A mechanism for the inhibition of fever by a virus.

Poxviruses encode proteins that block the activity of cytokines. Here we show that the study of such virulence factors can contribute to our understanding of not only virus pathogenesis but also the physiological role of cytokines. Fever is a nonspecific response to infection that contributes to host defense. Several cytokines induce an elevation of body temperature when injected into animals, but in naturally occurring fever it has been difficult to show that any cytokine has a critical role. We describe the first example of the suppression of fever by a virus and the molecular mechanism leading to it. Several vaccinia virus strains including smallpox vaccines express soluble interleukin 1 (IL-1) receptors, which bind IL-1 beta but not IL-1 alpha. These viruses prevent the febrile response in infected mice, whereas strains that naturally or through genetic engineering lack the receptor induce fever. Repair of the defective IL-1 beta inhibitor in the smallpox vaccine Copenhagen, a more virulent virus than the widely used vaccine strains Wyeth and Lister, suppresses fever and attenuates the disease. The vaccinia-induced fever was inhibited with antibodies to IL-1 beta. These findings provide strong evidence that IL-1 beta, and not other cytokines, is the major endogenous pyrogen in a poxvirus infection.

Suppression of glioblastoma angiogenicity and tumorigenicity by inhibition of endogenous expression of vascular endothelial growth factor.

The development of new capillary networks from the normal microvasculature of the host appears to be required for growth of solid tumors. Tumor cells influence this process by producing both inhibitors and positive effectors of angiogenesis. Among the latter, the vascular endothelial growth factor (VEGF) has assumed prime candidacy as a major positive physiological effector. Here, we have directly tested this hypothesis in the brain tumor, glioblastoma multiforme, one of the most highly vascularized human cancers. We introduced an antisense VEGF expression construct into glioblastoma cells and found that (i) VEGF mRNA and protein levels were markedly reduced, (ii) the modified cells did not secrete sufficient factors so as to be chemoattractive for primary human microvascular endothelial cells, (iii) the modified cells were not able to sustain tumor growth in immunodeficient animals, and (iv) the density of in vivo blood vessel formation was reduced in direct relation to the reduction of VEGF secretion and tumor formation. Moreover, revertant cells that recovered the ability to secrete VEGF regained each of these tumorigenic properties. These results suggest that VEGF plays a major angiogenic role in glioblastoma.

Arachidonate lipoxygenases as essential regulators of cell survival and apoptosis.

Arachidonic acid (AA) metabolites derived from both cyclooxygenase (COX) and lipoxygenase (LOX) pathways transduce a variety of signals related to cell growth. Here, we report that the AA LOX pathway also functions as a critical regulator of cell survival and apoptosis. Rat Walker 256 (W256) carcinosarcoma cells express 12-LOX and synthesize 12(S)- and 15(S)-hydroxyeicosatetraenoic acids as their major LOX metabolites. W256 cells transfected with 12-LOX-specific antisense oligonucleotide or antisense oligonucleotides directed to conserved regions of LOXs underwent time- and dose-dependent apoptosis. Likewise, treatment of W256 cells with various LOX but not COX inhibitors induced apoptotic cell death, which could be partially inhibited by exogenous 12(S)- or 15(S)-hydroxyeicosatetraenoic acids. The W256 cell apoptosis induced by antisense oligos and LOX inhibitors was followed by a rapid downregulation of bcl-2 protein, a dramatic decrease in the bcl-2/bax ratio, and could be suppressed by bcl-2 overexpression. In contrast, p53, which is wild type in W256 cells, did not undergo alterations during apoptosis induction. The results suggest that the LOX pathway plays an important physiological role in regulating apoptosis.

A multi-scale assessment of physiological processes in arctic tundra plants under natural and simulated climate change scenarios

Climate warming is predicted to cause an increase in the growing season by as much as 30% for regions of the arctic tundra. This will have a significant effect on the physiological activity of the vascular plant species and the ecosystem as a whole. The need to understand the possible physiological change within this ecosystem is confounded by the fact that research in this extreme environment has been limited to periods when conditions are most favorable, mid June–mid August. This study attempted to develop the most comprehensive understanding to date of the physiological activity of seven tundra plant species in the Alaskan Arctic under natural and lengthened growing season conditions. Four interrelated lines of research, scaling from cellular signals to ecosystem processes, set the foundation for this study. ^ I established an experiment looking at the physiological response of arctic sedges to soil temperature stress with emphasis on the role of the hormone abscisic acid (ABA). A manipulation was also developed where the growing season was lengthened and soils were warmed in an attempt to determine the maximum physiological capacity of these seven vascular species. Additionally, the physiological capacities of four evergreens were tested in the subnivean environment along with the potential role anthocyanins play in their activity. The measurements were scaled up to determine the physiological role of these evergreens in maintaining ecosystem carbon fluxes. ^ These studies determined that soil temperature differentials significantly affect vascular plant physiology. ABA appears to be a physiological modifier that limits stomatal processes when root temperatures are low. Photosynthetic capacity was limited by internal plant physiological mechanisms in the face of a lengthened growing season. Therefore shifts in ecosystem carbon dynamics are driven by changes in species composition and biomass production on a per/unit area basis. These studies also found that changes in soil temperatures will have a greater effect of physiological processes than would the same magnitude of change in air temperature. The subnivean environment exhibits conditions that are favorable for photosynthetic activity in evergreen species. These measurements when scaled to the ecosystem have a significant role in limiting the system's carbon source capacity. ^

