Preoperative localization and surgical margins in conservative breast surgery.

Breast-conserving surgery (BCS) is the treatment of choice for early breast cancer. The adequacy of surgical margins (SM) is a crucial issue for adjusting the volume of excision and for avoiding local recurrences, although the precise definition of an adequate margins width remains controversial. Moreover, other factors such as the biological behaviour of the tumor and subsequent proper systemic therapies may influence the local recurrence rate (LRR). However, a successful BCS requires preoperative localization techniques or margin assessment techniques. Carbon marking, wire-guided, biopsy clips, radio-guided, ultrasound-guided, frozen section analysis, imprint cytology, and cavity shave margins are commonly used, but from the literature review, no single technique proved to be better among the various ones. Thus, an association of two or more methods could result in a decrease in rates of involved margins. Each institute should adopt its most congenial techniques, based on the senologic equipe experience, skills, and technologies.

Persistent hiccup after surgical resection of a brainstem arteriovenous malformation: a case successfully treated with gabapentin during rehabilitation. Case report.

Persistent hiccup rarely occurs during rehabilitation, but its management can prove to be very difficult, particularly in presence of associated dysphagia, requiring longer hospitalization and higher risk of severe clinical complications. We present a case of persistent hiccup after surgical resection of a brainstem arteriovenous malformation successfully treated with gabapentin during rehabilitation.

The self-association coiled-coil domain of PML is sufficient for the oncogenic conversion of the retinoic acid receptor (RAR) alpha.

In acute promyelocytic leukemia (APL) the retinoic acid receptor alpha (RARα) becomes an oncogene through the fusion with several partners, mostly with promyelocytic leukemia protein (PML), all of which have in common the presence of a self-association domain. The new fusion proteins, therefore, differently from the wild-type RARα, which forms only heterodimers with retinoic X receptor alpha, are also able to homo-oligomerize. The presence of such a domain has been suggested to be crucial for the leukemogenic potential of the chimeric proteins found in APL blasts. Whether or not any self-association domain is sufficient to bestow a leukemogenic activity on RARα is still under investigation. In this work, we address this question using two different X-RARα chimeras, where X represents the coiled-coil domain of PML (CC-RARα) or the oligomerization portion of the yeast transcription factor GCN4 (GCN4-RARα). We demonstrate that in vitro both proteins have transforming potential, and recapitulate the main PML-RARα biological properties, but CC-RARα is uniquely able to disrupt PML nuclear bodies. Indeed, in vivo only the CC-RARα chimera induces efficiently APL in a murine transplantation model. Thus, the PML CC domain represents the minimal structural determinant indispensable to transform RARα into an oncogenic protein.

Blood parasites, total plasma protein and packed cell volume of small wild mammals trapped in three mountain ranges of the Atlantic Forest in Southeastern Brazil.

A study of blood parasites in small wild non-flying mammals was undertaken in three areas of the Atlantic Forest in Southeastern Brazil: Serra de Itatiaia, RJ, Serra da Bocaina, SP and Serra da Fartura, SP, from June 1999 to May 2001. A total of 450 animals (15 species) were captured in traps and it was observed in 15.5% of the blood smears the presence of Haemobartonella sp. and Babesia sp. in red blood cells. There was no statistically significant difference between parasited and non-parasited specimens regarding total plasma protein, packed cell volume and body weight, which strongly suggests that these specimens might be parasite reservoirs.

Blood parasites, total plasma protein and packed cell volume of small wild mammals trapped in three mountain ranges of the Atlantic Forest in Southeastern Brazil

A study of blood parasites in small wild non-flying mammals was undertaken in three areas of the Atlantic Forest in Southeastern Brazil: Serra de Itatiaia, RJ, Serra da Bocaina, SP and Serra da Fartura, SP, from June 1999 to May 2001. A total of 450 animals (15 species) were captured in traps and it was observed in 15.5 percent of the blood smears the presence of Haemobartonella sp. and Babesia sp. in red blood cells. There was no statistically significant difference between parasited and non-parasited specimens regarding total plasma protein, packed cell volume and body weight, which strongly suggests that these specimens might be parasite reservoirs.

