Relationships between non-occupational cadmium exposure and expression of nine cytochrome P450 forms in human liver and kidney cortex samples

This study was undertaken to assess associations between age, gender, cigarette smoke and non-workplace cadmium exposure, and liver pathology and inter-individual variation in cytochrome P450 (CYP) expression in human tissues. Autopsy specimens of twenty-eight Queensland residents whose ages ranged from 3 to 89 years were analyzed for the presence of nine CYP protein isoforms by immunoblotting. All subjects were Caucasians and their liver cadmium contents ranged from 0.11 to 3.95 kg/g wet weight, while their kidney cadmium contents were in the range of 2 to 63 mug/g wet weight. CYP1A2, CYP2A6, CYP2D6, CYP3A4, and CYP3A5 were detected in liver but not in kidney, and CYP1A1 and CYP1B1 were not found in liver or kidney. Lowered liver CYP2C8/19 protein contents were found to be associated with liver pathology. Importantly, we show elevated levels of CYP2C9 protein to be associated with cadmium accumulation in liver. No mechanism that explains this association is apparent, but there are two possibilities that require further study. One is that variation in CYP2C9 protein levels may be, in part, attributed to an individual's non-workplace exposure to cadmium, or an individual's CYP2C9 genotype may be a risk factor for cadmium accumulation. A positive correlation was found between liver CYP3A4 protein and subject age. Levels of liver CYPIA2 protein, but not other CYP forms, were increased in people more exposed to cigarette smoke, but there was no association between CYPIA2 protein and cadmium. CYP2A6 protein was found in all liver samples and CYP2A6 gene typing indicated the absence of CYP2A6 null allele (CYP2A6(D)) in this sample group, confirming very low prevalence of homozygous CYP2A6(D) in Caucasians. CYP2A6 gene types W/W, WIC, and CIC were not associated with variations in liver microsomal CYP2A6 protein. CYP2D6 protein was absent in all twenty-five kidney samples tested but was detectable in liver samples of all but two subjects, indicating the prevalence of the CYP2D6 null allele (CYP2D6(D)) in this sample group to be about 7%, typical of Caucasian populations. (C) 2001 Elsevier Science Inc. All rights reserved.

Distinction between familial and sporadic forms of colorectal cancer showing DNA microsatellite instability

Attempts to classify colorectal cancer into subtypes based upon molecular characterisation are overshadowed by the classical stepwise model in which the adenoma-carcinoma sequence serves as the morphological counterpart. Clarity is achieved when cancers showing DNA microsatellite instability (MSI) are distinguished as sporadic MSI-low (MSI-L), sporadic MSI-high (MSI-H) and hereditary non-polyposis colorectal cancer (HNPCC). Divergence of the 'methylator' pathway into MSI-L and MSI-H is at least partly determined by the respective silencing of MGMT and hMLH1. Multiple differences can be demonstrated between sporadic and familial (HNPCC) MSI-H colorectal cancer with respect to early mechanisms, evolution, molecular characterisation, demographics and morphology. By acknowledging the existence of multiple pathways, rapid advances in the fields of basic and translational research will occur and this will lead to improved strategies for the prevention, early detection and treatment of colorectal cancer. (C) 2002 Elsevier Science Ltd. All rights reserved.

Microevolution in island forms: the roles of drift and directional selection in morphological divergence of a passerine bird

