Comparative analysis of two immunohistochemical methods for antigen retrieval in the optical lobe of the honeybee Apis mellifera: Myosin-v assay

The present study compared two heating methods currently used for antigen retrieval (AR) immunostaining: the microwave oven and the steam cooker. Myosin-V, a molecular motor involved in vesicle transport, was used as a neuronal marker in honeybee Apis mellifera brains fixed in formalin. Overall, the steam cooker showed the most satisfactory AR results. At 100 degrees C, tissue morphology was maintained and revealed epitope recovery, while evaporation of the AR solution was markedly reduced; this is important for stabilizing the sodium citrate molarity of the AR buffer and reducing background effects. Standardization of heat-mediated AR of formalin-fixed and paraffin-embedded tissue sections results in more reliable immunostaining of the honeybee brain.

Antioxidant status and biomarkers of oxidative stress in bovine leukemia virus-infected dairy cows

Bovine leukemia virus (BLV) is among the most widespread livestock pathogens in many countries. Despite advances in understanding the pathogenesis of this disease, little is known about the involvement of oxidative stress. Therefore, this study examined the antioxidant status and the markers of oxidative stress in BLV-infected dairy cows. BLV infection was associated with an increase in triacylglycerol levels, a decrease in glutathione peroxidase (GSH-Px) activity and a tendency toward lower superoxide dismutase activity in the infected animals. No significant difference was observed in other markers of oxidative stress (i.e., conjugated dienes, hydroperoxides and malondialdehyde) in the infected animals compared to controls. A novel method for the analysis of oxidative stress, Z-scan based on the measurement of the mean-value of 9 in low density lipoprotein indicated that the infected animals had low-density lipoprotein particles that were slightly less modified than those from the healthy group. Thus, we conclude that BLV infection is associated with a selective decrease in GSH-Px activity without any alteration in the common plasma markers of oxidative stress. (C) 2011 Elsevier B.V. All rights reserved.

Apoptosis and necrosis of polymorphonuclear leukocytes in goat milk with high and low somatic cell counts

The purpose of the present trial was to compare the percentages of necrotic and apoptotic polymorphonuclear leukocytes (PMNL) in goat milk with low and high somatic cell count (SCC). Twenty eight milk samples were collected from 20 lactating goats, determined to be negative in bacteriological examination, and divided in three groups, according to their SCC: samples with SCC lower than 500 x 10(3) cells/mL; between 500 and 1500 x 10(3) cells/mL; and higher than 1500 x 10(3) cells/m L. SCC was performed in an automatic somatic cell counter. Apoptosis and necrosis were quantified using dual-color flow cytometry with fluorescein labeled annexin-V and propidium iodide (PI). Results of the present study showed a significant positive correlation between the percentage of the viable PMNL and milk SCC(r = 0.495, P=0.008), as well as a significant negative correlation between apoptotic PMNL and milk SCC(r = -0.486, P = 0.009). Results also pointed out lower PMNL viability rates due to higher apoptosis rates in milk samples with SCC lower than 5 x 10(5) cells/mL. (C) 2011 Elsevier B.V. All rights reserved.

gamma-Radiation induces apoptosis via sarcoplasmatic reticulum in guinea pig ileum smooth muscle cells

We investigated the effects of gamma-radiation on cells isolated from the longitudinal smooth muscle layer of the guinea pig ileum, a relatively radioresistant tissue. Single doses (up to 50 Gy) reduced the amount of sarcoplasmatic reticulum and condensed the myofibrils, as shown by electron microscopy 3 days post-irradiation. After that, contractility of smooth muscle strips was reduced. Ca(2+) handling was altered after irradiation, as shown in fura-2 loaded cells, with elevated basal intracellular Ca(2+), reduced amount of intrareticular Ca(2+), and reduced capacitive Ca(2+) entry. Radiation also induced apoptosis, judged from flow cytometry of cells loaded with proprium iodide. Electron microscopy showed that radiation caused condensation of chromatin in dense masses around the nuclear envelope, the presence of apoptotic bodies, fragmentation of the nucleus, detachment of cells from their neighbors, and reductions in cell volume. Radiation also caused activation of caspase 12. Apoptosis was reduced by the administration of the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone methyl ester (Z-VAD-FIVIK) during the 3 day period after irradiation, and by the chelator of intracellular Ca(2+), 1,2-bis(o-aminophenoxy)ethane-N,N,N`,N`-tetraacetic acid (BAPTA), from 1 h before until 2 h after irradiation. BAPTA also reduced the effects of radiation on contractility, basal intracellular Ca(2+), amount of intrareticular Ca(2+), capacitative Ca(2+) entry, and apoptosis. In conclusion, the effects of gamma radiation on contractility, Ca(2+) handling, and apoptosis appear due to a toxic action of intracellular Ca(2+). Ca(2+)-induced damage to the sarcoplasmatic reticulum seems a key event in impaired Ca(2+) handling and apoptosis induced by gamma-radiation. (c) 2008 Elsevier B.V. All rights reserved.