A presença de hemoparasitos em pequenos mamíferos silvestres não voadores foi pesquisada em animais de três áreas serranas do Sudeste brasileiro, pertencentes ao complexo da Serra do Mar e da Serra da Mantiqueira, nos Estados de São Paulo, Rio de Janeiro e Minas Gerais. Foram capturados 420 animais de 15 espécies, durante dois anos, dos quais, 15,5 por cento apresentaram Haemobartonella sp. e Babesia sp., observadas em lâmina de esfregaço sangüíneo no interior de suas hemácias. Os níveis de proteína total plasmática e de volume globular não apresentaram diferença estatisticamente significativa entre os indivíduos parasitados e não parasitados, assim como o peso corporal, o que sugere fortemente que esses animais possam ser reservatórios desses parasitos.

A redundant oncogenic potential of the retinoic receptor (RAR) alpha, beta and gamma isoforms in acute promyelocytic leukemia.

Alterations of the retinoic acid receptor (RAR)alpha locus are found in 100% of acute promyelocytic leukemia patients, where chromosomal translocations generate the promyelocytic leukemia (PML)-RARalpha chimeric protein. Here, we have investigated the biological properties of the other RAR isoforms (RARbeta and RARgamma), through the generation and characterization of artificial PML-RAR'x' fusion proteins. Surprisingly, we found that all of the RAR isoforms share an identical oncogenic potential in vitro, thus implying that the selection of the RARalpha locus in leukemia patients must occur--rather than through functional differences among the various RAR isoforms-as the consequence of the nuclear architecture of the different RAR loci.

Pattern of elevational distribution and richness of non volant mammals in Itatiaia National Park and its surroundings, in southeastern Brazil.

Itatiaia National Park (PNI) and its surroundings present a unique fauna due to different forest formations with well-defined climatic and vegetation bands. The Itatiaia massif has four vegetation types that follow an altitudinal gradient: lower montane forest, montane forest, upper montane forest, and Campos de Altitude. Hence, this region is ideal for studying geographical variation in biological diversity. The main objectives of this study were to report on nonvolant mammal species known to occur in Itatiaia National Park and its surroundings and to determine if their distributional pattern is related to elevation. A review of the literature and a complete survey of specimens deposited in museums, as well as small-mammal trapping were carried out in order to obtain a complete record of the species from the region. Precise locality data were obtained for all specimens recorded, allowing the inclusion of each collected or observed individual in an altitude and vegetational class. We made a direct ordination gradient of marsupial, primate, and rodent species abundance with the altitude. Sixty-nine mammal species were collected or reported for the Itatiaia massif, belonging to seven orders and 20 families. Of these, 33 species (47.8%) are included in the official list of threatened or believed-to-be threatened species in Rio de Janeiro State. The orders Rodentia, Carnivora, and Didelphimorphia had the highest species richness, with 25, 14, and 13 species respectively. When species were grouped according to the vegetation, 16 species occured in the lower montane, 56 in the montane forest, five in the upper montane, and 21 in the high-altitude fields (Campos de Altitude). The communities of marsupials, primates, and rodents have an ordination pattern related to the altitude. Species richness was higher between 500 m and 1,500 m above sea level in montane forest, which is in agreement with recent studies showing that species richness can reach its maximum at mid-elevations.

Pattern of elevational distribution and richness of non volant mammals in Itatiaia National Park and its surroundings, in Southeastern Brazil

