we're looking for the good."
Florence Sebern in her Virginia Village home. (Donna Bryson/Denverite)
A 1966 flyer gets off to a bold-font start.
"Attention Virginia Village Owners!! This 'House' Affects YOU!!"A picture showed a house perched on a steeply sloping lot. The Urbishes were among three couples listed as contacts. Sebern's mother, Mary Rita Urbish, died in 2014 at the age of 87. Her father, Ben Urbish, died two years later at 84. The couple was married 55 years.
In 1992, Sebern and her own young family moved in next door to the house where she grew up, her presence later allowing her parents to die in the three-bedroom home they'd bought for $12,500 when they married. Sebern found the flyer among her mother's papers. Sebern's sister, Celeste Orr, said their mother "never threw anything away. Literally everything was important to her."
The two sisters sat in a living room Sebern had decorated in warm colors. Across the street was a modest house just like hers and her parents' that a developer had owned and rented out when they were little girls. In 1966, the developer had a second home hauled to the corner lot with plans to rent it as well. That second house, which judging by the flyer photo had had a rough journey, was the offending structure. It was described in the flyer as smaller than the local covenants allowed and sitting on an undersized lot.
Celeste Orr looks through some of her mother's papers. (Donna Bryson/Denverite)
The Urbishes helped organize a lawsuit challenging the property owner's right to erect what we might today call an accessory dwelling unit. Neighbors were asked to donate to help cover legal costs and show their support by attending a Feb. 11, 1966 hearing at the City and County Building.
"The question is do we, as present homeowners, CARE ENOUGH TO SEE THAT THESE RESTRICTIVE COVENANTS REMAIN IN FORCE?" the flyer asked, the word "remain" not just capitalized but underlined.
"Apathy on our part can result in the setting of a precedent, which could allow even more undesirable violations in future years."
The suit failed to get the little house removed. It and its neighbor are still there, though no longer owned by the developer. The smaller house, surrounded now by towering spruces, sits at an angle to other homes lined up like clapboard soldiers on either side of the street.
The legal action did force the developer to move the little house further from the street to comply with setback rules. According to neighborhood lore, Sebern said, the angry developer spread word that he would have his revenge by finding tenants who would "disturb the peace."
Here's where the literary figure comes in.
The late black South African writer Es'kia Mphahlele is best known for "Down Second Avenue," which is both lyrical memoirs and passionate indictment of apartheid in his homeland. He came to the University of Denver in 1966 to pursue a doctorate in creative writing. It was Mphahlele, his wife Rebecca — who would earn a DU master's degree in social work and for a time was a caseworker for teenage mothers in Denver — and their children who rented the bigger of the developer's two houses opposite the Urbishes.
Denver at the time was segregated by practice if not by law. Sebern only heard years later that the developer may have believed the Mphahleles' race would irritate residents of a neighborhood that then, as it is now, was predominantly white.
Sebern's mother had discovered on a visit for a relative's wedding that Colorado's climate eased her asthma. At 20, then Mary Rita Barclay left rural Illinois with her younger sister to move to Denver. They traveled by train and hit Jim Crow at St. Louis. Sebern grew up on stories of her mother deliberately drinking from a fountain labeled "colored" and of her conviction that racism was unjust. Once in Denver, the Barclay sisters lived with relatives and found jobs. Mary Rita Barclay met Ben Urbish at a petroleum company where she did office work and he was an accountant.
Years later in Denver, Mary Rita Urbish welcomed the Mphahleles. Little Florence was just happy that new playmates were moving nearby. Sebern said she was oblivious to the tension over the house or any racial subtext. As the eldest of three girls, Sebern said she also was thrilled that the new neighbor kids were boys.
"Even out of strife, even out of uncertainty, we can get back to a new normal," Sebern said.
Es'kia and Rebecca Mphahlele had five children, three of whom were teens or young adults when the family moved to Denver. Sebern remembers only the youngest two, Dichaba, known as Chabi, who was about seven years old when she met him, and his brother Puso, who was about five.
An Urbish family 8 mm home movie, now transferred to digital, show Chabi, Puso, Sebern and her middle sister Mary Carol — Orr, just one year old at the time, is not in the shots. The four on camera were handling salamanders from nearby Cherry Creek with a childlike mixture of curiosity and mischief.
In another home movie, Chabi and Puso are among a gaggle of neighborhood kids at Sebern's sixth birthday party. In a community newspaper clipping among all the other documents that Mary Rita Urbish never threw away, Chabi, misspelled "Chubby," is just one in a list of guests — Teresa, Mary Eileen, Rickie — at the pow-wow themed party.
"The afternoon's activities ended with a "council fire" with her friends helping Florence blow out the birthday candles."
Sebern said: "We were friends. We were neighbors. We genuinely enjoyed each other."
Chabi Mphahlele, who grew up to be a musician, died in South Africa in 2015. Attempts to reach Puso Mphahlele were unsuccessful.
Florence Sebern views a home movie on her computer. (Donna Bryson/Denverite)
Es'kia Mphahlele doesn't figure much in the movies or Florence's memories.
It's unlikely the writer had time for neighborhood socializing. He had come to Denver intent on finishing his doctoral thesis in creative writing in two years, a goal he accomplished while also penning poetry, teaching, advising master's students and delivering public lectures.
The writer's thesis was the first draft of his novel "The Wanderers." The main character of that 1971 book, Timi Tabane, was, like Es'kia Mphahlele, an artist and teacher who longed for home over the decades politics forced him to travel the world. While written in Denver, the novel is set in the period and places before the Mphahleles arrived here.
Es'kia Mphahlele was banned from teaching because he opposed South Africa's Bantu Education Act of 1953, which relegated the country's black majority to separate and inferior education. He became an editor and writer for Johannesburg's famed "Drum" magazine. He covered such historic events as a mass women's march in 1956 against laws restricting the freedom of movement of black South Africans. In "Down Second Avenue," published in 1959, Es'kia Mphahlele said he wanted to write literature, not journalism, and feared for his children's future. In 1957, he wrote, "I was suddenly seized by a desire to leave South Africa for more sky to soar. I had been banned from teaching, and conditions were crushing me and I was shriveling in the acid of my bitterness; I was suffocating."
In exile, Es'kia Mphahlele did soar as a writer, teacher and cultural center administrator. But he also found exile took an emotional toll. He never seemed able to put down roots. Before heading to DU he had lived most recently in Kenya and earlier in France, Ghana, Senegal, Sierra Leone and Uganda and been a visiting professor at MIT. DU, courting him for the luster he could add to its literature program, granted him a fellowship and waived tuition.
"In Denver I had been as happy as one can be living in exile,"Es'kia Mphahlele later told his biographer N. Chabani Manganyi.
"I was in my element: attending courses in creative writing and working once more on a full-scale narrative that partially fulfilled my longstanding desire for imaginative writing. A creative slump had settled in during the years in Kenya, and in Denver I could make up for lost time."
Denver itself was no muse. Es'kia Mphahlele found the Rockies "dreary."
"I used to look at them and the snow in winter and wonder why so many Americans could feel ecstatic about them," he told Manganyi.