Immunoelectron microscopy provides evidence for the presence of mitochondrial heat shock 10-kDa protein (chaperonin 10) in red blood cells and a variety of secretory granules

Hsp10 (10-kDa heat shock protein, also known as chaperonin 10 or Cpn10) is a co-chaperone for Hsp60 in the protein folding process. This protein has also been shown to be identical to the early pregnancy factor, which is an immunosuppressive growth factor found in maternal serum. In this study we have used immunogold electron microscopy to study the subcellular localization of Hsp10 in rat tissues sections embedded in LR Gold resin employing polyclonal antibodies raised against different regions of human Hsp10. In all rat tissues examined including liver, heart, pancreas, kidney, anterior pituitary, salivary gland, thyroid, and adrenal gland, antibodies to Hsp10 showed strong labeling of mitochondria. However, in a number of tissues, in addition to the mitochondrial labeling, strong and highly specific labeling with the Hsp10 antibodies was also observed in several extramitochondrial compartments. These sites included zymogen granules in pancreatic acinar cells, growth hormone granules in anterior pituitary, and secretory granules in PP pancreatic islet cells. Additionally, the mature red blood cells which lack mitochondria, also showed strong reactivity with the Hsp10 antibodies. The observed labeling with the Hsp10 antibodies, both within mitochondria as well as in other compartments/cells, was abolished upon omission of the primary antibodies or upon preadsorption of the primary antibodies with the purified recombinant human Hsp10. These results provide evidence that similar to a number of other recently described mitochondrial proteins (viz., Hsp60, tumor necrosis factor receptor-associated protein- 1, P32 (gC1q-R) protein, and cytochrome c), Hsp10 is also found at a variety of specific extramitochondrial sites in normal rat tissue. These results raise important questions as to how these mitochondrial proteins are translocated to other compartments and their possible function(s) at these sites. The presence of these proteins at extramitochondrial sites in normal tissues has important implications concerning the role of mitochondria in apoptosis and genetic diseases.

Assault-related admissions to hospital in Central Australia

Objective: To determine the number of assault-related admissions to hospital in the Central Australia region of the Northern Territory over a six-year period. Design and setting: Retrospective analysis of all patients admitted to Alice Springs Hospital (ASH) and Tennant Creek Hospital (TCH) from July 1995 to June 2001, where the primary cause of injury was assault. Main outcome measures: Frequency of assault-related admission to hospital; demographic characteristics of the victims. Results: in the six years, there were 2449 assault-related admissions to ASH and 545 to TCH. Adults aged 25-34 years were most frequently hospitalised for assault, in a proportion greater than their proportion in the NT population, Females represented 59.7% of people admitted to ASH and 54.7% to TCH, greater than their proportion in the NT population. Aboriginals comprised 95.2% of ASH and 89.0% of TCH admissions, and were admitted in a significantly greater proportion than their proportion in the NT population (P < 0.001). The age-adjusted hospital admission rate resulting from assault has increased (P = 0.002) at an average rate of 1.6 (SE, 0.2) per 10 000 people per year. The proportion of assault-related admissions associated with alcohol has also increased significantly (P < 0.001). Conclusion: The frequency of assault-related admissions to hospital, especially among the Aboriginal population, suggests that this major public health issue is escalating.

Purification and characterisation of functional early pregnancy factor expressed in Sf9 insect cells and in Escherichia coli

Early pregnancy factor (EPF) is a secreted protein with growth regulatory and immunomodulatory properties. It is an extracellular form of the mitochondrial matrix protein chaperonin 10 (Cpn10), a molecular chaperone. An understanding of the mechanism of action of EPF and an exploration of therapeutic potential has been limited by availability of purified material. The present study was undertaken to develop a simple high-yielding procedure for preparation of material for structure/function studies, which could be scaled up for therapeutic application. Human EPF was expressed in Sf9 insect cells by baculovirus infection and in Escherichia coli using a heat inducible vector. A modified molecule with an additional N-terminal alanine was also expressed in E coli. The soluble protein was purified from cell lysates via anion exchange (negative-binding mode), cation exchange, and hydrophobic interaction chromatography, yielding similar to42 and 36 mg EPF from 300 ml bacterial and I L Sf9 cultures, respectively. The preparations were highly purified ( greater than or equal to99% purity on SDS-PAGE for the bacterial products and greater than or equal to97% for that of insect cells) and had the expected mass and heptameric structure under native conditions, as determined by mass spectrometry and gel permeation chromatography, respectively. All recombinant preparations exhibited activity in the EPF bioassay, the rosette inhibition test, with similar potency both to each other and to the native molecule. In two in vivo assays of immuno suppressive activity, the delayed-type hypersensitivity reaction and experimental autoimmune encephalomyelitis, the insect cell and modified bacterial products, both with N-terminal additions (acetylation or amino acid), exhibited similar levels of suppressive activity, but the bacterial product with no N-terminal modification had no effect in either assay. Studies by others have shown that N-terminal addition is not necessary for Cpn10 activity. By defining techniques for facile production of molecules with and without immunosuppressive properties, the present studies make it possible to explore mechanisms underlying the distinction between EPF and Cpn10 activity. (C) 2003 Elsevier Inc. All rights reserved.

