Effects of asbestos and silica on superoxide anion production in the guinea pig alveolar macrophage

Asbestos and silica are important industrial hazards. Exposure to these dusts can result in pulmonary fibrosis and, in the case of asbestos, cancer. Although the hazards of asbestos and silica exposure have long been known, the pathogenesis of dust-related disease is not well understood. Both silica and asbestos are thought to alter the function of the alveolar macrophage, but the nature of the biochemical alteration is unknown. Therefore, this study examined the effect of asbestos and silica on the activation pathway of the guinea pig alveolar macrophage. Activation of macrophages by physiological agents results in stimulation of phospholipase C causing phosphatidyl inositol turnover and intracellular calcium mobilization. Phosphatidyl inositol turnover produces diacylglycerol which activates protein kinase C causing superoxide anion production.^ Chrysotile stimulated alveolar macrophages to produce superoxide anion. This stimulation proceeded via phospholipase C, since chrysotile stimulated phosphatidyl inositol turnover and intracellular calcium mobilization. The possible involvement of a coupling protein was evaluated by pretreating cells with pertussis toxin. Pertussis toxin pretreatment partially inhibited chrysotile stimulation, suggesting that chrysotile activates a coupling protein in an non-classical manner. Potential binding sites for chrysotile stimulation were examined using a series of nine lectins. Chrysotile-stimulated superoxide anion production was blocked by pretreatment with lectins which bound to N-acetylglucosamine, but not by lectins which bound to mannose, fucose, or N-acetylgalactosamine. In addition, incubation with the N-acetylglucosamine polymer, chitin, inhibited chrysotile-stimulated superoxide anion production, suggesting that chrysotile stimulated superoxide anion production by binding to N-acetylglucosamine residues.^ On the other hand, silica did not stimulate superoxide anion production. The effect of silica on agonist stimulation of this pathway was examined using two stimulants of superoxide anion production, N-formyl-nle-leu-phe (FNLP, which stimulates through phospholipase C) and phorbol-12,13-dibutyrate (which directly activates protein kinase C). Sublethal doses of silica inhibited FNLP-stimulated superoxide anion production, but did not affect phorbol-12,13-dibutyrate-stimulated superoxide anion production, suggesting that the site of inhibition precedes protein kinase C. This inhibition was not due to cell membrane damage, since cell permeability to calcium-45 and rubidium-86 was not increased. It is concluded that chrysotile binds to N-acetylglucosamine residues on macrophage surface glycoproteins to stimulate the physiological pathway resulting in superoxide anion production. In contrast, silica does not stimulate superoxide anion production, but it did inhibit FNLP-stimulated superoxide anion production. ^

La hiperactividad y el empuje de la civilización

La clínica y el trabajo en las instituciones nos muestran cambios dramáticos en la subjetividad contemporánea. Las personas se encuentran expuestas a demandas sociales que tienen consecuencias funestas sobre ellas. Y que se manifiestan en grados diversos de hiperactividad, o actividad compulsiva, hasta el colapso subjetivo bajo la forma de depresión o irrupciones masivas de angustia. En los niños se verifica desde hace un tiempo un fenómeno al que se le ha dado un nombre Trastorno por Déficit Atencional con Hiperactividad, y cuyo tratamiento se confía al uso paradójico de estimulantes. En este artículo se relacionan las causas de este fenómeno, con las modalidades culturales que propician el reemplazo de la experiencia subjetiva por protocolos de acción predeterminados.

Growth and viability of macrophages continuously stimulated to produce nitric oxide

