bluuebunny/crossref_metadata_2025 huggingface dataset:
S100A8/A9 at low concentration promotes tumor cell growth via RAGE ligation and MAP kinase-dependent pathway
Authors: Ghavami Saeid , Rashedi Iran , Dattilo Brian M , Eshraghi Mehdi , Chazin Walter J , Hashemi Mohammad , Wesselborg Sebastian , Kerkhoff Claus , Los Marek
Published: March 2008

The complex formed by two members of the S100 calcium-binding protein family, S100A8/A9, exerts apoptosis-inducing activity against various cells, especially tumor cells. Here, we present evidence that S100A8/A9 also has cell growth-promoting activity at low concentrations. Receptor of advanced glycation end product (RAGE) gene silencing and cotreatment with a RAGE-specific blocking antibody revealed that this activity was mediated via RAGE ligation. To investigate the signaling pathways, MAPK phosphorylation and NF-κB activation were characterized in S100A8/A9-treated cells. S100A8/A9 caused a significant increase in p38 MAPK and p44/42 kinase phosphorylation, and the status of stress-activated protein kinase/JNK phosphorylation remained unchanged. Treatment of cells with S100A8/A9 also enhanced NF-κB activation. RAGE small interfering RNA pretreatment abrogated the S100A8/A9-induced NF-κB activation. Our data indicate that S100A8/A9-promoted cell growth occurs through RAGE signaling and activation of NF-κB.
Cytokines secreted by IL-2-activated lymphocytes induce endogenous nitric oxide synthesis and apoptosis in macrophages
Authors: Choi Kyoung-Seong , Song Eun-Kee , Yim Chang-Yeol
Published: March 2008

IL-2-activated killer (LAK) cells secrete inflammatory cytokines such as IFN-γ and TNF-α, which can induce NO synthesis (NOS). In this study, we investigated IL-2-activated lymphocyte-mediated macrophage apoptosis via NOS. LAK cells and their culture supernatants induced NOS in murine macrophages. NOS was markedly inhibited by blocking antibodies to IFN-γ and TNF-α, suggesting the key role of these lymphocyte cytokines in mediating NOS. Endogenous NO production inhibited macrophage proliferation and induced apoptosis in concordance with p53 accumulation and caspase-3 activation, processes that were inhibited by NG-monomethyl-l-arginine (a NOS inhibitor) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (a NO scavenger). Our study demonstrated a novel, noncontact-dependent mechanism of macrophage suppression by IL-2-activated lymphocytes: induction of growth inhibition and apoptosis of macrophages as a result of endogenous NOS induced by cytokines secreted from IL-2-activated lymphocytes.
Bayesian lithology and fluid prediction from seismic prestack data
Authors: Buland Arild , Kolbjørnsen Odd , Hauge Ragnar , Skjæveland Øyvind , Duffaut Kenneth
Published: March 2008

A fast Bayesian inversion method for 3D lithology and fluid prediction from prestack seismic data, and a corresponding feasibility analysis were developed and tested on a real data set. The objective of the inversion is to find the probabilities for different lithology-fluid classes from seismic data and geologic knowledge. The method combines stochastic rock physics relations between the elastic parameters and the different lithology-fluid classes with the results from a fast Bayesian seismic simultaneous inversion from seismic data to elastic parameters. A method for feasibility analysis predicts the expected modification of the prior probabilities to posterior probabilities for the different lithology-fluid classes. The feasibility analysis can be carried out before the seismic data are analyzed. Both the feasibility method and the seismic lithology-fluid probability inversion were applied to a prospect offshore Norway. The analysis improves the probability for gas sand from 0.1 to about 0.2–0.4 with seismic data.
Paired-Like Homeodomain Transcription Factors 1 and 2 Regulate Follicle-Stimulating Hormone β-Subunit Transcription through a Conserved cis-Element
Authors: Lamba Pankaj , Khivansara Vishal , D'Alessio Ana C. , Santos Michelle M. , Bernard Daniel J.
Published: March 2008

Paired-like homeodomain transcription factors (PITX) regulate the activity of pituitary hormone-encoding genes. Here, we examined mechanisms through which the family of PITX proteins control murine FSH β-subunit (Fshb) transcription. We observed that endogenous PITX1 and PITX2 isoforms from murine LβT2 gonadotrope cells could bind a highly conserved proximal cis-element. Transfection of PITX1 or PITX2C in heterologous cells stimulated both murine and human Fshb/FSHB promoter-reporter activities, and in both cases, mutation of the critical cis-element abrogated these effects. In homologous LβT2 cells, the same mutation decreased basal reporter activity and greatly reduced activin A-stimulated transcription from murine and human promoter-reporters. Transfecting dominant-negative forms of PITX1 or PITX2C or knocking down PITX1 or -2 expression by RNA interference in LβT2 cells inhibited murine Fshb transcription, confirming roles for endogenous PITX proteins. Both PITX1 and PITX2C interacted with Smad3 (an effector of the activin signaling cascade in these cells) in coprecipitation experiments, and the PITX binding site mutation greatly inhibited Smad2/3/4-stimulated Fshb transcription. In summary, both PITX1 and PITX2C regulate murine and human Fshb/FSHB transcription through a conserved cis-element in the proximal promoter. Furthermore, the data indicate both common and distinct mechanisms of PITX1 and PITX2C action.
Insulin-Like Growth Factor II Activates Phosphatidylinositol 3-Kinase-Protooncogenic Protein Kinase 1 and Mitogen-Activated Protein Kinase Cell Signaling Pathways, and Stimulates Migration of Ovine Trophectoderm Cells
Authors: Kim Jinyoung , Song Gwonhwa , Gao Haijun , Farmer Jennifer L. , Satterfield M. Carey , Burghardt Robert C. , Wu Guoyao , Johnson Greg A. , Spencer Thomas E. , Bazer Fuller W.
Published: March 2008