Es'kia Mphahlele had a reputation for being opinionated — particularly about what should and shouldn't be considered literature — and sometimes prickly and prone to provocative statements. Writer Gunnar Boklund, in a conversation Manganyi transcribes in his 1983 biography "Exiles and Homecomings," describes himself and other colleagues at DU at the time taking the opportunity of the thesis defense to give Es'kia Mphahlele some of his own medicine. Though they never doubted they would grant his doctorate, they questioned him sharply. Left out of their joke was the outside department representative, distinguished diplomat Josef Korbel. Korbel, father of Madeleine Albright, was the founder and first dean of DU's Graduate School of International Studies, which was renamed the Josef Korbel School of International Studies in 2008. Korbel died in 1977.
"We always asked the victim to depart while we discussed," Boklund recalled. "Then Korbel whispered: 'I must say I am shocked at the way you treated him. Don't you know that this man is a dangerous revolutionary?' "
Es'kia Mphahlele, who said he generally tried to keep his political opinions to himself at a time when Vietnam and civil rights were commanding U.S. headlines, expressed surprise that Korbel would have made such an assessment.
The writer's wife, meanwhile, strode into one of the most contentious issues of a contentious period.
In 1967 Colorado adopted the first law in the country liberalizing abortion regulations, six years before the United States Supreme Court found in Roe v. Wade that women have a constitutional right to have an abortion. Sebern said her mother was following the issue before the law changed and wanted to know more, but none of her neighbors were as interested. None except Rebecca Mphahlele, who suggested she and Mary Rita Urbish go the Capitol to listen to the legislators debate.
The front page of The Denver Post on March 10, 1967 described more than 150 people — "at least half of them women" — packed into a room on the third floor of the Capitol for a hearing of the House Committee on Health, Welfare and Institutions for a hearing that lasted three and a half hours.
Mary Rita Urbish, whose formal education had ended at high school, went on to be a leader in the pro-life movement. She helped found the group Colorado Right to Life a few months after that trip to the State House with Rebecca Mphahlele, who died in 2004. In 1988 Mary Rita Urbish was the lead plaintiff in a case that went to the Supreme Court of Colorado challenging rules that allowed public funds to be used to pay for abortions. The court ruled that it was constitutional to allow taxpayer money to be used to pay for abortions when medical conditions seriously threatened the life of a pregnant woman if her pregnancy came to term, but that it was unconstitutional to pay with public money when the issue was a potentially fatal medical condition of the unborn child. Mary Rita Urbish also was active in the state GOP and once spoke at the annual March for Life in Washington. Sebern, an office manager, also has been active in the right-to-life movement and politics.
Sebern said Rebecca Mphahlele gave her mother courage. When her mother told the story of the legislative hearing, Sebern said, "her emphasis was on Rebecca's willingness to go with her and willingness to help her understand a process that she didn't understand."
Once Es'kia Mphahlele had his PhD, he left with his family for a teaching post in Zambia. The Urbish home movies include a clip of Virginia Village neighbors seeing the Mphahleles off at the airport. The lighting is poor, cinematographically speaking, but the hugs and smiles are warm. The Mphahleles are weighed down with more hand luggage than we'd ever get away with today.
The next year, Es'kia Mphahlele was nominated for a Nobel literature prize.
After a short stint in Zambia, Es'kia Mphahlele returned to DU as an associate professor, this time buying a home in Belcaro. He told his biographer he encountered racism when he tried to buy and had to turn to a civil rights group for help. He moved to the University of Pennsylvania as a full professor in 1974.
Mphahlele, who once told a researcher that "an African cares very much where he dies and is buried," returned to South Africa with his family in 1977. Tensions with the apartheid government did not disappear, but he eventually took a post at Johannesburg's University of the Witwatersrand. He was the first black professor there and taught in an African literature department he created. He wrote two more novels and a second volume of memoirs, and lived to see apartheid end in 1994. He died in 2008; this December will be the centenary of his birth.
Sebern did not see the Mphahlele children during their second stay in Denver. She does not believe her mother ever reconnected with Rebecca Mphahlele. But her mother followed the Mphahleles' story. Mary Rita Urbish once told her daughter: "You know that they eventually went back and did great work."
Sebern said that 10 or 15 years ago she became curious about Es'kia Mphahlele and did some research on him on the internet. She has her mother's copy of "Down Second Avenue." It is dated 1971. Mary Rita Urbish must have found herself with questions about the author after he and his family left Virginia Village.
A quote from Eleanor Roosevelt was printed on a bookmark her mother had tucked inside her copy of Es'kia Mphahlele's best known work: "When you have decided what you believe, what you feel must be done, have the courage to stand alone and be counted."


Truth, Racial Healing & Transformation - Los Angeles (TRHT-LA) presents an interpretive Tongva history walk through Downtown Los Angeles. The walk attempts to re-envision the landscape as Yaangna - the principal ancestral village for Tongva and Gabrieleno communities prior to contact with European settlers and missionaries in the late 1700s. The walk offers a glimpse into indigenous worldviews and will highlight our current relationships with water and nature, public spaces, oral histories, land, and ancestry. Places to be visited include the Yaangna Plaque, Placita Olvera, Union Station, and other unacknowledged sites. The goal of the walk is to give voice to suppressed histories and inspire new futures for our city and communities.
Walk guided by Craig Torres, Cindi Alvitre, Julia Bogany and Desiree Martinez - local Tongva educators, historians, and cultural workers.
*The meeting place for the walk is in front of the Junipero Serra statue. The statue is located steps away from the Veteran War Flag Memorial at El Pueblo de Los Angeles.
Comfortable walking shoes and plenty of water are recommended.	- Capacity for this walk is 25 people. You must rsvp to attend.
In the event of rain, this walk will be rescheduled for a later date.
Perspectives shared on this walk represent particular worldviews and are not intended to be taken as definitive or comprehensive.


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Indication : This medication is an antiviral used to treat patients who have had flu symptoms, caused by the influenza virus, for less than 2 days. Tamiflu (oseltamivir) may also be used to prevent influenza in patients who has not yet had the symptoms but may have been exposed to the virus. This medication does not treat common colds.
Storage : This medication should be stored at room temperature. Keep the medication away from heat and moisture. Tamiflu liquid formula should be stored in the refrigerator. Do not freeze. Dispose unused liquid after 10 days.
Precaution : Before using this medicine, tell your health care providers about any of the following: if you are pregnant, planning to become pregnant, or breastfeeding; if you have had the nasal flu vaccine (FluMist) within the past two weeks; kidney, heart, or lung disease; a condition causing swelling or disorder of the brain; or any other serious disease or health problem.
Warnings : Do not use Tamiflu (oseltamivir) to replace your yearly flu shot because the Centers for Disease Control (CDC) recommends yearly flu shots that will help protect you from new strains of influenza viruses each year.
Contraindication : Do not take this medication if you are allergic to oseltamivir.
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This medication is an antiviral used to treat patients who have had flu symptoms, caused by the influenza virus, for less than 2 days. Tamiflu (oseltamivir) may also be used to prevent influenza in patients who has not yet had the symptoms but may have been exposed to the virus. This medication does not treat common colds.
