Transduction of CpG DNA-stimulated primary human B cells with bicistronic lentivectors.

Recently, using HIV-1-derived lentivectors, we obtained efficient transduction of primary human B lymphocytes cocultured with murine EL-4 B5 thymoma cells, but not of isolated B cells activated by CD40 ligation. Coculture with a cell line is problematic for gene therapy applications or study of gene functions. We have now found that transduction of B cells in a system using CpG DNA was comparable to that in the EL-4 B5 system. A monocistronic vector with a CMV promoter gave 32 +/- 4.7% green fluorescent protein (GFP)+ cells. A bicistronic vector, encoding IL-4 and GFP in the first and second cistrons, respectively, gave 14.2 +/- 2.1% GFP+ cells and IL-4 secretion of 1.3 +/- 0.2 ng/10(5) B cells/24 h. This was similar to results obtained in CD34+ cells using the elongation factor-1alpha promoter. Activated memory and naive B cells were transducible. After transduction with a bicistronic vector encoding a viral FLIP molecule, vFLIP was detectable by FACS or Western blot in GFP+, but not in GFP-, B cells, and 57% of sorted GFP+ B cells were protected against Fas ligand-induced cell death. This system should be useful for gene function research in primary B cells and development of gene therapies.

INTEGRAL and XMM-Newton observations towards the unidentified MeV source GRO J1411-64.

The COMPTEL unidentified source GRO J1411-64 was observed by INTEGRAL, and its central part, also by XMM-Newton. The data analysis shows no hint for new detections at hard X-rays. The upper limits in flux herein presented constrain the energy spectrum of whatever was producing GRO J1411-64, imposing, in the framework of earlier COMPTEL observations, the existence of a peak in power output located somewhere between 300-700 keV for the so-called low state. The Circinus Galaxy is the only source detected within the 4$\sigma$ location error of GRO J1411-64, but can be safely excluded as the possible counterpart: the extrapolation of the energy spectrum is well below the one for GRO J1411-64 at MeV energies. 22 significant sources (likelihood $> 10$) were extracted and analyzed from XMM-Newton data. Only one of these sources, XMMU J141255.6-635932, is spectrally compatible with GRO J1411-64 although the fact the soft X-ray observations do not cover the full extent of the COMPTEL source position uncertainty make an association hard to quantify and thus risky. The unique peak of the power output at high energies (hard X-rays and gamma-rays) resembles that found in the SED seen in blazars or microquasars. However, an analysis using a microquasar model consisting on a magnetized conical jet filled with relativistic electrons which radiate through synchrotron and inverse Compton scattering with star, disk, corona and synchrotron photons shows that it is hard to comply with all observational constrains. This and the non-detection at hard X-rays introduce an a-posteriori question mark upon the physical reality of this source, which is discussed in some detail.

Glomerular filtration rate: measure creatinine and height rather than cystatin C!

AIM: Inulin clearance (Cin) is the gold standard for assessing glomerular filtration rate (GFR). Other methods are based on the plasma creatinine concentration (Pcreat), creatinine clearance (Ccreat), the Haycock-Schwartz formula and the plasma concentration of cystatin C (PcysC), a 13 kDa basic protein produced at a constant rate by all nucleated cells. The present prospective study was thus designed to evaluate the reliability of PcysC as a marker of GFR in comparison with that of Pcreat, Ccreat and the Haycock-Schwartz formula, using Cin as the gold standard. METHODS: Ninety-nine children (51 m/48 f), with a median age of 8.3 y (1.0-17.9) were studied. Using a cut-off for Cin of 100 ml/min per 1.73 m2, 54 children (54.5%) had impaired GFR. Those with normal GFR were comparable for age, height, weight and body mass index. RESULTS: Logistic regression, ROC analysis and linear regression all showed that Ccreat was the best parameter to discriminate between impaired and normal GFR, followed by the Haycock-Schwartz formula, PcysC, and finally Pcreat, each one being significantly more predictive than the next. CONCLUSION: GFR is better assessed by the Haycock-Schwartz formula than by PcysC or Pcreat alone. It is therefore concluded that when urine collection is not possible, simply measuring the child's Pcreat and height is the best, easiest and cheapest way to assess GFR.

Prognostic value of fractional flow reserve: linking physiologic severity to clinical outcomes.