IGF-II, a potent stimulator of cellular proliferation, differentiation, and development, regulates uterine function and conceptus growth in several species. In situ hybridization analyses found that IGF-II mRNA was most abundant in the caruncular endometrial stroma of both cyclical and pregnant ewes. In the intercaruncular endometrium, IGF-II mRNA transitioned from stroma to luminal epithelium between d 14 and 20 of pregnancy. IGF-II mRNA was present in all cells of the conceptus but was particularly abundant in the yolk sac. Immunohistochemical analyses revealed that phosphorylated (p)-protooncogenic protein kinase 1, p-ribosomal protein S6 kinase, p-ERK1/2, and p-P38 MAPK proteins were present at low levels in a majority of endometrial cells but were most abundant in the nuclei of endometrial luminal epithelium and conceptus trophectoderm of pregnant ewes. In mononuclear trophectoderm cells isolated from d-15 conceptuses, IGF-II increased the abundance of p-pyruvate dehydrogenase kinase 1, p-protooncogenic protein kinase 1, p-glycogen synthase kinase 3B, p-FK506 binding protein 12-rapamycin associated protein 1, and p-ribosomal protein S6 kinase protein within 15 min, and the increase was maintained for 90 min. IGF-II also elicited a rapid increase in p-ERK1/2 and p-P38 MAPK proteins that was maximal at 15 or 30 min posttreatment. Moreover, IGF-II increased migration of trophectoderm cells. Collectively, these results support the hypothesis that IGF-II coordinately activates multiple cell signaling pathways critical to survival, growth, and differentiation of the ovine conceptus during early pregnancy.
Superoxide Destabilization of β-Catenin Augments Apoptosis of High-Glucose-Stressed Mesangial Cells
Authors: Lin Chun-Liang , Wang Jeng-Yi , Ko Jih-Yang , Surendran Kameswaran , Huang Yu-Ting , Kuo Yu-Hsia , Wang Feng-Sheng
Published: March 2008

Intense mesangial cell apoptosis contributes to the pathogenesis of diabetic nephropathy. Although reactive oxygen radicals and Wnt signaling components are potent regulators that modulate renal tissue remodeling and morphogenesis, cross-talk between oxidative stress and Wnt/β-catenin signaling in controlling high-glucose-impaired mesangial cell survival and renal function have not been tested. In this study, high glucose induced Ras and Rac1 activation, superoxide burst, and Wnt5a/β-catenin destabilization and subsequently promoted caspase-3 and poly (ADP-ribose) polymerase cleavage and apoptosis in mesangial cell cultures. The pharmacological and genetic suppression of superoxide synthesis by superoxide dismutase and diphenyloniodium, dominant-negative Ras (S17N), and dominant-negative Rac1 (T17N) abrogated high-glucose-induced glycogen synthase kinase (GSK-3β) activation and caspase-3 and poly (ADP-ribose) polymerase degradation. Inactivation of Ras and Racl also reversed Wnt/β-catenin expression and survival of mesangial cells. Stabilization of β-catenin by the transfection of stable β-catenin (Δ45) and kinase-inactive GSK-3β attenuated high-glucose-mediated mesangial cell apoptosis. Exogenous superoxide dismutase administration attenuated urinary protein secretion in diabetic rats and abrogated diabetes-mediated reactive oxygen radical synthesis in renal glomeruli. Immunohistological observation revealed that superoxide dismutase treatment abrogated diabetes-induced caspase-3 cleavage and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) and increased Wnt5a/β-catenin expression in renal glomeruli. Taken together, high glucose induced oxidative stress and apoptosis in mesangial cells. The Ras and Rac1 regulation of superoxide appeared to raise apoptotic activity by activating GSK-3β and inhibiting Wnt5a/β-catenin signaling. Controlling oxidative stress and Wnt/β-catenin signaling has potential for protecting renal tissue against the deleterious effect of high glucose.
Prokineticin 1 Signaling and Gene Regulation in Early Human Pregnancy
Authors: Evans Jemma , Catalano Rob D. , Morgan Kevin , Critchley Hilary O. D. , Millar Robert P. , Jabbour Henry N.
Published: March 2008

Prokineticin 1 (PROK1) is a recently described protein with a wide range of functions including tissue-specific angiogenesis, modulation of inflammatory responses, and regulation of hematopoiesis. The objective of this study was to investigate the role of PROK1 and prokineticin receptor 1 (PROKR1) in human endometrium during early pregnancy. PROK1 and PROKR1 expression is significantly elevated in first-trimester decidua, compared with nonpregnant endometrium. Expression of PROK1 and PROKR1 was localized in glandular epithelial and various cellular compartments within the stroma. To investigate the signaling pathways and target genes activated by PROK1, we generated an endometrial epithelial cell line stably expressing PROKR1 (Ishikawa PROKR1 cells). PROK1-PROKR1 interaction induced inositol phosphate mobilization and sequential phosphorylation of c-Src, epidermal growth factor receptor, and ERK 1/2. Gene microarray analysis on RNA extracted from Ishikawa PROKR1 cells treated with 40 nm PROK1 for 8 h revealed 49 genes to be differentially regulated. A number of these genes, including cyclooxygenase (COX)-2, leukemia inhibitory factor, IL-6, IL-8, and IL-11 are regulated in the endometrium during implantation and early pregnancy. We subsequently investigated the effect of PROK1 on expression of COX-2 in Ishikawa PROKR1 cells and first-trimester decidua. COX-2 mRNA and protein expression, and prostaglandin synthesis, were elevated in response to treatment with PROK1. Moreover, expression of COX-2 by PROK1 was dependent on activation of the Gq-phospholipase C-β-cSrc-epidermal growth factor receptor-MAPK/ERK kinase pathway. These data demonstrate that PROK1 and PROKR1 expression is elevated in human decidua during early pregnancy and that PROK1-PROKR1 interaction regulates expression of a host of implantation-related genes.
A Novel, Functional, and Highly Divergent Sex Hormone-Binding Globulin that May Participate in the Local Control of Ovarian Functions in Salmonids
Authors: Bobe Julien , Mahé Sophie , Nguyen Thaovi , Rime Hélène , Vizziano Denise , Fostier Alexis , Guiguen Yann
Published: March 2008

