Natural selection on fecundity variance in subdivided populations: kin selection meets bet hedging.

In a series of seminal articles in 1974, 1975, and 1977, J. H. Gillespie challenged the notion that the "fittest" individuals are those that produce on average the highest number of offspring. He showed that in small populations, the variance in fecundity can determine fitness as much as mean fecundity. One likely reason why Gillespie's concept of within-generation bet hedging has been largely ignored is the general consensus that natural populations are of large size. As a consequence, essentially no work has investigated the role of the fecundity variance on the evolutionary stable state of life-history strategies. While typically large, natural populations also tend to be subdivided in local demes connected by migration. Here, we integrate Gillespie's measure of selection for within-generation bet hedging into the inclusive fitness and game theoretic measure of selection for structured populations. The resulting framework demonstrates that selection against high variance in offspring number is a potent force in large, but structured populations. More generally, the results highlight that variance in offspring number will directly affect various life-history strategies, especially those involving kin interaction. The selective pressures on three key traits are directly investigated here, namely within-generation bet hedging, helping behaviors, and the evolutionary stable dispersal rate. The evolutionary dynamics of all three traits are markedly affected by variance in offspring number, although to a different extent and under different demographic conditions.

Role of MyD88 and Toll-like receptors 2 and 4 in the sensing of Parachlamydia acanthamoebae.

Parachlamydia acanthamoebae is a Chlamydia-related organism whose pathogenic role in pneumonia is supported by serological and molecular clinical studies and an experimental mouse model of lung infection. Toll-like receptors (TLRs) play a seminal role in sensing microbial products and initiating innate immune responses. The aim of this study was to investigate the roles of MyD88, TLR2, and TLR4 in the interaction of Parachlamydia with macrophages. Here, we showed that Parachlamydia entered bone-marrow derived macrophages (BMDMs) in a TLR-independent manner but did not multiply intracellularly. Interestingly, compared to live bacteria, heat-inactivated Parachlamydia induced the production of substantial amounts of tumor necrosis factor alpha (TNF), interleukin-6 (IL-6), and IL-12p40 by BMDMs and of TNF and IL-6 by peritoneal macrophages as well as RAW 264.7 and J774 macrophage cell lines. Cytokine production by BMDMs, which was partially inhibited upon trypsin treatment of Parachlamydia, was dependent on MyD88, TLR4, and, to a lesser extent, TLR2. Finally, MyD88(-/-), TLR4(-/-), and TLR2(-/-) mice were as resistant as wild-type mice to lung infection following the intratracheal instillation of Parachlamydia. Thus, in contrast to Chlamydia pneumoniae, Parachlamydia acanthamoebae weakly stimulates macrophages, potentially compensating for its low replication capacity in macrophages by escaping the innate immune surveillance.

Calcium Microdomains Control Exo-Endocytosis of Synaptic-Like Microvesicles in Astrocytes

During the last decade, the discovery that astrocytes possess a nonelectrical form of excitability (Ca21-excitability) that leads to the release of chemical transmitters, an activity called ''gliotransmission'', indicates that these cells may have additional important roles in brain function. Elucidating the stimulus-secretion coupling leading to the exocytic release of chemical transmitters (such as glutamate, Bezzi et al., Nature Neurosci, 2004) may therefore clarify i) whether astrocytes represent in full a new class of secretory cells in the brain and ii) whether they can participate to the fast brain signaling in the brain. Here by using a recently developed approach for studying vesicle recycling dynamics at synapses (Voglmaier et al., Neuron, 2006; Balaji and Ryan, PNAS, 2007) combined with epifluorescence and total internal reflection fluorescence (TIRF) imaging, we investigated the spatiotemporal characteristics of stimulus-secretion coupling leading glutamate exocytosis of synaptic-like microvesicles (SLMVs) in astrocytes. We performed the analysis at both the whole-cell and single-vesicle levels providing the first system for comparing exo-endocytic processes in astrocytes with those in neurons. Both the time course and modalities of secretion in astrocytes present more similarities to neurons then previously expected. We found that 1. the G-protein-coupled receptor (GPCR)-evoked exocytosis reached the maximum on a ms time scale and that 2. ER tubuli formed sub-micrometer domains beneath the plasma membrane in close proximity to exocytic vesicles, where fusion events were spatiotemporally correlated with fast Ca21 events.