Theory predicts that in small isolated populations random genetic drift can lead to phenotypic divergence; however this prediction has rarely been tested quantitatively in natural populations. Here we utilize natural repeated island colonization events by members of the avian species complex, Zosterops lateralis, to assess whether or not genetic drift alone is an adequate explanation for the observed patterns of microevolutionary divergence in morphology. Morphological and molecular genetic characteristics of island and mainland populations are compared to test three predictions of drift theory: (1) that the pattern of morphological change is idiosyncratic to each island; (2) that there is concordance between morphological and neutral genetic shifts across island populations; and (3) for populations whose time of colonization is known, that the rate of morphological change is sufficiently slow to be accounted for solely by genetic drift. Our results are not consistent with these predictions. First, the direction of size shifts was consistently towards larger size, suggesting the action of a nonrandom process. Second, patterns of morphological divergence among recently colonized populations showed little concordance with divergence in neutral genetic characters. Third, rate tests of morphological change showed that effective population sizes were not small enough for random processes alone to account for the magnitude of microevolutionary change. Altogether, these three lines of evidence suggest that drift alone is not an adequate explanation of morphological differentiation in recently colonized island Zosterops and therefore we suggest that the observed microevolutionary changes are largely a result of directional natural selection.

Catalytically active Dengue virus NS3 protease forms aggregates that are separable by size exclusion chromatography

An active form of the Dengue virus protease NS3 (CF40.Gly.NS3pro) was expressed in Escherichia coli. This construct consists of a critical 40 amino acid cofactor domain from NS2B fused to the N-terminal 184 amino acid protease domain of NS3 via a flexible, covalent linker (Gly(4)SerGly(4)). The recombinantly produced protein is soluble and has a hexa-histidine tag engineered at the N-terminus for ease of purification using metal affinity chromatography. However, the presence of lower molecular weight impurities after affinity chromatography indicated the need for additional purification steps. The consistent appearance of these impurities suggested that they may be the products of proteolysis and/or auto-proteolysis. The latter possibility was subsequently excluded by the observation of the same impurities in a purified, catalytically inactive form of the recombinant protease (CF40.Gly.NS3pro.SA). Further analysis indicated that these impurities may represent premature translation termination products. Regardless of their origin, they were shown to form various sized aggregates with full-length CF40.Gly.NS3pro that can be separated by size exclusion chromatography, yielding fractions of active protease of sufficient purity for crystallisation trials. The ultimate goal of these studies is to obtain a crystal structure of a catalytically active form of the Dengue virus NS3 protease for structure-based drug design. (C) 2002 Elsevier Science (USA). All rights reserved.

The trans-membrane protein p25 forms highly specialized domains that regulate membrane composition and dynamics

Trans-membrane proteins of the p24 family are abundant, oligomeric proteins predominantly found in cis-Golgi membranes. They are not easily studied in vivo and their functions are controversial. We found that p25 can be targeted to the plasma membrane after inactivation of its canonical KKXX motif (KK to SS, p25SS), and that p25SS causes the co-transport of other p24 proteins beyond the Golgi complex, indicating that wild-type p25 plays a crucial role in retaining p24 proteins in cis-Golgi membranes. We then made use of these observations to study the intrinsic properties of these proteins, when present in a different membrane context. At the cell surface, the p25SS mutant segregates away from both the transferrin receptor and markers of lipid rafts, which are enriched in cholesterol and glycosphingolipids. This suggests that p25SS localizes to, or contributes to form, specialized membrane domains, presumably corresponding to oligomers of p25SS and other p24 proteins. Once at the cell surface, p25SS is endocytosed, together with other p24 proteins, and eventually accumulates in late endosomes, where it remains confined to well-defined membrane regions visible by electron microscopy. We find that this p25SS accumulation causes a concomitant accumulation of cholesterol in late endosomes, and an inhibition of their motility - two processes that are functionally linked. Yet, the p25SS-rich regions themselves seem to-exclude not only Lamp1 but also accumulated cholesterol. One may envision that p25SS accumulation, by excluding cholesterol from oligomers, eventually overloads neighboring late endosomal membranes with cholesterol beyond their capacity (see Discussion). In any case, our data show that p25 and presumably other p24 proteins are endowed with the intrinsic capacity to form highly specialized domains that control membrane composition and dynamics. We propose that p25 and other p24 proteins control the fidelity of membrane transport by maintaining cholesterol-poor membranes in the Golgi complex.