Deregulated production of nitric oxide (NO) has been implicated in the development of certain human diseases, including cancer. We sought to assess the damaging potential of NO produced under long-term conditions through the development of a suitable model cell culture system. In this study, we report that when murine macrophage-like RAW264.7 cells were exposed continuously to bacterial lipopolysaccharide (LPS) or mouse recombinant interferon-γ (IFN-γ) over periods of 21–23 days, they continued to grow, but with doubling times 2 to 4 times, respectively, longer than the doubling time of unstimulated cells. Stimulated cells produced NO at rates of 30 to 70 nmol per million cells per day throughout the stimulation period. Within 24 hr after removal of stimulant, cells resumed exponential growth. Simultaneous exposure to LPS and IFN-γ resulted in decreased cell number, which persisted for 2 days after removal of the stimulants. Exponential growth was attained only after an additional 4 days. Addition of N-methyl-l-arginine (NMA), an NO synthase inhibitor, to the medium inhibited NO production by 90% of all stimulated cells, partially reduced doubling time of cells stimulated with LPS or IFN-γ, and partially increased viability and growth rates in those exposed to both LPS and IFN-γ. However, when incubated with LPS and IFN-γ at low densities both in the presence and in the absence of NMA, cells grew at a rate slower than that of unstimulated cells, with no cell death, and they resumed exponential growth 24 hr after removal of stimulants. Results from cell density experiments suggest that macrophages are protected from intracellularly generated NO; much of the NO damaging activity occurred outside of the producer cells. Collectively, results presented in this study suggest that the type of cellular toxicity observed in macrophages is markedly influenced by rate of exposure to NO: at low rates of exposure, cells exhibit slower growth; at higher rates, cells begin to die; at even higher rates, cells undergo growth arrest or die. The ability of RAW264.7 cells to produce NO over many cell generations makes the cell line a useful system for the study of other aspects of cellular damage, including genotoxicity, resulting from exposure to NO under long-term conditions.

Cellular responses to psychomotor stimulant and neuroleptic drugs are abnormal in mice lacking the D1 dopamine receptor

Stimulation of dopamine D1 receptors has profound effects on addictive behavior, movement control, and working memory. Many of these functions depend on dopaminergic systems in the striatum and D1–D2 dopamine receptor synergies have been implicated as well. We show here that deletion of the D1 dopamine receptor produces a neural phenotype in which amphetamine and cocaine, two addictive psychomotor stimulants, can no longer stimulate neurons in the striatum to express cFos or JunB or to regulate dynorphin. By contrast, haloperidol, a typical neuroleptic that acts preferentially at D2-class receptors, remains effective in inducing catalepsy and striatal Fos/Jun expression in the D1 mutants, and these behavioral and neural effects can be blocked by D2 dopamine receptor agonists. These findings demonstrate that D2 dopamine receptors can function without the enabling role of D1 receptors but that D1 dopamine receptors are essential for the control of gene expression and motor behavior by psychomotor stimulants.

Neurosecretory habituation in PC12 cells: modulation during parallel habituation.

PC12 cells habituate during repetitive stimulation with acetylcholine, bradykinin, or high potassium. Interspersing these stimulants did not affect the rate of habituation of the others, but it could modulate the amplitude of the norepinephrine secretion each could achieve. Stimulation with acetylcholine inhibited norepinephrine secretion caused by high potassium and bradykinin stimulation, while high potassium had no effect on acetylcholine or bradykinin, and bradykinin increased secretion caused by acetylcholine. Changes in norepinephrine secretion resulting from any of these stimulants correlated with changes in internal calcium levels. Cyclic AMP-, protein kinase C-, and calmodulin-dependent second messenger pathways all modulated norepinephrine secretion caused by acetylcholine and high potassium and showed a distinct hierarchy in their effectiveness. These data demonstrate that different receptor pathways can change the norepinephrine response of one another while not changing the levels of the molecules responsible for habituation.

Absenteísmo na equipe de enfermagem e suas implicações para a assistência prestada em unidades de urgência e emergência