Desempenho reprodutivo de porcas submetidas a infusões uterinas no início do estro

O objetivo deste trabalho foi avaliar o desempenho reprodutivo de porcas multíparas submetidas à infusão uterina de diferentes soluções, realizada no início do estro. Um total de 1.019 fêmeas foram controladas no período de verão (n=570) e inverno (n=449). Os animais foram submetidos a cinco tratamentos, que consistiram em infusão de plasma seminal (PS), sêmen morto (SM), solução de 17beta-estradiol (SE), solução fisiológica (SS) e um grupo-controle (CO). As fêmeas receberam três inseminações: a primeira, 8-12 horas após a detecção do estro, e as demais, nos turnos subseqüentes. Com relação à taxa de retorno ao estro e taxa de parto ajustada, não ocorreram diferenças entre os tratamentos (p>0,05). Na análise do modelo de regressão adotado para determinar o tamanho da leitegada, foi observada uma interação entre época do ano e tratamento (p<=0,01). No verão, as fêmeas que receberam infusão de PS apresentaram 0,89, 1,20, 1,34 e 2,31 leitões a mais, em relação aos tratamentos SE, SM, SS e CO (p<0,05), respectivamente. No inverno, o grupo submetido ao tratamento SS aumentou a produção em 1,25 e 0,91 leitões, respectivamente, em relação aos tratamentos SM e CO (p<0,05). Este trabalho demonstrou que a infusão uterina de PS foi eficiente somente no verão, para aumentar o número de leitões nascidos. Entretanto, são necessários estudos complementares esclarecendo o efeito deste tratamento em épocas do ano distintas e em diferentes ordens de parto.

Efeito do extrato aquoso de leucena na germinação e no desenvolvimento do milho

Foram desenvolvidos bioensaios, na Embrapa-Centro Nacional de Pesquisa de Milho e Sorgo (CNPMS), para avaliar o efeito de extratos aquosos, a frio e a quente, da parte aérea de leucena (Leucaena leucocephala (Lam.) De Wit.), sobre a germinação e o desenvolvimento das plantas de milho (Zea mays L.). Os extratos foram preparados na concentração 20% (p/v), e avaliados em solo e papel- germiteste, em casa de vegetação, e em papel-filtro em laboratório. O extrato obtido com água fria (EF) e aplicado ao solo não causou nenhum efeito fitotóxico sobre a germinação e o desenvolvimento das plantas de milho. O extrato obtido com água quente (EQ), quando aplicado em papel-germiteste ou papel-filtro, causou redução no comprimento da raiz seminal, mas não interferiu na germinação das sementes de milho. O comprimento da raiz seminal foi um indicador mais sensível aos efeitos do EQ do que a germinação.

Parâmetros escroto-testiculares e de sêmen em caprinos adultos submetidos à insulação escrotal

Ejaculados de seis bodes Moxotó e seis ½ sangue Moxotó-Pardo Alpina, adultos, mantidos em confinamento, foram avaliados antes e após a insulação do saco escrotal com bolsa de plástico, de parede dupla, durante 6,5 dias. A degeneração seminal ocorreu em todos os animais na 4ª semana após o início da insulação, ressaltando-se a redução na concentração, o aumento dos defeitos espermáticos e a redução do vigor celular, culminando com a necrospermia. A motilidade individual progressiva (MIP) atingiu os valores mais baixos na 3ª semana após o início da insulação, retornando aos valores normais entre a 8ª e 9ª semanas. Os defeitos espermáticos começaram a aumentar aos sete dias após o início da insulação escrotal. O volume apresentou oscilações atípicas e o aumento da temperatura escrotal afetou os parâmetros físicos e morfológicos do ejaculado de maneira consistente evidenciando-se o efeito negativo sobre a qualidade do sêmen.

Membrane interaction of polymyxin B and synthetic analogues studied in biomimetic systems: implications for antibacterial action

Membrane-active antimicrobial peptides, such as polymyxin B (PxB), are currently in the spotlight as potential candidates toovercome bacterial resistance. We have designed synthetic analogs ofPxB in order to determine the structural requirements for membraneaction. Since the mechanism of action of PxB involves interaction withboth the outer membrane and the cytoplasmic membrane of Gramnegative bacteria, we have used an approach based on mimicking theouter layers of these membranes using monolayers, Langmuir-Blodgettfilms and unilamelar vesicles, and applying a battery of biophysicalmethods in order to dissect the different events of membraneinteraction. Collectively, results indicate that the PxB analogues act inthe bacterial membrane by the same mechanism than PxB, and that cationic amphipathicity determines peptide activity.

Exovesicles from human activated dendritic cells fuse with resting dendritic cells, allowing them to present alloantigens.

