Caspofungin first-line therapy for invasive aspergillosis in allogeneic hematopoietic stem cell transplant patients: an European Organisation for Research and Treatment of Cancer study.

Caspofungin at standard dose was evaluated as first-line monotherapy of mycologically documented probable/proven invasive aspergillosis (IA) (unmodified European Organisation for Research and Treatment of Cancer/Mycosis Study Group criteria) in allogeneic hematopoietic SCT patients. The primary efficacy end point was complete or partial response at end of caspofungin treatment. Response at week 12, survival and safety were additional end points. Enrollment was stopped prematurely because of low accrual, with 42 enrolled and 24 eligible, giving the study a power of 85%. Transplant was from unrelated donors in 16 patients; acute or chronic GVHD was present in 15. In all, 12 patients were neutropenic (<500/microl) at baseline, 10 received steroids and 16 calcineurin inhibitors or sirolimus. Median duration of caspofungin treatment was 24 days. At the end of caspofungin therapy, 10 (42%) patients had complete or partial response (95% confidence interval: 22-63%); 1 (4%) and 12 (50%) had stable and progressing disease, respectively; one was not evaluable. At week 12, eight patients (33%) had complete or partial response. Survival rates at week 6 and 12 were 79 and 50%, respectively. No patient had a drug-related serious adverse event or discontinued because of toxicity. Caspofungin first-line therapy was effective and well tolerated in allogeneic hematopoietic SCT patients with mycologically documented IA.

An analysis of the benefit of using HEV genotype 3 antigens in detecting anti-HEV IgG in a European population.

BACKGROUND: The benefit of using serological assays based on HEV genotype 3 in industrialised settings is unclear. We compared the performance of serological kits based on antigens from different HEV genotypes. METHODS: Taking 20 serum samples from patients in southwest France with acute HEV infection (positive PCR for HEV genotype 3) and 550 anonymised samples from blood donors in southwest Switzerland, we tested for anti-HEV IgG using three enzyme immunoassays (EIAs) (MP Diagnostics, Dia.Pro and Fortress) based on genotype 1 and 2 antigens, and one immunodot assay (Mikrogen Diagnostik recomLine HEV IgG/IgM) based on genotype 1 and 3 antigens. RESULTS: All acute HEV samples and 124/550 blood donor samples were positive with ≥1 assay. Of PCR-confirmed patient samples, 45%, 65%, 95% and 55% were positive with MP Diagnostics, Dia.Pro, Fortress and recomLine, respectively. Of blood donor samples positive with ≥1 assay, 120/124 (97%), were positive with Fortress, 19/124 (15%) were positive with all EIAs and 51/124 (41%) were positive with recomLine. Of 11/20 patient samples positive with recomLine, stronger reactivity for HEV genotype 3 was observed in 1/11(9%), and equal reactivity for both genotypes in 5/11 (45.5%). CONCLUSIONS: Although recomLine contains HEV genotype 3, it has lower sensitivity than Fortress in acute HEV infection and fails to identify infection as being due to this genotype in approximately 45% of patients. In our single blood donor population, we observe wide variations in measured seroprevalence, from 4.2% to 21.8%, depending on the assay used.

Resistência de genótipos de feijão-de-corda ao pulgão-preto

O objetivo deste trabalho foi avaliar a resistência de genótipos de feijão-de-corda (Vigna unguiculata) ao pulgão-preto (Aphis craccivora). Foram realizados experimentos com e sem chance de escolha em casa de vegetação, na Universidade Federal do Ceará. O delineamento utilizado foi o de blocos ao acaso com vinte tratamentos, representados pelos genótipos 421-07-44, Chumbinho, Zebu, EPACE 10, Frade Preto, Inhumã, João Paulo II, Manteiguinha, Maranhão, Pitiúba, Quarenta Dias, Seridó, Sete Semanas, TVu 1037, TVu 1888, TVu 310, TVu 36, TVu 408P2, TVu 410 e VITA 7. Verificou-se que TVu 408P2, TVu 1037 e TVu 410 foram preteridos por adultos e ninfas do pulgão-preto, em ambos experimentos. Os genótipos TVu 408P2, TVu 410, TVu 36 e TVu 1037 apresentam resistência provavelmente do tipo antibiose ou antixenose. O genótipo 421-07-44 mostrou-se suscetível ao pulgão-preto.

