bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cloning, expression and activity ofcry1Iagene fromBacillus thuringiensisisolate
Authors: Li-Ming Dou , Lan-Lan Han , Jie Zhang , Kang-lai He , Kui-Jun Zhao , Da-Fang Huang , Fu-Ping Song 
Published: June 2008

AbstractA full-lengthcry1Iagene fragment, obtained by polymerase chain reaction (PCR) amplification with a pair of primers designed according tocry1Ia-type gene sequences and DNA fromBacillus thuringiensisas template, was introduced into the expression vector pET-21b and transformed intoEscherichia coliBL21(DE3). The molecular weight of the induced expression product was 81.2 kDa. The amino acid sequence of Cry1Ia was very different from sequences of 12 known Cry1Ia-type proteins. This gene was designated ascry1Ia8by the InternationalB. thuringiensisInsecticidal Protein Nomenclature Committee (accession number AF373207). The bioassay results indicated that Cry1Ia toxin protein showed distinct insecticidal activity againstOstrinia furnacalisandPlutella xylostellawith LC50of 0.268 μg/g and 2.227 μg/ml, respectively. It also had insecticidal activity againstLeguminivora glycinivorella, but not againstPyrrhalta aenescens. The novelcry1Ia8gene will be an important resource in constructing genetically engineered bacteria and transgenic plants for biocontrol of insect pests. It is also available for screening gene stacks to delay pest resistance.
Identification of Porcine haemagglutinating encephalomyelitis virus receptor in PK cell membranes
Authors: Hui-Jun Lu , Wen-Qi He , De-Guang Song , Li-Guo Liu , Ling-Zhu Chang , Zhi-Ping Li , Ke-Yan Chen , Feng Gao 
Published: June 2008

AbstractTo identify Porcine haemagglutinating encephalomyelitis virus (HEV) 67N receptor in porcine kidney (PK) cell membranes, the S1 protein of HEV was expressed in Pichia pastoris and purified by Ni2+ affinity chromatograph. Polyclonal antibodies to HEV were prepared by immunizing rabbits by injecting the purified S1 protein four times. After SDS–polyacrylamide gel electrophoresis (SDS–PAGE), the PK cell membrane proteins were transferred on to nitrocellulose membrane. A virus overlay protein binding assay (VOPBA) was performed using the recombinant S1 protein to identify the protein binding receptor, HEV-S1. The result showed that HEV-S1 protein bound to one band (about 90 kDa) in PK cell membranes. This result is very important for the study of the pathogenic mechanism of HEV.
Effects of donor sex and different cell treatments on development of yak–bovine interspecies nuclear transfer embryos
Authors: Ying Liu , Shi-En Zhu , Rong Li , Li-Li Wang , Hai-Ping Wang , Fang-Rong Ding , Jing Li , Song Li , Yun-Ping Dai 
Published: June 2008

AbstractThe purpose of this study was to evaluate the effects of donor sex, treatments of cell cycle synchronization and donor nuclei obtained from fresh or frozen–thawed conditions on developmental competence of yak–bovine interspecies nuclear transfer embryos. Bovine (Bos taurus) oocytes were used as recipients and yak (Bos grunniens) ear fibroblast cells were used as donors. Results indicated that the development rate of male blastocysts was higher than that of female (56.6% versus 39.5%, P<0.05), whereas cleavage and total cell number showed no difference between the two groups. No significant difference was observed in the development and quality of blastocysts with donor cells treated by serum starvation or contact inhibition, and there was no significant difference in embryo development with fresh or frozen–thawed donor cells, whereas the cleavage rate in the group of frozen–thawed cells was significantly lower than that of the fresh cell group (54.5% versus 78.2%, P<0.05). The results demonstrated that donor sex could impact the developmental competence of yak–bovine interspecies nuclear transfer embryos, whereas different treatments of cell cycle synchronization and freezing had little influence.
VLDL exits from the endoplasmic reticulum in a specialized vesicle, the VLDL transport vesicle, in rat primary hepatocytes
Authors: Siddiqi Shadab A. 
Published: June 2008