O Parque Nacional do Itatiaia e seu entorno apresenta fauna peculiar em razão de diferentes formações vegetais, consistindo em faixas definidas de clima e de vegetação. O maciço do Itatiaia tem quatro tipos de vegetação que seguem um gradiente altitudinal: Floresta Submontana, Floresta Montana, Floresta Alto-Montana e Campos de Altitude. Por isso, essa região é ideal para estudar variação geográfica na diversidade biológica. O principal objetivo deste estudo foi relatar as espécies de mamíferos não-voadores conhecidas para o Parque Nacional do Itatiaia e seu entorno e determinar se elas apresentam padrão de distribuição altitudinal. Foram realizados revisão completa da literatura e levantamento de espécimes depositados em museus, bem como esforços de captura de pequenos mamíferos, a fim de obter o levantamento completo das espécies da região. Dados precisos sobre as localidades foram obtidos para todos os espécimes levantados, permitindo a colocação de cada indivíduo coletado ou observado dentro de uma classe de altitude e de vegetação. Foi elaborado gradiente de ordenação direto da abundância de espécies de marsupial, de primatas e de roedores com a altitude. Foram coletadas ou relatadas 69 espécies de mamíferos para o maciço do Itatiaia, pertencendo a 7 ordens e 20 famílias. Dessas, 33 espécies (47,8%) estão incluídas na lista oficial de espécies ameaçadas de extinção, ou presumivelmente ameaçadas, do Estado do Rio de Janeiro. As ordens Rodentia, Carnivora e Didelphimorphia apresentaram as mais altas riquezas de espécies, com 25, 14 e 13 espécies, respectivamente. Quando agrupadas de acordo com a vegetação, 16 espécies ocorreram na Floresta Submontana, 56 ocorreram na Floresta Montana, 5, na Floresta Alto-Montana e 21, nos Campos de Altitude. As comunidades de marsupiais, primatas e roedores têm padrão de ordenação relacionado com a altitude. A riqueza de espécies foi maior entre 500 m e 1.500 m, na Floresta Montana, o que está de acordo com estudos recentes que mostram que a riqueza de espécies alcança seu valor máximo em elevações medianas.

Pattern of elevational distribution and richness of non volant mammals in Itatiaia National Park and its surroundings, in Southeastern Brazil

Itatiaia National Park (PNI) and its surroundings present a unique fauna due to different forest formations with well-defined climatic and vegetation bands. The Itatiaia massif has four vegetation types that follow an altitudinal gradient: lower montane forest, montane forest, upper montane forest, and Campos de Altitude. Hence, this region is ideal for studying geographical variation in biological diversity. The main objectives of this study were to report on nonvolant mammal species known to occur in Itatiaia National Park and its surroundings and to determine if their distributional pattern is related to elevation. A review of the literature and a complete survey of specimens deposited in museums, as well as small-mammal trapping were carried out in order to obtain a complete record of the species from the region. Precise locality data were obtained for all specimens recorded, allowing the inclusion of each collected or observed individual in an altitude and vegetational class. We made a direct ordination gradient of marsupial, primate, and rodent species abundance with the altitude. Sixty-nine mammal species were collected or reported for the Itatiaia massif, belonging to seven orders and 20 families. Of these, 33 species (47.8%) are included in the official list of threatened or believed-to-be threatened species in Rio de Janeiro State. The orders Rodentia, Carnivora, and Didelphimorphia had the highest species richness, with 25, 14, and 13 species respectively. When species were grouped according to the vegetation, 16 species occured in the lower montane, 56 in the montane forest, five in the upper montane, and 21 in the high-altitude fields (Campos de Altitude). The communities of marsupials, primates, and rodents have an ordination pattern related to the altitude. Species richness was higher between 500 m and 1,500 m above sea level in montane forest, which is in agreement with recent studies showing that species richness can reach its maximum at mid-elevations.

O Parque Nacional do Itatiaia e seu entorno apresenta fauna peculiar em razão de diferentes formações vegetais, consistindo em faixas definidas de clima e de vegetação. O maciço do Itatiaia tem quatro tipos de vegetação que seguem um gradiente altitudinal: Floresta Submontana, Floresta Montana, Floresta Alto-Montana e Campos de Altitude. Por isso, essa região é ideal para estudar variação geográfica na diversidade biológica. O principal objetivo deste estudo foi relatar as espécies de mamíferos não-voadores conhecidas para o Parque Nacional do Itatiaia e seu entorno e determinar se elas apresentam padrão de distribuição altitudinal. Foram realizados revisão completa da literatura e levantamento de espécimes depositados em museus, bem como esforços de captura de pequenos mamíferos, a fim de obter o levantamento completo das espécies da região. Dados precisos sobre as localidades foram obtidos para todos os espécimes levantados, permitindo a colocação de cada indivíduo coletado ou observado dentro de uma classe de altitude e de vegetação. Foi elaborado gradiente de ordenação direto da abundância de espécies de marsupial, de primatas e de roedores com a altitude. Foram coletadas ou relatadas 69 espécies de mamíferos para o maciço do Itatiaia, pertencendo a 7 ordens e 20 famílias. Dessas, 33 espécies (47,8%) estão incluídas na lista oficial de espécies ameaçadas de extinção, ou presumivelmente ameaçadas, do Estado do Rio de Janeiro. As ordens Rodentia, Carnivora e Didelphimorphia apresentaram as mais altas riquezas de espécies, com 25, 14 e 13 espécies, respectivamente. Quando agrupadas de acordo com a vegetação, 16 espécies ocorreram na Floresta Submontana, 56 ocorreram na Floresta Montana, 5, na Floresta Alto-Montana e 21, nos Campos de Altitude. As comunidades de marsupiais, primatas e roedores têm padrão de ordenação relacionado com a altitude. A riqueza de espécies foi maior entre 500 m e 1.500 m, na Floresta Montana, o que está de acordo com estudos recentes que mostram que a riqueza de espécies alcança seu valor máximo em elevações medianas.