Researchers' information needs in the bibliographic database : a literature review

This article is published online with Open Access and distributed under the terms of the Creative Commons Attribution Non-Commercial License.

O devir menor de Alice : linhas de escrita, linhas de vida. Sobre a aprendizagem da linguagem na educação infantil

As linhas de escrita que conformam o Devir Menor de Alice constituem-se a partir da necessidade de investigar o devir-criança dos corpos aprendentes no processo de aprendizagem da linguagem como potência capaz de engendrar, na imanência dos encontros educativos, um currículo que funcione como plano de constituição de um estilo singular de inscrição de si e do mundo no âmbito da educação infantil. Impulsionado por forças que se desdobram em um campo problemático que orienta um percurso investigativo de caráter cartográfico, o estudo busca problematizar: Por quais processos o problema da escrita pode ordenar um movimento expressivo de aprendizagem da linguagem? Em torno dessa problemática, concebe três questões centrais: (1) De que modo as práticas diferenciais de linguagem traçadas no movimento imanente do currículo deslocam de modo positivo o processo de aprendizagem da linguagem na educação infantil? (2) Do que trata concretamente o conceito de linguagem no movimento expressivo e de aprendizagem afetiva? (3) Por que é relevante abordar o movimento expressivo de aprendizagem da linguagem e por que fazer isso a partir do problema da escrita? Nesse processo investigativo, afirma que a aprendizagem da linguagem implica processos de subjetivação pelos quais a ideia do delírio, do sonho, do sonambulismo de Alice traz para a leitura e a escrita a necessária relação com a tradução: uma leitura que, ao invés de ler o real, o traduz com as forças intensivas do mundo, produzindo afecções nos corpos envolvidos (o leitor, o escritor, o texto, o próprio entorno) e fazendo variar sua potência; uma leitura que envolve as artistagens tradutórias de um agenciamento coletivo de enunciação pelo traçado de linhas de escrita e de vida; uma escrita como invenção: inscrição singular de um si-mundo; traçado desejante de criação. Pretende, portanto, defender que a aprendizagem da linguagem acontece como atividade expressiva quando há composição de um bloco de devir-aprendente por meio da criação de um estilo. Para tanto, recorre a algumas ferramentas conceituais produzidas por Gilles Deleuze (1925-1995) e Félix Guattari (1930-1992), Gilbert Simondon (1924-1989), Baruck Spinoza (1632-1677), Suely Rolnik (2006), Sandra Corazza (2013), Virginia Kastrup (1999) e Walter Kohan (2007), de modo a tentar intervir concretamente nas discussões em torno dos conceitos de aprendizagem, linguagem, tempo, signos, acontecimento, devir, escrita e estilo, seguindo pelo conceito de individuação e pelos movimentos de exploração intensiva dos meios cartografados nos encontros educativos estabelecidos em um Centro de Educação Infantil de Vitória.

The influence of light spectra, UV-A, and growth regulators on the in vitro seed germination of Senecio cineraria DC.

This study was carried out to investigate the effects of light spectra, additional UV-A, and different growth regulators on the in vitro germination of Senecio cineraria DC. Seeds were surface-sterilized and inoculated in MS medium to evaluate the following light spectra: white, white plus UV-A, blue, green, red or darkness. The maximum germinability was obtained using MS0 medium under white light (30%) and MS + 0.3 mg L-1 GA3 in the absence of light (30.5%). S. cineraria seeds were indifferent to light. Blue and green lights inhibited germination. Different concentrations of gibberellic acid (GA3) (0.1; 0.4; 0.6; 0.8; 1.0 and 2.0 mg L-1) and indole-3-acetic acid IAA (0.1; 0.3 and 1.0 mg L-1) were evaluated under white light and darkness. No concentration of GA3 enhanced seed germination percentage under white light. However, when the seeds were maintained in darkness, GA3 improved germination responses in all tested concentrations, except at 1.0 mg L-1. Under white light, these concentrations also increased the germination time and reduced germination rate. Germination rate, under light or darkness, was lower using IAA compared with GA3.

