Reprogramming of human fibroblasts to induced pluripotent stem cells under xeno-free conditions

The availability of induced pluripotent stem cells (iPSCs)has created extraordinary opportunities for modeling andperhaps treating human disease. However, all reprogrammingprotocols used to date involve the use of products of animal origin. Here, we set out to develop a protocol to generate and maintain human iPSC that would be entirelydevoid of xenobiotics. We first developed a xeno-free cellculture media that supported the long-term propagation of human embryonic stem cells (hESCs) to a similar extent as conventional media containing animal origin products or commercially available xeno-free medium. We also derivedprimary cultures of human dermal fibroblasts under strictxeno-free conditions (XF-HFF), and we show that they can be used as both the cell source for iPSC generation as well as autologous feeder cells to support their growth. We also replaced other reagents of animal origin trypsin, gelatin, matrigel) with their recombinant equivalents. Finally, we used vesicular stomatitis virus G-pseudotyped retroviral particles expressing a polycistronic construct encoding Oct4, Sox2, Klf4, and GFP to reprogram XF-HFF cells under xeno-free conditions. A total of 10 xeno-free humaniPSC lines were generated, which could be continuously passaged in xeno-free conditions and aintained characteristics indistinguishable from hESCs, including colonymorphology and growth behavior, expression of pluripotency-associated markers, and pluripotent differentiationability in vitro and in teratoma assays. Overall, the resultspresented here demonstrate that human iPSCs can be generatedand maintained under strict xeno-free conditions and provide a path to good manufacturing practice (GMP) applicability that should facilitate the clinical translation of iPSC-based therapies.

A protocol describing the genetic correction of somatic human cells and subsequent generation of IPS cell

The generation of patient-specific induced pluripotent stem cells (iPSCPSCPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSCPSCPSC technology can be used to generate disease-free progenitor cells of potential interest for autologous cell therapy. We explain a protocol for the reproducible generation of genetically corrected iPSCPSCPSCs starting from the skin biopsies of Fanconi anemia patients using retroviral transduction with OCT4, SOX2 and KLF4. Before reprogramming, the fibroblasts and/or keratinocytes of the patients are genetically corrected with lentiviruses expressing FANCA. The same approach may be used for other diseases susceptible to gene therapy correction. Genetically corrected, characterized lines of patient-specific iPSCPSCPSCs can be obtained in 4–5 months.

Iowa Gender Wage Study, 1999

In 1993, Iowa Workforce Development (then the Department of Employment Services) conducted a survey to determine if there was a gender gap in wages paid. The results of that survey indicated that women were paid 68 cents per dollar paid to males. We felt a need to determine if this relationship of wages paid to each gender has changed since the 1993 study. In 1999, the Commission on the Status of Women requested that Iowa Workforce Development conduct research to update the 1993 information. A survey, cosponsored by the Commission on the Status of Women and Iowa Workforce Development, was conducted in 1999. The results of the survey showed that women earned 73 percent of what men earned when both jobs were considered. (The survey asked respondents to provide information on a primary job and a secondary job.) The ratio for the primary job was 72 percent, while the ratio for the secondary job was 85 percent. Additional survey results detail the types of jobs respondents had, the types of companies for which they worked and the education and experience levels. All of these characteristics can contribute to these ratios. While the large influx of women into the labor force may be over, it is still important to look at such information to determine if future action is needed. We present these results with that goal in mind. We are indebted to those Iowans, female and male, who voluntarily completed the survey. This study was completed under the general direction of Judy Erickson. The report was written by Shazada Khan, Teresa Wageman, Ann Wagner, and Yvonne Younes with administrative and technical assistance from Michael Blank, Margaret Lee and Gary Wilson. The Iowa State University Statistical Lab provided sampling advice, data entry and coding and data analysis.

Comparação de procedimentos de quantificação de caulinita e gibbsita na fração argila desferrificada de Latossolos

A acurada determinação dos teores dos minerais que constituem a fração argila dos Latossolos pode contribuir para o aprimoramento de seu uso e manejo. Neste trabalho, teores de caulinita e gibbsita da fração argila desferrificada de Latossolos foram determinados por calorimetria exploratória diferencial (CED) e pela aplicação do método de Rietveld a dados de difratometria de raios X (DRX), e comparados com seus respectivos teores calculados pelo método da alocação. Possivelmente, diferenças cristalográficas entre a caulinita e a gibbsita utilizadas na calibração da CED e suas respectivas contrapartidas encontradas nas amostras analisadas tenham contribuído para a menor exatidão dos teores de caulinita e de gibbsita determinados pelo referido método térmico. Os teores de gibbsita das argilas desferrificadas quantificados com a abordagem DRX-Rietveld foram praticamente idênticos aos calculados por alocação, ao passo que os de caulinita foram similares aos respectivos teores determinados com o método de Rietveld.

