bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gq-dependent signalling by the lysophosphatidic acid receptor LPA3 in gastric smooth muscle: reciprocal regulation of MYPT1 phosphorylation by Rho kinase and cAMP-independent PKA
Authors: Sriwai Wimolpak , Zhou Huiping , Murthy Karnam S. 
Published: April 2008

The present study characterized the signalling pathways initiated by the bioactive lipid, LPA (lysophosphatidic acid) in smooth muscle. Expression of LPA3 receptors, but not LPA1 and LPA2, receptors was demonstrated by Western blot analysis. LPA stimulated phosphoinositide hydrolysis, PKC (protein kinase C) and Rho kinase (Rho-associated kinase) activities: stimulation of all three enzymes was inhibited by expression of the Gαq, but not the Gαi, minigene. Initial contraction and MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation induced by LPA were abolished by inhibitors of PLC (phospholipase C)-β (U73122) or MLCK (myosin light-chain kinase; ML-9), but were not affected by inhibitors of PKC (bisindolylmaleimide) or Rho kinase (Y27632). In contrast, sustained contraction, and phosphorylation of MLC20 and CPI-17 (PKC-potentiated inhibitor 17 kDa protein) induced by LPA were abolished selectively by bisindolylmaleimide. LPA-induced activation of IKK2 {IκB [inhibitor of NF-κB (nuclear factor κB)] kinase 2} and PKA (protein kinase A; cAMP-dependent protein kinase), and degradation of IκBα were blocked by the RhoA inhibitor (C3 exoenzyme) and in cells expressing dominant-negative mutants of IKK2(K44A) or RhoA(N19RhoA). Phosphorylation by Rho kinase of MYPT1 (myosin phosphatase targeting subunit 1) at Thr696 was masked by phosphorylation of MYPT1 at Ser695 by PKA derived from IκB degradation via RhoA, but unmasked in the presence of PKI (PKA inhibitor) or C3 exoenzyme and in cells expressing IKK2(K44A). We conclude that LPA induces initial contraction which involves activation of PLC-β and MLCK and phosphorylation of MLC20, and sustained contraction which involves activation of PKC and phosphorylation of CPI-17 and MLC20. Although Rho kinase was activated, phosphorylation of MYPT1 at Thr696 by Rho kinase was masked by phosphorylation of MYPT1 at Ser695 via cAMP-independent PKA derived from the NF-κB pathway.
The guanine-nucleotide-exchange factor BopE from Burkholderia pseudomallei adopts a compact version of the Salmonella SopE/SopE2 fold and undergoes a closed-to-open conformational change upon interaction with Cdc42
Authors: Upadhyay Abhishek , Wu Huan-Lin , Williams Christopher , Field Terry , Galyov Edouard E. , van den Elsen Jean M. H. , Bagby Stefan 
Published: April 2008

BopE is a type III secreted protein from Burkholderia pseudomallei, the aetiological agent of melioidosis, a severe emerging infection. BopE is a GEF (guanine-nucleotide-exchange factor) for the Rho GTPases Cdc42 (cell division cycle 42) and Rac1. We have determined the structure of BopE catalytic domain (amino acids 78–261) by NMR spectroscopy and it shows that BopE78–261 comprises two three-helix bundles (α1α4α5 and α2α3α6). This fold is similar to that adopted by the BopE homologues SopE and SopE2, which are GEFs from Salmonella. Whereas the two three-helix bundles of SopE78–240 and SopE269–240 form the arms of a ‘Λ’ shape, BopE78–261 adopts a more closed conformation with substantial interactions between the two three-helix bundles. We propose that arginine and proline residues are important in the conformational differences between BopE and SopE/E2. Analysis of the molecular interface in the SopE78–240–Cdc42 complex crystal structure indicates that, in a BopE–Cdc42 interaction, the closed conformation of BopE78–261 would engender steric clashes with the Cdc42 switch regions. This implies that BopE78–261 must undergo a closed-to-open conformational change in order to catalyse guanine nucleotide exchange. In an NMR titration to investigate the BopE78–261–Cdc42 interaction, the appearance of additional peaks per NH for residues in hinge regions of BopE78–261 indicates that BopE78–261 does undergo a closed-to-open conformational change in the presence of Cdc42. The conformational change hypothesis is further supported by substantial improvement of BopE78–261 catalytic efficiency through mutations that favour an open conformation. Requirement for closed-to-open conformational change explains the 10–40-fold lower kcat of BopE compared with SopE and SopE2.
Rapid and extensive uptake and activation of hydrophobic triphenylphosphonium cations within cells
Authors: Ross Meredith F. , Prime Tracy A. , Abakumova Irina , James Andrew M. , Porteous Carolyn M. , Smith Robin A. J. , Murphy Michael P. 
Published: April 2008