This medication should be stored at room temperature. Keep the medication away from heat and moisture. Tamiflu liquid formula should be stored in the refrigerator. Do not freeze. Dispose unused liquid after 10 days.
Before using this medicine, tell your health care providers about any of the following: if you are pregnant, planning to become pregnant, or breastfeeding; if you have had the nasal flu vaccine (FluMist) within the past two weeks; kidney, heart, or lung disease; a condition causing swelling or disorder of the brain; or any other serious disease or health problem.
This medication is an antiviral used to treat patients who have had flu symptoms, caused by the influenza virus, for less than 2 days. Tamiflu (oseltamivir) may also be used to prevent influenza in patients who has not yet had the symptoms but may have been exposed to the virus. This medication does not treat common colds. This medication should be stored at room temperature. Keep the medication away from heat and moisture. Tamiflu liquid formula should be stored in the refrigerator. Do not freeze. Dispose unused liquid after 10 days.Read the directions on the prescription label. Do not take larger doses than prescribed by your doctor. Do not take this medication longer than recommended by your doctor. Take this medication as directed on the label or by your pharmacist, doctor, or other healthcare provider. As soon as flu symptoms appear (ie. Chills, fever, muscle aches, runny or stuffy nose, and sore throat), begin taking Tamiflu (oseltamivir) as soon as possible. If you were prescribed with Tamiflu capsules, take with a full glass of water (8 ounces). If you were prescribed with Tamiflu oral suspension (liquid formula), shake well before you measure your dose with a measuring syringe. Do not use a regular table spoon to measure your dose. Tamiflu (oseltamivir) can be taken with or without food. Take Tamiflu (oseltamivir) with milk or food if you notice the medicine upsets your stomach. Patients taking Tamiflu (oseltamivir) to treat flu symptoms should take the medication every 12 hours for 5 days. Patents taking Tamiflu (oseltamivir) for flu prevention should take the medication every 24 hours for 10 days or as prescribed by your doctor. Take Tamiflu (oseltamivir) for the entire length of time as directed by your doctor even when your symptoms improve because the infection may not be completed treated. Contact your doctor if your symptoms do not improve or becomes worse. Do not take this medication if you are allergic to oseltamivir. Before using this medicine, tell your health care providers about any of the following: if you are pregnant, planning to become pregnant, or breastfeeding; if you have had the nasal flu vaccine (FluMist) within the past two weeks; kidney, heart, or lung disease; a condition causing swelling or disorder of the brain; or any other serious disease or health problem.Do not use Tamiflu (oseltamivir) to replace your yearly flu shot because the Centers for Disease Control (CDC) recommends yearly flu shots that will help protect you from new strains of influenza viruses each year.Severe: Hallucinations, sudden confusion, delirium, self-injury, or unusual behavior especially in children.Common: Nosebleed; vomiting, diarrhea, nausea; eye redness or discomfort; dizziness, headache; cough or other respirator symptoms; or sleep problems (insomnia).Other: None., DoctorSolve is a reputable pharmacy that offers affordable prices on quality Tamiflu medication.
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text:
BMC Genomics
Generation and analysis of expressed sequence tags from the ciliate protozoan parasite Ichthyophthirius multifiliis
Jason W Abernathy1,
Peng Xu1,
Ping Li1,
De-Hai Xu2,
Huseyin Kucuktas1,
Phillip Klesius2,
Covadonga Arias1 &
Zhanjiang Liu1
BMC Genomics volume 8, Article number: 176 (2007) Cite this article
The ciliate protozoan Ichthyophthirius multifiliis (Ich) is an important parasite of freshwater fish that causes 'white spot disease' leading to significant losses. A genomic resource for large-scale studies of this parasite has been lacking. To study gene expression involved in Ich pathogenesis and virulence, our goal was to generate expressed sequence tags (ESTs) for the development of a powerful microarray platform for the analysis of global gene expression in this species. Here, we initiated a project to sequence and analyze over 10,000 ESTs.
We sequenced 10,368 EST clones using a normalized cDNA library made from pooled samples of the trophont, tomont, and theront life-cycle stages, and generated 9,769 sequences (94.2% success rate). Post-sequencing processing led to 8,432 high quality sequences. Clustering analysis of these ESTs allowed identification of 4,706 unique sequences containing 976 contigs and 3,730 singletons. These unique sequences represent over two million base pairs (~10% of Plasmodium falciparum genome, a phylogenetically related protozoan). BLASTX searches produced 2,518 significant (E-value < 10-5) hits and further Gene Ontology (GO) analysis annotated 1,008 of these genes. The ESTs were analyzed comparatively against the genomes of the related protozoa Tetrahymena thermophila and P. falciparum, allowing putative identification of additional genes. All the EST sequences were deposited by dbEST in GenBank (GenBank: EG957858–EG966289). Gene discovery and annotations are presented and discussed.
This set of ESTs represents a significant proportion of the Ich transcriptome, and provides a material basis for the development of microarrays useful for gene expression studies concerning Ich development, pathogenesis, and virulence.
The ciliate protozoan Ichthyophthirius multifiliis (Ich)is one of the most devastating pathogens. It infects fish skin and gills, and causes white spot diseases in many species of freshwater fish worldwide, which leads to significant losses in the aquaculture industry. The ciliate parasite has three main life-cycle stages: the reproductive tomont, the infective theront, and a parasitic trophont [1–3]. The mature trophont drops off the host to become the tomont where it attaches to a substrate, and undergoes multiple divisions to produce hundreds to thousands of tomites within a cyst. Tomites bore their way through the cyst into water, and differentiate into theronts that infect fish. Once they burrow into the fish epithelium, theronts become trophonts that feed and mature in the host.
In spite of great losses caused by Ich to the aquaculture industry, molecular studies of the parasite have been scarce [see a recent review [4]]. Limited studies have concentrated on immune responses of the host and factors affecting them [5–11]. One of the difficulties for the studies of Ich is the problem involved in long-term maintenance of Ich isolates. Ich isolates appear to lose infectivity or become senescent after a certain number of passages [12–15]. Most often a significant decrease in infectivity is observed after about 50 passages [15]. Not only the infectivity decreases with higher numbers of passages, but also the development of the parasite as measured by the period required for trophonts to emerge from fish [15].
The Ich senescence phenomenon is interesting not only as a developmental biology issue, but also as a potential research system to study the virulence factors involved in the parasite pathogenesis. Assuming the life cycles of Ich and its infectivity are controlled by gene products, then it would be of great interest to learn what genes are involved in the loss of infectivity, and in the slowing down of its development. However, as very limited molecular information is available from Ich, in-depth research is limited by the lack of information and the lack of genomic resources.
