Identification and minisequencing-based discrimination of SHV beta-lactamases in nosocomial infection-associated Klebsiella pneumoniae in Brisbane, Australia

Extended-spectrum beta-lactamases (ESBLs) are active against oxyimino cephalosporins and monobactams. Twenty-one Klebsiella pneumoniae isolates obtained between 1991 and 1995 at the Princess Alexandra Hospital in Brisbane, Australia, were subject to amplification and sequencing of the SHV beta-lactamase-encoding genes. Thirteen strains were phenotypically ESBL positive. Of these, six strains carried the bla(SHV-2a) gene and seven strains carried the bla(SHV-12) gene. Eight strains were phenotypically ESBL negative. Of these, seven strains carried the non-ESBL bla(SHV-11) gene and one strain carried the non-ESBL bla(SHV-1) gene. There was complete correspondence between the ESBL phenotype and the presence or absence of an ESBL-encoding gene(s). In addition, it was determined that of the 13 ESBL-positive strains, at least 4 carried copies of a non-ESBL-encoding gene in addition to the bla(SHV-2a) or bla(SHV12) gene. A minisequencing-based assay was developed to discriminate the different SHV classes. This technique, termed first-nucleotide change, involves the identification of the base added to a primer in a single-nucleotide extension reaction. The assay targeted polymorphisms at the first bases of codons 238 and 240 and reliably discriminated ESBL-positive strains from ESBL-negative strains and also distinguished strains carrying bla(SHV-2a) from strains carrying bla(SHV-12). In addition, this method was used to demonstrated an association between the relative copy numbers of bla(SHV) genes in individual strains and the levels of antibiotic resistance.

Monitoring and isolation of blood dendritic cells from apheresis products in healthy individuals: a platform for cancer immunotherapy

The fundamental role of dendritic cells (DC in initiating and directing the primary immune response is well established. Furthermore, it is now accepted that DC may be useful in new vaccination strategies for preventing certain malignant and infectious diseases. As blood DC (BDC physiology differs from that of the DC homologues generated in vitro from monocyte precursors, it is becoming more relevant to consider BDC for therapeutic interventions. Until recently, protocols for the isolation of BDC were laborious and inefficient; therefore, their use for investigative cancer immunotherapy is not widespread. In this study, we carefully documented BDC counts, yields and subsets during apheresis (Cobe Spectra), the initial and essential procedure in creating a BDC isolation platform for cancer immunotherapy. We established that an automated software package (Version 6,0 AutoPBPC) provides an operator-independent reliable source of motionuclear cells (MNC for BDC preparation. Further, we observed that BDC might be recovered in high yields, often greater than 100% relative to the number of circulating BDC predicted by blood volume. An average of 66 million (range, 17-179) BDC per 10-1 procedure were obtained, largely satisfying the needs for immunization. Higher yields were possible on total processed blood volumes of 151. BDC were not activated by the isolation procedure and, more importantly, both BDC subsets (CD11c(+)CD123(low) and CD11c(-)CD123(high)) were equally represented. Finally, we established that the apheresis product could be used for antibody-based BDC immunoselection and demonstrated that fully functional BDC can be obtained by this procedure. (C) 2002 Published by Elsevier Science B.V.

Terminally ill cancer patients' wish to hasten death

This exploratory study investigated factors associated with the wish to hasten death among a sample of terminally ill cancer patients. Semi-structured interviews conducted on a total of 72 hospice and home palliative care patients were subjected to qualitative analysis using QSR-NUDIST. The main themes to emerge suggested that patients with a high wish to hasten death had greater concerns with physical symptoms and psychological suffering, perceived themselves to be more of a burden to others, and experienced higher levels of demoralization, while also reporting less confidence in symptom control, fewer social supports, less satisfaction with life experiences and fewer religious beliefs when compared with patients who had a moderate or no wish to hasten death. The implications of these findings will be discussed.

Expectations of the effects of drinking on couple relationship functioning: An assessment of women in distressed relationships who consume alcohol at harmful levels

Based on a cognitive-social learning model of alcohol use, it was hypothesised that women with both alcohol and relationship problems would endorse more positive expectations of the effects of alcohol consumption on their relationship and would report lower relational efficacy than women without relationship or alcohol problems. Measures of relationship-referent alcohol expectancies and relational efficacy were completed by 174 married women with both alcohol and relationship problems (n = 20), alcohol problems alone (it = 26), relationship problems alone (n = 30), or neither problem (n = 98). Women without either alcohol or relationship problems strongly rejected expectations of enhanced relationship functioning (e.g., enhanced intimacy, increased emotional expression) following alcohol consumption, whereas women with both alcohol and relationship problems were ambivalent about these positive expectations. Women with both problems also reported lower relational efficacy than the other groups of women. Negative expectations about the effect of alcohol consumption on relationships in women with low relational efficacy may inhibit harmful drinking. (C) 2002 Elsevier Science Ltd. All rights reserved.

