bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protein kinase C is an important signaling mediator associated with motility of intact sea urchin spermatozoa
Authors: White Daniel , de Lamirande Eve , Gagnon Claude 
Published: November 2007

SUMMARYNumerous kinases and phosphatases are most likely implicated in sperm motility initiation and maintenance. Data on these signaling molecules were mostly obtained from studies conducted on in vitrodemembranated–reactivated sperm models but are not necessarily representative of the in vivo situation. We therefore investigated the effect of a variety of cell-permeable chemicals, mostly kinase inhibitors,on the motility initiation and maintenance of intact sea urchin spermatozoa. Among the 20 substances tested, the protein kinase C (PKC) inhibitor chelerythrine was the most potent, arresting motility at concentrations starting from 1.5–2 μmol l–1. Motility was also inhibited by two other PKC inhibitors as well as staurosporine. Furthermore,these inhibitors prevented the motility-associated increase in phosphorylation of at least four PKC substrates. These phospho-PKC target proteins, as assessed with an antibody specific to phosphorylated motifs of PKC substrates,were found to be associated with the flagellum, either in the Triton X-100 soluble portion or the axoneme (Triton X-100 insoluble). A phosphorylated PKC-like enzyme was also detected by immunoblotting in the flagellum, as well as a significant 50 kDa PKC cleavage product. Taken together, the data strongly indicate for the first time that, in vivo, which means on intact spermatozoa, PKC is a key signaling mediator associated with the maintenance of sea urchin sperm motility.
Lung ventilation contributes to vertical lymph movement in anurans
Authors: Hedrick Michael S. , Drewes Robert C. , Hillman Stanley S. , Withers Philip C. 
Published: November 2007

SUMMARY
Anurans (frogs and toads) generate lymphatic fluid at 10 times the rate in mammals, largely as a consequence of their very `leaky' vasculature and high interstitial compliance. Lymph is ultimately pumped into the venous system by paired, dorsally located lymph hearts. At present, it is unclear how lymphatic fluid that accumulates in central body subcutaneous lymph sacs is moved to the anterior and posterior lymph hearts in the axillary regions and how lymph is moved, against gravity, to the dorsally located lymph hearts. In this study,we tested the hypothesis that lung ventilation, through its consequent effects on lymph sac pressure, contributes to the vertical movement of lymphatic fluid in the cane toad (Chaunus marinus) and the North American bullfrog(Lithobates catesbeiana). We measured pressure in the dorsal, lateral and subvertebral lymph sacs of anesthetized cane toads and bullfrogs during artificial lung inflation and deflation. We also measured pressure in the subvertebral lymph sac, which adheres to the dorsal surface of the lungs,simultaneously with brachial (forelimb) and pubic (posterior) sac pressure during ventilation in freely behaving animals. There were highly significant(P&lt;0.001) relationships between lung pressure and lymph sac pressures (r2=0.19–0.72), indicating that pulmonary pressure is transmitted to the highly compliant lymph sacs that surround the lungs. Subvertebral sac pressure of resting animals was not significantly different between L. catesbeiana (518±282 Pa) and C. marinus (459±111 Pa). Brachial sac compliance (ml kPa–1 kg–1) also did not differ between the two species (33.6±5.0 in L. catesbeiana and 37.0±9.4 in C. marinus). During expiration (lung deflation), reductions in expanding subvertebral sac pressure are communicated to the brachial lymph sac. Changes in brachial and pubic lymph sac pressures were correlated almost entirely during expiration rather than inspiration. The change in brachial sac pressure during expiration was 235±43 Pa for C. marinus and 215±50 Pa for L. catesbeiana, which is of sufficient magnitude to move lymph the estimated 0.5–1.0 cm vertical distance from the forelimb to the vicinity of the anterior lymph hearts. We suggest that lymph is moved during expiration to the subvertebral sac from anterior and posterior lymph sacs. During lung inflation, increased lymph sac pressure moves lymph to axillary regions, where lymph hearts can return lymph to the vascular space. Consequently, pulmonary ventilation has an important role for lymph movement and, hence, blood volume regulation in anurans.
Comparative thyroidology: thyroid gland location and iodothyronine dynamics in Mozambique tilapia (Oreochromis mossambicus Peters) and common carp (Cyprinus carpio L.)
Authors: Geven Edwin J. W. , Nguyen Nha-Khanh , van den Boogaart Marcel , Spanings F. A. Tom , Flik Gert , Klaren Peter H. M. 
Published: November 2007