The role of a Christian church in the career development of its congregants

The purpose of this study was to examine the role of a Christian church in the career development of its congregants. Contemporary theorists in the 21st century view career development as the totality of an individual's life, and the literature revealed that 85% of Americans claim the practice of Christianity as a major life role. Therefore, an understanding of the church's role in the lives of its congregants is essential when conceptualizing career development theories. Traditional and contemporary theories formed the framework for this examination, which was guided by four research questions: How do congregants of a local church view its contribution to their career development; how do church leaders characterize the potential of the church for making a contribution to the career development of congregants; how useful are church sermon concepts to the career development of congregants; how do church programs and activities contribute to the career development of congregants? A Christian church in South Florida was the study's site, as it was identified as a church which focused on career development. Basic interpretive qualitative inquiry was used to collect and analyze three data sources: interviews, sermon recordings, and church documents. Twenty-four participants were interviewed using two interview guides to elicit perspectives of 15 congregants and 9 church leaders. The interviews and 13 sermon recordings were transcribed and analyzed. Church documents were categorized and analyzed for evidence of career development programs and activities. The findings revealed that the church played the following role in the participants' life career development: empowerment, guidance for life, learning and development, safety and support, and servant-leadership. As a result of their church participation, and through the learning and development from programs and activities, participants developed an awareness of their identity, purpose, and meaning for their lives. These constructs supported their interactions within the environments of home, work, school, and community. This holistic perspective revealed that an integration of traditional and contemporary career development theories was necessary to conceptualize the role of this Christian church in the career development of its congregants.

Síntese química e avaliação das propriedades antibacterianas e antiparasitárias de análogos de peptídeos antimicrobianos de anuros

Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2015.

Linking thermal imaging to physiological indicators in Carica papaya L. under different watering regimes

Water deficit is the most limiting factor for yield and fruit-quality parameters in papaya crop (Carica papaya L.), deficit-irrigation (DI) strategies offering a feasible alternative to manage limiting water resources. When DI is applied, it is crucial to assess the physiological status of the crop in order to maintain the plant within a threshold value of water stress so as no to affect yield or fruit-quality parameters. The aim of this work was to evaluate the feasibility of thermal imaging in young papaya plants to assess the physiological status of this crop when it is subjected to different DI regimes, studying the relationships between the changes in leaf temperature (Tleaf) and in the major physiological parameters (i.e., stomatal conductance to water vapor, gs; transpiration, E; and net photosynthesis, An). The trial was conducted in a greenhouse from March to April of 2012. Plants were grown in pots and subjected to four irrigation treatments: (1) a full irrigation treatment (control), maintained at field capacity; (2) a partial root-zone drying treatment, irrigated with 50% of the total water applied to control to only one side of roots, alternating the sides every 7 days; (3) a regulated deficit irrigation (50% of the control, applied to both sides of plant); (4) and a non-irrigated treatment, in which irrigation was withheld from both sides of the split root for 14 days, followed by full irrigation until the end of the study. Significant relationships were found between Tleaf and major physiological variables such as gs, E and An. Additionally, significant relationships were found between the difference of leaf-to-air temperature (ΔTleaf–air) and gas-exchange measurements, which were used to establish the optimum range of ΔTleaf–air as a preliminary step to the crop-water monitoring and irrigation scheduling in papaya, using thermal imaging as the main source of information. According to the results, we conclude that thermal imaging is a promising technique to monitor the physiological status of papaya during drought conditions.