A cDNA encoding for a novel rainbow trout SHBG was identified and characterized. Phylogenetic analysis showed that this novel SHBG, named SHBGb, was a highly divergent paralog of the classical SHBG (SHBGa) form previously known in vertebrates including zebrafish, seabass, and rainbow trout. Using all available sequences, no SHBGb-like sequence could be identified in any fish species besides Atlantic salmon. Rainbow trout SHBGa and SHBGb share only 26% sequence identity at the amino acid level and exhibit totally distinct tissue distribution, thus demonstrating a functional shift of SHBGb. Indeed, shbga mRNA was predominantly expressed in liver and spleen but could not be detected in the ovary, whereas shbgb had a predominant ovarian expression but could not be detected in liver. Despite its high divergence, rainbow trout SHBGb expressed in COS-7 cells could bind estradiol and testosterone with high affinity and specificity. Both rainbow trout shbgb mRNA and proteins were localized to the granulosa cells of vitellogenic ovarian follicles, whereas SHBGb immunoreactivity was also found in theca cells. Finally, shbgb ovarian mRNA expression exhibited a significant drop between late vitellogenesis and oocyte maturation at a time when ovarian aromatase (cyp19a) gene expression and estradiol circulating levels exhibited a dramatic decrease. Together, these observations show that SHBGb is a functional and highly divergent SHBG paralog probably arising from a salmonid-specific duplication of the shbg gene.
Molecular Characterization of V59E NIS, a Na+/I− Symporter Mutant that Causes Congenital I− Transport Defect
Authors: Reed-Tsur Mia D. , De la Vieja Antonio , Ginter Christopher S. , Carrasco Nancy
Published: March 2008

I− is actively transported into thyrocytes via the Na+/I− symporter (NIS), a key glycoprotein located on the basolateral plasma membrane. The cDNA encoding rat NIS was identified in our laboratory, where an extensive structure/function characterization of NIS is being conducted. Several NIS mutants have been identified as causes of congenital I− transport defect (ITD), including V59E NIS. ITD is characterized by low thyroid I− uptake, low saliva/plasma I− ratio, hypothyroidism, and goiter and may cause mental retardation if untreated. Studies of other ITD-causing NIS mutants have revealed valuable information regarding NIS structure/function. V59E NIS was reported to exhibit as much as 30% of the activity of wild-type NIS. However, this observation was at variance with the patients’ phenotype of total lack of activity. We have thoroughly characterized V59E NIS and studied several amino acid substitutions at position 59. We demonstrated that, in contrast to the previous report, V59E NIS is inactive, although it is properly targeted to the plasma membrane. Glu and all other charged amino acids or Pro at position 59 also yielded nonfunctional NIS proteins. However, I− uptake was rescued to different degrees by the other substitutions. Although the Km values for Na+ and I− were not altered in these active mutants, we found that the structural requirement for NIS function at position 59 is a neutral, helix-promoting amino acid. This result suggests that the region that contains V59 may be involved in intramembrane helix-helix interactions during the transport cycle without being in direct contact with the substrates.
Enhancement of nortriptyline penetration through human epidermis: influence of chemical enhancers and iontophoresis
Authors: Merino Virginia , Micó-Albiñana Teresa , Nácher Amparo , Díez-Sales Octavio , Herráez Marina , Merino-Sanjuán Matilde
Published: March 2008

Different known percutaneous chemical enhancers and iontophoresis have been tested in-vitro to study their ability to increase transdermal absorption of nortriptyline hydrochloride (20 mg mL−1). The chemicals 1-dodecanol, Span 20, Azone, (R)-(+)-limonene or isopropyl myristate were used as an overnight pretreatment at 5% (w/w) in ethanol. Furthermore, isopropyl myristate (20%, w/w) and propylene glycol (15%, w/w) were tested in the same vehicle. Iontophoresis was applied directly to the nortriptyline hydrochloride donor solution for three different concentrations (20, 2 and 0.5 mgmL−1). The chemical enhancers slightly increased the nortriptyline transdermal flux but iontophoresis was more efficient. In this case, nortriptyline transdermal flux was concentration dependent, having a higher flux when the concentration was lowered. Therefore, iontophoresis was the most suitable technique to increase transdermal absorption of nortriptyline and it could be an alternative method to provide therapeutic concentrations of this drug in smoking cessation treatment.
Effect of chitosan crosslinking on bitterness of artemether using response surface methodology
Authors: Shah Punit P , Mashru Rajashree C , Thakkar Arti R , Badhan Atul C
Published: March 2008