Cuidados de enfermería en el mantenimiento del donante de órganos con muerte encefálica. Los grandes desconocidos

España es líder en donaciones mundialmente, gracias al “Spanish Model”. En este trabajo se da un repaso a todos los ámbitos que envuelven la donación de órganos para su ulterior trasplante: Diagnóstico médico de muerte encefálica, fisiopatología desarrollada tras la muerte encefálica, pero el objetivo principal es desarrollar un plan de cuidados enfermero para el mantenimiento del posible donante de órganos

Hepatitis G virus infection in primary Sjögren's syndrome: analysis in a series of 100 patients.

OBJECTIVE To determine the prevalence and clinical significance of hepatitis G virus (HGV) infection in a large cohort of patients with primary Sjögren¿s syndrome (SS). PATIENTS AND METHODS The study included 100 consecutive patients (92 female and eight male), with a mean age of 62 years (range 31¿80) that were prospectively visited in our unit. All patients fulfilled the European Community criteria for SS and underwent a complete history, physical examination, as well as biochemical and immunological evaluation for liver disease. Two hundred volunteer blood donors were also studied. The presence of HGV-RNA was investigated in the serum of all patients and donors. Aditionally, HBsAg and antibodies to hepatitis C virus were determined. RESULTS Four patients (4%) and six volunteer blood donors (3%) presented HGV-RNA sequences in serum. HGV infection was associated with biochemical signs of liver involvement in two (50%) patients. When compared with primary SS patients without HGV infection, no significant differences were found in terms of clinical or immunological features. HCV coinfection occurs in one (25%) of the four patients with HGV infection. CONCLUSION The prevalence of HGV infection in patients with primary SS is low in the geographical area of the study and HCV coinfection is very uncommon. HGV infection alone does not seen to be an important cause of chronic liver injury in the patients with primary SS in this area.

Proliferative and Osteogenic Differentiation Potentials of Human Fetal Bone Cells

BACKGROUND. Human primary fetal bone cells (hFBC) are being characterized for use in bone tissue regeneration. Unlike human mesenchymal stem cells (hMSC), hFBC are partially differentiated with high expansion and regeneration potential. To date, proliferative and osteoblastic differentiation capacities of fetal bone cells remain poorly examined. The goal of this study was to define an environmental culture conditions for optimal proliferation and production of extracellular bone matrix leading to efficient bone repair. METHODS. Human primary FBC derived from our dedicated, consistent banks of bone cells comprising several fetal donors. For proliferation study, monolayer cultures of both cell types were expanded in DMEM or α- MEM media. Osteoblastic differentiation potentials of both hFBC and hMSC were evaluated through RT-PCR. Regulation of osteogenic differentiation by protein ligands Wnt3a and Wnt5a was studied by ALP enzymatic activity measurement. RESULTS. Evaluation of the proliferation rate demonstrated that hFBC proliferated more rapidly in α-MEM medium. Regarding growth factors that could stimulate cell proliferation rate, we observed that PDGF, FGF2 and Wnt3a had positive effects on proliferation of hFBC. Gene expression analysis demonstrated a higher expression of runx2 in hFBC cultured in basal conditions, which was was similar than that was observed in hMSC in osteoinductive culture conditions. Expression of sox9 was very low in hBFC and hMSC, compared to expression observed in fetal cartilage cells. Looking at osteogenic differentiation capacity, ALP activity was positively regulated byWnt5awhen hFBCwere cultured inα-MEM, but not in DMEM. Conversely, Wnt3a was shown to block the effect of osteogenic inductors on differentiation of both cell types. CONCLUSION. Data presented in this study indicate that the proliferation and differentiation of fetal and mesenchymal stem cells is optimal in α- MEM. Evidence for a pre-differentiated state of hBFC was given by extracellular matrix spontaneous mineralization as well as by higher ALP activity levels observed for these cells in baseline culture conditions, in comparison with hMSC. As we showed that, in vitro, hFBC express a higher capacity to differentiate in osteoblasts, they represent an attractive and promising prospect for fundamental research, and specifically for a new generation of skeletal tissue engineering.