Mitochondria-targeted molecules comprising the lipophilic TPP (triphenylphosphonium) cation covalently linked to a hydrophobic bioactive moiety are used to modify and probe mitochondria in cells and in vivo. However, it is unclear how hydrophobicity affects the rate and extent of their uptake into mitochondria within cells, making it difficult to interpret experiments because their intracellular concentration in different compartments is uncertain. To address this issue, we compared the uptake into both isolated mitochondria and mitochondria within cells of two hydrophobic TPP derivatives, [3H]MitoQ (mitoquinone) and [3H]DecylTPP, with the more hydrophilic TPP cation [3H]TPMP (methyltriphenylphosphonium). Uptake of MitoQ by mitochondria and cells was described by the Nernst equation and was ∼5-fold greater than that for TPMP, as a result of its greater binding within the mitochondrial matrix. DecylTPP was also taken up extensively by cells, indicating that increased hydrophobicity enhanced uptake. Both MitoQ and DecylTPP were taken up very rapidly into cells, reaching a steady state within 15 min, compared with ∼8 h for TPMP. This far faster uptake was the result of the increased rate of passage of hydrophobic TPP molecules through the plasma membrane. Within cells MitoQ was predominantly located within mitochondria, where it was rapidly reduced to the ubiquinol form, consistent with its protective effects in cells and in vivo being due to the ubiquinol antioxidant. The strong influence of hydrophobicity on TPP cation uptake into mitochondria within cells facilitates the rational design of mitochondria-targeted compounds to report on and modify mitochondrial function in vivo.
Urocortin 2 combined with angiotensin-converting enzyme inhibition in experimental heart failure
Authors: Rademaker Miriam T. , Charles Christopher J. , Nicholls M. Gary , Richards A. Mark 
Published: April 2008

Ucn2 (urocortin 2) is a recently discovered peptide with therapeutic potential in heart failure. As any new treatment is likely to be used in conjunction with standard ACEI (angiotensin-converting enzyme inhibitor) therapy, it is important that the combined effects of these agents are assessed. In the present study, we investigated the effects of Ucn2 and an ACEI (captopril) administered for 3 h, both separately and together, in eight sheep with pacing-induced heart failure. Ucn2 and captopril alone both increased CO (cardiac output; Ucn2&gt;captopril) and decreased arterial pressure (captopril&gt;Ucn2), left atrial pressure (Ucn2&gt;captopril) and peripheral resistance (Ucn2=captopril) relative to controls. Compared with either treatment alone, combined treatment further improved CO and reduced peripheral resistance and cardiac preload, without inducing further falls in blood pressure. In contrast with the marked increase in plasma renin activity observed with captopril alone, Ucn2 administration reduced renin activity, whereas the combined agents resulted in intermediate renin levels. All active treatments decreased circulating levels of aldosterone (Ucn2+captopril&gt;Ucn2=captopril), endothelin-1 and the natriuretic peptides (Ucn2+captopril=Ucn2&gt;captopril), whereas adrenaline (epinephrine) fell only with Ucn2 (Ucn2+captopril=Ucn2), and vasopressin increased during captopril alone. Ucn2, both separately and in conjunction with captopril, increased urine output, sodium and creatinine excretion and creatinine clearance. Conversely, captopril administered alone adversely affected these renal indices. In conclusion, co-treatment with Ucn2 and an ACEI in heart failure produced significantly greater improvements in haemodynamics, hormonal profile and renal function than achieved by captopril alone. These results indicate that dual treatment with these two agents is beneficial.
Proposals for the collection of multifrequency acoustic data
Authors: Korneliussen Rolf J. , Diner Noel , Ona Egil , Berger Laurent , Fernandes Paul G. 
Published: April 2008