This work examines the influence of various process parameters on artemether entrapped in crosslinked chitosan microparticles for masking bitterness. A central composite design was used to optimize the experimental conditions for bitterness masking. Critical parameters such as the amounts of artemether, chitosan and crosslinking agent have been studied to evaluate how they affect responses such as incorporation efficiency, particle size and drug release at pH 6.8. The desirability function approach has been used to find the best compromise between the experimental results. The optimized microparticles were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. Bitterness score was evaluated by human gustatory sensation test. Multiple linear regression analysis revealed that the crosslinking of chitosan significantly affects incorporation efficiency, particle size and drug release at pH 6.8. The bitterness score of microparticles was decreased to 0, compared with 3+ for pure artemether. The proposed method completed masked the bitter taste of artemether.
Pimaradienoic acid inhibits vascular contraction and induces hypotension in normotensive rats
Authors: Tirapelli Carlos R , Filho Mario dos Anjos Neto , Bonaventura Daniella , Melo Mirian C C , Ambrosio Sergio R , de Oliveira Ana M , Bendhack Lusiane M , da Costa Fernando B
Published: March 2008

The present investigation was designed to investigate the effect of the diterpene ent-pimara-8(14),15-dien-19-oic acid (pimaradienoic acid, PA) on smooth muscle extracellular Ca2+ influx. To this end, the effect of PA on phenylephrine- and KCl-induced increases in cytosolic calcium concentration ([Ca2+]c), measured by the variation in the ratio of fluorescence intensities (R340/380 nm) of Fura-2, was analysed. Whether bolus injection of PA could induce hypotensive responses in conscious normotensive rats was also evaluated. PA inhibited the contraction induced by phenylephrine (0.03 or 10 μmol L−1) and KCl (30 or 90 μmol L−1) in endothelium-denuded rat aortic rings in a concentration dependent manner. Pre-treatment with PA (10, 100, 200 μmol L−1) attenuated the contraction induced by CaCl2 (0.5 nmol L−1 or 2.5 μmol L−1) in denuded rat aorta exposed to Ca2+-free medium containing phenylephrine (0.1 μmol L−1) or KCl (30 μmol L−1). Interestingly, the inhibitory effect displayed by PA on CaCl2-induced contraction was more pronounced when KCl was used as the stimulant. Phenylephrine- and KCl-induced increases in [Ca2+]c were inhibited by PA. Similarly, verapamil, a Ca2+-channel blocker, also inhibited the increase in [Ca2+]c induced by either phenylephrine or KCl. Finally, bolus injection of PA (1–15 mg kg−1) produced a dose-dependent decrease in mean arterial pressure in conscious normotensive rats. The results provide the first direct evidence that PA reduces vascular contractility by reducing extracellular Ca2+ influx through smooth muscle cellular membrane, a mechanism that could mediate the hypotensive response induced by this diterpene in normotensive rats.
Pharmacological effect of capsaicin on rat avoidance behaviours elicited by sine-wave electrical stimulation of different frequencies by Neurometer
Authors: Fujiuchi Akiyoshi , Toga Tetsuo
Published: March 2008

The Neurometer is a diagnostic device for measuring the perception and threshold of transcutaneous stimulation. It has been used in patients to selectively activate Aβ-, Aδ- and C-fibres in the primary afferents at different stimulus frequencies (2000, 250 and 5 Hz, respectively). In this study, we investigated use of the Neurometer to selectively activate nerves in conscious rats. The behavioural endpoint of paw withdrawal was used to measure the current threshold (CT). This behaviour was elicited by a lower stimulus current than other behaviours evoked by Neurometer stimulation and caused only mild stress in rats. Repeated topical application of capsaicin (four doses of 100 μg) or systemic administration of the capsaicin analogue resiniferatoxin (30 or 300 μg kg−1) increased the CT value for this behaviour at 5 Hz stimulation but not at 2000 Hz or 250 Hz. This change in CT at 5 Hz is probably due to C-fibre desensitization by the pharmacological treatments. The combination of 5 Hz sine-wave stimulation with a Neurometer and the observation of paw withdrawal behaviour make it possible to perform preclinical studies of C-fibres in animals as an alternative to the use of high- and low-rate heating of the paw.
Anti-inflammatory effects of red pepper (Capsicum baccatum) on carrageenan- and antigen-induced inflammation
Authors: Spiller Fernando , Alves Márcia K , Vieira Sílvio M , Carvalho Toni A , Leite Carlos E , Lunardelli Adroaldo , Poloni José A , Cunha Fernando Q , de Oliveira Jarbas R
Published: March 2008

Inflammation is a pivotal component of a variety of diseases, such as atherosclerosis and tumour progression. Various naturally occurring phytochemicals exhibit anti-inflammatory activity and are considered to be potential drug candidates against inflammation-related pathological processes. Capsicum baccatum L. var. pendulum (Willd.) Eshbaugh (Solanaceae) is the most consumed species in Brazil, and its compounds, such as capsaicinoids, have been found to inhibit the inflammatory process. However, the anti-inflammatory effects of C. baccatum have not been characterized. Thus, this study was designed to evaluate the effects of C. baccatum juice in animal models of acute inflammation induced by carrageenan and immune inflammation induced by methylated bovine serum albumin. Pretreatment (30 min) of rats with pepper juice (0.25–2.0 g kg−1) significantly decreased leucocyte and neutrophil migration, exudate volume and protein and LDH concentration in pleural exudates of a pleurisy model. This juice also inhibited neutrophil migration and reduced the vascular permeability on carrageenan-induced peritonitis in mice. C. baccatum juice also reduced neutrophil recruitment and exudate levels of pro-inflammatory cytokines TNF-α and IL-1β in mouse inflammatory immune peritonitis. Furthermore, we demonstrated that the main constituent of C. baccatum juice, as extracted with chloroform, is capsaicin. In agreement with this, capsaicin was able to inhibit the neutrophil migration towards the inflammatory focus. To our knowledge, this is the first demonstration of the anti-inflammatory effect of C. baccatum juice and our data suggest that this effect may be induced by capsaicin. Moreover, the anti-inflammatory effect induced by red pepper may be by inhibition of pro-inflammatory cytokine production at the inflammatory site.
Topical anti-inflammatory activity of Eugenia brasiliensis Lam. (Myrtaceae) leaves
Authors: Pietrovski Evelise Fernandes , Magina Michele Debiasi Alberton , Gomig Franciane , Pietrovski Caroline Fernandes , Micke Gustavo Amadeu , Barcellos Michele , Pizzolatti Moacir Geraldo , Cabrini Daniela Almeida , Brighente Inês Maria Costa , Otuki Michel Fleith
Published: March 2008