High rate of detection of primary aldosteronism, including surgically treatable forms, after 'non-selective' screening of hypertensive patients

Background Wide testing of the aldosterone: renin ratio among hypertensive individuals has revealed primary aldosteronism to be common, with most patients normokalaemic. Some investigators, however, have reported aldosterone-producing adenoma to be rare among patients so detected. Objective To test the hypothesis that differences among reported studies in the rate of detection of aldosterone-producing adenoma (as opposed to bilateral adrenal hyperplasia) reflect differences in the procedures used for diagnosis of primary aldosteronism, and the methods used to identify aldosterone-producing adenomas. Methods In the newly established Princess Alexandra Hospital Hypertension Unit (PAHHU), we used procedures developed by Greenslopes Hospital Hypertension Unit (which reports that more than 30% of patients with primary aldosteronism have aldosterone-producing adenomas) to diagnose primary aldosteronism and determine the subtype. All patients with an increased aldosterone: renin ratio (measured after correction for hypokalaemia and while the patient was not receiving interfering medications) underwent fludrocortisone suppression testing to confirm or exclude primary aldosteronism; if they were positive, they underwent genetic testing to exclude glucocorticoid-remediable aldosteronism before adrenal venous sampling was used to differentiate lateralizing from bilateral primary aldosteronism. Results This approach allowed PAHHU to diagnose, within 2 years, 54 patients [only seven (13%) hypokalaemic] with primary aldosteronism. All tested negative for glucocorticoid-remediable aldosteronism. Aldosterone production was lateralized to one adrenal in 15 patients (31%; only six hypokalaemic) and was bilateral in 34 (69%; all normokalaemic) of 49 patients who underwent adrenal venous sampling. Among patients with lateralizing adrenal hyperplasia, computed tomography revealed an ipsilateral mass in only six and a contralateral lesion in one. Fourteen patients underwent unilateral adrenalectomy, which cured the hypertension in seven and improved it in the remainder. In patients with bilateral primary aldlosteronism, hypertension responded to spironolactone (112.5-50 mg/ day) or amiloride (2.5-10 mg/day). Conclusion When performed with careful regard to confounding factors, measurement of the aldosterone: renin ratio in all hypertensive individuals, followed by fludrocortisone suppression testing to confirm or exclude primary aldosteronism and adrenal venous sampling to determine the subtype, can result in the detection of significant numbers of patients with specifically treatable or potentially curable hypertension. (C) 2003 Lippincott Williams Wilkins.

Opening windows into the mind: mothers' preferences for mental state explanations and children's theory of mind

A new questionnaire, the Maternal Mental State Input Inventory (MMSII) was created to measure mothers' preferences for introducing and elaborating on mental states in conversation with their young children. In two studies, the questionnaire was given to mothers of young children, and the children's theory of mind (ToM) development was assessed with standard tasks. In both studies, the questionnaire exhibited good internal reliability, and significant correlations emerged between mothers' self-reported preferences for elaborated, explanatory talk about the mental states and children's theory of mind performance. Further, mothers' conversational preferences, as measured by the MMSII, were the best predictors of children's theory of mind development when relevant control variables were included in the analyses. These results converge with naturalistic observational research that has demonstrated links between mothers' conversational styles and their children's theory of mind. They go further in suggesting that mothers' tendencies toward elaborated, explanatory talk about a range of mental states is particularly beneficial to children's theory of mind development. (C) 2003 Elsevier Inc. All rights reserved.

Eastwards / westwards: which direction for gender studies in the 21st century?