Linking thermal imaging to physiological indicators in Carica papaya L. under different watering regimes

Water deficit is the most limiting factor for yield and fruit-quality parameters in papaya crop (Carica papaya L.), deficit-irrigation (DI) strategies offering a feasible alternative to manage limiting water resources. When DI is applied, it is crucial to assess the physiological status of the crop in order to maintain the plant within a threshold value of water stress so as no to affect yield or fruit-quality parameters. The aim of this work was to evaluate the feasibility of thermal imaging in young papaya plants to assess the physiological status of this crop when it is subjected to different DI regimes, studying the relationships between the changes in leaf temperature (Tleaf) and in the major physiological parameters (i.e., stomatal conductance to water vapor, gs; transpiration, E; and net photosynthesis, An). The trial was conducted in a greenhouse from March to April of 2012. Plants were grown in pots and subjected to four irrigation treatments: (1) a full irrigation treatment (control), maintained at field capacity; (2) a partial root-zone drying treatment, irrigated with 50% of the total water applied to control to only one side of roots, alternating the sides every 7 days; (3) a regulated deficit irrigation (50% of the control, applied to both sides of plant); (4) and a non-irrigated treatment, in which irrigation was withheld from both sides of the split root for 14 days, followed by full irrigation until the end of the study. Significant relationships were found between Tleaf and major physiological variables such as gs, E and An. Additionally, significant relationships were found between the difference of leaf-to-air temperature (ΔTleaf–air) and gas-exchange measurements, which were used to establish the optimum range of ΔTleaf–air as a preliminary step to the crop-water monitoring and irrigation scheduling in papaya, using thermal imaging as the main source of information. According to the results, we conclude that thermal imaging is a promising technique to monitor the physiological status of papaya during drought conditions.

Cardioprotective effect of ornitho-kinin in an anesthetized, open-chest chicken model of acute coronary occlusion

The generation of bradykinin (BK; Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) in blood and kallidin (Lys-BK) in tissues by the action of the kallikrein-kinin system has received little attention in non-mammalian vertebrates. In mammals, kallidin can be generated by the coronary endothelium and myocytes in response to ischemia, mediating cardioprotective events. The plasma of birds lacks two key components of the kallikrein-kinin system: the low molecular weight kininogen and a prekallikrein activator analogous to mammalian factor XII, but treatment with bovine plasma kallikrein generates ornitho-kinin [Thr6,Leu8]-BK. The possible cardioprotective effect of ornitho-kinin infusion was investigated in an anesthetized, open-chest chicken model of acute coronary occlusion. A branch of the left main coronary artery was reversibly ligated to produce ischemia followed by reperfusion, after which the degree of myocardial necrosis (infarct size as a percent of area at risk) was assessed by tetrazolium staining. The iv injection of a low dose of ornitho-kinin (4 µg/kg) reduced mean arterial pressure from 88 ± 12 to 42 ± 7 mmHg and increased heart rate from 335 ± 38 to 402 ± 45 bpm (N = 5). The size of the infarct was reduced by pretreatment with ornitho-kinin (500 µg/kg infused over a period of 5 min) from 35 ± 3 to 10 ± 2% of the area at risk. These results suggest that the physiological role of the kallikrein-kinin system is preserved in this animal model in spite of the absence of two key components, i.e., low molecular weight kininogen and factor XII.

Hypoxia Inducible Factor-Dependent Regulation of Angiogenesis by Nitro-Fatty Acids

Objective-Nitro-fatty acids (NO(2)-FAs) are emerging as a new class of cell signaling mediators. Because NO(2)-FAs are found in the vascular compartment and their impact on vascularization remains unknown, we aimed to investigate the role of NO(2)-FAs in angiogenesis. Methods and Results-The effects of nitrolinoleic acid and nitrooleic acid were evaluated on migration of endothelial cell (EC) in vitro, EC sprouting ex vivo, and angiogenesis in the chorioallantoic membrane assay in vivo. At 10 mu mol/L, both NO(2)-FAs induced EC migration and the formation of sprouts and promoted angiogenesis in vivo in an NO-dependent manner. In addition, NO(2)-FAs increased intracellular NO concentration, upregulated protein expression of the hypoxia inducible factor-1 alpha (HIF-1 alpha) transcription factor by an NO-mediated mechanism, and induced expression of HIF-1 alpha target genes, such as vascular endothelial growth factor, glucose transporter-1, and adrenomedullin. Compared with typical NO donors such as spermine-NONOate and deta-NONOate, NO(2)-FAs were slightly less potent inducers of EC migration and HIF-1 alpha expression. Short hairpin RNA-mediated knockdown of HIF-1 alpha attenuated the induction of vascular endothelial growth factor mRNA expression and EC migration stimulated by NO(2)-FAs. Conclusion-Our data disclose a novel physiological role for NO(2)-FAs, indicating that these compounds induce angiogenesis in an NO-dependent mechanism via activation of HIF-1 alpha. (Arterioscler Thromb Vasc Biol. 2011;31:1360-1367.)