Eugenia brasiliensis Lam., a plant from the south of Brazil, is used in the popular medicine for rheumatism treatment. This study reports that topical application of hydroalcoholic extract, fractions and isolated compounds from E. brasiliensis caused an inhibition of ear oedema in response to topical application of croton oil on the mouse ear. For oedema inhibition, the estimated ID50 values (dose reducing the inflammatory response by 50% relative to the control value) for hydroalcoholic extract and fractions (hexane, ethyl acetate and dichloromethane) were 0.17, 0.29, 0.13 and 0.14 mg/ear, respectively, with inhibition of 79 ± 7%, 87 ± 6%, 88 ± 5% and 96 ± 2%, respectively. Isolated phenolic compounds (quercetin, catechin and gallocatechin) were also effective in inhibiting the oedema (inhibition of 61 ± 5%, 66 ± 2% and 37 ± 9%, respectively). Moreover, both extract and isolated compounds caused inhibition of polymorphonuclear cells influx (inhibition of 85 ± 6%, 81 ± 5%, 73 ± 6% and 76 ± 6%, respectively). The histological analysis of the ear tissue clearly confirmed that the extract and compounds of E. brasiliensis inhibited the influx of polymorphonuclear cells to mouse ear skin after application of croton oil. Furthermore, hydroalcoholic extract was also effective in inhibiting the arachidonic acid-mediated mouse ear oedema (ID50 value was 1.94 mg/ear and inhibition of 60 ± 7%). Therefore, these results consistently support the notion that E. brasiliensis possesses topical anti-inflammatory activity.
Vascular myorelaxing activity of isolates from South African Hyacinthaceae partly mediated by activation of soluble guanylyl cyclase in rat aortic ring preparations
Authors: Fusi Fabio , Ferrara Antonella , Koorbanally Chantal , Crouch Neil R , Mulholland Dulcie A , Sgaragli Giampietro
Published: March 2008

The vasorelaxing effect of isolates (compounds 1, 2, 3, and 4 (homoisoflavanones), compound 5 (sesquiterpenoid), compounds 6 and 7 (bufadienolides)) from the South African Hyacinthaceae has been assessed using rat aortic ring preparations. Compounds 2, 3, and 4 inhibited the tonic contraction induced by both 60 mm K+ (K60) and phenylephrine, compound 3 being the most potent. Compounds 5, 6, and 7 caused a modest concentration-dependent relaxation, whereas compound 1 was ineffective. Under K25- or K60-induced depolarization, compound 3 displayed antispasmodic effects not reversed by tetraethylammonium. Under precontraction induced with phenylephrine, compound 3 shifted to the left the concentration-relaxation curves of either isoprenaline or sodium nitroprusside. 1 H-[1,2,4] oxidazolol [4,3-a] quinoxalin-1-one shifted to the right the concentration-relaxation curve of compound 3, while 3′-isobutyl-1-methylxanthine had no effect. In the absence of extracellular Ca2+, compound 3 (estimated pIC50 = 4.66) and ryanodine reduced the response to phenylephrine. Phenylephrine-stimulated influx of extracellular Ca2+ was markedly reduced when tissues were pretreated with compound 3 (pIC50 = 5.14) or nifedipine, but stimulated by ryanodine. Compound 3 partially antagonized the contraction induced by phorbol 12-myristate-13-acetate. To our knowledge, this has been the first account describing the vasodilating activity of homoisoflavonoids: compound 3 proved an effective vasorelaxing agent, partly acting via the activation of soluble guanylyl cyclase.
The anti-inflammatory modulatory role of Solidago chilensis Meyen in the murine model of the air pouch
Authors: Liz Rafael , Vigil Silvana Virginia Gagliotti , Goulart Stella , Izabel Maria , Moritz Goularte , Schenkel Eloir Paulo , Fröde Tânia Silvia
Published: March 2008

The aim of this study was to investigate the anti-inflammatory efficacy of an aqueous extract (AE), and its butanolic (BuOH) and aqueous residual (AR) fractions, derived from the rhizome of Solidago chilensis in inflammation caused by carrageenan in mice. Solidago chilensis Meyen rhizome was extracted using hot water at 90°C under infusion. The extract was filtered and lyophilized. Part of the aqueous extract was fractionated with n-BuOH, resulting in butanolic (BuOH) and aqueous residual (AR) fractions. Adult Swiss mice were used in the in-vivo experiments. We evaluated the effect of rhizome aqueous extract of Solidago chilensis and these two derived fractions on the inflammation induced by carrageenan in the mouse model of the air pouch. The aqueous extract and its derived fractions significantly inhibited leucocytes, neutrophils, exudation, myeloperoxidase and adenosine deaminase activity, as well as nitric oxide, interleukin-1 beta (IL-1β), neutrophil chemokine (KC) and tumour necrosis factor-alpha (TNF-α) levels (P < 0.05). Indometacin and dexamethasone inhibited all the studied inflammatory parameters (P < 0.01) with the exceptions that indometacin did not inhibit TNF-α levels and dexamethasone did not inhibit KC levels (P > 0.05). These results indicate that Solidago chilensis has a significant anti-inflammatory action on acute inflammatory responses and that its inhibitory activity may be due not only to the inhibition of pro-inflammatory mediators, but also to the inhibition of leucocyte infiltration.
The stepwise specification of embryonic stem cells to hematopoietic fate is driven by sequential exposure to Bmp4, activin A, bFGF and VEGF
Authors: Pearson Stella , Sroczynska Patrycja , Lacaud Georges , Kouskoff Valerie
Published: March 2008