BACKGROUND: Fractional flow reserve (FFR) has become an established tool for guiding treatment, but its graded relationship to clinical outcomes as modulated by medical therapy versus revascularization remains unclear. OBJECTIVES: The study hypothesized that FFR displays a continuous relationship between its numeric value and prognosis, such that lower FFR values confer a higher risk and therefore receive larger absolute benefits from revascularization. METHODS: Meta-analysis of study- and patient-level data investigated prognosis after FFR measurement. An interaction term between FFR and revascularization status allowed for an outcomes-based threshold. RESULTS: A total of 9,173 (study-level) and 6,961 (patient-level) lesions were included with a median follow-up of 16 and 14 months, respectively. Clinical events increased as FFR decreased, and revascularization showed larger net benefit for lower baseline FFR values. Outcomes-derived FFR thresholds generally occurred around the range 0.75 to 0.80, although limited due to confounding by indication. FFR measured immediately after stenting also showed an inverse relationship with prognosis (hazard ratio: 0.86, 95% confidence interval: 0.80 to 0.93; p < 0.001). An FFR-assisted strategy led to revascularization roughly half as often as an anatomy-based strategy, but with 20% fewer adverse events and 10% better angina relief. CONCLUSIONS: FFR demonstrates a continuous and independent relationship with subsequent outcomes, modulated by medical therapy versus revascularization. Lesions with lower FFR values receive larger absolute benefits from revascularization. Measurement of FFR immediately after stenting also shows an inverse gradient of risk, likely from residual diffuse disease. An FFR-guided revascularization strategy significantly reduces events and increases freedom from angina with fewer procedures than an anatomy-based strategy.

Accurate calculations of intermolecular interaction energies using explicitly correlated coupled cluster wave functions and a dispersion-weighted MP2 method.

Explicitly correlated coupled-cluster calculations of intermolecular interaction energies for the S22 benchmark set of Jurecka, Sponer, Cerny, and Hobza (Chem. Phys. Phys. Chem. 2006, 8, 1985) are presented. Results obtained with the recently proposed CCSD(T)-F12a method and augmented double-zeta basis sets are found to be in very close agreement with basis set extrapolated conventional CCSD(T) results. Furthermore, we propose a dispersion-weighted MP2 (DW-MP2) approximation that combines the good accuracy of MP2 for complexes with predominately electrostatic bonding and SCS-MP2 for dispersion-dominated ones. The MP2-F12 and SCS-MP2-F12 correlation energies are weighted by a switching function that depends on the relative HF and correlation contributions to the interaction energy. For the S22 set, this yields a mean absolute deviation of 0.2 kcal/mol from the CCSD(T)-F12a results. The method, which allows obtaining accurate results at low cost, is also tested for a number of dimers that are not in the training set.

Oikeuskulttuuri, Lakimiesyhdistys ja WSLT

Puheenvuoro Oikeuskulttuurin maisema -tilaisuudessa Säätytalolla 5.6.1997

The role of bradykinin B(1) and B(2) receptors for secondary brain damage after traumatic brain injury in mice.

Inflammatory mechanisms are known to contribute to the pathophysiology of traumatic brain injury (TBI). Since bradykinin is one of the first mediators activated during inflammation, we investigated the role of bradykinin and its receptors in posttraumatic secondary brain damage. We subjected wild-type (WT), B(1)-, and B(2)-receptor-knockout mice to controlled cortical impact (CCI) and analyzed tissue bradykinin as well as kinin receptor mRNA and protein expression up to 48 h thereafter. Brain edema, contusion volume, and functional outcome were assessed 24 h and 7 days after CCI. Tissue bradykinin was maximally increased 2 h after trauma (P<0.01 versus sham). Kinin B(1) receptor mRNA was upregulated up to four-fold 24 h after CCI. Immunohistochemistry showed that B(1) and B(2) receptors were expressed in the brain and were significantly upregulated in the traumatic penumbra 1 to 24 h after CCI. B(2)R(-/-) mice had significantly less brain edema (-51% versus WT, 24 h; P<0.001), smaller contusion volumes ( approximately 50% versus WT 24 h and 7 d after CCI; P<0.05), and better functional outcome 7 days after TBI as compared with WT mice (P<0.05). The present results show that bradykinin and its B(2) receptors play a causal role for brain edema formation and cell death after TBI.