The movement of VLDL [very-LDL (low-density lipoprotein)] from the ER (endoplasmic reticulum) to the Golgi is required for its eventual secretion from hepatocytes and represents a potential target in controlling elevated concentrations of its metabolite LDL, the major determinant of atherosclerosis. To study this process, an in vitro ER-budding assay was developed to examine the generation of the VTV (VLDL transport vesicle) and PTV (protein transport vesicles) using ER isolated from [14C]TAG (triacylglycerol) and [3H]protein-labelled primary rat hepatocytes. VTVs do not contain albumin, as determined by immunoblots. VTVs were distributed in light-density fractions, whereas PTVs were mainly in the mid-portion of the sucrose gradient. Electron microscopy revealed that VTVs were larger (∼100–120 nm) in size than PTVs (∼55–70 nm). ER from 0.4 mM OA (oleic acid)-treated hepatocytes budded VTVs of a lighter density as compared with VTVs budded from ER of 0.1 mM or 0.004 mM OA-treated hepatocytes. The generation of VTVs from rat hepatic ER required cytosol, ATP, Sar1 (a GTPase) and incubation at 37 °C. Proteinase K treatment did not degrade the VTV cargo protein, apoB100 (apolipoprotein 100), indicating that VTVs were sealed. Immunoblots showed that VTV concentrated apoB100, Sar1 and rSec22b, and excluded albumin and calnexin. VTVs were shown to fuse with cis-Golgi and delivered their cargo to the Golgi lumen, as determined by in vitro fusion, and acquired endoglycosidase H resistance. These results suggest that a new ER-derived transport vesicle (VTV) has been identified and characterized which transports nascent VLDL from the hepatic ER to the Golgi.
Translational regulation of lipoprotein lipase in adipocytes: depletion of cellular protein kinase Cα activates binding of the C subunit of protein kinase A to the 3′-untranslated region of the lipoprotein lipase mRNA
Authors: Unal Resat , Pokrovskaya Irina , Tripathi Preeti , Monia Brett P. , Kern Philip A. , Ranganathan Gouri 
Published: June 2008

Adipose LPL (lipoprotein lipase) plays an important role in regulating plasma triacylglycerols and lipid metabolism. We have previously demonstrated that PKCα (protein kinase Cα) depletion inhibits LPL translation in 3T3-F442A adipocytes. Using in vitro translation experiments, the minimum essential region on the 3′UTR (3′-untranslated region) of LPL mRNA required for the inhibition of translation was identified as the proximal 39 nt. These results were confirmed by RNase protection analysis using cytoplasmic proteins isolated from the adipocytes treated with PKCα antisense oligomers and the LPL 3′UTR transcript (LPL 3′UTR nt: 1512–1640). The protein components involved in this RNA-binding interaction from PKCα depletion were passed through an affinity column containing a sequence of the LPL 3′UTR and, after Western blotting, the RNA-binding proteins were identified as the catalytic and the regulatory subunits of PKA (protein kinase A), Cα and RIIβ, and AKAP (A-kinase-anchoring protein) 121. This RNA inhibitory complex consisted of the same RNA-binding proteins that have been identified previously as mediators of LPL translational inhibition by PKA activation, suggesting that PKCα depletion inhibits LPL translation through PKA activation. In additional experiments, PKC depletion by prolonged PMA treatment or PKCα antisense oligomers resulted in an increase in PKA activity in 3T3-F442A adipocytes, comparable with PKA activation with adrenaline (epinephrine) treatment. These results demonstrate that LPL translational inhibition occurs through an RNA-binding complex involving PKA subunits and AKAP121, and this complex can be activated either through traditional PKA activation methods or through the depletion of PKCα.
Catecholaminergic polymorphic ventricular tachycardia-related mutations R33Q and L167H alter calcium sensitivity of human cardiac calsequestrin
Authors: Valle Giorgia , Galla Daniela , Nori Alessandra , Priori Silvia G. , Gyorke Sandor , de Filippis Vincenzo , Volpe Pompeo 
Published: June 2008

