Heterologous expression and purification of recombinant allophycocyanin in marine Streptomyces sp isolate M097

Allophycocyanin is one of the most important marine active peptides. Previous studies suggested that recombinant allophycocyanin (rAPC) could remarkably inhibit the S-180 carcinoma in mice, indicating its potential pharmaceutical uses. Based on intergeneric conjugal transfer, heterologous expression of rAPC was first achieved in marine Streptomyces sp. isolate M097 through inserting the apc gene into the thiostrepton-induced vector pIJ8600. The transformation frequency for this system was approximately 10(-4) exconjugants/recipient. In the transformed Streptomyces sp. isolate M097, the yield of purified rAPC could amount to about 38 mg/l using a simple purification protocol, and HPLC analysis showed that the purity of the protein reached about 91.5%. In vitro activity tests also revealed that the purified rAPC had effective scavenging abilities on superoxide and hydroxyl radicals. This would widen the usefulness of the marine Streptomyces as a host to express the rAPC and to offer industrial strain for the production of rAPC.

attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector

An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048.

Possible suppression of exogenous beta-1,3-glucanase gene gluc78 on rice transformation and growth

Three genes encoding for fungal cell wall degrading enzymes (CWDE), ech42, nag7O and gluc78 from the biocontrol fungus Trichoderma atroviride were transformed into rice mediated by Agrobacterium tumefaciens singly and in all possible combinations. A total of more than 1800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. Our data indicated that gluc78 gene had negative effects on transformation frequency and plant growth. Some regenerated plants with gluc78 gene were stunted; spontaneously produced brown specks; could not tassel. The combination with either one of the two other genes (ech42, nag70) present in the same T-DNA region reduced the negative effect of gluc78 on plant growth. These results indicated that expression of several genes in one T-DNA region interfered with each other and expression of exogenous gene in recipient plant was a complex behavior. (c) 2007 Published by Elsevier Ireland Ltd.

成年女儿照料老年父母的心理体验及关系质量作用的研究

Previous researches about family caregiving revealed that caregiving has both negative and positive effects on caregivers’ well-being. Based on Lawton’s two-factor model, this study aims at examining how caring for old parents would affect adult daughters’ psychological well-being. According to Lawton, objective stressors as caregiving would arouse two different kinds of caregivers’ subjective appraisal, i.e., negative appraisal and positive appraisal, which in turn correlate with the negative and positive dimensions of caregivers’ psychological well-being, respectively. There were two main purposes of this study: a) to verify both the negative and positive paths in the two-factor model and their relatively independence; and b) to examine the effects of relationship quality between caregiver and care-recipient on those paths. The results are as follows: 1) Caregiving stressors have significant positive predictive effect on caregivers’ negative appraisal, but have no direct effect on caregivers’ positive appraisal. 2) Caregivers’ negative appraisal has significant positive predictive effect on their negative emotional experience, while caregivers’ positive appraisal has significant positive predictive effect on their positive emotional experience. 3) Certain dimensions of relationship quality, including the Appreciation and General Appraisal, have significant negative predictive effect on caregivers’ negative appraisal, and have significant positive predictive effect on caregivers’ positive appraisal. 4) The Appreciation dimension of relationship quality moderates the path from caregiving demands to caregivers’ burden; and the General Appraisal of relationship quality moderates the path from caregivers’ positive appraisal to life satisfaction. Based on the above results, the researcher concluded that a) both the negative path and positive path exist in caregiving process, and they are relatively independent from each other; and b) relationship quality does moderate certain paths in the model. Meanwhile, the main effect of relationship quality on caregivers’ experience is also significant and more remarkable. This study attempts to explain these results in terms of coping resources. Both relationship quality and many other factors might be explained as resources that caregivers utilize to cope with stress of caregiving. With more resources, caregivers tend to appraise more positively, and less negatively, and vice versa. However, the resources which might affect caregivers’ positive appraisal, as well as the ways they work, may be different from that affect caregivers’ negative appraisal.

The Power of Partnerships in Global Governance

Abrahamsen, Rita, 'The Power of Partnerships in Global Governance', Third World Quarterly (2004) 25(8) pp.1453-1467 RAE2008

The ecological effects of exotic disease resistance genes introgressed into British gooseberries

Warren, J. and James, P. (2006). The ecological effects of exotic disease resistance genes introgressed into British gooseberries. Oecologia 147(1),69-75. RAE2008

Factors affecting the survival of sheep embryos after transfer within a MOET program

F. Baria, M. Khalid, W. Haresign, A. Murray and B. Merrell (2003). Factors affecting the survival of sheep embryos after transfer within a MOET program. Theriogenology, 59 (5/6), 1265-1275. Sponsorship: DEFRA RAE2008