Abstract
Korneliussen, R. J., Diner, N., Ona, E., Berger, L., and Fernandes, P. G. 2008. Proposals for the collection of multifrequency acoustic data. – ICES Journal of Marine Science, 65: 982–994. Acoustic surveys are used to estimate the abundance and distribution of many fish species, and have been based traditionally on data collected at a single acoustic frequency. Although it has been known for some time that the use of additional frequencies can provide information on the nature of the acoustic target, the knowledge and technology required to combine the so-called “multifrequency data” in an appropriate manner has been limited. The use of several transducers of different frequencies is now common on board research vessels and fishing vessels, so multifrequency data are often collected. In order for these data to be combined appropriately, their physical and spatial characteristics from each frequency should be as similar as possible. We detail the requirements deemed necessary to collect multifrequency data in an appropriate manner. They can be stringent and may not always be achievable, so we also consider the consequences of combining acoustic data originating in transducers with varying degrees of spatial separation and with different beam widths.
Plasma matrix metalloproteinase‐7 as a metastatic marker and survival predictor in patients with renal cell carcinomas
Authors: Ramankulov Azizbek , Lein Michael , Johannsen Manfred , Schrader Mark , Miller Kurt , Jung Klaus 
Published: April 2008

We evaluated the clinical usefulness of plasma matrix metalloproteinase‐7 (MMP‐7) as a diagnostic and prognostic biomarker in patients with renal cell carcinoma (RCC). MMP‐7 was quantified in plasma of 50 healthy subjects and 97 RCC patients using a Fluorokine MultiAnalyte Profiling assay. RCC patients were stratified into the following groups: without metastases (N0M0; n = 39), with lymph nodes (N1M0; n = 13), and with distant metastases (M1; n = 45). Diagnostic performance of MMP‐7 was analyzed by the receiver operating characteristics (ROC) curve. Kaplan–Meier analysis and the Cox regression model were used to estimate the impact of MMP‐7 on the cancer‐specific survival outcome of RCC patients. MMP‐7 was significantly higher in both metastatic groups N1M0 and M1 (medians, 3.82 and 3.34 µg/L) compared to N0M0 group or controls (medians, 1.85 and 1.64 µg/L; all P < 0.001). In ROC analysis, the area under the ROC curve of MMP‐7 was 0.80 in the detection of metastases in RCC (P < 0.0001). In the Kaplan–Meier analysis, patients with MMP‐7 above the 95th percentile of controls showed less favorable survival rates compared to those with normal MMP‐7 (log‐rank test, 15.7; P < 0.0001). High MMP‐7 was associated with cancer‐related mortality estimated by univariate Cox regression (risk ratio, 4.34, 95% CI, 1.12–10.6; P = 0.032). The multivariate Cox regression model determined MMP‐7 (risk ratio, 2.70, 95% CI, 1.39–5.24; P = 0.003) and metastases (risk ratio, 5.81, 95% CI, 2.77–12.2; P < 0.0001) as independent determinants of cancer‐related survival outcomes. In conclusion, increased plasma MMP‐7 could be related to metastatic disease and poor prognosis in patients with RCC. (Cancer Sci 2008; 99: 1188–1194)
Functional analysis of single amino‐acid mutations in the thrombopoietin‐receptor Mpl underlying congenital amegakaryocytic thrombocytopenia
Authors: Tijssen Marloes R. , Di Summa Franca , Van Den Oudenrijn Sonja , Zwaginga Jaap Jan , Van Der Schoot C. Ellen , Voermans Carlijn , De Haas Masja 
Published: April 2008

SummaryCongenital amegakaryocytic thrombocytopenia (CAMT) is a rare disorder that presents with severe thrombocytopenia and absence of megakaryocytes in the bone marrow. The disease may develop into bone marrow aplasia. Genetic defects in the gene encoding the thrombopoietin (Tpo) receptor, MPL, are the cause of this disease. In a previous study, we identified four missense mutations in CAMT patients, predicting Arg102Pro, Pro136His, Arg257Cys and Pro635Leu. To investigate whether these mutations result in defective Tpo‐binding and/or signalling, full‐length wildtype and mutant MPL were transduced into K562 cells. Expression levels and the ability to activate the mitogen‐activated protein kinase, Janus kinase‐signal transducer and activator of transcription and phosphoinositide‐3 kinase pathways upon Tpo‐binding were studied. The results predicted that MPL carrying the P136H or P635L mutation was not properly expressed, whereas the R102P and R257C mutations resulted in impaired signal transduction. Our results indicate that a severe clinical course may be expected when these mutations lead to absent Mpl expression or signalling in CAMT patients with missense mutations.
Developing idiotype vaccines for lymphoma: from preclinical studies to phase III clinical trials
Authors: Park Hyun Jun , Neelapu Sattva S. 
Published: April 2008