Eastwards / Westwards: Which Direction for Gender Studies in the XXIst Century? is a collection of essays which focus on themes and methods that characterize current research into gender in Asian countries in general. In this collection, ideas derived from Gender Studies elsewhere in the world have been subjected to scrutiny for their utility in helping to describe and understand regional phenomena. But the concepts of Local and Global – with their discoursive productions – have not functioned as a binary opposition: localism and globalism are mutually constitutive and researchers have interrogated those spaces of interaction between the ‘self’ and the ‘other’, bearing in mind their own embeddedness in social and cultural structures and their own historical memory. Contributors to this collection provided a critical transnational perspective on some of the complex effects of the dynamics of cultural globalization, by exploring the relation between gender and development, language, historiography, education and culture. We have also given attention to the ideological and rhetorical processes through which gender identity is constructed, by comparing textual grids and patterns of expectation. Likewise, we have discussed the role of ethnography, anthropology, historiography, sociology, fiction, popular culture and colonial and post-colonial sources in (re)inventing old/new male/female identities, their conversion into concepts and circulation through time and space. This multicultural and trans-disciplinary selection of essays is totally written in English, fully edited and revised, therefore, it has a good potential for an immediate international circulation. This project may trace new paths and issues for discussion on what concerns the life, practices and narratives by and about women in Asia, as well as elsewhere in the present day global experience. Academic readership: Researchers, scholars, educators, graduate and post-graduate students, doctoral students and general non-fiction readers, with a special interest in Gender Studies, Asia, Colonial and Post-Colonial Literature, Anthropology, Cultural Studies, History, Historiography, Politics, Race, Feminism, Language, Linguistics, Power, Political and Feminist Agendas, Popular Culture, Education, Women’s Writing, Religion, Multiculturalism, Globalisation, Migration. Chapter summary: 1. “Social Gender Stereotypes and their Implication in Hindi”, Anjali Pande, Jawaharlal Nehru University, New Delhi, India. This essay looks at the subtle ways in which gender identities are constructed and reinforced in India through social norms of language use. Language itself becomes a medium for perpetuating gender stereotypes, forcing its speakers to confirm to socially defined gender roles. Using examples from a classroom discussion about a film, this essay will highlight the underlying rigid male-female stereotypes in Indian society with their more obvious expressions in language. For the urban woman in India globalisation meant increased economic equality and exposure to changed lifestyles. On an individual level it also meant redefining gender relations and changing the hierarchy in man-­woman relationships. With the economic independence there is a heightened sense of liberation in all spheres of social life, a confidence to fuzz the rigid boundaries of gender roles. With the new films and media celebrating this liberated woman, who is ready to assert her sexual needs, who is ready to explode those long held notions of morality, one would expect that the changes are not just superficial. But as it soon became obvious in the course of a classroom discussion about relationships and stereotypes related to age, the surface changes can not become part of the common vocabulary, for the obvious reason that there is still a vast gap between the screen image of this new woman and the ground reality. Social considerations define the limits of this assertiveness of women, whereas men are happy to be liberal within the larger frame of social sanctions. The educated urban woman in India speaks in favour of change and the educated urban male supports her, but one just needs to scratch the surface to see the time tested formulae of gender roles firmly in place. The way the urban woman happily balances this emerging promise of independence with her gendered social identity, makes it necessary to rethink some aspects of looking at gender in a gradually changing, traditional society like India. 2. “The Linguistic Dimension of Gender Equality”, Alissa Tolstokorova, Kiev Centre for Gender Information and Education, Ukraine. The subject-matter of this essay is gender justice in language which, as I argue, may be achieved through the development of a gender-related approach to linguistic human rights. The last decades of the 20th century, globally marked by a “gender shift” in attitudes to language policy, gave impetus to the social movement for promoting linguistic gender equality. It was initiated in Western Europe and nowadays is moving eastwards, as ideas of gender democracy progress into developing countries. But, while in western societies gender discrimination through language, or linguistic sexism, was an issue of concern for over three decades, in developing countries efforts to promote gender justice in language are only in their infancy. My argument is that to promote gender justice in language internationally it is necessary to acknowledge the rights of women and men to equal representation of their gender in language and speech and, therefore, raise a question of linguistic rights of the sexes. My understanding is that the adoption of the Universal Declaration of Linguistic Rights in 1996 provided this opportunity to address the problem of gender justice in language as a human rights issue, specifically as a gender dimension of linguistic human rights. 