Desempenho de bezerros aleitados com soro de queijo em associação ao colostro

O objetivo deste trabalho foi avaliar o desempenho de bezerros mestiços, aleitados com soro de queijo em associação ao colostro. Foram utilizados 24 bezerros Holandês x Zebu, distribuídos em delineamento inteiramente casualizado, com três tratamentos e oito repetições. Os tratamentos consistiram de: leite integral, 50% de leite integral + 50% de soro de queijo, e 30% de colostro + 70% de soro de queijo. O desenvolvimento dos bezerros foi avaliado até os 60 dias de idade, por meio de pesagens semanais e mensurações corporais. Para a estimativa de consumo, foram feitas medições diárias de consumo da dieta sólida, pelo método oferta/sobra. O desempenho econômico foi avaliado pelo cálculo de indicadores de custos e receitas, e de medidas de resultados econômicos. Não houve influência da dieta líquida sobre o consumo de matéria seca e sobre as médias de medidas corporais dos bezerros. Foram observadas diferenças significativas no ganho de peso dos animais na segunda e na quarta semanas de vida. A dieta com colostro apresentou o maior lucro adicional em comparação ao tratamento controle. A associação do soro de queijo ao colostro é a opção mais vantajosa economicamente, já que a substituição de 100% do leite por esses subprodutos não prejudicou o desempenho dos animais.

Características de carcaça e qualidade da carne de novilhas alimentadas com silagem de capim-marandu

O objetivo deste trabalho foi avaliar as características de carcaça e da carne de novilhas ¾ Zebu x Holandês submetidas a dieta com substituição da silagem de sorgo por silagem de capim-marandu. Utilizou-se o delineamento experimental inteiramente casualizado. Os tratamentos consistiram da substituição da fração volumosa da dieta de 100, 70 e 30% de silagem de sorgo por, respectivamente, 100, 70 e 30% de silagem de capim-marandu. Foi adicionado concentrado na proporção de 1,2% de peso corporal, e as dietas foram isoproteicas. A substituição da silagem de sorgo pela silagem de capim-marandu aumentou a perda de água por cozimento até o ponto máximo de 49,98% de substituição, elevou a força de cisalhamento e a capacidade de retenção de água da carne, mas não influenciou os pesos e os rendimentos das carcaças, os percentuais de cortes, as medidas de carcaça, a área de olho de lombo, o pH das carcaças, a espessura de gordura e as características de cor de músculo e gordura. A substituição da silagem de sorgo por silagem de capim-marandu diminui o ganho de peso de corpo vazio e reduz a maciez da carne de novilhas ¾ Zebu x Holandês, mas não afeta as características quantitativas da carcaça.

Proteomics of methylene blue photo-treated plasma before and after removal of the dye by an absorbent filter.