SummaryTherapeutic vaccines for B‐cell non‐Hodgkin lymphoma (NHL) using the clonal tumour immunoglobulin idiotype (Id) have been under development for more than three decades. A major obstacle for rapid progress in the field has been that the Id vaccine is patient‐specific and required the generation of a custom‐made product. The manufacturing issues were recently overcome by advances in hybridoma and recombinant DNA technology which facilitated the completion of several phase I and II clinical trials. The strong immunogenicity and apparent clinical benefit observed on the early phase studies led to the initiation of three randomized phase III clinical trials that are also nearing completion. This review will focus on the development of Id vaccines before and after the introduction of rituximab for the treatment of B‐cell NHL and also discuss potential strategies to enhance the efficacy of active immunotherapy in the future.
Results of bone marrow examination in 275 patients with histological features that suggest an indolent type of cutaneous B‐cell lymphoma
Authors: Senff Nancy J. , Kluin‐Nelemans Hanneke C. , Willemze Rein 
Published: April 2008

SummaryWhether or not bone marrow biopsies should be performed routinely in patients with skin lesions that show histological features consistent with an indolent B‐cell lymphoma [marginal zone lymphoma (MZL) or follicle centre lymphoma (FCL)] has been debated. As no studies have addressed this question for this group of lymphomas, we evaluated the results of bone marrow biopsy examination in 275 patients with histological features consistent with MZL (n = 82) or FCL (n = 193) first presenting in the skin. In the MZL group, two of 82 patients (2%) showed bone marrow involvement, which was the only extracutaneous localization in one of these patients. In the FCL group, 22 of 193 patients (11%) had bone marrow involvement and was the only extracutaneous localization in nine of them. FCL patients with skin lesions and a positive bone marrow had a significantly worse prognosis when compared with patients with only skin lesions (5‐year disease‐specific survival 63% vs. 95%; P = 0·001). These results indicate that bone marrow investigation is essential for staging patients with a FCL first presenting in the skin. Bone marrow examination appears to have limited value in patients with MZL presenting in the skin.
Triiodothyronine stimulates hepatocyte proliferation in two models of impaired liver regeneration
Authors: Columbano A. , Simbula M. , Pibiri M. , Perra A. , Deidda M. , Locker J. , Pisanu A. , Uccheddu A. , Ledda‐Columbano G. M. 
Published: April 2008

Abstract.  Objectives: Liver regeneration is attenuated in old age and is substantially slower after 90% than after 70% partial hepatectomy (PH). We have previously demonstrated that the proliferative response to a primary mitogen is intact in aged mice, indicating that impaired liver regeneration is not due to loss of proliferative capacity. Here, we have investigated whether mitogenic effects of triiodothyronine (T3) could reverse the impaired regeneration of ageing or 90% hepatectomy, in the rat. Materials and methods: T3 (20 µg/100 g body weight) was administered to 14‐month‐old rats subjected to 70% PH or to young rats subjected to 90% PH. Cell‐proliferative capacity was determined by bromodeoxyuridine incorporation and microscopy and changes of cell cycle‐related proteins were analysed by Western blot analysis. Results: Treatment of old intact rats with T3 increased cyclin D1 expression that was followed by an enhanced proliferative response, the labelling index (LI), being 7.8% versus 1.3% of controls. T3 given before 70% PH stimulated regenerative response (LI was 10.8% versus 2.28%), and expression of cyclin D1 and proliferating cell nuclear antigen (PCNA) 24 h after PH. Pre‐treatment with T3 also improved the regenerative response of the liver after 90% hepatectomy (LI was 27.9% versus 14.2%). Conclusions: These findings show in principle that mitogen‐induced hyperplasia could be applied to human therapy in patients with reduced regenerative capacity or massive loss of hepatocytes.
Allergenicity and cross‐reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach
Authors: Luengo O. , Mollá R. , Gámez C. , Cardona V. , López E. , Sastre B. , Waisel Y. , Belmonte J. , Cadahia Á. , Lahoz C. , Del Pozo V. 
Published: April 2008