Trata-se de um estudo descritivo e exploratório, com abordagem qualitativa dos dados. Objetivou-se analisar a percepção da equipe de enfermagem sobre as condições geradoras de absenteísmo e suas implicações na assistência nas unidades de Urgência e Emergência (UE) das cinco distritais de saúde no município de Ribeirão Preto/SP. Os sujeitos foram profissionais da equipe de enfermagem (enfermeiro, auxiliar e técnico de enfermagem) que atuam nestas unidades. Foram selecionados 2 profissionais de cada categoria, a partir dos critérios de inclusão do estudo, sem considerar sexo, faixa etária e tempo de trabalho no serviço, totalizando 30 participantes. Os dados foram coletados por meio de entrevistas individuais semiestruturadas, conduzidas a partir de um roteiro norteador composto pelas variáveis (Processo de Gestão de Recursos Humanos (RH); Condição de Trabalho em Equipe e Qualidade do Cuidado Prestado) abordadas no estudo. Para análise dos dados utilizou-se análise de conteúdo, modalidade temática. Após análise dos dados, as categorias encontradas foram: TEMA 1 - Gerenciamento, organização e enfrentamento para a operacionalização do trabalho de enfermagem (Subtema 1 - Operacionalização da escala de trabalho frente ao desafio do quantitativo da equipe de enfermagem na unidade de UE; Subtema 2 - Reorganização do trabalho e a perspectiva dos trabalhadores frente à mudança para as 30h/semanais e a terceirização do serviço; Subtema 3 - Tempo de permanência do profissional no serviço; Subtema 4 - Comunicação como ferramenta para desenvolver o trabalho em equipe e gerenciar conflitos); TEMA 2 - Condições impostas ao trabalhador e sua influência no desenvolvimento do trabalho (Subtema 1 - Plano de carreira e salário como estimulantes para desenvolvimento do trabalho; Subtema 2 - Vínculo empregatício: vantagens e desvantagens; Subtema 3 - Educação permanente e sua importância para desenvolvimento do trabalho; Subtema 4 - Influência da estrutura física, materiais e equipamentos no cuidado) e TEMA 3 - Avaliação do serviço e da assistência prestada. No que diz respeito ao quantitativo de enfermagem disposto nas unidades, todos os entrevistados relatam que é um quantitativo razoável e que, em alguns momentos, se sentem sobrecarregados quando ocorrem ausências não previstas. Ao se tratar da terceirização das unidades estatutárias, relata-se que não houve comunicação prévia do evento e é visível a insegurança e frustração por parte dos entrevistados. Ressalta-se que a unidade terceirizada não sofreu mudanças em sua rotina. A rotatividade é presente nestas unidades de UE, sendo maior em determinada unidade e ocorre por inúmeros motivos, dentre eles, aposentadoria, transferência para Unidade Básica de Saúde (UBS), conflitos na equipe e/ou com pacientes, dentre outros. Todos os entrevistados sugerem que a comunicação é fundamental para o desenvolvimento do trabalho em equipe e é através dela que os conflitos possam vir a ser resolvidos. Neste momento, percebe-se, a partir das falas, que a comunicação é diferente entre as unidades e, portanto, existem níveis diferentes de conflitos entre as unidades. O município não possui um plano de carreira efetivo, portanto os entrevistados demonstram desmotivação para buscar novos conhecimentos. Quanto ao salário, estes têm a visão de que é razoável, sendo considerado elevado em relação às demais instituições de saúde do município, porém, defasado em relação à categoria profissional. Os profissionais terceirizados relatam uma certa insatisfação por trabalhar da mesma forma que os estatutários, recebendo um menor salário e sem os mesmos benefícios, o que nos leva à categoria vínculo empregatício, onde a estabilidade é abordada com visões positivas e negativas. Ao se tratar da visão negativa, os entrevistados sugerem que muitos colegas não sabem lidar com esse benefício, se ausentando do trabalho ou trabalhando de uma forma não adequada, prejudicando a rotina do serviço. No que tange à educação permanente, temos a diferença mais gritante do estudo, visto que os entrevistados estatutários relatam que não possuem a disponibilização, através da prefeitura, de cursos de atualização, capacitação e constante aprendizado enquanto que os terceirizados relatam atualizações constantes e apoio por parte da instituição com a qual eles estão vinculados. É unânime que todos os entrevistados consideram que a estrutura física, materiais e equipamentos interferem diretamente no cuidado. Ao serem questionados em relação à avaliação do cuidado prestado, eles o consideram bom, podendo ser melhor caso fossem disponibilizadas condições de trabalho mais adequadas. Considera-se o estudo como um possível instrumento de avaliação dos serviços prestados em unidade de UE, bem como das condições de trabalho fornecidas ao trabalhador e sua satisfação profissional

Drug abuse - game without winners. A basic handbook for commanders. Second edition.