SUMMARY
In teleosts, the thyroid gland is mostly found in the subpharyngeal region. However, in several species thyroid follicles are found in, for example,heart, head kidney and kidney. Such heterotopic thyroid follicles are active,and considered to work in concert with the subpharyngeal thyroid. In Mozambique tilapia (Oreochromis mossambicus) thyroid activity is,indeed, restricted to the subpharyngeal region; in common carp (Cyprinus carpio) the functional endocrine thyroid is associated with renal tissues. The subpharyngeal follicles of carp comprise only 10% of the total thyroid tissue, and these follicles neither accumulate iodide nor synthesize or secrete thyroid hormones to a significant degree. Although the shape and size of carp subpharyngeal and renal follicles vary, the epithelial cell height of the thyrocytes and thyroxine immunoreactivity do not differ, which suggests that the activity of the carp subpharyngeal thyroid follicles is dormant. Differences in thyroid physiology between the two fish species were further assessed at the level of peripheral thyroid hormone metabolism. Carp clears plasma of thyroid hormones faster than tilapia does. Furthermore, a significant amount of conjugated thyroid hormones was observed in the plasma of tilapia, which was preceded by the occurrence of thyroid hormone conjugates in the subpharyngeal region and coincides with the appearance of conjugates in the surrounding water. Apparently, plasma thyroid hormone conjugates in tilapia originate from the thyroid gland and function in the excretion of thyroid hormones. Our data illustrate the variability in teleostean thyroidology, an important notion for those studying thyroid physiology.
Mudskippers brood their eggs in air but submerge them for hatching
Authors: Ishimatsu Atsushi , Yoshida Yu , Itoki Naoko , Takeda Tatsusuke , Lee Heather J. , Graham Jeffrey B. 
Published: November 2007

SUMMARYIntertidal mudflats are highly productive ecosystems that impose severe environmental challenges on their occupants due to tidal oscillations and extreme shifts in habitat conditions. Reproduction on mudflats requires protection of developing eggs from thermal and salinity extremes,O2 shortage, dislodgement by currents, siltation and predation. Mudskippers are air-breathing, amphibious fishes, and one of few vertebrates that reside on mudflats. They lay their eggs in mud burrows containing extremely hypoxic water, raising the question of how the eggs survive. We found that the Japanese mudskipper Periophthalmus modestus deposits its eggs on the walls of an air-filled chamber within its burrow. To ensure adequate O2 for egg development, the burrow-guarding male mudskipper deposits mouthfuls of fresh air into the egg chamber during each low tide, a behaviour that can be upregulated by egg-chamber hypoxia. When egg development is complete the male, on a nocturnal rising tide, removes the egg-chamber air and releases it outside the burrow. This floods the egg chamber and induces egg hatching. Thus, P. modestus has developed a reproductive strategy that allows it to nurture eggs in this severe habitat rather than migrating away from the mudflat. This requires that mudskipper eggs be specialized to develop in air and that the air-breathing capacity of the egg-guarding male be integrated in a complex behavioural repertoire that includes egg guarding, ferrying air to and from the egg chamber, and sensing O2 levels therein, all in concert with the tidal cycle.
An investigation of the effects of lorry suspension performance on road maintenance costs
Authors: Costanzi M , Cebon D 
Published: November 2007

A mathematical model of the interaction between a fleet of heavy vehicles and a spraysealed road surface is developed and validated using field performance data. Key features of the model include: (a) modelling the dynamic performance of the vehicle fleet using an ensemble of ‘quarter car’ vehicle models; (b) careful accounting for the ‘spatial repeatability’ of tyre forces; (c) use of accelerated pavement performance test data to model the evolution of the road surface profile; (d) surface maintenance intervention based on permanent deformation (rutting), potholing and excessive surface roughness. The model is used to predict long-term road maintenance intervention costs and to compare the predicted road maintenance costs for various suspension scenarios in the vehicle fleet. The simulation results indicate that conversion of the heavy vehicle fleet from conventional leaf spring suspensions to ‘road-friendly’ suspensions would results in a reduction in road maintenance costs per tonne-km of 14 per cent. Increasing the freight load by 3.0 tonnes per vehicle would reduce this benefit to approximately 1 per cent. If the fleet was to have 50 per cent poorly maintained shock absorbers, the simulations show an increase in road maintenance expenditures per tonne-km: about 5 per cent higher than conventional suspensions at the lower weights and 21 per cent higher at the higher weights.
Roughness in lubricated rolling contact: The dry contact limit
Authors: Olver A. V. , Dini D 
Published: November 2007