Foucault’s Perspective of Performance Appraisal in Public Administration

The main purpose of performance appraisal in organizations is, or should be, to improve the engagement, learning process and progress of the employees and to align individual with team and organizational performance. However, performance appraisal can also be interpreted as an instrument of normalization, discipline and surveillance. This study thus aims to explore and discuss the complex schema of intrinsic and extrinsic objectives of performance appraisal system of the Portuguese public organizations (SIADAP - Performance Evaluation Integrated System). We have developed an exploratory and qualitative case study to capture appraisers and appraises perceptions. The data were analyzed in light of foucauldian theories. According to the qualitative data, namely the discourses of the appraisers and their subordinates, the SIADAP is seen as an instrument of control and dominance that aims to introduce political rationalities, limiting the career progression of the employees. Though some key points of Foucault’s perspective were identified, foucauldian framework revealed some limitations to capture all the complexity inherent to performance appraisal. This study opens new perspectives about the SIADAP and can be of major importance as far as political reflection about performance appraisal in public organizations is concerned.

Innovation through virtual communities of Practice: motivation and constraints in the knowledge-creation process

Communities of Practice are places which provide a sound basis for organizational learning, enabling knowledge creation and acquisition thus improving organizational performance, leveraging innovation and consequently increasing competitively. Virtual Communities of Practice (VCoP‟s) can perform a central role in promoting communication and collaboration between members who are dispersed in both time and space. The ongoing case study, described here, aims to identify both the motivations and the constraints that members of an organization experience when taking part in the knowledge creating processes of the VCoP‟s to which they belong. Based on a literature review, we have identified several factors that influence such processes; they will be used to analyse the results of interviews carried out with the leaders of VCoP‟s in four multinationals. As future work, a questionnaire will be developed and administered to the other members of these VCoP‟s

Virtual communities of practice: investigating motivations and constraints in the processes of knowledge creation and transfer

With accelerated market volatility, faster response times and increased globalization, business environments are going through a major transformation and firms have intensified their search for strategies which can give them competitive advantage. This requires that companies continuously innovate, to think of new ideas that can be transformed or implemented as products, processes or services, generating value for the firm. Innovative solutions and processes are usually developed by a group of people, working together. A grouping of people that share and create new knowledge can be considered as a Community of Practice (CoP). CoP’s are places which provide a sound basis for organizational learning and encourage knowledge creation and acquisition. Virtual Communities of Practice (VCoP's) can perform a central role in promoting communication and collaboration between members who are dispersed in both time and space. Nevertheless, it is known that not all CoP's and VCoP's share the same levels of performance or produce the same results. This means that there are factors that enable or constrain the process of knowledge creation. With this in mind, we developed a case study in order to identify both the motivations and the constraints that members of an organization experience when taking part in the knowledge creating processes of VCoP's. Results show that organizational culture and professional and personal development play an important role in these processes. No interviewee referred to direct financial rewards as a motivation factor for participation in VCoPs. Most identified the difficulty in aligning objectives established by the management with justification for the time spent in the VCoP. The interviewees also said that technology is not a constraint.

Virtual communities of practice: examining the motivations and constraints in their knowledge creation and knowledge transfer preocesses

Paper accepted for the OKLC 2009 - International Conference on Organizational Learning, Knowledge and Capabilities (26-28th, April 2009, Amsterdam, the Netherlands).

Factors related with adiponectinemia in obese and normal-weight women and with its variation in weight loss programs

Objective: To assess different factors influencing adiponectinemia in obese and normal-weight women; to identify factors associated with the variation (Δ) in adiponectinemia in obese women following a 6-month weight loss program, according to surgical/non-surgical interventions. Methods: We studied 100 normal-weight women and 112 obese premenopausal women; none of them was on any medical treatment. Women were characterized for anthropometrics, daily macronutrient intake, smoking status, contraceptives use, adiponectin as well as IL-6 and TNF-α serum concentrations. Results: Adiponectinemia was lower in obese women (p < 0.001), revealing an inverse association with waist-to-hip ratio (p < 0.001; r = –0.335). Normal-weight women presented lower adiponectinemia among smokers (p = 0.041); body fat, waist-to-hip ratio, TNF-α levels, carbohydrate intake, and smoking all influence adiponectinemia (r 2 = 0.436). After weight loss interventions, a significant modification in macronutrient intake occurs followed by anthropometrics decrease (chiefly after bariatric procedures) and adiponectinemia increase (similar after surgical and non-surgical interventions). After bariatric intervention, Δ adiponectinemia was inversely correlated to Δ waist circumference and Δ carbohydrate intake (r 2 = 0.706). Conclusion: Anthropometrics, diet, smoking, and TNF-α levels all influence adiponectinemia in normal-weight women, although explaining less than 50% of it. In obese women, anthropometrics modestly explain adiponectinemia. Opposite to non-surgical interventions, after bariatric surgery adiponectinemia increase is largely explained by diet composition and anthropometric changes.