SummaryBackground
The genus Senecio is the largest genus of the family Asteraceae (Compositae).The allergenicity of Senecio has not been assessed previously.Objective
The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance.Methods
Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one‐dimensional electrophoresis (SDS‐PAGE) and two‐dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross‐reactivity with other pollen.Results
Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human‐allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two‐dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross‐reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies.Conclusion
S. jacobea constitute a newly discovered allergenic source. It shows cross‐reactivity with other members of the Asteraceae plant family as well as with P. judaica.
Cyclooxygenase‐2 gene polymorphisms in an Australian population: association of the −1195G > A promoter polymorphism with mild asthma
Authors: Shi J. , Misso N. L. , Kedda M.‐A. , Horn J. , Welch M. D. , Duffy D. L. , Williams C. , Thompson P. J. 
Published: April 2008

SummaryBackground
Cyclooxygenase (COX)‐2 is an inducible enzyme responsible for catalysing the formation of prostaglandins (PGs) in settings of inflammation. Single nucleotide polymorphisms (SNPs) of the COX‐2 gene may influence gene transcription and PG production in the asthmatic airway.Objective
To evaluate the frequencies of COX‐2 SNPs in an Australian Caucasian population, and determine potential associations between common COX‐2 promoter SNPs and asthma, asthma severity and aspirin‐intolerant asthma (AIA).Methods
The frequencies of 25 COX‐2 SNPs were determined in a random population (n=176). The SNPs with a minor allele frequency of >10% were then studied in asthmatic (n=663), non‐asthmatic controls (n=513) and AIA subjects (n=58). Genotype, allele and haplotype associations were assessed. Functional assessment of SNPs was performed by transfection into HeLa cells measured using the luciferase dual‐reporter assay system.Results
Eighteen COX‐2 SNPs were not detected, five were rare and two promoter SNPs, −1195G>A (rs689465), and −1290A>G (rs689466), were further studied. The A allele of the −1195 SNP was present at a significantly higher frequency among all asthmatic subjects (P=0.012). Over 60% of the asthmatic individuals were −1195A homozygotes compared with 54.6% of the control subjects (odds ratio, 1.35; 95% CI, 1.06–1.72, P=0.03). After classifying for severity, the mild asthmatics represented 64.6% of −1195AA individuals, the highest of all the asthma groups compared with 54.6% of the control subjects (odds ratio, 1.5; 95% CI, 1.12–2.02, P=0.02). The −1290A/−1195G/−765G haplotype was associated with a reduced incidence of asthma (odds ratio, 0.76; 95% CI, 0.61–0.95, P=0.017).Conclusion
The −1195G>A polymorphism appears to be associated with asthma, and in particular with mild asthma.
Secondary hyperparathyroidism in primary osteoporosis and osteopenia: optimizing calcium and vitamin D intakes to levels recommended by expert panels may not be sufficient for correction
Authors: Yendt Edmund R. , Kovacs Katherine A. , Jones Glenville 
Published: April 2008

SummaryObjective  To compare biochemical variables, renal function and calcium and vitamin D intakes in euparathyroid and hyperparathyroid patients with primary osteoporosis and osteopenia and describe the measures necessary to normalize serum PTH in the patients with secondary hyperparathyroidism.Design and patients  We reviewed the charts of normocalcemic patients with primary osteoporosis and osteopenia first seen during the years 1991–2003 and identified 75 with elevated serum PTH levels at baseline. These patients were compared to all the 143 euparathyroid patients first seen in 1998 and 1999. Patients were restudied after 1 year and we attempted to follow patients with secondary hyperparathyroidism until PTH levels became normal.Measurements  At baseline serum PTH, ionized calcium, inorganic phosphate, alkaline phosphatase, creatinine, a complete blood count and serum 25 hydroxy vitamin D were measured in the early morning fasting state. These tests were repeated at follow up.Results  In one‐third of the hyperparathyroid patients, the standard baseline treatment failed to correct the secondary hyperparathyroidism necessitating extraordinary measures including unusually large doses of vitamin D (i.e. 50 000 IU vitamin D2 twice weekly) or the substitution of calcium citrate for calcium carbonate as a calcium supplement.Conclusion  Large doses of vitamin D are frequently necessary to suppress secondary hyperparathyroidism in patients with primary osteoporosis and osteopenia. This suggests that vitamin D metabolism may be altered in some of these patients.