Two missense mutations, R33Q and L167H, of hCASQ2 (human cardiac calsequestrin), a protein segregated to the lumen of the sarcoplasmic reticulum, are linked to the autosomal recessive form of CPVT (catecholaminergic polymorphic ventricular tachycardia). The effects of these mutations on the conformational, stability and Ca2+ sensitivity properties of hCASQ2, were investigated. Recombinant WT (wild-type) and mutant CASQ2s were purified to homogeneity and characterized by spectroscopic (CD and fluorescence) and biochemical (size-exclusion chromatography and limited proteolysis) methods at 500 and 100 mM KCl, with or without Ca2+ at a physiological intraluminal concentration of 1 mM; Ca2+-induced polymerization properties were studied by turbidimetry. In the absence of Ca2+, mutations did not alter the conformation of monomeric CASQ2. For L167H only, at 100 mM KCl, emission fluorescence changes suggested tertiary structure alterations. Limited proteolysis showed that amino acid substitutions enhanced the conformational flexibility of CASQ2 mutants, which became more susceptible to tryptic cleavage, in the order L167H&gt;R33Q&gt;WT. Ca2+ at a concentration of 1 mM amplified such differences: Ca2+ stabilized WT CASQ2 against urea denaturation and tryptic cleavage, whereas this effect was reduced in R33Q and absent in L167H. Increasing [Ca2+] induced polymerization and precipitation of R33Q, but not that of L167H, which was insensitive to Ca2+. Based on CASQ2 models, we propose that the Arg33→Gln exchange made the Ca2+-dependent formation of front-to-front dimers more difficult, whereas the Leu167→His replacement almost completely inhibited back-to-back dimer interactions. Initial molecular events of CPVT pathogenesis begin to unveil and appear to be different depending upon the specific CASQ2 mutation.
Insulin action on glucose transporters through molecular switches, tracks and tethers
Authors: Zaid Hilal , Antonescu Costin N. , Randhawa Varinder K. , Klip Amira 
Published: June 2008

Glucose entry into muscle cells is precisely regulated by insulin, through recruitment of GLUT4 (glucose transporter-4) to the membrane of muscle and fat cells. Work done over more than two decades has contributed to mapping the insulin signalling and GLUT4 vesicle trafficking events underpinning this response. In spite of this intensive scientific research, there are outstanding questions that continue to challenge us today. The present review summarizes the knowledge in the field, with emphasis on the latest breakthroughs in insulin signalling at the level of AS160 (Akt substrate of 160 kDa), TBC1D1 (tre-2/USP6, BUB2, cdc16 domain family member 1) and their target Rab proteins; in vesicle trafficking at the level of vesicle mobilization, tethering, docking and fusion with the membrane; and in the participation of the cytoskeleton to achieve optimal temporal and spatial location of insulin-derived signals and GLUT4 vesicles.
Registration of Down syndrome in the Medical Birth Registry of Norway: Validity and time trends
Authors: MELVE KARI KLUNGSØYR , LIE ROLV TERJE , SKJAERVEN ROLV , VAN DER HAGEN CARL BIRGER , GRADEK GYRI AASLAND , JONSRUD CHRISTOPHER , BRAATHEN GEIR J. , IRGENS LORENTZ M. 
Published: June 2008

AbstractObjective. To validate Down syndrome registration in the Medical Birth Registry of Norway (MBRN), 2001–2005, and study time trends and geographical differences in Down syndrome prevalence,1967–2005. Design/setting. Population‐based cohort study, Norway. Population. 2.3 million pregnancies and births registered in the MBRN, 1967–2005. Methods. We linked data from the MBRN during 2001–2005 with data from Norway's four laboratories of medical genetics. We calculated sensitivity and positive predictive values (PPV) of the MBRN registration overall, and by background variables. Prevalence rates from 1967 to 2005, overall and regional, were presented graphically as smoothed lowess estimates, crude and standardized for maternal age. Time trends were evaluated, adjusting for maternal age by logistic regression. Main outcome measures. Sensitivity, PPV, and prevalence rates. Results. Five hundred and seventy‐six verified cases of Down syndrome gave a prevalence of 2.0 per 1,000 among 288,213 births and terminations in the MBRN during 2001–2005. Of verified cases, 470 (81.6%) were registered with Down syndrome in the MBRN, while 470 (90.2%) of 521 MBRN‐registered cases were verified. Sensitivity was higher in the Northern (93.1%; p = 0.005) and Middle (90.6%; p = 0.02) region relative the Southern (76.3%), higher for mothers ≥35 years (92.9%) than younger ones (86.1%; p = 0.01), and higher for live births (88.8%) relative stillbirths (55.6%; p<0.001). When adjusting for maternal age, there were no significant time trends in prevalence rates from 1967 to 2005. Regional differences over time were found, probably representing reporting differences. Conclusions. Validity of registration in the MBRN was satisfactory during 2001–2005. Increasing prevalence rates over time were explained by increasing maternal age.
MMP‐2, TIMP‐1, E‐cadherin, and β‐catenin expression in endometrial serous carcinoma compared with low‐grade endometrial endometrioid carcinoma and proliferative endometrium
Authors: SHACO‐LEVY RUTHY , SHARABI SHALOM , PIURA BENJAMIN , SION‐VARDY NETTA 
Published: June 2008