EST analysis is one of the most effective means for gene discoveries, gene expression profiling, and functional genome studies [16–23]. It is also one of the most efficient ways for the identification of differentially expressed genes [24–28]. In order to provide genomic resources for the analysis of differentially expressed genes at different developmental stages of the Ich parasite, and for the analysis of genes differentially expressed when infectivity is being lost, the objectives of this study were to create cDNA libraries suitable for the analysis of expressed sequence tags (ESTs) and to generate an EST resource for Ich to allow cDNA-based design of microarrays for the study of gene expression in relation to the passages and development of the parasite. Before this work, there were only 511 Ich sequences in the GenBank dbEST (release 100606) [29]. A brief examination of these existing EST sequences indicated that a large proportion of them were trophont only reads, histones, ribosomal proteins, and immobilization antigen-related sequences. Here we report sequencing of 10,368 Ich EST clones, and generation of 8,432 high quality EST sequences. This EST resource should provide the material basis for the development of microarrays for Ich, and serve as a platform for its functional genomic studies including the development and pathogenesis of Ich, and the host-parasite interactions.
Generation of the Ich ESTs
As summarized in Table 1, a total of 10,368 clones were sequenced from a normalized Ich library made from pooled cells from all three life cycle stages: theront, tomont, and trophont. Readable sequences were generated with 9,769 clones (94.2% sequencing success rate). After base calling, sequences were processed by using Phred [30, 31] to eliminate low quality sequences below Q20. Sequences passing Q20 were uploaded into Vector NTI Advance 10 (Invitrogen, Carlsbad, CA) for vector trimming and removal of sequences with very short inserts (<100 bp). The post-sequencing processing resulted in 8,432 high quality sequences.
Table 1 A summary of the EST analysis.
The processed sequences were subjected to cluster analysis using Vector NTI to evaluate sequence redundancies. Of the 8,432 sequences, 4,702 sequences fell within 976 contigs while 3,730 sequences were singletons. On average, each contig contained 4.8 sequences. Taken together, the 976 contigs and the 3,730 singletons made up 4,706 unique sequences (Table 1).
The Ich genome expression appeared to be extremely polarized with a few genes expressed at very high levels. In spite of normalization, transcripts from a few genes were sequenced at very high frequencies. The top 20 contigs with the largest number of ESTs are summarized in Table 2. Of the top 20 most abundantly sequenced transcripts, four of them were detected over 0.5% of total sequences. Of these, the most abundantly sequenced EST cluster, cluster 276 with 764 ESTs, accounted for 7.36% of all sequenced clones. BLASTX searches indicated that this transcript was most similar to a hypothetical protein, TTHERM_02141640, from Tetrahymena thermophila. The second most abundantly sequenced transcript was cluster 60 with 119 ESTs. It was identified as a transcript most similar to a hypothetical protein, TTHERM_02641280, from T. thermophila. The functions of these hypothetical proteins are unknown at present. These are two transcripts sequenced at exceptionally high frequencies. Obviously, the presence of such abundant transcripts suggested a failure in the normalization processes. However, it is puzzling to us because we believe the overall normalization processes may have worked based on several other observations: 1) the overall gene discovery rate (unique sequences over all sequences analyzed) was 55.8%, a reasonable rate for the sequencing depth of approximately 10,000 clones; 2) most other anticipated highly expressed genes such as ribosomal protein genes, actin genes, tubulin genes, and dynein genes were not detected at high levels. Nonetheless, we believe that this information is relevant and important as these genes should be the subject for additional subtraction for further EST sequencing in this species. In addition, such information should provide some basic picture about the most abundantly expressed genes in the parasite. As these hypothetical protein genes are transcribed at such high levels, they may be crucially important for the growth and development, or other life-cycle processes of the parasite.
Table 2 The most abundant ESTs detected from the EST sequencing
This work demonstrated that pooling of samples from all three stages of Ich life cycle followed by normalization was an effective way to reduce common messages across all three life stages. As one would expect, many structural genes would be expressed highly abundantly in all stages of the life cycle. In addition to making savings economically, pooling of samples allowed very effective normalization of these common transcripts without going through three rounds of normalization. This is consistent with our previous experience for the generation of a large number of catfish and oyster ESTs [32–36]. It is obvious that the pooling of samples from three developmental stages made it impossible to provide information concerning expression profiling in relation to developmental stages. However, such information would not be highly meaningful in normalized cDNA libraries where the major focus was to develop EST resources, rather than expression profiling. The other limitation caused by construction of a pooled cDNA library is the loss of sequencing flexibility as to the number of clones to be sequenced from each developmental stage library if they had been separately constructed.
The Ich transcribed sequences are highly A/T-rich, similar to the situation in T. thermophila. Our unique sequences combined contain 2.18 megabases, approximately 10% of the genomic sequence size of the related protozoan Plasmodium falciparum, and 2.1% of the T. thermophila genomic sequence. As Ich is a ciliate and most closely related to Tetrahymena, this EST resource should represent a good sample of the transcribed fraction of the Ich genome for the estimation of its genome contents as compared with Tetrahymena. Based on the EST sequences, the average G+C content of Ich transcribed sequences was found to be 33.4%, even more A/T-rich than those of the closely related hymenostome T. thermophila, which has an average G+C content of 38% at protein coding regions [37]. The entire genome of T. thermophila was much more A/T-rich than the transcribed fraction, with a G+C content of only 22% [38]. It is highly probable that the Ich genome is also highly A/T-rich. To further the analysis, we found approximately 1% of the unique ESTs sequenced contained simple sequence repeats. The majority of the simple sequence repeats were of di-nucleotide repeats (68.8%) with AC and AG repeats being the majority. Tri-nucleotide and tetra-nucleotide repeats accounted for 23.7% and 7.5% of the identified microsatellites, respectively (Table 3).
Table 3 A summary of simple sequence repeats identified from the Ich ESTs. Percentages indicated in the parentheses are percentage of each type of repeat among all repeats
The putative identities of the sequenced ESTs were assessed using BLASTX searches against the non-redundant (NR) database in GenBank [39]. All the search results are summarized in supplemental Table 1. Of the 4,706 unique ESTs, 2,518 (53.5%) had significant (E-value < 10-5) hits. The remaining 2,188 (46.5%) EST sequences were not similar to any known sequences. Additional searches using the Swiss-Prot database resulted in putative identities for six additional unknown ESTs (Supplemental Table 1).
Identification of putative secretory proteins
Secretory proteins have been shown to be an important component in many biological processes, including pathogenesis of parasites [40–42]. We therefore searched for transcripts with putative signal peptides (suggestive of peptides of secretory proteins) within the EST set using the program SignalP 3.0 [43]. We found 314 ESTs with signal peptides, representing 6.7% of the unique sequences. Of these, 180 (3.8%) were from ESTs with no significant (E-value < 10-5) BLASTX hit to the NR database in GenBank (Supplemental Table 1).
Comparative analysis to related taxa
The parasite Ich is phylogenetically placed between the protozoan's Plasmodium falciparum and Tetrahymena thermophila. Previous studies using 18S rDNA, histone genes, and I-antigens [44–46] suggested that Ich was more related to T. thermophila than to P. falciparum. Furthermore, T. thermophila and Ich share the ciliate nuclear genetic code, while P. falciparum uses the standard genetic code for translation. As the entire genome sequence of P. falciparum is available and the macronuclear sequencing project of T. thermophila was just recently completed, we made comparative BLAST analyses against both genome sequences.