A difficulty with the standard fast Fourier transform (FFT) perturbation model of roughness in lubricated rolling contacts is that it does not necessarily converge towards the elastic case as the film thickness is reduced; rather it leads to a situation in which all the roughness is completely flattened. This is rarely the case for real engineering surfaces. Here, it is shown that this difficulty can be avoided by carrying out a Fourier transform of the elastostatically flattened roughness and using the resulting (complex) amplitude as the low-film thickness limit of each Fourier component in the elastohydrodynamic lubrication (EHL) analysis. Results give a plausible convergence to the elastostatic solution, which is nevertheless consistent with the expected near-full-film EHL behaviour and which becomes identical to the earlier model for roughness that, statically, can be fully flattened. As expected, hydrodynamic action persists at the finest scale, even for very thin films.
Arsenic Speciation Analysis in Human Saliva
Authors: Yuan Chungang , Lu Xiufen , Oro Nicole , Wang Zhongwen , Xia Yajuan , Wade Timothy J , Mumford Judy , Le X Chris 
Published: November 2007

Abstract
Background: Determination of arsenic species in saliva is potentially useful for biomonitoring of human exposure and studying arsenic metabolism. Arsenic speciation in saliva has not been reported previously.
Methods: We separated arsenic species in saliva using liquid chromatography (LC) and quantified them by inductively coupled plasma mass spectrometry. We further confirmed the identities of arsenic species by LC coupled with electrospray ionization tandem mass spectrometry. These methods were successfully applied to the determination of arsenite (AsIII), arsenate (AsV), and their methylation metabolites, monomethylarsonic acid (MMAV), and dimethylarsinic acid (DMAV), in &gt;300 saliva samples collected from people who were exposed to varying concentrations of arsenic.
Results: The mean (range) concentrations (μg/L) in the saliva samples from 32 volunteers exposed to background levels of arsenic were AsIII 0.3 [not detectable (ND) to 0.7], AsV 0.3 (ND to 0.5), MMAV 0.1 (ND to 0.2), and DMAV 0.7 (ND to 2.6). Samples from 301 people exposed to increased concentrations of arsenic in drinking water showed detectable AsIII in 99%, AsV in 98%, MMAV in 80%, and DMAV in 68% of samples. The mean (range) concentrations of arsenic species in these saliva samples were (in μg/L) AsIII 2.8 (0.1–38), AsV 8.1 (0.3–120), MMAV 0.8 (0.1–6.0), and DMAV 0.4 (0.1–3.9). Saliva arsenic correlated with drinking water arsenic. Odds ratios for skin lesions increased with saliva arsenic concentrations. The association between saliva arsenic concentrations and the prevalence of skin lesions was statistically significant (P &lt;0.001).
Conclusions: Speciation of AsV, AsIII, MMAV, and DMAV in human saliva is a useful method for monitoring arsenic exposure.
Latent MMP-9 is bound to TIMP-1 before secretion
Authors: Roderfeld Martin , Graf Jürgen , Giese Bernd , Salguero-Palacios Rebeca , Tschuschner Annette , Müller-Newen Gerhard , Roeb Elke 
Published: November 2007

AbstractExpression patterns of matrix metalloproteinase-9 (MMP-9) and its specific inhibitor, tissue inhibitor of metalloproteinases-1 (TIMP-1), are closely correlated with physiological and pathological processes characterized by the degradation and accumulation of the extracellular matrix (ECM). Both, activated MMP-9 and pro-MMP-9 can bind to TIMP-1, and most cell types secrete MMP-9 in complex with TIMP-1. Utilizing immunofluorescence, we observed intracellular co-localization of MMP-9 and TIMP-1 in stimulated human fibrosarcoma cells. In the present study we searched for the origin of the complex formation between the latent enzyme and its specific inhibitor on a subcellular level. Fluorescence resonance energy transfer (FRET) between the fluorescently labeled enzyme and its inhibitor in co-transfected cells were measured. MMP-9 and TIMP-1 were fused to cyan (CFP) and yellow (YFP) variants of the green fluorescent protein and transiently expressed in human hepatoma cells. The intracellular distribution of fluorescently labeled TIMP-1 and MMP-9 was analyzed by confocal laser scanning microscopy. Intracellular complex formation in the Golgi apparatus was verified, demonstrating FRET between MMP-9-CFP and TIMP-1-YFP. Our data provide evidence that the proMMP-9-TIMP-1 complex is already present in the Golgi apparatus. This may be of significance for a number of intracellular and extracellular biochemical processes involving proMMP-9. However, the magnitude and functional relevance of this finding remain unknown.
Defining the extended substrate specificity of kallikrein 1-related peptidases
Authors: Borgoño Carla A. , Gavigan Julie-Ann , Alves Juliano , Bowles Ben , Harris Jennifer L. , Sotiropoulou Georgia , Diamandis Eleftherios P. 
Published: November 2007