Dendritic cells (DCs) can release microvesicles, but the latter's numbers, size, and fate are unclear. Fluorescently labeled DCs were visualized by laser-scanning microscopy. Using a Surpass algorithm, we were able to identify and quantify per cell several hundred microvesicles released from the surface of stimulated DCs. We show that most of these microvesicles are not of endocytic origin but result from budding of the plasma membrane, hence their name, exovesicle. Using a double vital staining, we show that exovesicles isolated from activated DCs can fuse with the membrane of resting DCs, thereby allowing them to present alloantigens to lymphocytes. We concluded that, within a few hours from their release, exovesicles may amplify local or distant adaptive immunological response.

Fas and Fas ligand in embryos and adult mice: ligand expression in several immune-privileged tissues and coexpression in adult tissues characterized by apoptotic cell turnover.

The cell surface receptor Fas (FasR, Apo-1, CD95) and its ligand (FasL) are mediators of apoptosis that have been shown to be implicated in the peripheral deletion of autoimmune cells, activation-induced T cell death, and one of the two major cytolytic pathways mediated by CD8+ cytolytic T cells. To gain further understanding of the Fas system., we have analyzed Fas and FasL expression during mouse development and in adult tissues. In developing mouse embryos, from 16.5 d onwards, Fas mRNA is detectable in distinct cell types of the developing sinus, thymus, lung, and liver, whereas FasL expression is restricted to submaxillary gland epithelial cells and the developing nervous system. Significant Fas and FasL expression were observed in several nonlymphoid cell types during embryogenesis, and generally Fas and FasL expression were not localized to characteristic sites of programmed cell death. In the adult mouse, RNase protection analysis revealed very wide expression of both Fas and FasL. Several tissues, including the thymus, lung, spleen, small intestine, large intestine, seminal vesicle, prostate, and uterus, clearly coexpress the two genes. Most tissues constitutively coexpressing Fas and FasL in the adult mouse are characterized by apoptotic cell turnover, and many of those expressing FasL are known to be immune privileged. It may be, therefore, that the Fas system is implicated in both the regulation of physiological cell turnover and the protection of particular tissues against potential lymphocyte-mediated damage.

Avaliação de nove linhagens de milho em cruzamentos dialélicos quanto à tolerância ao alumínio

Avaliaram-se nove linhagens endogâmicas de milho e seus cruzamentos quanto à tolerância ao alumínio (Al), em um dialelo incompleto. Foram utilizados os índices fenotípicos de comprimento relativo de raiz seminal (CRRS), comprimento líquido de raiz seminal (CLRS) e coloração por hematoxilina, determinados após sete dias de crescimento em solução nutritiva contendo 222 mimoles L-1 de Al. Os resultados dos cruzamentos dialélicos demonstraram que os efeitos aditivos foram mais importantes que os efeitos não-aditivos para os três índices avaliados. As linhagens L13, L724, L723 e L16 seriam as mais indicadas para a obtenção de híbridos visando tolerância ao Al, por apresentarem os melhores valores de capacidade geral de combinação e por participarem dos cruzamentos de melhor capacidade específica de combinação. Uma correlação de 0,76 foi observada entre os índices CLRS e a coloração por hematoxilina, de 0,63 entre CLRS e CRRS e de 0,27 entre CRRS e coloração por hematoxilina. A coloração por hematoxilina demonstrou ser uma valiosa ferramenta para programas de melhoramento de milho que visam a seleção de genótipos com maior tolerância ao alumínio.

Assessing inter-rater reliability when the raters are fixed: two concepts and two estimates.

Intraclass correlation (ICC) is an established tool to assess inter-rater reliability. In a seminal paper published in 1979, Shrout and Fleiss considered three statistical models for inter-rater reliability data with a balanced design. In their first two models, an infinite population of raters was considered, whereas in their third model, the raters in the sample were considered to be the whole population of raters. In the present paper, we show that the two distinct estimates of ICC developed for the first two models can both be applied to the third model and we discuss their different interpretations in this context.

Membrane interaction of polymyxin B and synthetic analogues studied in biomimetic systems: implications for antibacterial action

Membrane-active antimicrobial peptides, such as polymyxin B (PxB), are currently in the spotlight as potential candidates toovercome bacterial resistance. We have designed synthetic analogs ofPxB in order to determine the structural requirements for membraneaction. Since the mechanism of action of PxB involves interaction withboth the outer membrane and the cytoplasmic membrane of Gramnegative bacteria, we have used an approach based on mimicking theouter layers of these membranes using monolayers, Langmuir-Blodgettfilms and unilamelar vesicles, and applying a battery of biophysicalmethods in order to dissect the different events of membraneinteraction. Collectively, results indicate that the PxB analogues act inthe bacterial membrane by the same mechanism than PxB, and that cationic amphipathicity determines peptide activity.