The tBLASTx or BLASTX searches of Ich ESTs against the T. thermophila and P. falciparum genomes are summarized in Supplemental Table 2, and are presented in Figure 1. As expected based on the phylogenetic relationships, more Ich ESTs were similar to the genome sequences of T. Thermophila than to that of P. falciparum. Of the 4,706 Ich ESTs, 1,759 sequences were similar (E-value < 10-5) to the T. thermophila genome sequences; whereas 817 were similar to the P. falciparum genome sequences. In total, 695 ESTs were similar to both T. thermophila and P. falciparum genomes, and thus are common to all three protists.
Venn diagram summary of sequence comparisons of the Ich ESTs with Tetrahymena thermophila and Plasmodium falciparum genomes. A total of 4,706 unique Ich ESTs were used as queries yielding 1,759 significant (E-value < 10-5) hits to the T. thermophila genome, and 817 to the P. falciparum genome. A total of 695 sequences were ESTs with common hits to both genomes.
Of the 1,759 significant hits against the T. thermophila genome, 1,673 had been identified with a putative identity using BLASTX searches against the NR database, while the tBLASTx searches against the T. thermophila genome allowed identification of putative identities for additional 86 unique ESTs. Similarly, BLASTX searches against the P. falciparum genome allowed identification of 9 additional ESTs. Taken together, the BLAST searches against these two genomes allowed putative identities of 95 additional unique ESTs, bringing the total number of ESTs with significant similarities to known genes to 2,613.
Such genome searches also revealed that of the 2,518 ESTs that had significant hits in BLASTX searches against the NR database, 845 had no significant hits to the Tetrahymena genome. Clearly, these ESTs were similar to sequences of organisms other than the ciliate Tetrahymena.
These results clearly suggest conservation of a large fraction of gene sequences among the three protozoa parasites, with a higher level of conservation between Ich and the T. thermophila genome than between the Ich genome and the P. falciparum genome; although a significant fraction of gene sequences are also shared between the genomes of T. thermophila and P. falciparum. The results of this comparative analysis are compatible with existing phylogenetic analyses using several molecular markers such as the 18S rDNA, histone genes, and the I-antigens. Obviously, use of a large set of sequences should provide a greater confidence concerning genome evolution. The comparative analysis suggested that the EST resource generated from this study should be useful for phylogenetic analysis and studies concerning genome evolution.
The unique Ich sequences were compared to annotations through the Gene Ontology Consortium [47] using the automated software Blast2GO [48]. We were able to obtain GO terms for 1,008 unique sequences using this method. Of these, 304 were contigs and 704 were singletons. Sequence descriptions, gene ontology (GO) and enzyme commission (EC) numbers are summarized in Supplemental Table 3. There were 258 sequences with both GO terms and EC numbers.
Gene ontology graphs using percentages of 2nd level GO terms are presented in Figure 2 under the categories of cellular components (Fig. 2A), molecular functions (Fig. 2B), and biological processes (Fig. 2C). Of the cellular component GO terms, 45% and 26% were related directly with cellular and organelle components, respectively. In the category of molecular functions, the vast majority were involved in catalytic activity (41%) and binding activities (39%). Under the category of biological processes, 45% were involved in physiological processes; 43% were involved in cellular processes, 6% in regulation of biological processes, 4% in response to stimuli, and 2% in development (Figure 2).
Pie charts of 2nd level gene ontology (GO) terms. Overall, 1,008 unique sequences were annotated using the Blast2GO software and included in the graphs. Each of the three GO categories is presented including cellular component (a), molecular function (b), and biological process (c).
We have produced 8,432 high quality I. multifiliis EST sequences. Sequence analysis indicated the presence of 4,706 unique sequences in the EST set. This should represent a significant fraction of the Ich genes, although the exact gene number of the parasite is unknown at present. The majority of the unique EST sequences had similarities to known genes, making them more amenable to functional analysis. The EST sequences should enhance the effectiveness of molecular studies, especially for gene expression profiling and the analysis of genes involved in virulence and infectivity. Microarrays can now be designed using either cDNA microarray or oligo-based platforms using the EST information. Additionally, the cluster and redundancy information should be useful for further subtraction of the most abundant transcripts included in the cDNA library, making further EST analysis in the parasite more effective.
The source of mRNA for this analysis was derived and expanded from a single parasite cloned from the infected fish. The source of the original Ichthyophthirius multifiliis was isolated from an infected fish obtained from a local pet shop and the parasite was transmitted to channel catfish held in a 50-l glass aquarium at the USDA-ARS Aquatic Animal Health Research Laboratory, Auburn, AL. The transmission of I. multifiliis was achieved through co-habitation of the infected fish with two fingerlings of channel catfish (3 inches in size). When the two catfish were infected, the symptoms of Ich, white spots, started to emerge when trophonts were collected by scraping with a glass slide. Channel catfish infected with maturing trophonts were rinsed in dechlorinated water and the skin was gently scraped to dislodge the parasites. Trophonts were harvested by filtering through a 0.22 μ m filter to remove fish skin. The trophonts were placed into a Petri dish to allow them to develop into theronts that were used to infect 8 fish each for the collection of trophonts, toments, and theronts, respectively. Trophonts were directly collected from the skin surface of the 8 infected fish. To collect tomonts and theronts, trophonts isolated from fish were placed in Petri dishes and allowed to attach. After replacing the water in the Petri dishes with fresh dechlorinated water to remove contaminating mucus, the trophonts were incubated at 24°C for 8 h to harvest tomonts (32–128 cells/cyst) or 24 h to harvest theronts. Trophonts, tomonts and theronts were washed with PBS (pH 7.4), concentrated with a centrifuge (Beckman Coulter, Inc., Miami, FL) at 228 × g for 5 min and discarded supernatant. After washing 3 times with PBS, parasite samples from the three life stages were stored in liquid nitrogen and used for the isolation of RNA for the construction of normalized cDNA library.
RNA isolation
Total RNA was isolated from the samples using the TRIzol reagent method from Invitrogen (Carlsbad, CA, USA) according to manufacturer's instructions. Briefly, samples of tomont, theront, and trophont were resuspended after thawing on ice, and 100 μl each were combined in a sterile tube to provide a total of 300 μl of Ich samples with equal fractions from each of its three life stages. As the major objective of this study was to generate EST resources with maximal efficiency of gene discovery, a pooled sample followed with normalization would allow inclusion of all transcripts in the library while reducing cost for library construction and increasing gene discovery rate. Three milliliters of TRIzol reagent was added to the sample tube. Cells were lysed by repetitively pipetting up and down. RNA was isolated following the manufacturer's protocol. The RNA pellet was resuspended in 100 μl of RNase-free double distilled water and divided into 25 μl aliquots. RNA aliquots were checked for quality using agarose gel electrophoreses containing formaldehyde.