The differentiation of embryonic stem (ES) cells offers a powerful approach to study mechanisms implicated in cell fate decision. A major hurdle, however,is to promote the directed and efficient differentiation of ES cells toward a specific lineage. Here, we define in serum-free media the minimal factor requirement controlling each step of the differentiation process, resulting in the production of highly enriched hematopoietic progenitors. Four factors -Bmp4, activin A, bFGF (Fgf2) and VEGF (VegfA) - are sufficient to drive the selective and efficient differentiation of mouse ES cells to hematopoiesis. Each of these factors appears to regulate a step of the process: Bmp4 promotes the very efficient formation of mesoderm; bFGF and activin A induce the differentiation of these mesodermal precursors to the hemangioblast fate; and VEGF is required for the production of fully committed hematopoietic progenitors. The stimulation of mesodermal precursors by bFGF and activin A switches on very rapidly the hematopoietic program, allowing us to dissect the molecular events leading to the formation of the hemangioblast. Runx1,Scl (Tal1) and Hhex expression is upregulated within 3 hours of stimulation, whereas upregulation of Lmo2 and Fli1is observed later. Interestingly, increased expression levels of genes such as cMyb, Pu.1 (Sfpi1), Gata1 and Gata2 are not observed at the onset of hemangioblast commitment. This stepwise control of differentiation is extremely efficient, giving rise to a very high frequency of hematopoietic precursors, and provides an optimal system for understanding the molecular machineries involved in blood progenitor commitment.
Life-cycle-generation-specific developmental processes are modified in theimmediate uprightmutant of the brown algaEctocarpus siliculosus
Authors: Peters Akira F. , Scornet Delphine , Ratin Morgane , Charrier Bénédicte , Monnier Annabelle , Merrien Yves , Corre Erwan , Coelho Susana M. , Cock J. Mark
Published: March 2008

Development of the sporophyte and gametophyte generations of the brown alga E. siliculosus involves two different patterns of early development,which begin with either a symmetric or an asymmetric division of the initial cell, respectively. A mutant, immediate upright (imm), was isolated that exhibited several characteristics typical of the gametophyte during the early development of the sporophyte generation. Genetic analyses showed that imm is a recessive, single-locus Mendelian factor and analysis of gene expression in this mutant indicated that the regulation of a number of life-cycle-regulated genes is specifically modified in immmutant sporophytes. Thus, IMM appears to be a regulatory locus that controls part of the sporophyte-specific developmental programme, the mutant exhibiting partial homeotic conversion of the sporophyte into the gametophyte,a phenomenon that has not been described previously.
Drosophilaoptic lobe neuroblasts triggered by a wave of proneural gene expression that is negatively regulated by JAK/STAT
Authors: Yasugi Tetsuo , Umetsu Daiki , Murakami Satoshi , Sato Makoto , Tabata Tetsuya
Published: March 2008

Neural stem cells called neuroblasts (NBs) generate a variety of neuronal and glial cells in the central nervous system of the Drosophilaembryo. These NBs, few in number, are selected from a field of neuroepithelial(NE) cells. In the optic lobe of the third instar larva, all NE cells of the outer optic anlage (OOA) develop into either NBs that generate the medulla neurons or lamina neuron precursors of the adult visual system. The number of lamina and medulla neurons must be precisely regulated because photoreceptor neurons project their axons directly to corresponding lamina or medulla neurons. Here, we show that expression of the proneural protein Lethal of scute [L(1)sc] signals the transition of NE cells to NBs in the OOA. L(1)sc expression is transient, progressing in a synchronized and ordered `proneural wave' that sweeps toward more lateral NEs. l(1)sc expression is sufficient to induce NBs and is necessary for timely onset of NB differentiation. Thus, proneural wave precedes and induces transition of NE cells to NBs. Unpaired (Upd), the ligand for the JAK/STAT signaling pathway,is expressed in the most lateral NE cells. JAK/STAT signaling negatively regulates proneural wave progression and controls the number of NBs in the optic lobe. Our findings suggest that NBs might be balanced with the number of lamina neurons by JAK/STAT regulation of proneural wave progression, thereby providing the developmental basis for the formation of a precise topographic map in the visual center.
The effect of hypoxia on gill morphology and ionoregulatory status in the Lake Qinghai scaleless carp, Gymnocypris przewalskii
Authors: Matey Victoria , Richards Jeffrey G. , Wang Yuxiang , Wood Chris M. , Rogers Joe , Davies Rhiannon , Murray Brent W. , Chen X.-Q. , Du Jizeng , Brauner Colin J.
Published: March 2008