Methylene blue (MB) and light are used for virus inactivation of plasma for transfusion. However, the presence of MB has been the subject of concern, and efforts have been made to efficiently remove the dye after photo-treatment. For this study, plasma was collected by apheresis from 10 donors (group A), then treated using the MacoPharma THERAFLEX procedure (MB; 1 microM, and light exposure; 180 J/cm(2)) (group B), and finally filtered in order to remove the dye (group C). Proteins were analyzed by two-dimensional electrophoresis, and peptides showing modifications were characterized by mass spectrometry. Clottable and antigenic fibrinogen levels, as well as fibrin polymerization time were measured. Analyses of the gels focused on a region corresponding to pI between 4.5 and 6.5, and M(r) from 7000 to 58 000. In this area, 387 +/- 47 spots matched, and four of these spots presented significant modifications. They corresponded to changes of the gamma-chain of fibrinogen, of transthyretin, and of apolipoprotein A-I, respectively. A decrease of clottable fibrinogen and a prolongation of fibrin polymerization time were observed in groups B and C. Removal of MB by filtration was not responsible for additional protein alterations. The effect of over-treatment of plasma by very high concentrations of MB (50 microM) in association with prolonged light exposure (3 h) was also analyzed, and showed complex alterations of most of the plasma proteins, including fibrinogen gamma-chain, transthyretin, and apolipoprotein A-I. Our data indicates that MB treatment at high concentration and prolonged illumination severely injure plasma proteins. By contrast, at the MB concentration used to inactivate viruses, damages are apparently very restricted.

Forage production of elephant grass under intermittent stocking

The objective of this work was to evaluate the dry matter production of elephant grass (Pennisetum purpureum) genotypes, managed under intermittent stocking. A completely randomized design was used, with two genotypes and three replicates. The treatments consisted of factorial combinations (2x2x2) of genotypes ('BRS Kurumi' and the clone CNPGL 00‑1‑3), two light interception levels (LI) at the onset of grazing (90 and 95%), and two post‑grazing canopy heights (30 and 50 cm). A total of 24 Holstein x Zebu crossbred heifers were used. The stocking density varied in order to finish the grazing periods in two days. The interval between the defoliation, based on 95% LI, resulted in a higher leaf mass per grazing cycle. The post‑grazing height of 30 cm did not affect the number of grazing cycles but provided a greater herbage accumulation rate. The cultivar BRS Kurumi has higher pasture growth, lower rest period, and greater number of grazing cycles, which results in increased forage production in the growing season.

Extracellular-signal-regulated kinase 5 modulates the antioxidant response by transcriptionally controlling Sirtuin 1 expression in leukemic cells.

Cancer cell metabolism differs from that of non-transformed cells in the same tissue. This specific metabolism gives tumor cells growing advantages besides the effect in increasing anabolism. One of these advantages is immune evasion mediated by a lower expression of the mayor histocompatibility complex class I molecules. The extracellular-signal-regulated kinase-5 regulates both mayor histocompatibility complex class I expression and metabolic activity. However, the mechanisms underlying are largely unknown. We show here that extracellular-signal-regulated kinase-5 regulates the transcription of the NADH(+)-dependent histone deacetylase silent mating type information regulation 2 homolog 1 (Sirtuin 1) in leukemic Jurkat T cells. This involves the activation of the transcription factor myocyte enhancer factor-2 and its binding to the sirt1 promoter. In addition, extracellular-signal-regulated kinase-5 is required for T cell receptor-induced and oxidative stress-induced full Sirtuin 1 expression. Extracellular-signal-regulated kinase-5 induces the expression of promoters containing the antioxidant response elements through a Sirtuin 1-dependent pathway. On the other hand, down modulation of extracellular-signal-regulated kinase-5 expression impairs the anti-oxidant response. Notably, the extracellular-signal-regulated kinase-5 inhibitor BIX02189 induces apoptosis in acute myeloid leukemia tumor cells without affecting T cells from healthy donors. Our results unveil a new pathway that modulates metabolism in tumor cells. This pathway represents a promising therapeutic target in cancers with deep metabolic layouts such as acute myeloid leukemia.

Telomere length dynamics in normal individuals and in patients with hematopoietic stem cell-associated disorders.