Normalized library construction
The Creator Smart cDNA Library Construction Kit from Clontech (Mountain View, CA) and components from the TRIMMER-DIRECT Kit from Evrogen (Moscow, Russia) were used for the construction of the normalized cDNA library. Total RNA concentration was checked on a spectrophotometer and 1 μg RNA was combined with 1 μl of SMART IV oligonucleotide (Clontech) and 1 μl CDS-3M adapter (Evrogen) for first-strand cDNA synthesis. The reaction was incubated at 72°C for 2 min followed by immediate cooling on ice for 2 min. Next, 2 μl of 5× first strand buffer, 1 μl of DTT (20 mM), 1 μl of dNTP mix (10 mM), and 1 μl of PowerScript reverse transcriptase were added to the tube and incubated at 42°C for 1 h in a thermal cycler (PTC-100, Bio-Rad, Hercules, CA) then placed on ice. The SMART cDNA cloning system allows the enrichment of full-length cDNA through the use of a 5'-linker with 3'-GGG tails. Reverse transcriptase has terminal transferase activity that preferentially adds three additional Cs at the end of first strand cDNA. As a result, the first strand cDNA is able to base pair with the 5'-linker with 3'-GGG tails. Once base paired, the reverse transcriptase would switch the template and extend into the linker sequences allowing PCR amplification of full-length cDNA using a single primer (the 5'-linker has the same sequences as the linker containing poly T used for the synthesis of the first strand cDNA). Truncated cDNAs are not able to base pair with the 5'-linker and, therefore, get lost in the PCR amplification of the full-length cDNA.
The first strand cDNA was initially amplified by long-distance PCR (LD-PCR) using hot-start amplification. For the reaction, the following were combined in a reaction tube: 1.5 μl of the first strand cDNA, 60 μl of sterile deionized water, 7.5 μl of 10× Advantage 2 PCR buffer, 1.5 μl of 50× dNTP mix, 3 μl of 5' PCR primer and 1.5 μl of 50× Advantage 2 polymerase mix. The tube was mixed and briefly centrifuged and added to a pre-heated (95°C) thermal cycler. Cycle settings were 95°C for 1 min followed by 19 cycles of 95°C for 7 s, 66°C for 20 s, and 72°C for 5.5 min. The product was analyzed on a 1.1% agarose gel to determine the sizes and amount of the cDNA products before proceeding to the next step. The LD-PCR reaction was purified and eluted in 30 μl of sterile Nanopure water using the QIAquick PCR Purification Kit (Qiagen, Valencia, CA).
For the normalization procedure, the TRIMMER-DIRECT Kit from Evrogen (Moscow, Russia) was used. This system is specially developed to normalize cDNA enriched with full length sequences [49, 50]. The cDNA from the LD-PCR was quantified [~100 ng/μl] and 1 μl was mixed with 1 μl of the 4× hybridization buffer and 2 μl of sterile water. The mix was overlaid with mineral oil and incubated for 3 min at 98°C followed by 4 h at 70°C. Then, 5 μl of 2× DSN buffer (preheated to 70°C) and 0.25 Kunitz units of DSN enzyme were added and incubated at 70°C for 20 min. The DSN enzyme specifically degrades double-stranded molecules. The reaction was inactivated by adding 10 μl of DSN stop solution, and sterile water added to a final volume of 40 μl.
Following normalization, two rounds of PCR were performed using 1 μl of the normalization reaction as template. A shorter primer M1 (first 23 bases of the SMART IV oligonucleotide) was used in the first round of PCR with 15 amplification cycles using the same thermal cycling parameters as above; and an even shorter primer M2 (first 20 bases of the SMART IV oligonucleotide) was used in the second round of PCR for 15 amplification cycles of 95°C for 7 s, 64°C for 20 s, and 72°C for 5.5 min. Products were checked on a 1.1% agarose gel. The PCR products were quantified and 3 μg were used for treatment with proteinase K. All the subsequent procedures including proteinase K treatment, restriction digestion with Sfi I, size fractionation, and ligation followed the manufacture's instructions (Clontech). The cDNA was ligated to the pDNR-LIB vector. Electroporation (MicroPulser, Bio-Rad, Hercules, CA) was performed using DH12S electrocompetent cells following supplier's instructions (Invitrogen). A total of approximately 700,000 primary recombinant clones were obtained, and the library was amplified, titered, and stored in glycerol stocks in a -80°C freezer.
Plasmid isolation and EST sequencing
Independent colonies were picked and grown for 20 h at 37°C in 1.2 ml LB broth containing 30 μg/ml chloramphenicol. Plasmid DNA was isolated using the Perfectprep Plasmid 96 Vacuum Direct Bind Kit from Eppendorf (Westbury, NY). Plasmids were stored at -20°C until usage. The cDNA inserts were directionally sequenced from the 5'-end of the cDNAs using universal M13(-21) primer and the BigDye terminator sequencing kit version 3.1 from Applied Biosystems (Foster City, CA) on a 3130XL DNA analyzer (Applied Biosystems).
Base calling was performed using the Phred program [30, 31] at quality cut-off set at Phred 20. Raw sequences were then imported into the Vector NTI Advance 10 software (Invitrogen) and were subjected to trimming of vector sequences and 5'adapter sequences using default settings. Afterwards, poly (A) tails were trimmed where necessary and sequences less than 100 bases were removed. Contigs were built in Vector NTI ContigExpress using default settings. All unique sequences were compared to the GenBank database using BLASTX in the non-redundant (NR), Swiss-Prot, and Plasmodium falciparum 3D7 genome database. For comparison to the Tetrahymena thermophila SB210 genome, tBLASTx was used. The cut-off for sequence similarity used was E-value < 10-5 for all analyses. Ciliate nuclear translation code was used in the BLAST searches. Search results from genome comparisons were summarized using a Venn diagram.
Gene ontology (GO) annotations were assigned using the program Blast2GO [48]. BLASTX results were loaded into the program and the default settings were used to assign GO terms to all unique sequences. From these annotations, pie charts were made using 2nd level GO terms based on biological process, molecular function, and cellular component.
Putative secretory proteins and signal peptides were identified using both neural networks and hidden Markov model methods in SignalP 3.0 [43]. All 4,706 unique ESTs were used as the tester sequences. Open reading frames were predicted using both OrfPredictor [51] and BLASTX, with ciliate nuclear genetic code for ESTs of known genes and just OrfPredictor with unknown ESTs. The resulting deduced protein sequences from the ORFs were uploaded into SignalP 3.0. Sequences were identified as putatively secretory, predicted with signal peptides if both D-score in the neural network model and prediction probability in the hidden Markov model were significant.
Total lengths of all ESTs, G+C% content and simple repetitive elements were estimated using the Repeatmasker program [52].
All Ich EST sequences were submitted to the dbEST database of NCBI. Continuous accession numbers are from EG957858–EG966289.
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This project was supported by a Specific Cooperative Agreement with USDA ARS Aquatic Animal Health Laboratory under the Contract Number 58-6420-5-030, and in part by CSREES from a grant of USDA NRI Animal Genome Tools and Resources Program (award # 2006-35616-16685) to support the first year stipend of JWA. We are grateful for an equipment grant from the National Research Initiative Competitive Grant no. 2005-35206-15274 from the USDA Cooperative State Research, Education, and Extension Service.