3. “The Rebirth of an Old Language: Issues of Gender Equality in Kazakhstan”, Maria Helena Guimarães, Polytechnic Institute of Porto, Portugal. The existing language situation in Kazakhstan, while peaceful, is not without some tension. We propose to analyze here some questions we consider relevant in the frame of cultural globalization and gender equality, such as: free from Russian imperialism, could Kazakhstan become an easy prey of Turkey’s “imperialist dream”? Could these traditionally Muslim people be soon facing the end of religious tolerance and gender equality, becoming this new old language an easy instrument for the infiltration in the country of fundamentalism (it has already crossed the boarders of Uzbekistan), leading to a gradual deterioration of its rich multicultural relations? The present structure of the language is still very fragile: there are three main dialects and many academics defend the re-introduction of the Latin alphabet, thus enlarging the possibility of cultural “contamination” by making the transmission of fundamentalist ideas still easier through neighbour countries like Azerbaijan, Uzbekistan and Turkmenistan (their languages belong to the same sub-group of Common Turkic), where the Latin alphabet is already in use, and where the ground for such ideas shown itself very fruitful. 4. “Construction of Womanhood in the Bengali Language of Bangladesh”, Raasheed Mahmood; University of New South Wales, Sydney. The present essay attempts to explore the role of gender-based language differences and of certain markers that reveal the status accorded to women in Bangladesh. Discrimination against women, in its various forms, is endemic in communities and countries around the world, cutting across class, race, age, and religious and national boundaries. One cannot understand the problems of gender discrimination solely by referring to the relationship of power or authority between men and women. Rather one needs to consider the problem by relating it to the specific social formation in which the image of masculinity and femininity is constructed and reconstructed. Following such line of reasoning this essay will examine the nature of gender bias in the Bengali language of Bangladesh, holding the conviction that as a product of social reality language reflects the socio-cultural behaviour of the community who speaks it. This essay will also attempt to shed some light on the processes through which gender based language differences produce actual consequences for women, who become exposed to low self-esteem, depression and systematic exclusion from public discourse. 5. “Marriage in China as an expression of a changing society”, Elisabetta Rosado David, University of Porto, Portugal, and Università Ca’Foscari, Venezia, Italy. In 29 April 2001, the new Marriage Law was promulgated in China. The first law on marriage was proclaimed in 1950 with the objective of freeing women from the feudal matrimonial system. With the second law, in 1981, values and conditions that had been distorted by the Cultural Revolution were recovered. Twenty years later, a new reform was started, intending to update marriage in the view of the social and cultural changes that occurred with Deng Xiaoping’s “open policy”. But the legal reform is only the starting point for this case-study. The rituals that are followed in the wedding ceremony are often hard to understand and very difficult to standardize, especially because China is a vast country, densely populated and characterized by several ethnic minorities. Two key words emerge from this issue: syncretism and continuity. On this basis, we can understand tradition in a better way, and analyse whether or not marriage, as every social manifestation, has evolved in harmony with Chinese culture. 6. “The Other Woman in the Portuguese Colonial Empire: The Case of Portuguese India”, Maria de Deus Manso, University of Évora, Portugal. This essay researches the social, cultural and symbolic history of local women in the Portuguese Indian colonial enclaves. The normative Portuguese overseas history has not paid any attention to the “indigenous” female populations in colonial Portuguese territories, albeit the large social importance of these social segments largely used in matrimonial and even catholic missionary strategies. The first attempt to open fresh windows in the history of this new field was the publication of Charles Boxer’s referential study about Women in lberian Overseas Expansion, edited in Portugal only after the Revolution of 1975. After this research we can only quote some other fragmentary efforts. In fact, research about the social, cultural, religious, political and symbolic situation of women in the Portuguese colonial territories, from the XVI to the XX century, is still a minor historiographic field. In this essay we discuss this problem and we study colonial representations of women in the Portuguese Indian enclaves, mainly in the territory of Goa, using case studies methodologies. 7. “Heading East this Time: Critical Readings on Gender in Southeast Asia”, Clara Sarmento, Polytechnic Institute of Porto, Portugal. This essay intends to discuss some critical readings of fictional and theoretical texts on gender condition in Southeast Asian countries. Nowadays, many texts about women in Southeast Asia apply concepts of power in unusual areas. Traditional forms of gender hegemony have been replaced by other powerful, if somewhat more covert, forms. We will discuss some universal values concerning conventional female roles as well as the strategies used to recognize women in political fields traditionally characterized by male dominance. Female empowerment will mean different things at different times in history, as a result of culture, local geography and individual circumstances. Empowerment needs to be perceived as an individual attitude, but it also has to be facilitated at the macro­level by society and the State. Gender is very much at the heart of all these dynamics, strongly related to specificities of historical, cultural, ethnic and class situatedness, requiring an interdisciplinary transnational approach.