A functional assessment methodology for alcohol dependent patients undergoing rehabilitative treatments.

PURPOSE: We propose a functional assessment approach for patients with alcoholic dependence of working age undergoing aerobic training. The background is the WHO indication (ICIDH-2) to use measurable 'activities' as a means to assess the individual 'participation' in social life which also implies work capacity. Defining sustainable energetic levels for the individual is an important issue for both the quantification of an effective training and the evaluation of possible improvements following training. METHODS: Fifty-six 'alcohol dependent' patients, as defined by DSM IV (Diagnostic and Statistical Manual of Mental Disorders), admitted to our Unit in a 16 month-period participated in the study. Eighteen healthy subjects served as controls (Group C). Out of all the 56 patients, 33 (Group A) underwent an aerobic training and 23 subjects (Group N) underwent the same pharmacological and psychological therapy but without aerobic training. Patients were assigned to the treatment (A) or no treatment (N) group according to a 'quasi-experimental' design (i.e. temporal selection criteria). The evaluation protocol consisted of submaximal symptom-limited tests. The tests consisted of bouts of 'basic' activities (walking, lifting, arm-work) to be performed at different intensities. We estimated the total energetic work (TW) performed in the tests by means of formulas available in the literature. The maximal energetic intensity (EI) reached during the tests was also estimated and expressed in MET (multiple of the basal metabolism). RESULTS: Significant differences in work capacity were observed between patients and healthy subjects at baseline. Group A significantly increased TW after rehabilitation, while Group N did not increment their performance at the re-test. CONCLUSIONS: The proposed approach could be useful in the functional assessment of deconditioned subjects with alcohol dependence in working age, and could monitor the changes in work capacity following training.

A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.

The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation.

Red cell physiology.

The erythrocyte, the ultimate product of mammalian erythroid maturation, appears as a highly specialized and, paradoxically, a very simplified cell. In fact it lacks a nucleus and the intracellular organelles are essentially designed to transport oxygen from the outer environment to respiring tissues through the sophisticated functional properties of intraerythrocytic hemoglobin. In this respect, since oxygen transport is such a vital process, the red cell has often been considered, in an excess of oversimplification, as merely a "biological bag' enveloping a viscous solution of concentrated hemoglobin and containing only those few enzymes which are needed to maintain the cell functionally active. However, within the cell a number of different processes are contemporaneously going on, hemoglobin acts as an oxygen and carbon dioxide transporter, glycolysis and the pentose phosphate shunt are devoted to the production of ATP and NADPH, respectively, and membrane organization provides the cell with a good deformability, allowing it to cross narrow splenic capillaries and channels without any appreciable damage for several weeks of activity. All these processes are, indeed, highly integrated and concur to define a complex scenery centered on the oxygenation-deoxygenation cycle of hemoglobin. Within this emerging scheme, hemoglobin appears to display, besides the basic function of oxygen transport, several other biological functions which are driven by the oxygen-linked conformational transition and whose relative importance, in the economy of the cell and of the organism, is not easy to qualify. Some of these aspects are described and discussed.

Hemolytic anemias due to disorders of red cell membrane skeleton.