Erosão hídrica associada a algumas variáveis hidrológicas em pomar de maçã submetido a diferentes manejos do solo

O manejo do solo em cultivos perenes, como pomares, influencia a erosão hídrica. O presente estudo avaliou as perdas de solo e água sob chuva simulada em um Latossolo, em pomar de maçã, entre os meses de agosto de 2007 e abril de 2008, na estação experimental de fruticultura de clima temperado da Embrapa Uva e Vinho, em Vacaria (RS). O trabalho foi conduzido em parcelas experimentais de 3,5 x 11 m, sob chuvas com 1 h de duração e intensidade constante ao longo delas, com variação de 70 a 88 mm h-1 entre uma chuva e outra. Os sistemas de manejo estudados foram: 1) capina manual sob a copa das plantas e solo coberto com gramíneas e leguminosas no restante da área (ST); 2) aveia não dessecada, em que as sementes foram incorporadas ao solo, com capina manual em toda a área, dois meses antes do início dos testes de chuva (AN); 3) aveia dessecada quimicamente sete dias antes do início dos testes de chuva, em que as sementes foram incorporadas ao solo com enxada rotativa em toda a área, dois meses antes do início dos testes (AD); e 4) solo sem cobertura, em que a vegetação, após ter sido dessecada, foi removida da superfície do solo em toda a área com capina manual, um dia antes de iniciar os testes de chuva (SC). A forma de manejo da superfície do solo e o número de chuvas influenciaram a erosão hídrica; as perdas de solo variaram amplamente, enquanto as perdas de água apresentaram menor variação. Apesar da mobilização do solo para implantação de aveia, os tratamentos AN e AD com esse cultivo mostraram a mesma eficácia de controle da erosão em relação ao tratamento ST, em que o solo foi mobilizado apenas sob a copa das plantas. A eliminação da cobertura do solo no tratamento SC aumentou expressivamente as perdas de solo em relação aos tratamentos com cobertura; no que se refere às perdas de água, a diferença foi menor. O tempo de ocorrência do escoamento superficial influenciou a perda de solo; observou-se aumento dessa variável com o aumento do tempo de enxurrada até certo momento, a partir do qual diminuiu, independentemente do tratamento; a perda de água aumentou até certo momento e estabilizou. Houve relação inversa entre razão de perda de solo e razão de perda de água, independentemente do tipo de cobertura do solo e do sistema de manejo sob as macieiras. O modelo exponencial ajustou-se a essa relação.

Erosão hídrica em campo nativo sob diversos manejos: perdas de água e solo e de fósforo, potássio e amônio na água de enxurrada

Os campos nativos do sul do Brasil são utilizados para o pastejo de bovinos e ovinos. Após o verão, no início da estação fria, a fitomassa excedente do pastejo seca, sendo comum sua queima para facilitar o rebrote da pastagem na primavera. No entanto, a queima da fitomassa mineraliza nutrientes e deixa o solo descoberto, potencializando a erosão hídrica. Este trabalho teve o objetivo de quantificar as perdas totais de água e solo e os teores de P, K e NH4+ na água da enxurrada, em campo nativo sobre um Latossolo Bruno aluminoférrico típico. Foram estudados os tratamentos: campo nativo sem queima e sem adubo; campo nativo sem queima e com superfosfato triplo; campo nativo com queima e sem adubo; e campo nativo com queima e com superfosfato triplo. Sobre as parcelas, com 11 m de comprimento, na direção do declive, e 3,5 m de largura, aplicou-se uma chuva simulada com intensidade de 75 mm h-1 e duração de 3 h. A queima influenciou a erosão hídrica e as perdas de nutrientes. A queima da fitomassa do campo nativo diminuiu os tempos de início e pico da enxurrada e a infiltração de água no solo, bem como aumentou a taxa máxima de enxurrada, as perdas de água e solo por erosão hídrica e os teores e as perdas totais de P, K e NH4+ na água de enxurrada, em relação à ausência da queima. A concentração de sedimentos na enxurrada foi maior no tratamento com queima da fitomassa do que no sem queima. O maior e o menor valor da concentração de sedimentos ocorreram em menor tempo de duração do escoamento superficial no tratamento com queima do que no tratamento sem queima, contribuindo para que a perda total de solo fosse 8,9 vezes maior com a queima do que sem a queima do campo. O teor e a perda total de P na água da enxurrada foram maiores no tratamento com adubo fosfatado do que no tratamento sem adubo, tanto no tratamento com queima da fitomassa quanto no sem queima. Para o K e o NH4+, os teores e as perdas totais foram maiores no tratamento com queima do que no sem queima. O modelo exponencial do tipo y = ae-bx ajustou-se aos teores de P, K e NH4+ na água da enxurrada, em relação ao tempo de ocorrência do escoamento superficial (p < 0,01).