Trypanosoma cruzi prevalence and clinical forms in blood donor candidates in Brazil

The prevalence and clinical forms of Trypanosoma cruzi were evaluated among blood donor candidates attended at a general hospital in Rio de Janeiro, Brazil, from January 1997 to April 1999. The investigation was done by means of the indirect hemagglutination test and was confirmed via ELISA. Data were collected from clinical examinations, conventional electrocardiogram, chest radiography and echocar-diography. The results showed that despite Trypanosoma cruzi prevalence of 1.17% (128 patients), mainly in males aged 40 years or over, 70.8% of these patients, mainly males aged 19 to 39 years, demonstrated abnormalities that allowed the diagnosis of cardiopathy and/or esophagopathy. This once again corroborates the importance of Trypanosoma cruzi infection in urban centers.

Development of electrochemical methods for determination of tramadol - analytical application to pharmaceutical dosage forms

A square-wave voltammetric (SWV) method and a flow injection analysis system with amperometric detection were developed for the determination of tramadol hydrochloride. The SWV method enables the determination of tramadol over the concentration range of 15-75 µM with a detection limit of 2.2 µM. Tramadol could be determined in concentrations between 9 and 50 µM at a sampling rate of 90 h-1, with a detection limit of 1.7 µM using the flow injection system. The electrochemical methods developed were successfully applied to the determination of tramadol in pharmaceutical dosage forms, without any pre-treatment of the samples. Recovery trials were performed to assess the accuracy of the results; the values were between 97 and 102% for both methods.

Cloud computing sobre a plataforma Windows Azure

A utilização massiva da internet e dos serviços que oferece por parte do utilizador final potencia a evolução dos mesmos, motivando as empresas a apostarem no desenvolvimento deste tipo de soluções. Requisitos como o poder de computação, flexibilidade e escalabilidade tornam-se cada vez mais indissociáveis do desenvolvimento aplicacional, o que leva ao surgimento de paradigmas como o de Cloud Computing. É neste âmbito que surge o presente trabalho. Com o objetivo de estudar o paradigma de Cloud Computing inicia-se um estudo sobre esta temática, onde é detalhado o seu conceito, a sua evolução histórica e comparados os diferentes tipos de implementações que suporta. O estudo detalha posteriormente a plataforma Azure, sendo analisada a sua topologia e arquitetura, detalhando-se os seus componentes e a forma como esta mitiga alguns dos problemas mencionados. Com o conhecimento teórico é desenvolvido um protótipo prático sobre esta plataforma, em que se exploram algumas das particularidades da topologia e se interage com as principais redes sociais. O estudo culmina com uma análise sobre os benefícios e desvantagens do Azure e através de um levantamento das necessidades da empresa, determinam-se as oportunidades que a utilização da plataforma poderá proporcionar.

Análise descritiva das práticas e contextos em fisioterapia respiratória nos hospitais centrais públicos de Luanda

Mestrado em Fisioterapia