Ligand-induced endocytosis and nuclear localization of angiotensin II receptors expressed in CHO cells

A construct (AT1R-NF) containing a "Flag" sequence added to the N-terminus of the rat AT1 receptor was stably expressed in Chinese hamster ovary cells and quantified in the cell membrane by confocal microscopy after reaction with a fluorescein-labeled anti-Flag monoclonal antibody. Angiotensin II bound to AT1R-NF and induced endocytosis with a half-time of 2 min. After 60-90 min, fluorescence accumulated around the cell nucleus, suggesting migration of the ligand-receptor complex to the nuclear membrane. Angiotensin antagonists also induced endocytosis, suggesting that a common step in the transduction signal mechanism occurring after ligand binding may be responsible for the ligand-receptor complex internalization.

The N-terminal 33 amino acid domain of Siva-1 is sufficient for nuclear localization

Siva-1 induces apoptosis in multiple pathological processes and plays an important role in the suppression of tumor metastasis, protein degradation, and other functions. Although many studies have demonstrated that Siva-1 functions in the cytoplasm, a few have found that Siva-1 can relocate to the nucleus. In this study, we found that the first 33 amino acid residues of Siva-1 are required for its nuclear localization. Further study demonstrated that the green fluorescent protein can be imported into the nucleus after fusion with these 33 amino acid residues. Other Siva-1 regions and domains showed less effect on Siva-1 nuclear localization. By site-mutagenesis of all of these 33 amino acid residues, we found that mutants of the first 1-18 amino acids affected Siva-1 nuclear compartmentalization but could not complete this localization independently. In summary, we demonstrated that the N-terminal 33 amino acid residues were sufficient for Siva-1 nuclear localization, but the mechanism of this translocation needs additional investigation.

Localization of metastasis within the sentinel lymph node biopsies: a predictor of additional axillary spread of breast cancer?

PURPOSE: To explore the relationship between morphological characteristics and histologic localization of metastasis within sentinel lymph nodes (SLN) and axillary spread in women with breast cancer. METHODS: We selected 119 patients with positive SLN submitted to complete axillary lymph node dissection from July 2002 to March 2007. We retrieved the age of patients and the primary tumor size. In the primary tumor, we evaluated histologic and nuclear grade, and peritumoral vascular invasion (PVI). In SLNs we evaluated the size of metastasis, their localization in the lymph node, number of foci, number of involved lymph nodes, and extranodal extension. RESULTS: Fifty-one (42.8%) patients had confirmed additional metastasis in non-sentinel lymph nodes (NLSN). High histologic grade, PVI, intraparenchymatous metastasis, extranodal neoplastic extension and size of metastasis were associated with positive NLSN. SLN metastasis affecting the capsule were associated to low risk incidence of additional metastasis. After multivariate analysis, PVI and metastasis size in the SLN remained as the most important risk factors for additional metastasis. CONCLUSIONS:The risk of additional involvement of NSLN is higher in patients with PVI and it increases progressively according the histologic localization in the lymph node, from capsule, where the afferent lymphatic channel arrives, to the opposite side of capsule promoting the extranodal extension. Size of metastasis greater than 6.0 mm presents higher risk of additional lymph node metastasis.

Oxidative stress: molecular perception and transduction of signals triggering antioxidant gene defenses

Molecular oxygen (O2) is the premier biological electron acceptor that serves vital roles in fundamental cellular functions. However, with the beneficial properties of O2 comes the inadvertent formation of reactive oxygen species (ROS) such as superoxide (O2·-), hydrogen peroxide, and hydroxyl radical (OH·). If unabated, ROS pose a serious threat to or cause the death of aerobic cells. To minimize the damaging effects of ROS, aerobic organisms evolved non-enzymatic and enzymatic antioxidant defenses. The latter include catalases, peroxidases, superoxide dismutases, and glutathione S-transferases (GST). Cellular ROS-sensing mechanisms are not well understood, but a number of transcription factors that regulate the expression of antioxidant genes are well characterized in prokaryotes and in yeast. In higher eukaryotes, oxidative stress responses are more complex and modulated by several regulators. In mammalian systems, two classes of transcription factors, nuclear factor kB and activator protein-1, are involved in the oxidative stress response. Antioxidant-specific gene induction, involved in xenobiotic metabolism, is mediated by the "antioxidant responsive element" (ARE) commonly found in the promoter region of such genes. ARE is present in mammalian GST, metallothioneine-I and MnSod genes, but has not been found in plant Gst genes. However, ARE is present in the promoter region of the three maize catalase (Cat) genes. In plants, ROS have been implicated in the damaging effects of various environmental stress conditions. Many plant defense genes are activated in response to these conditions, including the three maize Cat and some of the superoxide dismutase (Sod) genes.