Mode of access: Internet.

A comparison of drug use in driver fatalities and similarly exposed drivers. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Untersuchungen über die Wirkung von Convallamarin bei Hunden.

Inaug.-diss.-Hannover, 1912.

Dissertatio academica, sistens inebriantia ... /

Also published in Linné's Amoenitates academicae, v. 6. ed. 1, 1763; ed. 2, 1789, p. 180-196; and Gilbert's Systema plantarum Europe, v. 6, 1786, p. 399-414.

Subtypes of Illicit Drug Users: A Latent Class Analysis of Data From an Australian Twin Sample

This article applies methods of latent class analysis (LCA) to data on lifetime illicit drug use in order to determine whether qualitatively distinct classes of illicit drug users can be identified. Self-report data on lifetime illicit drug use (cannabis, stimulants, hallucinogens, sedatives, inhalants, cocaine, opioids and solvents) collected from a sample of 6265 Australian twins (average age 30 years) were analyzed using LCA. Rates of childhood sexual and physical abuse, lifetime alcohol and tobacco dependence, symptoms of illicit drug abuse/dependence and psychiatric comorbidity were compared across classes using multinomial logistic regression. LCA identified a 5-class model: Class 1 (68.5%) had low risks of the use of all drugs except cannabis; Class 2 (17.8%) had moderate risks of the use of all drugs; Class 3 (6.6%) had high rates of cocaine, other stimulant and hallucinogen use but lower risks for the use of sedatives or opioids. Conversely, Class 4 (3.0%) had relatively low risks of cocaine, other stimulant or hallucinogen use but high rates of sedative and opioid use. Finally, Class 5 (4.2%) had uniformly high probabilities for the use of all drugs. Rates of psychiatric comorbidity were highest in the polydrug class although the sedative/opioid class had elevated rates of depression/suicidal behaviors and exposure to childhood abuse. Aggregation of population-level data may obscure important subgroup differences in patterns of illicit drug use and psychiatric comorbidity. Further exploration of a 'self-medicating' subgroup is needed.

Protein kinase C activation and its role in kidney disease

Protein kinase C (PKC) comprises a superfamily of isoenzymes, many of which are activated by cofactors such as diacylglycerol and phosphatidylserine. In order to be capable of activation, PKC must first undergo a series of phosphorylations. In turn, activated PKC phosphorylates a wide variety of intracellular target proteins and has multiple functions in signal transduced cellular regulation. A role for PKC activation had been noted in several renal diseases, but two that have had most investigation are diabetic nephropathy and kidney cancer. In diabetic nephropathy, an elevation in diacylglycerol and/or other cofactor stimulants leads to an increase in activity of certain PKC isoforms, changes that are linked to the development of dysfunctional vasculature. The ability of isoform-specific PKC inhibitors to antagonize diabetes-induced vascular disease is a new avenue for treatment of this disorder. In the development and progressive invasiveness of kidney cancer, increased activity of several specific isoforms of PKC has been noted. It is thought that this may promote the kidney cancer's inherent resistance to apoptosis, in natural regression or after treatments, or it may promote the invasiveness of renal cancers via cellular differentiation pathways. In general, however, a more complete understanding of the functions of individual PKC isoforms in the kidney, and development or recognition of specific inhibitors or promoters of their activation, will be necessary to apply this knowledge for treatment of cellular dysregulation in renal disease.