The Fish Molecular Genetic and Biotechnology Laboratory, Department of Fisheries and Allied Aquacultures and Program of Cell and Molecular Biosciences, Aquatic Genomics Unit, Auburn University, AL, 36849, USA
Jason W Abernathy, Peng Xu, Ping Li, Huseyin Kucuktas, Covadonga Arias & Zhanjiang Liu
Aquatic Animal Health Research Laboratory, Agricultural Research Service, United States Department of Agriculture, Post Office Box 952, Auburn, AL, 36831, USA
De-Hai Xu & Phillip Klesius
Jason W Abernathy
Peng Xu
Ping Li
De-Hai Xu
Huseyin Kucuktas
Phillip Klesius
Covadonga Arias
Zhanjiang Liu
Correspondence to Zhanjiang Liu.
JWA, CA and ZJL planned the experiment and drafted the manuscript. HK, PL and PX provided technical assistance including cDNA library construction and sequencing support. PK and DX provided technical support and valuable Ich knowledge along with sample preparations. All were involved in final manuscript editing.
Additional file 1: Supplemental Table 1, Excel spreadsheet; Table of I. multifiliis unique EST sequences; Provided information includes I. multifiliis BLASTX top hits to the non-redundant database in GenBank with unique EST name and accession numbers. Also included are significant protein domain comparisons to the Swiss-Prot database. Putative secretory proteins are highlighted. (XLS 801 KB)
12864_2006_889_MOESM2_ESM.xls
Additional file 2: Supplemental Table 2, Excel spreadsheet; Summary of BLAST searches of the Ich ESTs against Tetrahymena thermophila and Plasmodium falciparum genomes. Provided information includes I. multifiliis BLASTX top hits to the non-redundant database in GenBank with unique EST name, tBLASTx top hits to the T. thermophila genome, and BLASTX top hits to the P. falciparum genome sequences. This table correlates with the Venn diagram in figure 1. (XLS 400 KB)
Additional file 3: Supplemental Table 3, Excel spreadsheet; Table of gene ontology (GO) profiles; Provided information includes unique EST name, accession numbers, BLASTX top hit, GO identification numbers and enzyme commission (EC) numbers. (XLS 190 KB)
Abernathy, J.W., Xu, P., Li, P. et al. Generation and analysis of expressed sequence tags from the ciliate protozoan parasite Ichthyophthirius multifiliis. BMC Genomics 8, 176 (2007). https://doi.org/10.1186/1471-2164-8-176
DOI: https://doi.org/10.1186/1471-2164-8-176
Enzyme Commission
Infected Fish
BLASTX Search
Normalize cDNA Library
Submission enquiries: bmcgenomics@biomedcentral.com

redpajama_set_name: RedPajamaCommonCrawl


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Gobert was really checking out in the beginning. The electrical gremlins got him this time. Oh well, it's wait 'til next year for Ducati.
However, if the Duc looked this good on a fast, high horsepower track, I'm eager to see how gobert does once things get a bit twistier.
Honda's decision not to update the RC51 is a marketing, rather than engineering decision. As they proved today, the twin is certainly capable of putting down power that will compete with any of the 4s, and the traditional VTwin user-friendly powerband is very well suited to tire management as well.
However, Honda doesn't ultimately race to win (although that certainly is important to them) but rather to sell bikes. Honda sportbike twins have never been big market place successes, although many have been highly regarded by enthusiasts. For example, the Hawk 650GT is a real cult classic, and good used ones still sell for about what they listed new, but they never were marketplace successes. The 996 Hawk, and later the RCC51 also did not click with the market.
Honda is as much a prisoner of their own marketing success as are Harley and Ducati. The Honda sportbike image is that of a refined 4 (or more) cylinder (V or inline). They have very successfully marketed that since their 60s racing successes (6 cyl 250s and 5 cyl 125s!) and the CB750. Now, when they try market a Vtwin sportbike, their customer base reacts about like Harleys would if they tried to sell an inline 4 cruiser!
Suzuki has been much more successful in selling their sporting V-Twins for some reason, largely because they have carved out a unique market segment for them, rather than trying to market them as pure sports bikes -- the TL1000 was also a market-place dud.
So far as I know, there is no support for the theory that Ducati will introduce a street V-4 (although they actually DID make one in the past -- intended for the police market). Even if they were to win the MotoGP championship with one, I doubt that the market wants a V4 Ducati.
Aprilia producing a 3 makes more sense because the Aprilia brand is not so strongly identified with the V-twin powerplant.
Now Bob stop it with all that logical, fact-based reasoning. You know ol' KP is about pure myopia and nothing more. Silly Bob, when will you learn.
I'm interested in your thoughts about the speed of the Duc today. Gobert looked really fast until it pulled a Ducati on him.
OK, I spoke too soon. Tommy Hayden just put the Kwacker 636 at the top of the SuperStock podium! Nice show.
This is a class that the AMA needs to kill (or to do what WSB did, change it to 1000cc) because it is almost totally a Suzuki GSX-R 750 Cup. Only other bikes are a few 600s (and now 636) that move up a class to compete.
Excellent Post. I agree with your premise that Honda races to sell bikes. Thus the V-Twin will be discarded as soon as a suitable inline or mult cylinder V comes along.
Yep but being in the same position I would of made the same decision. i.e. He has won the supersport crown before and he isn't going to race the 600 during the season.
It's a matter of focus.
Thats a pretty good smack back. I personally think that the EBOZ is the only chance for the inline crowd this year. I think you have to admit that the focus of liter bikes has not been racing until recently. The big four will come out with liter (inline 4s) bikes targeted for racing. The GSXR1000 is not refined now. But given time the next generation will be.
From what I understand, Gobert's crew discovered a problem with the brakes on his bike immediately prior to the start, and they went with their "B" bike instead. In fact, I read that he had to start from pit road because his bike wasn't ready for the start.
The bike is certainly fast enough to win, and Gobert is also, albeit having a rep as being somewhat erratic -- hopefully that is in the past.
I think where Ducati falls down in the AMA is that they do not have the quality team that Honda have. Merlin Plumlee, the crew chief for BBoz (and Nicky last year) has worked with both Honda and Kawasaki at both WSB and AMA for many years and is widely regarded as one of the best in the entire world (he also helped my build my first racebike), and the other Honda teams, as well as the Yoshi Suzuki teams are nearly as professional. These bikes very seldom have any mechanical failures.
The Ducati Austin team is the former Harley Davidson team, and while seemingly pretty good, are just not at the same level. They are also fairly new to running Ducatis, as this is only their second year.
Pegram's Dream Team Racing was pretty impressive for a new team. Be interesting to see how they do as the season progresses.
Long time Formula Extreme racer Mark Miller just announced via e-mail that he has decided to retire.
2001 Erion Honda rider Mark Miller officially retires from AMA/Pro roadracing.
"I have been the luckiest man on the planet to be able to achieve a boyhood dream of being a full-time, paid professional roadracer for six years. Additionally, I've been fortunate to have worked in a motorcycle racing community with some of the most incredible and interesting people in the world.