Nuclear calcium signaling: a cell within a cell

Calcium (Ca2+) is a versatile second messenger that regulates a wide range of cellular functions. Although it is not established how a single second messenger coordinates diverse effects within a cell, there is increasing evidence that the spatial patterns of Ca2+ signals may determine their specificity. Ca2+ signaling patterns can vary in different regions of the cell and Ca2+ signals in nuclear and cytoplasmic compartments have been reported to occur independently. No general paradigm has been established yet to explain whether, how, or when Ca2+ signals are initiated within the nucleus or their function. Here we highlight that receptor tyrosine kinases rapidly translocate to the nucleus. Ca2+ signals that are induced by growth factors result from phosphatidylinositol 4,5-bisphosphate hydrolysis and inositol 1,4,5-trisphosphate formation within the nucleus rather than within the cytoplasm. This novel signaling mechanism may be responsible for growth factor effects on cell proliferation.

Átomos na integração: a aproximação Brasil-Argentina no campo nuclear e a construção do Mercosul

Este trabalho tenciona ser uma reflexão sobre a importância da aproximação entre o Brasil e a Argentina no campo nuclear, na segunda metade da década de 80 e no início dos anos 90, como fundamento para a construção de um relacionamento novo que suplantou uma rivalidade histórica entre ambos. O processo de aproximação encetado com um propósito aparentemente econômico, tinha uma dimensão estratégica. As repercussões desse esforço iam além dos interesses bilaterais imediatos e se refletiram na busca de uma nova inserção internacional dos dois países, num momento de transformações profundas nos planos interno e internacional. Através do programa de integração Brasil-Argentina (que mais tarde também envolveu o Uruguai) e do MERCOSUL buscou-se estabelecer uma arquitetura político-jurídica, que criou uma realidade que extrapola o campo econômico e projeta-se no terreno político.

A integração bilateral Brasil-Argentina: tecnologia nuclear e Mercosul

O presente trabalho focaliza o complexo processo de integração bilateral Brasil-Argentina, iniciado formalmente na década de 80, desdobrado em sua dupla abrangência: a) cooperação pacífica da tecnologia nuclear; b) constituição de um mercado comum. A evolução desse processo conduziu-se através de ciclos fundamentais: primeira etapa, antecedentes de aproximação; segunda etapa, os decisivos instrumentos da ampla integração; terceira etapa, solidificação de duas bases, uma de nível tecnológico nuclear, pactuando um sistema sui generis de salvaguardas e a vigência do Tratado de Tlatelolco (emendado), e, outra, de nível econômico pelo Tratado de Assunção, instituindo o Mercosul.

Brazil and the nuclear issues in the years of the Luiz Inácio Lula da Silva government (2003-2010)

The present article is an analysis of Brazilian nuclear diplomacy and of the international relevance of the national nuclear program adopted during the two Presidential terms of Luiz Inácio Lula da Silva. Particular attention will be given to the Brazilian position in the agreement on nuclear non-proliferation, to the role of mediation between Iran and the nuclear powers, and finally to the national ambitions concerning the industrial and military uses of the nuclear technologies.

Comparative study on the localization of adult Schistosoma mansoni worms in albino mice anesthetized with pentobarbital sodium, ether or chlorophorm

The effect of anesthetic drugs on the localization of adult worms in albino mice was compared. The animals with 56 days of infection were anesthetized with pentobarbital sodium, ether or chlorophorm. Perfusion was carried out immediately after, recovering the worms and classifying them in relation to their localization on the liver or portal vein and the mesenteric veins. Our results showed that pentobarbital sodium produced a greater displacement of the worms to the liver (89%) than ether (76%) and chlorophorm (34%) did, when compared to the control group (22%). The difference between pentobarbital sodium and ether was significant (p < 0.05). We suggest that anesthetic drugs may not be used in studies on the distribution of adult worms in several hosts.

PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp.

In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs.

Ressonância nuclear magnética no diagnóstico de pseudoaneurisma de ventrículo esquerdo. Antigo problema, nova opção

Paciente feminina, 67 anos, internada por infarto agudo do miocárdio de parede lateral, com edema agudo de pulmão e evolução para choque cardiogênico nas primeiras horas. Ecocardiograma transesofágico e ressonância nuclear magnética confirmaram o diagnóstico de pseudoaneurisma de ventrículo esquerdo. A paciente foi submetida a tratamento cirúrgico com sucesso.