An ambiguity-based theory of the linguistic verbal joke in English

Instytut Filologii Angielskiej

KONSTYTUCYJNE PRAWO DO OCHRONY ZDROWIA W ŚWIETLE DERYWACYJNEJ KONCEPCJI WYKŁADNI

W niniejszym artykule autor przedstawił sposób odtworzenia normy prawnej zawierającej prawo do ochrony zdrowia na podstawie przepisu prawnego art. 68 ust. 1 Konstytucji RP. Przełożenie przepisu prawnego na normę prawną odbyło się zgodnie z założeniami derywacyjnej koncepcji wykładni, której istotą jest uzyskanie równoznaczności pomiędzy przepisem a odtworzoną z niego normą. W tym celu konieczne było przeprowadzenie wszystkich trzech faz wykładni: porządkującej, rekonstrukcyjnej i percepcyjnej. Niniejszy artykuł ukazuje, jak przebiegają poszczególne czynności interpretacyjne podejmowane w oparciu o dyrektywy derywacyjnej koncepcji wykładni. Pozwala to zobaczyć w szczegółowy sposób przebieg procesu przekształcania przepisu prawnego w normę prawną. Rezultatem przeprowadzonej wykładni było uzyskanie dostatecznie jednoznacznej normy prawnej. Na tej podstawie wykazano, że prawo do ochrony zdrowia jest zasadą prawa, ponieważ możliwe było odniesienie treści normy prawnej do określonych kryteriów. Ponadto, norma prawna odtworzona na podstawie przepisu konstytucyjnego zawiera wszystkie elementy, przede wszystkim określa adresata i nakazane zachowanie. Pozwala to wskazać sytuacje prawne, jakie wyznacza odtworzona w toku wykładni norma prawna, czyli obowiązek, uprawnienie i prawo podmiotowe. Postrzeganie konstytucyjnego prawa do ochrony zdrowia jako normy prawnej umożliwia także znacznie szersze i głębsze rozpatrzenie aspektów jego obowiązywania. W tym kontekście szczególnie przydatna jest derywacyjna koncepcja wykładni prawa.

Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents

Folliculogenesis is a complex process regulated by various paracrine and autocrine factors. In vitro growth systems of primordial and preantral follicles have been developed for future use of immature oocytes, as sources of fertilizable oocytes and for studying follicular growth and oocyte maturation mechanisms. Rodents were often chosen for in vitro follicular culture research and a lot of factors implicated in folliculogenesis have been identified using this model. To date, the mouse is the only species in which the whole process of follicular growth, oocyte maturation, fertilization and embryo transfer into recipient females was successfully performed. However, the efficiency of in vitro culture systems must still be considerably improved. Within the follicle, numerous events affect cell proliferation and the acquisition of oocyte developmental competency in vitro, including interactions between the follicular cells and the oocyte, and the composition of the culture medium. Effects of the acting factors depend on the stage of follicle development, the culture system used and the species. This paper reviews the action of endocrine, paracrine factors and other components of culture medium on in vitro growth of preantral follicles in rodents.

Invasive fungal infections among organ transplant recipients: results of the Transplant-Associated Infection Surveillance Network (TRANSNET).

BACKGROUND: Invasive fungal infections (IFIs) are a major cause of morbidity and mortality among organ transplant recipients. Multicenter prospective surveillance data to determine disease burden and secular trends are lacking. METHODS: The Transplant-Associated Infection Surveillance Network (TRANSNET) is a consortium of 23 US transplant centers, including 15 that contributed to the organ transplant recipient dataset. We prospectively identified IFIs among organ transplant recipients from March, 2001 through March, 2006 at these sites. To explore trends, we calculated the 12-month cumulative incidence among 9 sequential cohorts. RESULTS: During the surveillance period, 1208 IFIs were identified among 1063 organ transplant recipients. The most common IFIs were invasive candidiasis (53%), invasive aspergillosis (19%), cryptococcosis (8%), non-Aspergillus molds (8%), endemic fungi (5%), and zygomycosis (2%). Median time to onset of candidiasis, aspergillosis, and cryptococcosis was 103, 184, and 575 days, respectively. Among a cohort of 16,808 patients who underwent transplantation between March 2001 and September 2005 and were followed through March 2006, a total of 729 IFIs were reported among 633 persons. One-year cumulative incidences of the first IFI were 11.6%, 8.6%, 4.7%, 4.0%, 3.4%, and 1.3% for small bowel, lung, liver, heart, pancreas, and kidney transplant recipients, respectively. One-year incidence was highest for invasive candidiasis (1.95%) and aspergillosis (0.65%). Trend analysis showed a slight increase in cumulative incidence from 2002 to 2005. CONCLUSIONS: We detected a slight increase in IFIs during the surveillance period. These data provide important insights into the timing and incidence of IFIs among organ transplant recipients, which can help to focus effective prevention and treatment strategies.

Unrecognized pretransplant and donor‐derived cryptococcal disease in organ transplant recipients.