AbstractObjective. To investigate the expression of MMP‐2, TIMP‐1, E‐cadherin and β‐catenin in endometrial serous carcinoma (ESC), low‐grade endometrial endometrioid carcinoma (EEC), and proliferative endometrium. Methods. We performed an immunohistochemical study on 14 cases of ESC, 15 cases of low‐grade EEC, and 10 cases of proliferative endometrium. Results. Compared with low‐grade EEC, ESC showed significantly increased MMP‐2 and TIMP‐1 expression, as well as decreased membranous β‐catenin staining. E‐cadherin expression was significantly lower in ESC and EEC as compared with proliferative endometrium. Conclusions. We suggest that MMP‐2 and TIMP‐1 expression and loss of β‐catenin have a role in the pathogenesis and progression of ESC. Decreased E‐cadherin may have an important role in the development of both ESC and EEC. Furthermore, the dissimilarities in MMP‐2, TIMP‐1, E‐cadherin and β‐catenin expressions in ESC compared with EEC may be responsible, along with other factors, for their different biological behavior.
Prevalence and correlates of jugular injections among injection drug users
Authors: HODA ZIA , KERR THOMAS , LI KATHY , MONTANER JULIO S. G. , WOOD EVAN 
Published: June 2008

AbstractBackground. Jugular injection of drugs has been reported, although little is known about the prevalence of and risk factors associated with this behaviour. We evaluated factors associated with jugular injection among a cohort of injection drug users (IDU) in Vancouver, Canada. Methods. We used univariate statistics and logistic regression to examine factors associated with jugular injection among participants in the Vancouver Injecting Drug Users Study (VIDUS), a large prospective cohort study of IDU recruited through snowball sampling methods in Vancouver, Canada. Findings. Between December 2004 and November 2005, 780 IDU were followed up as part of VIDUS and 198 (25%) reported jugular injection in the previous 6 months. In multivariate analyses, factors associated independently with jugular injection included: being of the female gender [adjusted odds ratio (aOR) = 1.72, 95% confidence interval (CI): 1.14–2.59; p = 0.010], daily heroin use (aOR = 2.89, 95% CI: 1.93–4.34; p < 0.001), daily cocaine use (aOR = 1.76, 95% CI: 1.12–2.76; p = 0.014], requiring help injecting (aOR = 4.44, 95% CI: 2.64–7.46; p < 0.001), and involvement in the sex‐trade (aOR = 2.71, 95% CI: 1.6–4.55; p < 0.001). Interpretation. Reporting a history of jugular injecting was alarmingly high in the cohort and was associated with several identifiable demographic and drug‐using characteristics. Given previous reports demonstrating the risk of infection and vascular trauma due to this behaviour, these populations should be considered seriously as a target for safer injecting education.
A cross‐sectional survey of young people attending a music festival: associations between drug use and musical preference
Authors: LIM MEGAN S. C. , HELLARD MARGARET E. , HOCKING JANE S. , AITKEN CAMPBELL K. 
Published: June 2008

AbstractIntroduction and Aims. Drug use is becoming normalised among young Australian people involved in music sub‐cultures. We aimed to determine prevalences of illicit drug use in this population and associations between preferences for different music genres and recent use of particular illicit drugs. Design and Methods. A cross‐sectional questionnaire of young people (aged 16–29 years) attending a music festival. Results. Of 939 respondents, 46% had used illicit drugs (principally cannabis) in the past month, a significantly higher proportion than among respondents to the 2004 National Drug Strategy Household Survey (18%). Participants who favoured dance/house or rap music were more likely to have used illicit drugs recently than the remainder of the sample, while those who favoured pop or alternative music were less likely to have used drugs in the past month. Discussion and Conclusions. These data suggest that music festival attendees use illicit drugs more commonly than their age‐matched cohort in the general community, and that music festivals venues (particularly those that cater for dance/house and rap) would be appropriate places for interventions to promote safer drug use.
Patients' help‐seeking behaviours for health problems associated with methadone and buprenorphine treatment
Authors: WINSTOCK ADAM R. , LEA TOBY , SHERIDAN JANIE 
Published: June 2008