Kaunokirjallisuus ja henkilönnimet

Väitöksenalkajaisesitelmä Åbo Akademissa 4. huhtikuuta 2003

Improved diagnosis of periprosthetic joint infection by multiplex PCR of sonication fluid from removed implants.

The microbiological diagnosis of periprosthetic joint infection (PJI) is crucial for successful antimicrobial treatment. Cultures have limited sensitivity, especially in patients receiving antibiotics. We evaluated the value of multiplex PCR for detection of microbial DNA in sonication fluid from removed orthopedic prostheses. Cases of PJI in which the prosthesis (or part of it) was removed were prospectively included. The removed implant was sonicated, and the resulting sonication fluid was cultured and subjected to multiplex PCR. Of 37 PJI cases (17 hip prostheses, 14 knee prostheses, 4 shoulder prostheses, 1 elbow prosthesis, and 1 ankle prosthesis), pathogens were identified in periprosthetic tissue in 24 (65%) cases, in sonication fluid in 23 (62%) cases, and by multiplex PCR in 29 (78%) cases. The pathogen was detected in 5 cases in sonication fluid only (Propionibacterium acnes in all cases; none of these patients had previously received antibiotics) and in 11 cases by multiplex PCR only (all of these patients had previously received antibiotics). After exclusion of 8 cases caused by P. acnes or Corynebacterium species, which cannot be detected due to the absence of specific primers in the PCR kit, sonication cultures were positive in 17 cases and multiplex PCR sonication cultures were positive in 29 cases (59% versus 100%, respectively; P < 0.01). Among 19 cases (51%) receiving antibiotics, multiplex PCR was positive in all 19 (100%), whereas sonication cultures grew the organism in 8 (42%) (P < 0.01). Multiplex PCR of sonication fluid is a promising test for diagnosis of PJI, particularly in patients who previously received antibiotics. With modified primer sets, multiplex PCR has the potential for further improvement of the diagnosis of PJI.

The Dream of Gerontius (Cardinal Newman) op. 38

Digitoitu 5. 1. 2008.

Short story

Digitoitu 14. 1. 2009

Människa – moral – miljö : en undersökning av livsåskådningar i finländskt miljötänkande

I Människa – moral – miljö undersöks syner på människa och moral i finländskt miljötänkande. Den pågående brytningen mellan modernitet och senmodernitet avspeglas i olika former av nutida finländskt miljötänkande. I avhandlingen analyseras fyra miljötänkares texter: teologen Seppo Kjellberg, biologen Pentti Linkola, miljöfilosofen Leena Vilkka och filosofen Georg Henrik von Wright. Alla fyra formerna av tänkande uttrycker försök att komma till rätta med miljöproblemen i sent 1900-tal. Vart och ett av försöken vill erbjuda en väg ut ur den moderna världens dilemma. Men samtliga försök som studerats i avhandlingen uppvisar oförlösta spänningar. De flesta försök till nytänkande uppvisar senmoderna drag. T.ex. delar alla fyra uppfattningen att det vetenskapliga tänkandets utveckling och dess sätt att umgås med naturen skapar problem som inte kan lösas enbart med tekniskt vetenskapliga medel. Men i samtliga analyserade tankesystem är bundenheten till viktiga drag i moderniteten fortfarande stark. T.ex. när det gäller moraluppfattning kan man spåra en syn där vägen till moralisk kunskap liknar vägen till faktakunskap. Det tolkas i avhandlingen som ett uttryck för moderna tankestrukturer, där tron på möjligheten att heltäckande beskriva verkligheten är stark, liksom tilltron till att man med förnuftets hjälp (ensamt) kan tänka ut och motivera de rätta vägarna till ett moraliskt förhållningssätt gentemot den övriga naturen. De moderna tendenserna i materialet tycks i samtliga fall göra försöken att lösgöra sig från det som åstadkommer miljöproblemen om intet. Miljöproblemen är en följd av modernitetstänkandet och de kan därför knappast lösas utan en medveten brottning och uppgörelse med flera av de drag som konstituerar den moderna världen och den moderna kulturen.