Goldfish and crucian carp at low temperature exhibit plasticity in gill morphology during exposure to hypoxia to enhance gas exchange. Hypoxia-induced changes in gill morphology and cellular ultrastructure of the high altitude scaleless carp from Lake Qinghai, China, were investigated to determine whether this is a general characteristic of cold water carp species. Fish were exposed to acute hypoxia (0.3 mg O2 l–1) for 24 h followed by 12 h recovery in normoxic water (6 mg O2l–1 at 3200 m altitude), with no mortality. Dramatic alterations in gill structure were initiated within 8 h of hypoxia and almost complete by 24 h, and included a gradual reduction of filament epithelial thickness (>50%), elongation of respiratory lamellae, expansion of lamellar respiratory surface area (>60%) and reduction in epithelial water–blood diffusion distance (<50%). An increase in caspase 3 activity in gills occurred following 24 h exposure to hypoxia, indicating possible involvement of apoptosis in gill remodeling. Extensive gill mucous production during hypoxia may have been part of a general stress response or may have played a role in ion exchange and water balance. The large increase in lamellar surface area and reduction in diffusion distance presumably enhances gas transfer during hypoxia (especially in the presence of increased mucous production) but comes with an ionoregulatory cost, as indicated by a 10 and 15% reduction in plasma [Na+] and [Cl–],respectively, within 12–24 h of hypoxia. Within 12 h of hypoxia exposure, `wavy-convex'-mitochondria rich cells (MRCs) with large apical crypts and numerous branched microvilli were transformed into small`shallow-basin' cells with a flattened surface. As the apical membrane of MRCs is the site for active ion uptake from the water, a reduction in apical crypt surface area may have contributed to the progressive reduction in plasma[Na+] and [Cl–] observed during hypoxia. The changes in the macro- and ultra-structure of fish gills, and plasma[Na+] and [Cl–] during hypoxia were reversible,showing partial recovery by 12 h following return to normoxia. Although the large morphological changes in the gill observed in the scaleless carp support the hypothesis that gill remodeling during hypoxia is a general characteristic of cold water carp species, the reduced magnitude of the response in scaleless carp relative to goldfish and crucian carp may be a reflection of their more active lifestyle or because they reside in a moderately hypoxic environment at altitude.
Kinematics and power requirements of ascending and descending flight in the pigeon (Columba livia)
Authors: Berg Angela M. , Biewener Andrew A.
Published: March 2008

Ascending or descending locomotion involves a change in potential energy(PE) and a corresponding change in power requirement. We sought to test whether the mechanical power required for steady ascending or descending flight is a simple sum of the power required for level flight and the power necessary for potential energy change. Pigeons (Columba livia) were trained to fly at varying angles of ascent and descent (60°, 30°,0°, –30°, –60°), and were recorded using high-speed video. Detailed three-dimensional kinematics were obtained from the recordings, allowing analysis of wing movement. Aerodynamic forces and power requirements were then estimated from kinematic data. As expected,`PE flight power' increased significantly with angle of flight (0.234 W deg.–1), though there appeared to be a limit on the amount of PE that the birds could gain or dissipate per wingbeat. We found that the total power output for flight at various angles was not different from the sum of power required for level flight and the PE rate of change for a given angle, except for the steep –60° descent. The total power for steep descent was higher than this sum because of a higher induced power due to the bird's deceleration and slower flight velocity. Aerodynamic force estimates during mid-downstroke did not differ significantly in magnitude or orientation among flight angles. Pigeons flew fastest during–30° flights (4.9±0.1 m s–1) and slowest at 60° (2.9±0.1 m s–1). Although wingbeat frequency ranged from 6.1 to 9.6 Hz across trials, the variation was not significant across flight angles. Stroke plane angle was more horizontal, and the wing more protracted, for both +60° and –60° flights, compared with other flight path angles.
The effect of food temperature on postprandial metabolism in albatrosses
Authors: Battam H. , Chappell M. A. , Buttemer W. A.
Published: March 2008

Heat generated by the specific dynamic action (SDA) associated with feeding is known to substitute for the thermoregulatory costs of cold-exposed endotherms; however, the effectiveness of this depends on food temperature. When food is cooler than core body temperature, it is warmed by body heat and,consequently, imposes a thermoregulatory challenge to the animal. The degree to which this cost might be `paid' by SDA depends on the relative timing of food heating and the SDA response. We investigated this phenomenon in two genera of endotherms, Diomedea and Thalassarche albatrosses,by measuring postprandial metabolic rate following ingestion of food at body temperature (40°C) and cooler (0 and 20°C). This permitted us to estimate potential contributions to food warming by SDA-derived heat, and to observe the effect of cold food on metabolic rate. For meal sizes that were∼20% of body mass, SDA was 4.22±0.37% of assimilated food energy,and potentially contributed 17.9±1.0% and 13.2±2.2% of the required heating energy of food at 0°C for Diomedea and Thalassarche albatrosses, respectively, and proportionately greater quantities at higher food temperatures. Cold food increased the rate at which postprandial metabolic rate increased to 3.2–4.5 times that associated with food ingested at body temperature. We also found that albatrosses generated heat in excess by more than 50% of the estimated thermostatic heating demand of cold food, a probable consequence of time delays in physiological responses to afferent signals.
The effect of thermal history on the susceptibility of reef-building corals to thermal stress
Authors: Middlebrook Rachael , Hoegh-Guldberg Ove , Leggat William
Published: March 2008