Loss of skeletal muscle in cancer: biochemical mechanisms

Patients with cancer often undergo a specific loss of skeletal muscle mass, while the visceral protein reserves are preserved. This condition known as cachexia reduces the quality of life and eventually results in death through erosion of the respiratory muscles. Nutritional supplementation or appetite stimulants are unable to restore the loss of lean body mass, since protein catabolism is increased mainly as a result of the activation of the ATP-ubiquitin-dependent proteolytic pathway. Several mediators have been proposed. An enhanced protein degradation is seen in skeletal muscle of mice administered tumour necrosis factor (TNF), which appears to be mediated by oxidative stress. There is some evidence that this may be a direct effect and is associated with an increase in total cellular-ubiquitin-conjugated muscle proteins. Another cytokine, interleukin-6 (IL-6), may play a role in muscle wasting in certain animal tumours, possibly through both lysosomal (cathepsin) and non-lysosomal (proteasome) pathways. A tumour product, proteolysis-inducing factor (PIF) is produced by cachexia-inducing murine and human tumours and initiates muscle protein degradation directly through activation of the proteasome pathway. The action of PIF is blocked by eicosapentaenoic acid (EPA), which has been shown to attenuate the development of cachexia in pancreatic cancer patients. When combined with nutritional supplementation EPA leads to accumulation of lean body mass and prolongs survival. Further knowledge on the biochemical mechanisms of muscle protein catabolism will aid the development of effective therapy for cachexia.

Analysis of Transglutaminase-2 in macrophage function

Programmed cell death, apoptosis, is a highly regulated process that removes damaged or unwanted cells in vivo and has significant immunological implications. Defective clearance of apoptotic cells by macrophages (professional phagocytes) is known to result in chronic inflammatory and autoimmune disease. Transglutaminase-2 (TG2) is a Ca2+-dependent protein crosslinking enzyme known to play an important role in apoptotic cell clearance by macrophages through an ill-defined mechanism. Several studies have implicated TG2 in the apoptosis programme e.g. raised TG2 levels in cells undergoing apoptosis; increased cell death with down-regulation of TG2; up-regulation of TG2 in monocytes upon differentiation into macrophages. Defective clearance of apoptotic cells by TG2 null mice has been described though in this context the role of TG2 is yet to be elucidated. Here we aim to characterise the role of TG2 in macrophage function with a focus on apoptotic cell clearance. THP-1 monocytes were induced to differentiate to macrophage-like cells by three different stimulants and were analysed for the presence of TG2. Macrophage-apoptotic cell interaction studies in the presence and absence of irreversible TG2 inhibitors resulted in significant inhibition of interaction indicating a possible role for TG2 in apoptotic cell clearance. TG2 was expressed at the macrophage cell surface and its association with ß3 integrin expression suggests the possible link between TG2 and ß3 integrins. Our current findings suggest that TG2 had got a crucial but yet to be defined role in apoptotic cell clearance.

The control of mitosis in mammalian tissues

The question of which factors are central in determining whether a cell will undertake a new round of mitosis or will decycle has been examined in the isolated thymic lymphocyte model. Such cell populations possess both in vivo and in vitro a subpopulation of quiescent lymphoblasts which may be induced to reinitiate their mitotic programme. In the intact animal the major determinant of proliferative activity is the plasma ionised calcium concentration. However it has been established in culture that a variety of hormones, ions, cyclic nucleotides, plant lectins and ionophores may like calcium elicit a mitogenic response. These agents do not appear however to initiate DNA synthesis in an identical fashion. Rather there are two distinct intracellular mitogenic axes. The first axis includes a number of adenylate cyclase stimulants, cyclic AMP, phosphodiesterase inhibitors and magnesium ions. It was found that all these mitogens required extracellular magnesium ions to exhibit their stimulatory capacity. This dichotomy in mitogenic activity was further emphasised by the observation that these mitogens are all inhibited by testosterone, whilst the magnesium-independent mitogens were insensitive to this androgen. Indeed this second group of stimulatory factors required the presence of calcium ions in the extracellular milieu for activity, and were, in contrast to the magnesium-dependent mitogens inhibited by the presence of oestradiol in the culture. By examining the interrelationships between these various mitogens and inhibitors it has been possible to propose a mechanism to describe the activation process in the thymocyte. Studies of the metabolism of cyclic nucleotides, membrane potential and transmembrane ion fluxes indicate that there may be a complex relationship between membrane fluidity, ion balance and cyclic nucleotide levels which may individually or in concert promote the initiation of DNA synthesis. A number of possible mechanisms are discussed to account for these observations.