In light of the recent WTC disasters, my final race weekend at VIR will be especially memorable to me not only as my final pro race, but because it was our first pro race back since the attacks of Sept.11.
Next stop for my career, at least short-term, is to release a home video production with an insider's look at motorcycle roadracing. Release date is set for March 2002, and it's tentatively titled "Superbike Life".


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MAD TOAST LIVE!
Entries from July 2010
Episode 143 - Montana Skies
July 20th, 2010 · Comments
Jennifer and Jonathan Adams create a "new acoustic" sound with the pairing of electric cello and classical guitar. Their originals have been featured on NPR and the Travel Channel. Using a wide range of effects, their musical wizardry combines to create a sound that is truly remarkable--we're thrilled to have them back on the show!
http://www.montanaskiesmusic.ning.com
Tags: Music · Live Music
Episode 142 - Ida Jo
IDA JO
Courage. Strength. Devotion. Ida Jo's debut album "Providence" is about all those things and is available now on iTunes, CDBaby, and Amazon. Nominated for several Madison Area Music Awards, including String Instrumentalis of the Year, as well as Pop Album, Pop Song, and R&B Song for her work with Mike Droho & The Compass Rose, Ida Jo will showcase her own fiddle and songwriting chops on Mad Toast Live, as well as the talented young women's chorus she leads, called "Midnight Voices".
idajoandtheshow.com
Episode 141 - Tim Seisser Trio
(Episode 2 of 2)
TIM SEISSER TRIO
A staff writer for Bass Musician Magazine, and with influences ranging from Jaco Pastorius and Victor Wooten to Ray Brown and Paul Chambers, Tim Seisser is one of Chicago's premier electric bassists. He's been on the show before, but he's back with his own combo–a must see for all jazz lovers! In 2008, Tim released his first album as a leader entitled "Wire and Wood". It received high acclaim from jazz reviewers and has been played on multiple Chicago area jazz stations including WDCB and WNUA.
www.timseisser.com
A staff writer for Bass Musician Magazine, and with influences ranging from Jaco Pastorius and Victor Wooten to Ray Brown and Paul Chambers, Tim Seisser is one of Chicago's premier electric bassists. He's been on the show before, but he's back with his own combo--a must see for all jazz lovers! In 2008, Tim released his first album as a leader entitled "Wire and Wood". It received high acclaim from jazz reviewers and has been played on multiple Chicago area jazz stations including WDCB and WNUA.
Copyright © 2011 Chris Wagoner. All rights reserved.


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Ouachita Stories
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Ouachita Circle Magazine
Ouachita faculty, staff honored for combined 780 years of service
November 18, 2020 - Rachel Gaddis
Ouachita Baptist University recently recognized 36 faculty and staff members in celebration of their milestone anniversaries working for the university. Together they have served a total of 780 years at the start of the 2020-2021 academic year, with six faculty and staff members celebrating 35 years or more.
This year's longest-serving honorees are Janice Ford and Kevin Sullivan. Ford, professor and government documents librarian, celebrated 45 years of service and Sullivan, facilities management, was recognized for 40 years.
In her time at Ouachita, Ford has worked with government documents, specifically the McClellan Collection, and securing interlibrary loans for the Riley-Hickingbotham Library.
"Janice has always had the library's good as her lodestar," said Ray Granade, director of library services and professor of history. "Second, she has always been driven to learn, improve her skills and thereby provide better service to our patrons. As a librarian of learning, diligence and integrity, particularly her ability to procure through interlibrary loan even obscure and rare materials that our patrons need, Janice has embodied the reason that Ouachita has invested in professional librarians."
Sullivan, in addition to his many projects for facilities management over the years, installs the senior sidewalks on Ouachita's campus each year.
"He not only has installed every one of them, but he built the complete assemble of the sandblaster that we use to put the names in the concrete. There is nothing he can't fix or figure out," said John Hardman, director of facilities management. "Kevin is the ultimate professional in everything that he does."
Those celebrating 35 years of service to Ouachita include: Steve Hennagin, professor of mathematics; Sherri Phelps, director of human resources; Doug Reed, associate vice president for academic affairs, Herbert and LaDelle Moody Professor of Pre-Law Studies and director of Maddox Public Affairs Center; and Ruby Williams, facilities management.
"The dedication of Ouachita faculty and staff matters – they are investing years of their lives in students," said Ouachita President Ben Sells. "Beyond these personal influences, their collective service over the years also has contributed to the university's increased enrollment, highest graduation rate and, this year, to bringing students back to campus for in-person learning during a pandemic.
"Many parents over the years have entrusted their students to Ouachita because of our faculty and staff's love for students, as well as the excellence they exhibit in their fields," he added. "Ouachita is pleased to recognize these 36 colleagues for their faithful service."
Other faculty and staff members honored include:
30 Years: Nancy Hardman, Edna and Freda Linn Professor of Communication Sciences and Pre-Medical Studies; Deborah Lee, facilities management; Craig Ward, men's tennis coach and coordinator of tennis.
25 Years: Robin Green, facilities management; Laurie Huneycutt, School of Fine Arts administrative assistant; Glenda Secrest, professor of music; Molly Wallace, director of health services; and Kim Ward, tennis center manager.
20 Years: Dennis Brashier, facilities management; Detri Brech, Charles S. and Elma Grey Goodwin Holt Professor of Dietetics and Pre-Medical Studies; Kevin Cornelius, professor of physics; Larry Dyess, facilities management; Kent Faught, professor of management and Holder of the Jay and Lynn Heflin Chair of Business; Tracey Knight, executive assistant to the president; and Martha Womack, Campus Ministries administrative assistant.
15 Years: James Files, associate professor of accounting; Melinda Fowler, IT software developer; Tim Harrell, director of Student Life; Kevin Herrington, network services administrator; Jonathan Hughes, facilities management; Benny Johnson, facilities management; Wesley Kluck, vice president for student development and university physician; Susan Monroe, staff accompanist; Tara Pritchard, Social Sciences administrative assistant; and Diane Runyan, Elrod Center administrative assistant.
10 Years: Ashlee Giles, director of enrollment initiatives and senior admissions counselor; Bethany Hicks, associate professor of history; Elizabeth Kelly, associate professor of sociology and Elrod Center applied research associate; René Zimny, assistant professor of Art and Design; and Mike Pritchard, facilities management.
Ouachita announces students named to Fall 2021 President's List
Ouachita announces students named to Fall 2021 Dean's List
Ouachita receives initial approval for 4-year BSN program in Arkadelphia
Ouachita's Dr. Nathan Reyna receives national Innovation in Education Award
Ouachita to host ceramics exhibit by Logan Hunter and Hannah May
From the "Ouachitonian": Dr. Wesley Kluck


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Local Dunkirk NY Landlord & Tenant Lawyers
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At The Law Office of Rachel K. Marrero in Buffalo, New York, we are fully prepared to help you with the important and sensitive legal challenges affecting your family, your property and your future. To that end, our law firm offers exceptional representation and support in all areas of family law and real estate law, and we serve a diverse range of clients throughout Erie, Niagara, Chautauqua and...
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