AbstractIntroduction and Aims. Clients in opioid substitution therapy often have considerable unmet health‐care needs. The current study aimed to explore health problems related to opioid substitution therapy among clients on methadone and buprenorphine treatment. Design and Methods. A self‐complete, cross‐sectional survey conducted among 508 patients receiving methadone and buprenorphine treatment at community pharmacies in New South Wales (NSW), Australia. Results. The most common problems for which participants had ever sought help were dental (29.9%), constipation (25.0%) and headache (24.0%). The most common problems for which participants would currently like help were dental (41.1%), sweating (26.4%) and reduced sexual enjoyment (24.2%). There were no significant differences between those currently on methadone and those currently on buprenorphine for any of the health problems explored, nor differences for gender or treatment duration. Participants on methadone doses 100 mg or above were significantly more likely to want help currently for sedation. Discussion and Conclusions. The considerable unmet health care needs among participants in this study suggest that treatment providers should consider improving the detection and response to common health problems related to opioid substitution therapy.
Alcohol and suicide in eastern Europe
Authors: LANDBERG JONAS 
Published: June 2008

AbstractIntroduction and Aims. The aim of this paper was to estimate how suicide rates in seven eastern European countries are affected by changes in population drinking and to put the results into a comparative perspective. Design and Methods. The analysis included data on annual suicide mortality rates and per capita consumption for the post‐war period from: Russia, Belarus, Poland, Hungary, Bulgaria, the former Czechoslovakia and the former German Democratic Republic (GDR). Overall and gender‐specific models were estimated using the Box–Jenkins technique for time–series analysis. The estimates were pooled into two groups, i.e. spirits countries (Russia, Belarus and Poland) and non‐spirits countries (Hungary, Bulgaria, former Czechoslovakia and former GDR). Results. All countries obtained positive alcohol effect estimates. The effects on the overall population were largest in the spirits countries, where a 1‐litre increase in per capita consumption was associated with an increase in overall suicide rates of 5.7–7.5%. The effects were somewhat smaller in the non‐spirits countries, 2.7–4.7%. The estimates for males were larger, but showed the same national variations as the overall population estimates. The female estimates were generally smaller than for men and did not differ between the two country groups. Discussion and Conclusions. The results suggest that per capita consumption matters for suicide mortality in these eastern European countries, but that the strength of the relationship is contingent upon the drinking culture, so that it tends to be stronger in countries with detrimental drinking patterns.
Reactivity of Brain Parenchymal Arterioles after Ischemia and Reperfusion
Authors: CIPOLLA MARILYN J. , BULLINGER LISA V. 
Published: June 2008

ABSTRACTObjective: We investigated the effect of ischemia and reperfusion on the vasoactive function of penetrating brain parenchymal arterioles under pressurized conditions.Methods: Parenchymal arterioles (<50 μm in diameter) from within the middle cerebral artery territory were dissected from male Wistar rats that were either nonischemic control (n = 16) or ischemic for one hour and reperfused for 24 hours (n = 16) by temporary filament occlusion of the middle cerebral artery. Arterioles were mounted on glass cannulas within an arteriograph chamber that allowed for the measurement of lumen diameter and control over intravascular pressure.Results: After one hour of equilibration at 10 mmHg, spontaneous myogenic tone developed in both groups of animals, constricting control arterioles from 69 ± 9 to 49 ± 11 μm (29.5 ± 10.2%) and ischemic arterioles from 66 ± 9 to 45 ± 11 μm (33.1 ± 14.1%); p > 0.05. Contraction to the nitric oxide synthase inhibitor nitro‐L‐arginine (10–4M) was significantly diminished in ischemic arterioles, constricting only 3.2 ± 3.3 vs. 15.6 ± 12.5% in control arterioles (p = 0.017). Both groups dilated to nifedipine; however, the response was significantly diminished after ischemia. The EC50 for nifedipine in control arterioles was 3.54 ± 0.11 vs. 9.90 ± 0.71 nM for ischemic arterioles (p = 0.024).Conclusions: These findings demonstrate that functional changes occur in brain parenchymal arterioles after ischemia and reperfusion, a result that may significantly influence stroke outcome by altering blood flow to an ischemic region.