Generation of CTL responses using Kunjin replicon RNA

The Kunjin replicon was used to express a polytope that consisted of seven hepatitis C virus cytotoxic T lymphocyte epitopes and one influenza cytotoxic T lymphocyte epitope for vaccination studies. The self-replicating nature of, and expression from, the ribonucleic acid was confirmed in vitro . Initial vaccinations with one dose of Kun-Poly ribonucleic acid showed that an influenza-specific cytotoxic T lymphocyte response was elicited more efficiently by intradermal inoculation compared with intramuscular delivery. Two micrograms of ribonucleic acid delivered in the ear pinnae of mice was sufficient to elicit a detectable cytotoxic T lymphocyte response 10 days post-vaccination. Further vaccination studies showed that four of the seven hepatitis C virus cytotoxic T lymphocyte epitopes were able to elicit weak cytotoxic T lymphocyte responses whereas the influenza epitope was able to elicit strong, specific cytotoxic T lymphocyte responses following three doses of Kun-Poly ribonucleic acid. These studies vindicate the use of the Kunjin replicon as a vector to deliver encoded proteins for the development of cell-mediated immune responses.

Amino acids 1-20 of the hepatitis C virus (HCV) core protein specifically inhibit HCV IRES- dependent translation in Hep G2 cells, and inhibit both HCV IRES- and cap-dependent translation in HuH7 and CV-1 cells.

A self-modulating mechanism by the hepatitis C virus (HCV) core protein has been suggested to influence the level of HCV replication, but current data on this subject are contradictory. We examined the effect of wild-type and mutated core protein on HCV IRES- and cap-dependent translation. The wild-type core protein was shown to inhibit both IRES- and cap-dependent translation in an in vitro system. This effect was duplicated in a dose-dependent manner with a synthetic peptide representing amino acids 1-20 of the HCV core protein. This peptide was able to bind to the HCV IRES as shown by a mobility shift assay. In contrast, a peptide derived from the hepatitis B virus (HBV) core protein that contained a similar proportion of basic residues was unable to inhibit translation or bind the HCV IRES. A recombinant vaccinia-HCV core virus was used to examine the effect of the HCV core protein on HCV IRES-dependent translation in cells and this was compared with the effects of an HBV core-recombinant vaccinia virus. In CV-1 and HuH7 cells, the HCV core protein inhibited translation directed by the IRES elements of HCV, encephalomyocarditis virus and classical swine fever virus as well as cap-dependent translation, whereas in HepG2 cells, only HCV IRES-dependent translation was affected. Thus, the ability of the HCV core protein to selectively inhibit HCV IRES-dependent translation is cell-specific. N-terminal truncated (aa 1-20) HCV core protein that was expressed from a novel recombinant vaccinia virus in cells abrogated the inhibitory phenotype of the core protein in vivo, consistent with the above in vitro data.

Nerve growth factor overexpression and autocrine loop in breast cancer cells

We show here that nerve growth factor (NGF), the canonical neurotrophic factor, is synthesized and released by breast cancer cells. High levels of NGF transcript and protein were detected in breast cancer cells by reverse transcription-PCR, Western blotting, ELISA assay and immunohistochemistry. Conversely, NGF production could not be detected in normal breast epithelial cells at either the transcriptional or protein level. Confocal analysis indicated the presence of NGF within classical secretion vesicles. Breast cancer cell-produced NGF was biologically active, as demonstrated by its ability to induce the neuronal differentiation of embryonic neural precursor cells. Importantly, the constitutive growth of breast cancer cells was strongly inhibited by either NGF-neutralizing antibodies or K-252a, a pharmacological inhibitor of NGF receptor TrkA, indicating the existence of an NGF autocrine loop. Together, our data demonstrate the physiological relevance of NGF in breast cancer and its potential interest as a marker and therapeutic target.

Left ventricular dyssynchrony predicts benefit of cardiac resynchronization therapy in patients with end-stage heart failure before pacemaker implantation

We evaluated patients with end-stage heart failure who have a high likelihood of response to cardiac resynchronization therapy (biventricular pacing). It appears that 20% of patients do not respond to this expensive therapy despite the use of selection criteria (dilated cardiomyopathy, heart failure, New York Heart Association class II or IV, left ventricular election fraction 120 ms). The presence of left ventricular dys-synchrony is needed to result in improvement after cardiac resynchronization therapy. (C)2003 by Excerpta Medica, Inc.

Qualidade tecnológica do arroz branco polido submetido à irradiação gama (Co60)

A aplicação da radiação ionizante, com o propósito de garantir a segurança dos grãos, tem sido apresentada como alternativa eficaz e segura ao uso de defensivos agrícolas e prática promissora, a ser utilizada para estender a vida útil e reduzir as perdas das safras de grãos. Contudo, assim como outros tratamentos de conservação de alimentos, a irradiação pode alterar sua qualidade. Este trabalho foi realizado com o objetivo de avaliar a qualidade de cocção do arroz submetido a diferentes doses de irradiação gama Co60 (0,0 kGy, 6,5 kGy e 7,5 kGy). Observou-se que o arroz irradiado com 7,5 kGy apresentou um tempo de cozimento significativamente maior (18,21 min.) que o do arroz irradiado na dose 6,5 kGy (17,11 min) e 0,0 kGy (16,83 min.). Os valores médios de expansão do volume do arroz não irradiado foi de 365,65%; já, para o arroz irradiado, os valores foram menores, apresentando-se com 322,19%. Quanto à perda de sólidos solúveis, essa foi significativamente maior no arroz branco polido irradiado (17,0%), do que no arroz não irradiado (10,35%). Embora a irradiação gama tenha influenciado em alguns parâmetros de cocção do arroz, refletindo-se em maior tempo de cozimento, para a irradiação na dose 7,5 kGy, no menor coeficiente de expansão do volume e na maior perda de sólidos solúveis, os valores encontrados assemelham-se aos valores referenciados, na literatura, como desejáveis para a qualidade deste cereal.

PÁSCOA, CRUZADOS E GENOCÍDIOS (M/18)

“Acredita-se que mais de cem milhões de cristãos enfrentam perseguições diariamente em todo o mundo por causa da sua fé em Jesus Cristo”, Eric Metaxas, “comparou a atitude da maior parte das lideranças cristãs com o silêncio dos cristão no tempo da Alemanha nazi”, Metaxas escreveu uma biografia de Dietrich Bonhoeffer, torturado e morto pelos nazis – 1906/45, foi um teólogo pastor luterano e membro da resistência anti-nazi, tendo estado envolvido com o Almirante Canaris da Abwehr para assassinar Hitler. Bonhoeffer, que foi um crítico das perseguições aos judeus, tinha por lema: “O silêncio diante do mal é o próprio mal”. § "It is believed that over one hundred million Christians face persecution every day around the world because of their faith in Jesus Christ," Eric Metaxas, "compared the attitude of most Christian leaders with Christian of silence in the German time Nazi, "Metaxas wrote a biography of Dietrich Bonhoeffer, tortured and killed by the Nazis - 1906/45, was a pastor Lutheran theologian and member of the anti-Nazi resistance, having been involved with Admiral Canaris of the Abwehr to assassinate Hitler. Bonhoeffer, who was a critic of the persecution of Jews, had a motto: "Silence in the face of evil is evil itself."