AbstractHuman kallikrein 1-related peptidases (KLKs) form a subfamily of 15 extracellular (chymo)tryptic-like serine proteases. KLKs 4, 5, 13 and 14 display altered expression/activity in diverse pathological conditions, including cancer. However, their distinct (patho)physiological roles remain largely uncharacterized. As a step toward distinguishing their proteolytic functions, we attempt to define their primary and extended substrate specificities and identify candidate biological targets. Heterologously expressed KLKs 4, 5, 13 and 14 were screened against fluorogenic 7-amino-4-carbamoylmethylcoumarin positional scanning-synthetic combinatorial libraries with amino acid diversity at the P1–P4 positions. Our results indicate that these KLKs share a P1 preference for Arg. However, each KLK exhibited distinct P2–P4 specificities, attributable to structural variations in their surface loops. The preferred P4–P1 substrate recognition motifs based on optimal subsite occupancy were as follows: VI-QSAV-QL-R for KLK4; YFWGPV-RK-NSFAM-R for KLK5; VY-R-LFM-R for KLK13; and YW-KRSAM-HNSPA-R for KLK14. Protein database queries using these motifs yielded many extracellular targets, some of which represent plausible KLK substrates. For instance, cathelicidin, urokinase-type plasminogen activator, laminin and transmembrane protease serine 3 were retrieved as novel putative substrates for KLK4, 5, 13 and 14, respectively. Our findings may facilitate studies on the role of KLKs in (patho)physiology and can be used in the development of selective KLK inhibitors.
Characterization and comparative 3D modeling of CmPI-II, a novel ‘non-classical’ Kazal-type inhibitor from the marine snail Cenchritis muricatus (Mollusca)
Authors: González Yamile , Pons Tirso , Gil Jeovanis , Besada Vladimir , Alonso-del-Rivero Maday , Tanaka Aparecida S. , Araujo Mariana S. , Chávez María A. 
Published: November 2007

Abstract
The complete amino acid sequence obtained by electrospray ionization tandem mass spectrometry of the proteinase inhibitor CmPI-II isolated from Cenchritis muricatus is described. CmPI-II is a 5480-Da protein with three disulfide bridges that inhibits human neutrophil elastase (HNE) (K
i 2.6±0.2 nM), trypsin (K
i 1.1±0.9 nM), and other serine proteinases such as subtilisin A (K
i 30.8±1.2 nM) and pancreatic elastase (K
i 145.0±4.4 nM); chymotrypsin, pancreatic and plasma kallikreins, thrombin and papain are not inhibited. CmPI-II shares homology with the Kazal-type domain and may define a new group of ‘non-classical’ Kazal inhibitors according to its CysI-CysV disulfide bridge position. The 3D model of CmPI-II exhibits similar secondary structure characteristics to Kazal-type inhibitors and concurs with circular dichroism experiments. A 3D model of the CmPI-II/HNE complex provides a structural framework for the interpretation of its experimentally determined K
i value. The model shows both similar and different contacts at the primary binding sites in comparison with the structure of turkey ovomucoid third domain (OMTKY3)/HNE used as template. Additional contacts calculated at the protease-inhibitor interface could also contribute to the association energy of the complex. This inhibitor represents an exception in terms of specificity owing to its ability to strongly inhibit elastases and trypsin.
Novel expression of kallikreins, kallikrein-related peptidases and kinin receptors in human pleural mesothelioma
Authors: Chee Jessica , Singh Jasmine , Naran Anupam , Misso Neil L. , Thompson Philip J. , Bhoola Kanti D. 
Published: November 2007