During the past 10 years, knowledge of the composition, function and supramolecular assembly of the red cell membrane has been greatly expanded by progress in molecular and cell biology. Detailed information on the organization of membrane cytoskeletal proteins and their molecular characterization has allowed us to correlate a number of protein abnormalities with clinical symptoms that are peculiar to hereditary hemolytic anemias (HHA). In particular, three general principles emerge that can help us to understand the pathogenetic mechanisms of HHA: (a) protein-protein and protein-lipid interactions greatly influence the correct assembly of the membrane skeleton; (b) the red blood cell (RBC) membrane skeleton mostly determines the shape (discocyte), deformability (rheologic properties) and durability (half-life and resistence to shear stress) of the erythrocytes; (c) changes in cytoskeletal composition and/or organization can produce alterations in all of the above properties, and therefore they are responsible for the onset of the hemolytic damage.

The effect of magnesium on glycolysis of permeabilized Ehrlich ascites tumor cells.

We have previously observed that extracellular Mg2+ influences the phosphofructokinase (PFK) activity of intact Ehrlich Ascites tumour cells (EATC). In this study we have investigated the mechanism by which Mg2+ modulates this key glycolytic enzyme in EATC made permeable to the cation by either digitonin or dextran sulphate. Results showed that when Mg2+ is freely permeable to the cytosol, the in vivo PFK activity, calculated as FDP/G6P ratio, is not increased as it is in intact cells. We also observed that in permeabilized cells Mg2+ determines the increase of glucose 6 phosphate (G6P), fructose 1,6 bisphosphate (FDP) and lactate production. We hypothesize that extracellular Mg2+ regulates PFK and glycolysis in these neoplastic cells not by entering the cytosol but by a specific interaction with the plasma membrane.

Magnesium in normal and neoplastic cell proliferation: state of the art on in vitro data.

Information about the involvement of Mg2+ in all biochemical processes that participate in cell proliferation is reviewed in order to define the role of this divalent cation in normal and pathological growth. The lack of conclusive data about cell Mg2+ homeostasis does not suggest any definitive model for its role in the control of cell proliferation. On the other hand, new important information about its absolute requirement in crucial steps of cell activation that can, beside other functions, trigger cell division, strongly support the involvement of Mg2+ in the control of cell proliferation. Studies on the growth of cells in vitro, however, while confirming the indispensible requirement for Mg2+ in extracellular media, do not completely clarify the mechanism(s) or the exact phase/point of the cell cycle where Mg2+ exerts its regulation. Furthermore, the observation that tumour cells grown in culture are influenced by external divalent cations confirms the involvement of Mg2+ in cancer as well as in normal cell proliferation. The proposed explanations (theories, hypotheses) are described and discussed.

The effect of Mg2+ upon 6-phosphofructokinase activity in Ehrlich ascites tumor cells in vivo.

The effect of Mg2+ addition to intact Ehrlich ascites tumor cells (EATC) has been investigated. A decrease of glucose 6-phosphate (G6P) content and an increase of fructose 1,6-diphosphate (FDP) content are detected in glucose utilizing EATC incubated with increasing Mg2+ concentrations (from 0 to 5.0 mM). The strong enhancement of FDP/G6P ratio is taken as evidence for in vivo stimulation of phosphofructokinase 1 (PFK) (ATP:D-fructose-6-phosphate 1-phosphotransferase; EC 2.7.1.11). A similar effect can be observed when glucose is replaced by fructose as the glycolytic substrate. Stimulation of PFK is paralleled by substantial depletion of ATP. Cytochalasin B prevents the observed phenomena. Cell total Mg increases by about 15% when EATC are incubated with 5 mM Mg2+. The overall data show that extracellular Mg2+ may modulate glycolytic flux in EATC in vivo. Implications and significance of these phenomena in the regulation of cancer cell metabolic features are discussed.

Impairment of microsomal calcium sequestration activity upon superoxide dismutase depletion in rat liver.

We have studied the Ca2+-accumulation activity of microsomal vesicles isolated from the liver of rats held for from 2 to 8 weeks on a copper-deficient diet. With this treatment that deeply modifies fatty acid composition, microsomal membranes show progressively lower Ca2+ sequestration. The activity can be fully restored upon physiological copper supply to the depleted animals. The determination of kinetic parameters of microsomal Ca2+ uptake shows that copper deficiency affects mainly the apparent velocity, leaving unaffected the apparent affinity of the pump for Ca2+. Many similarities were found between this model and the Morris hepatomas with different growth rate. The data support the hypothesis that the oxidative stress imposed on the cell by the loss of superoxide dismutase can influence many cell features, with different implications in the regulation of several biological and biochemical functions.