BACKGROUND: Cryptococcosis occurring ≤30 days after transplantation is an unusual event, and its characteristics are not known. METHODS: Patients included 175 solid-organ transplant (SOT) recipients with cryptococcosis in a multicenter cohort. Very early-onset and late-onset cryptococcosis were defined as disease occurring ≤30 days or >30 days after transplantation, respectively. RESULTS: Very early-onset disease developed in 9 (5%) of the 175 patients at a mean of 5.7 days after transplantation. Overall, 55.6% (5 of 9) of the patients with very early-onset disease versus 25.9% (43 of 166) of the patients with late-onset disease were liver transplant recipients (P = .05). Very early cases were more likely to present with disease at unusual locations, including transplanted allograft and surgical fossa/site infections (55.6% vs 7.2%; P < .001). Two very early cases with onset on day 1 after transplantation (in a liver transplant recipient with Cryptococcus isolated from the lung and a heart transplant recipient with fungemia) likely were the result of undetected pretransplant disease. An additional 5 cases involving the allograft or surgical sites were likely the result of donor‐acquired infection. CONCLUSIONS: A subset of SOT recipients with cryptococcosis present very early after transplantation with disease that appears to occur preferentially in liver transplant recipients and involves unusual sites, such as the transplanted organ or the surgical site. These patients may have unrecognized pretransplant or donor-derived cryptococcosis.

Characterization and analysis of the exogenous application of selected phytohormones on C. reinhardtii metabolism

Gemstone Team Genes to Fuels

Interleukin-15 receptor blockade in non-human primate kidney transplantation.

BACKGROUND: Interleukin (IL)-15 is a chemotactic factor to T cells. It induces proliferation and promotes survival of activated T cells. IL-15 receptor blockade in mouse cardiac and islet allotransplant models has led to long-term engraftment and a regulatory T-cell environment. This study investigated the efficacy of IL-15 receptor blockade using Mut-IL-15/Fc in an outbred non-human primate model of renal allotransplantation. METHODS: Male cynomolgus macaque donor-recipient pairs were selected based on ABO typing, major histocompatibility complex class I typing, and carboxy-fluorescein diacetate succinimidyl ester-based mixed lymphocyte responses. Once animals were assigned to one of six treatment groups, they underwent renal transplantation and bilateral native nephrectomy. Serum creatinine level was monitored twice weekly and as indicated, and protocol biopsies were performed. Rejection was defined as a increase in serum creatinine to 1.5 mg/dL or higher and was confirmed histologically. Complete blood counts and flow cytometric analyses were performed periodically posttransplant; pharmacokinetic parameters of Mut-IL-15/Fc were assessed. RESULTS: Compared with control animals, Mut-IL-15/Fc-treated animals did not demonstrate increased graft survival despite adequate serum levels of Mut-IL-15/Fc. Flow cytometric analysis of white blood cell subgroups demonstrated a decrease in CD8 T-cell and natural killer cell numbers, although this did not reach statistical significance. Interestingly, two animals receiving Mut-IL-15/Fc developed infectious complications, but no infection was seen in control animals. Renal pathology varied widely. CONCLUSIONS: Peritransplant IL-15 receptor blockade does not prolong allograft survival in non-human primate renal transplantation; however, it reduces the number of CD8 T cells and natural killer cells in the peripheral blood.

RECORDING PROJECT: THE COMPLETE (PUBLISHED) PIANO WORKS OF ROBERT HELPS

This recording represents the complete solo piano works of Robert Helps (1928-2001). As of this writing (March, 2008), approx.120 minutes of Helps' solo piano music has been published, all of which is included on the Digital Media (CD). This project includes the following works: Trois Hommages, Quartet, Nocturne, Valse Mirage, In Retrospect, Three Etudes, Portrait, Three Etudes for the Left Hand, Starscape, Recollections, Shall We Dance and Image. (His few remaining pieces are officially "pending publication" and are therefore not included in this project.) Robert Helps, American pianist and composer, enjoyed a successful career on both fronts, teaching at such institutions as San Francisco Conservatory, Stanford University, the University of California, Berkeley, the New England Conservatory, the Manhattan School of Music and Princeton University. Helps, never the recipient of a university or conservatory degree, received private instruction from pianist Abby Whiteside and composer Roger Sessions. His recording of the Sessions' Sonatas is considered to be their benchmark performance. As a composer, he received commission and awards from the American Academy of Arts and Letters, the Ford Foundation, the Guggenheim Foundation and the National Endowment for the Arts. Helps' compositions were anachronistic in style: his compositional style ranges from Post-Impressionism, Neo­ Romanticsim and early 20th century Atonalism, although he never engaged in serial practices. Since his death in 2001, the Robert Helps Trust has been established at the University of South Florida. Funds are being used to support the continued publishing of his scores. The Robert Helps International Composition Competition and Festival was established in 2005.