The mutualistic relationship between corals and their unicellular dinoflagellate symbionts (Symbiodinium sp.) is a fundamental component within the ecology of coral reefs. Thermal stress causes the breakdown of the relationship between corals and their symbionts (bleaching). As with other organisms, this symbiosis may acclimate to changes in the environment, thereby potentially modifying the environmental threshold at which they bleach. While a few studies have examined the acclimation capacity of reef-building corals, our understanding of the underlying mechanism is still in its infancy. The present study focused on the role of recent thermal history in influencing the response of both corals and symbionts to thermal stress, using the reef-building coral Acropora aspera. The symbionts of corals that were exposed to 31°C for 48 h (pre-stress treatment) 1 or 2 weeks prior to a 6-day simulated bleaching event (when corals were exposed to 34°C) were found to have more effective photoprotective mechanisms. These mechanisms included changes in non-photochemical quenching and xanthophyll cycling. These differences in photoprotection were correlated with decreased loss of symbionts, with those corals that were not prestressed performing significantly worse, losing over 40% of their symbionts and having a greater reduction in photosynthetic efficiency. These results are important in that they show that thermal history, in addition to light history, can influence the response of reef-building corals to thermal stress and therefore have implications for the modeling of bleaching events. However, whether acclimation is capable of modifying the thermal threshold of corals sufficiently to cope as sea temperatures increase in response to global warming has not been fully explored. Clearly increases in sea temperatures that extend beyond 1–2°C will exhaust the extent to which acclimation can modify the thermal threshold of corals.
Persistence of motor unit and muscle fiber types in the presence of inactivity
Authors: Roy Roland R. , Pierotti David J. , Garfinkel Alan , Zhong Hui , Baldwin Kenneth M. , Edgerton V. Reggie
Published: March 2008

The clarity of categorizing skeletal muscle fibers in individual motor units into phenotypes based on quantitative single fiber enzyme activities and as a function of neuromuscular activity level was examined. Neuromuscular activity was eliminated in adult cat hindlimb muscles by spinal cord isolation(SI), i.e. complete spinal cord transection at a low thoracic and a high sacral level with bilateral dorsal rhizotomy between the transection sites. One motor unit was isolated via ventral root teasing procedures from the tibialis anterior (TA) muscle of each hindlimb in control and SI cats, and physiologically tested and glycogen depleted through repetitive stimulation;fibers comprising each motor unit were visualized through glycogen staining. Each motor unit was composed of fibers of the same myosin immunohistochemical type. Myofibrillar adenosine triphosphatase, succinate dehydrogenase andα-glycerophosphate dehydrogenase activities were determined for a sample of motor unit and non-motor unit fibers, providing a measure of three enzyme activities often used to characterize fiber phenotype within a single unit. Although normal enzyme activities were altered after 6 months of inactivity,the relationships among the three enzymes were largely maintained. These data demonstrate that it is not the diversity in any single enzyme property but the profile of several metabolic pathways that underlies the significance of fiber phenotypes. These profiles must reflect a high level of coordination of expression of selected combinations of genes. Although neuromuscular activity level influences fiber phenotype, the present results demonstrate that activity-independent mechanisms remain important sources of the control of phenotype establishment in the near absence of activity.
Jumping in a wingless stick insect,Timema chumash(Phasmatodea,Timematodea, Timematidae)
Authors: Burrows Malcolm
Published: March 2008

The stick insect Timema chumash belongs to a sub-order of the phasmids that is thought to have diverged early from other stick insects, and which is restricted to the southwest of North America. It jumps by rapidly extending the tibiae of both its hind legs simultaneously from an initially fully flexed position, unlike any other stick insect that has been described. The hind legs are 1.5 times longer than the front and middle legs, but still represent only half the length of its body, and the femoro-tibial joints show few specialisations for jumping. In its best jumps, the wingless body is accelerated in 12 ms to a take-off velocity of 0.9 m s–1 and experiences an acceleration of 75 m s–2, the equivalent of 8 g. This performance requires an energy expenditure of 19 μJ,generates a power output of 1.6 mW and exerts a force of 3.6 mN. The jump propels the body forward a distance of 80 mm from a mean take-off angle of 39°. Heights of 20 mm were also achieved. Elevation of the jump was controlled by the initial position of the hind legs; when the hind tibiae and femora projected above the dorsal outline of the body the jump was forwards,when parallel with the long axis of the body the jump was backwards and could result in somersaulting. The jumping movements would appear to displace Timema in different directions away from a potential predator.
Matched regulation of gastrointestinal performance in the Burmese python,Python molurus
Authors: Cox Christian L. , Secor Stephen M.
Published: March 2008

In Burmese pythons fasting and feeding cause dramatic regulation of gastric acid production and intestinal nutrient uptake. Predictably, other components of their gastrointestinal tract are similarly regulated with each meal. We therefore assessed the matched regulation of gastrointestinal performance by comparing the postprandial activities and capacities of gastric (pepsin),pancreatic (amylase and trypsin) and intestinal (aminopeptidase-N and maltase)enzymes, and intestinal nutrient uptake. Tissue samples were collected from pythons fasted and at 0.25, 0.5, 1, 2, 3, 4, 6, 10 and 15 days following their consumption of rodent meals equaling 25% of snake body mass. With feeding,pythons experience no significant change in stomach mass, whereas both the pancreas and small intestine doubled in mass. Feeding also triggered a depletion of gastric mucosal pepsinogen, a respective 5.7- and 20-fold increase in the peak activities of pancreatic trypsin and amylase, and a respective 2.3- and 5.5-fold increase in the peak activities of intestinal maltase and aminopeptidase-N. Enzyme activities peaked between 2 and 4 days postfeeding and returned to fasting levels by day 10. Independent of digestive stage, python intestine exhibited a proximal to distal decline in enzyme activity. For both sugars and proteins, intestinal capacities for enzyme activity were significantly correlated with nutrient uptake capacities. The concomitant postprandial upregulation of tissue morphology, intestinal nutrient transport rates and enzyme activities illustrate, for the python, the matched regulation of their gastrointestinal performance with each meal.