Couple communication and female problem drinking: A behavioral observation study

Couples with alcohol and relationship problems often report poor communication, yet little is known about the communication of maritally distressed couples in which the woman abuses alcohol (MDWA couples). Compared with maritally distressed couples without alcohol problems (MDNA) and couples with neither problem (NDNA), MDWA couples showed a distinctive pattern of negative communication. Similar to MDNA men, MDWA men spoke negatively to their partners but listened positively to their partners much like NDNA men. MDWA women listened negatively, much as MDNA women did, but spoke positively, like NDNA women did. The interactions of MDWA couples can be characterized as a male-demand-female-withdraw pattern, which is a gender reversal of the female-demand-male-withdraw pattern often observed in MDNA couples.

Charting the course of ovarian development in vertebrates

The decision of the embryonic gonad to differentiate as either a testis or an ovary is a critical step in vertebrate development. The molecular basis of this decision has been the focus of much study, particularly over the past decade. Here we contrast the knowledge of early gonadal development and the switch to testis differentiation with the lack of molecular understanding of ovarian development at early stages. We review current knowledge regarding mechanisms of ovarian morphogenesis and propose a model for the hierarchical control of development of the fetal ovary, incorporating the few genes already known to be important and several signals or factors that are hypothesised to exist in the early ovary.

Linkage and Association Analysis of Radiation Damage Repair Genes XRCC3 and XRCC5 with Nevus Density in Adolescent Twins

Previous studies have shown that a deficiency in DNA damage repair is associated with increased cancer risk, and exposure to UV radiation is a major risk factor for the development of malignant melanoma. High density of common nevi (moles) is a major risk factor for cutaneous melanoma. A nevus may result from a mutation in a single UV-exposed melanocyte which failed to repair DNA damage in one or more critical genes. XRCC3 and XRCC5 may have an effect on nevus count through their function as components of DNA repair processes that may be involved directly or indirectly in the repair of DNA damage due to UV radiation. This study aims to test the hypothesis that the frequency of flat or raised moles is associated with polymorphism at or near these DNA repair genes, and that certain alleles are associated with less efficient DNA repair, and greater nevus density. Twins were recruited from schools in south eastern Queensland and were examined close to their 12th birthday. Nurses examined each individual and counted all moles on the entire body surface. A 10cM genome scan of 274 families (642 individuals) was performed and microsatellite polymorphisms in XRCC3 and adjacent to XRCC5 were also typed. Linkage and association of nevus count to these loci were tested simultaneously using a structural-equation modeling approach implemented in MX. There is weak evidence for linkage of XRCC5 to a QTL influencing raised mole count, and also weak association. There is also weak evidence for association between flat mole count and XRCC3. No tests were significant after correction for testing multiple alleles, nor were any of the tests for total association significant. If variation in XRCC3 or XRCC5 influences UV sensitivity, and indirectly affects nevus density, then the effects are small.

Dilemas contempor??neos das pol??ticas p??blicas

O presente ensaio trata de temas identificados como dilemas de pol??ticas p??blicas: a transversalidade e o compartilhamento da gest??o das pol??ticas de prote????o do meio ambiente. O objeto espec??fico de an??lise deste texto situa-se na incorpora????o da vari??vel ambiental nas pol??ticas setoriais para o avan??o do desenvolvimento sustent??vel.

Perfil intra-empreendedor: um estudo inicial em funcion??rios da Infraero-sede

Considerando os conceitos de intra-empreendedorismo e sua import??ncia para as organiza????es atualmente, realizou-se pesquisa juntos aos ocupantes do cargo de Profissional de Servi??os Aeroportu??rios (PSA), na sede da Empresa Brasileira de Infra-Estrutura Aeroportu??ria (Infraero), buscando verificar se eles possuem ou n??o perfil intra-empreendedor. Utilizou-se um question??rio, composto de 28 quest??es referentes ??s necessidades e habilidades indispens??veis ao desenvolvimento do intra-empreendedorismo, para avaliar o perfil dos funcion??rios, bem como levantar as caracter??sticas que apareceram com maior e menor freq????ncia na amostra selecionada. Como resultado, concluiu-se que a maioria dos empregados pesquisados possui perfil intra-empreendedor, por??m necessitando do desenvolvimento de outras caracter??sticas, tais como necessidade de independ??ncia e de autorealiza????o, tamb??m imprescind??veis. Foram sugeridas, tamb??m, a????es que podem ser adotadas na organiza????o para incentivar os trabalhadores a exercer suas capacidades empreendedoras.

An overview of domestic aspects in US climate policy

The article shows how domestic aspects influence the United States national and international climate policy. To accomplish the task, the authors analyzes the discussions when Bill Clinton was ruling the country, a time during which global discussions were forwarded. The paper recalls the debate in the Bush administration and the growing polarization since Barack Obama took office.