The telomere length in nucleated peripheral blood (PB) cells indirectly reflects the mitotic history of their precursors: the hematopoietic stem cells (HSCs). The average length of telomeres in PB leukocytes can be measured using fluorescence in situ hybridization and flow cytometry (flow FISH). We previously used flow FISH to characterize the age-related turnover of HSCs in healthy individuals. In this review, we describe results of recent flow FISH studies in patients with selected hematopoietic stem cell-associated disorders: chronic myelogenous leukemia (CML) and several bone marrow failure syndromes. CML is characterized by a marked expansion of myeloid Philadelphia chromosome positive (Ph+) cells. Nevertheless, nonmalignant (Ph-) HSCs typically coexist in the bone marrow of CML patients. We analyzed the telomere length in > 150 peripheral blood leukocytes (PBLs) and bone marrow samples of patients with CML as well as samples of Ph- T-lymphocytes. Compared to normal controls, the overall telomere fluorescence in PBLs of patients with CML was significantly reduced. However, no telomere shortening was observed in Ph- T-lymphocytes. Patients in late chronic phase (CP) had significantly shorter telomeres than those assessed earlier in CP. Our data suggest that progressive telomere shortening is correlated with disease progression in CML. Within the group of patients with bone marrow failure syndromes, we only found significantly shortened telomeres (compared to age-adjusted controls) in granulocytes from patients with aplastic anemia (AA). Strikingly, the telomere length in granulocytes from AA patients who had recovered after immunosuppressive therapy (recAA) did not differ significantly from controls, whereas untreated patients and nonresponders with persistent severe pancytopenia (sAANR) showed marked and significant telomere shortening compared to healthy donors and patients with recAA. Furthermore, an inverse correlation between age-adjusted telomere length and peripheral blood counts was found in support of a model in which the degree of cytopenia and the amount of telomere shortening are correlated. These results support the concept of extensive proliferation of HSCs in subgroups of AA patients and suggest a potential use of telomere-length measurements as a prognostic tool in this group of disorders as well.

Biotechnology in Lausanne: the Rh D project.

Hemolytic disease of the newborn is an often fatal condition of some newborn babies due to the immunogenicity of their Rh D positive erythrocytes in the Rh D negative mother. This condition can be prevented by injecting anti-Rh D antibodies. The current source of these antibodies is blood from immunized human donors. In order to avoid problems with limited supply and donor safety, the Rh D project was set up to develop recombinant monoclonal anti-Rh D antibodies as a possible replacement. In a multidisciplinary collaboration between the Zentrallaboratorium Blutspendedienst (ZlB) of the Swiss Red Cross, the Center of Biotechnology of the University and the EPFL (CBUE), and the Institute of Chemical and Biochemical Engineering (EPFl), co-funded by the Swiss National Science Foundation and ZLB, a candidate monoclonal anti-Rh D antibody has been selected, expressed in CHO cells, and a manufacturing process for large-scale production has been developed.

Isolation and in vitro chondrogenic potential of human foetal spine cells.

Cell therapy for nucleus pulposus (NP) regeneration is an attractive treatment for early disc degeneration as shown by studies using autologous NP cells or stem cells. Another potential source of cells is foetal cells. We investigated the feasibility of isolating foetal cells from human foetal spine tissues and assessed their chondrogenic potential in alginate bead cultures. Histology and immunohistochemistry of foetal tissues showed that the structure and the matrix composition (aggrecan, type I and II collagen) of foetal intervertebral disc (IVD) were similar to adult IVD. Isolated foetal cells were cultured in monolayer in basic media supplemented with 10% Fetal Bovine Serum (FBS) and from each foetal tissue donation, a cell bank of foetal spine cells at passage 2 was established and was composed of around 2000 vials of 5 million cells. Gene expression and immunohistochemistry of foetal spine cells cultured in alginate beads during 28 days showed that cells were able to produce aggrecan and type II collagen and very low level of type I and type X collagen, indicating chondrogenic differentiation. However variability in matrix synthesis was observed between donors. In conclusion, foetal cells could be isolated from human foetal spine tissues and since these cells showed chondrogenic potential, they could be a potential cell source for IVD regeneration.