AbstractMalignant mesothelioma is an aggressive cancer of the pleura that is causally related to exposure to asbestos fibres. The kallikrein serine proteases [tissue (hK1) and plasma (hKB1) kallikreins, and kallikrein-related peptidases (KRP/hK2–15)] and the mitogenic kinin peptides may have a role in tumourigenesis. However, it is not known whether hK1, hKB1, KRP/hK proteins or kinin receptors are expressed in pleural mesotheliomas. The expression of hK1, hKB1, KRP/hK2, 5, 6, 7, 8 and 9, and kinin B1and B2receptors was assessed in archived selected normal tissue and mesothelioma tumour sections by immunoperoxidase and immunofluorescence labelling. hK1, hKB1 and kinin B1and B2receptors were expressed in malignant cells of the epithelioid and sarcomatoid components of biphasic mesothelioma tumour cells. The percentage of cells with cytoplasmic and nuclear labelling and the intensity of labelling were similar for hK1, hKB1 and the kinin receptors. KRP/hK2, 6, 8 and 9 were also expressed in the cytoplasm and nuclei of mesothelioma cells, whereas KRP/hK5 and hK7 showed predominantly cytoplasmic localisation. This is a first report, but further studies are required to determine whether these proteins have a functional role in the pathogenesis of mesothelioma and/or may be potential biomarkers for pleural mesothelioma.
Cellular localization of MAGI-1 caspase cleavage products and their role in apoptosis
Authors: Ivanova Saška , Repnik Urška , Banks Lawrence , Turk Vito , Turk Boris 
Published: November 2007

Abstract
MAGI-1 is a membrane-associated guanylate kinase (MAGUK) protein present at adherent and tight junctions, where it acts as a structural and signaling scaffold. During apoptosis, MAGI-1 is cleaved by caspases at Asp761 into N- and C-terminal cleavage products, allowing further dismantling of the cell junctions, one of the key features of apoptosis. Here, we investigated the cellular distribution and possible proapototic role of MAGI-1 caspase cleavage products. Full-length MAGI-1 exhibited submembrane localization, while the N-terminal caspase cleavage product of MAGI-1 is translocated to the cytosol and the C-terminal caspase cleavage product accumulates in the nucleus. When overexpressed in MDCK cells, both N- and C-terminal MAGI-1 caspase cleavage products exhibited minor proapoptotic activity, although their role in apoptosis is probably more passive.
Activity of ulilysin, an archaeal PAPP-A-related gelatinase and IGFBP protease
Authors: Tallant Cynthia , García-Castellanos Raquel , Marrero Aniebrys , Canals Francesc , Yang Yongzheng , Reymond Jean-Louis , Solà Maria , Baumann Ulrich , Gomis-Rüth F. Xavier 
Published: November 2007

Abstract
Human growth and development are conditioned by insulin-like growth factors (IGFs), which have also implications in pathology. Most IGF molecules are sequestered by IGF-binding proteins (IGFBPs) so that exertion of IGF activity requires disturbance of these complexes. This is achieved by proteolysis mediated by IGFBP proteases, among which the best characterised is human PAPP-A, the first member of the pappalysin family of metzincins. We have previously identified and studied the only archaeal homologue found to date, Methanosarcina acetivorans ulilysin. This is a proteolytically functional enzyme encompassing a pappalysin catalytic domain and a pro-domain involved in maintenance of latency of the zymogen, proulilysin. Once activated, the protein hydrolyses IGFBP-2 to -6 and insulin chain β in vitro. We report here that ulilysin is also active against several other substrates, viz (azo)casein, azoalbumin, and extracellular matrix components. Ulilysin has gelatinolytic but not collagenolytic activity. Moreover, the proteolysis-resistant skeletal proteins actin and elastin are also cleaved, as is fibrinogen, but not plasmin and α1-antitrypsin from the blood coagulation cascade. Ulilysin develops optimal activity at pH 7.5 and strictly requires peptide bonds preceding an arginine residue, as determined by means of a novel fluorescence resonance energy transfer assay, thus pointing to biotechnological applications as an enzyme complementary to trypsin.
Cysteine proteases: destruction ability versus immunomodulation capacity in immune cells
Authors: Zavašnik-Bergant Tina , Turk Boris 
Published: November 2007

Abstract
Cysteine proteases (cathepsins) play a pivotal role in various physiological processes, as well as in several diseases. In the immune response, maturation of major histocompatibility class II (MHC II) molecules and processing of antigens for further presentation by MHC II is tightly linked to the enzymes of the endosomal/lysosomal system, of which cysteine proteases constitute a major proportion. Furthermore, the process of autophagy provides access for cytosolic antigens to proteolysis by lysosomal cathepsins and subsequent MHC II presentation. Other specific functions of proteolytic enzymes associated with the immune response, such as activation of granzymes by cathepsin C in T-lymphocytes, are introduced and covered in this review.