Sodium/hydrogen exchanger NHA2 is critical for insulin secretion in β-cells.

NHA2 is a sodium/hydrogen exchanger with unknown physiological function. Here we show that NHA2 is present in rodent and human β-cells, as well as β-cell lines. In vivo, two different strains of NHA2-deficient mice displayed a pathological glucose tolerance with impaired insulin secretion but normal peripheral insulin sensitivity. In vitro, islets of NHA2-deficient and heterozygous mice, NHA2-depleted Min6 cells, or islets treated with an NHA2 inhibitor exhibited reduced sulfonylurea- and secretagogue-induced insulin secretion. The secretory deficit could be rescued by overexpression of a wild-type, but not a functionally dead, NHA2 transporter. NHA2 deficiency did not affect insulin synthesis or maturation and had no impact on basal or glucose-induced intracellular Ca(2+) homeostasis in islets. Subcellular fractionation and imaging studies demonstrated that NHA2 resides in transferrin-positive endosomes and synaptic-like microvesicles but not in insulin-containing large dense core vesicles in β-cells. Loss of NHA2 inhibited clathrin-dependent, but not clathrin-independent, endocytosis in Min6 and primary β-cells, suggesting defective endo-exocytosis coupling as the underlying mechanism for the secretory deficit. Collectively, our in vitro and in vivo studies reveal the sodium/proton exchanger NHA2 as a critical player for insulin secretion in the β-cell. In addition, our study sheds light on the biological function of a member of this recently cloned family of transporters.

Identification and staining of distinct populations of secretory organelles in astrocytes.

Increasing evidence indicates that astrocytes, the most abundant glial cell type in the brain, respond to an elevation in cytoplasmic calcium concentration ([Ca(2+)]i) by releasing chemical transmitters (also called gliotransmitters) via regulated exocytosis of heterogeneous classes of organelles. By this process, astrocytes exert modulatory influences on neighboring cells and are thought to participate in the control of synaptic circuits and cerebral blood flow. Studying the properties of exocytosis in astrocytes is a challenge, because the cell biological basis of this process is incompletely defined. Astrocytic exocytosis involves multiple populations of secretory vesicles, including synaptic-like microvesicles (SLMVs), dense-core granules (DCGs), and lysosomes. Here we summarize the available information for identifying individual populations of secretory organelles in astrocytes, including DCGs, SLMVs, and lysosomes, and present experimental procedures for specifically staining such populations.

Distribution of neuropeptide Y Y1 receptors in rodent peripheral tissues.

Using a sensitive immunohistochemical technique, the localization of neuropeptide Y (NPY) Y1-receptor (Y1R)-like immunoreactivity (LI) was studied in various peripheral tissues of rat. Wild-type (WT) and Y1R-knockout (KO) mice were also analyzed. Y1R-LI was found in small arteries and arterioles in many tissues, with particularly high levels in the thyroid and parathyroid glands. In the thyroid gland, Y1R-LI was seen in blood vessel walls lacking alpha-smooth muscle actin, i.e., perhaps in endothelial cells of capillaries. Larger arteries lacked detectable Y1R-LI. A distinct Y1R-immunoreactive (IR) reticulum was seen in the WT mouse spleen, but not in Y1R-KO mouse or rat. In the gastrointestinal tract, Y1R-positive neurons were observed in the myenteric plexus, and a few enteroendocrine cells were Y1R-IR. Some cells in islets of Langerhans in the pancreas were Y1R-positive, and double immunostaining showed coexistence with somatostatin in D-cells. In the urogenital tract, Y1R-LI was observed in the collecting tubule cells of the renal papillae and in some epithelial cells of the seminal vesicle. Some chromaffin cells of adrenal medulla were positive for Y1R. The problem of the specificity of the Y1R-LI is evaluated using adsorption tests as well as comparisons among rat, WT mouse, and mouse with deleted Y1R. Our findings support many earlier studies based on other methodologies, showing that Y1Rs on smooth muscle cells of blood vessels mediate NPY-induced vasoconstriction in various organs. In addition, Y1Rs in other cells in parenchymal tissues of several organs suggest nonvascular effects of NPY via the Y1R.