Sequence and tissue distribution of chicken cellular nucleic acid binding protein cDNA

We sequenced cDNAs coding for chicken cellular nucleic acid binding protein (CNBP). Two slightly different variations of the open reading frame were found, each of which translates into a protein with seven zinc finger domains. The longest transcript contains an in-frame insert of 3 bp. The sequence conservation between chick CNBP cDNAs with human, rat and mouse CNBP cDNAs is extreme, especially in the coding region, where the deduced amino acid sequence identity with human, rat and mouse CNBP is 99%. CNBP-like transcripts were also found in various tissues from insect, shrimp, fish and lizard. Regions with remarkable nucleotide conservation were also found in the 3' untranslated region, indicating important functions for these regions. Quantitative reverse transcription polymerase chain reaction (RT-PCR) indicated that in the chick, CNBP is present in all tissues examined in approximately equal ratios to total RNA. RT-PCR of total RNA isolated from different phyla indicate CNBP-like proteins art widespread throughout the animal kingdom. The extraordinary level of conservation suggests an important physiological role for CNBP. (C) 1997 Elsevier Science Inc.

Nitric oxide inhibition of the rat olfactory cyclic nucleotide-gated cation channel

The effects of nitric oxide (NO) and other cysteine modifying agents were examined on cyclic nucleotide-gated (CNG) cation channels from rat olfactory receptor neurons. The NO compounds, S-nitroso-cysteine (SNC) and 3-morpholino-sydnonomine (SIN-1), did not activate the channels when applied for up to 10 min. The cysteine alkylating agent, N-ethylmaleimide (NEM), and the oxidising agent, dithionitrobensoate (DTNB), were also without agonist efficacy. Neither SNC nor DTNB altered the cAMP sensitivity of the channels. However, 2-min applications of SIN-1, SNC and DTNB inhibited the cAMP-gated current to approximately 50% of the control current level. This inhibition showed no spontaneous reversal for 5 min but was completely reversed by a 2-min exposure to DTT. The presence of cAMP protected the channels against NO-induced inhibition. These results indicate that inhibition is caused by S-nitrosylation of neighboring sulfhydryl groups leading to sulfhydryl bond formation. This reaction is favored in the closed channel state. Since recombinantly expressed rat olfactory alpha and beta CNG channel homomers and alpha/beta heteromers are activated and not inhibited by cysteine modification, the results of this study imply the existence of a novel subunit or tightly bound factor which dominates the effect of cysteine modification in the native channels. As CNG channels provide a pathway for calcum influx, the results may also have important implications for the physiological role of NO in mammalian olfactory receptor neurons.

Peroxisome proliferator-activated receptors in tumorigenesis: Targets of tumour promotion and treatment

The peroxisome proliferator-activated receptors (PPAR) are ligand-activated transcription factors. There are three genes that code for the PPAR isoforms: PPAR alpha, PPAR beta and PPAR gamma. In the present review, studies characterizing the various PPAR isoforms are discussed. Peroxisome proliferator-activated receptor alpha has been implicated in the lipid-lowering effects of the fibrate drugs. Peroxisome proliferator-activated receptor gamma has a clear role in adipocyte differentiation and is therapeutically targeted by the thiazolidinedione drugs for the treatment of type II diabetes. The physiological role of PPAR beta is less well understood but, as described in the present review, recent studies have implicated it with a role in colon cancer. In the present review, particular attention is focused on the role of PPAR in the regulation of expression of proteins associated with cell cycle control and tumorigenesis.

Mutant GABA(A) receptor gamma 2-subunit in childhood absence epilepsy and febrile seizures

Epilepsies affect at least 2% of the population at some time in life, and many forms have genetic determinants(1,2). We have found a mutation in a gene encoding a GABA, receptor subunit in a large family with epilepsy. The two main phenotypes were childhood absence epilepsy (CAE) and febrile seizures (FS), There is a recognized genetic: relationship between FS and CAE, yet the two syndromes have different ages of onset, and the physiology of absences and convulsions is distinct. This suggests the mutation has age-dependent effects on different neuronal networks that influence the expression of these clinically distinct, but genetically related, epilepsy phenotypes. We found that the mutation in GABRG2 (encoding the gamma2-subunit) abolished in vitro sensitivity to diazepam, raising the possibility that endozepines do in fact exist and have a physiological role in preventing seizures.

Localization of a brain sulfotransferase, SULT4A1, in the human and rat brain: An immunohistochemical study

Cytosolic sulfotransferases are believed to play a role in the neuromodulation of certain neurotransmitters and drugs. To date, four cytosolic sulfotransferases have been shown to be expressed in human brain. Recently, a novel human brain sulfotransferase has been identified and characterized, although its role and localization in the brain are unknown. Here we present the first immunohistochemical (IHC) localization of SULT4A1 in human brain using an affinity-purified polyclonal antibody raised against recombinant human SULT4A1. These results are supported and supplemented by the IHC localization of SULT4A1 in rat brain. In both human and rat brains, strong reactivity was found in several brain regions, including cerebral cortex, cerebellum, pituitary, and brainstem. Specific signal was entirely absent on sections for which preimmune serum from the corresponding animal, processed in the same way as the postimmune serum, was used in the primary screen. The findings from this study may assist in determining the physiological role of this SULT isoform.