bluuebunny/crossref_metadata_2025_split huggingface dataset:
In situ gelling and mucoadhesive polymer vehicles for controlled intranasal delivery of plasmid DNA
Authors: Park Jeong‐Sook , Oh Yu‐Kyoung , Yoon Ho , Kim Jung Mogg , Kim Chong‐Kook 
Published: August 2002

AbstractNasal administration of plasmid DNA is emerging as a new route of delivery for therapeutic genes and DNA vaccines. To improve the intranasal absorption of plasmid DNA, we designed delivery systems composed of in situ gelling and mucoadhesive polymers. Poloxamers (Pol) were used to provide in situ gelling property. Polycarbophil (PC) or polyethylene oxide (PEO) was used as mucoadhesive polymers. The gelation temperatures of the formulations slightly decreased by the mucoadhesive polymers, but not by plasmid DNA. The in vitro release of plasmid DNA from the gels followed Fickian diffusion. The absorption of plasmid DNA varied with the contents and type of mucoadhesive polymers. Of vehicles, Pol/PC 0.2% showed the highest absorption with an area under the curve value 11‐fold higher than saline, the conventional vehicle. The nasal retention of plasmid DNA was highly prolonged by mucoadhesive polymers. At 3 h postdose, the nasal tissue levels of plasmid DNA given in Pol/PC and Pol/PEO 0.8% were 10‐ and 40‐fold higher relative to saline. The histopathology of nasal tissues was not altered after repeated dosing over 2 weeks. The mRNA expression of plasmid DNA delivered by Pol or Pol/PEO 0.4% was observed in the nasal tissues. These results indicate that the nasal absorption of plasmid DNA can be effectively and safely enhanced by using in situ gelling and mucoadhesive polymer‐based vehicles. © 2001 Wiley Periodicals, Inc. J Biomed Mater Res 59: 144–151, 2002
Paired comparison of apheresis platelet function after storage in two containers
Authors: Zingsem Jürgen , Glaser Anke , Zimmermann Robert , Weisbach Volker , Kalb Rainer , Ruf Andreas , Eckstein Reinhold 
Published: August 2002

AbstractPlatelet quality after storage strongly depends on the pre‐storage quality as well as on the storage conditions determined by the storage container. In this paired study, we evaluated two different containers (MedSep CLX™ and Delmed DPL‐110). The Fresenius AS104 cell separator was used to prepare 17 platelet concentrates that were split and distributed into the containers to be compared. Cell counts, blood gas analysis, morphological scores, glucose and lactate levels, platelet activation, and platelet aggregation were measured before splitting at the day of preparation and after storage at day 3 and day 5. At day 3, there was no significant difference between the two bags apart from increased lactate and decreased pCO2 concentrations in the CLX™ bags. At day 5 there were significantly higher lactate concentrations, pO2 levels, and aggregation after stimulation in the CLX™ group, while the glucose and pCO2 concentrations were significantly lower in these platelet concentrates as compared to the DPL‐110 group. However, these parameters did not influence the functional parameters tested. While the platelet quality decreased during storage in all bags, the functional changes were nearly identical in both bags tested. We conclude that both bags are equivalent for 5‐day storage of platelet concentrates. J. Clin. Apheresis. 16:10–14, 2001. © 2001 Wiley‐Liss, Inc.
Cortisol regulates the expression of Notch in osteoblasts
Authors: Pereira Rosa M.R. , Delany Anne M. , Durant Deena , Canalis Ernesto 
Published: August 2002

AbstractGlucocorticoids have important effects on osteoblastic replication, differentiation, and function, and the Notch family of receptors is considered to play a role in osteoblastic cell differentiation. We postulated that cortisol could regulate Notch and Notch ligand expression in osteoblastic cells, providing an additional mechanism by which glucocorticoids could regulate osteoblastic differentiation. We examined the expression and regulation of Notch1, 2, 3, and 4 and their ligands Jagged 1 and 2 and Delta 1 and 3 by cortisol in cultures of osteoblastic MC3T3‐E1 cells. Cortisol caused a time‐dependent increase in Notch1 and 2 mRNA levels in MC3T3 cells. Notch3 and 4 were not detected in the presence or absence of cortisol. MC3T3 cells expressed Delta 1 and Jagged 1 but not Jagged 2 or Delta 3 mRNAs, and cortisol did not have a substantial effect on the expression of any of these ligands. Cortisol increased the rate of Notch1 and 2 transcription and, in transcriptionally arrested cells, did not modify the decay of the transcripts, indicating a transcriptional level of control. In conclusion, cortisol stimulates Notch1 and 2 transcription in osteoblasts. Since Notch signaling appears to play a negative role in osteoblastic differentiation, its increased expression could be relevant to the actions of cortisol in bone. J. Cell. Biochem. 85: 252–258, 2002. © 2002 Wiley‐Liss, Inc.
Human papillomavirus‐16 E7 protein inhibits the DNA interaction of the TATA binding transcription factor
Authors: Maldonado Edio , Cabrejos María Eugenia , Banks Lawrence , Allende Jorge E. 
Published: August 2002

AbstractPrevious studies have shown that the HPV‐16 E7 protein interacts with TBP. This interaction was found to take place through residues in the carboxy terminal half of E7, mutation of which resulted in weaker transforming activity. In addition, binding of E7 to TBP was found to be increased following protein kinase CK2 (casein kinase II) phosphorylation of E7, and mutation of this CK2 site also reduces E7's transforming activity. To date, however, there is no information on the effects of E7 upon TBP function. In order to address this we have performed a series of assays to investigate the effects of E7 upon the ability of human and S. pombe TBP to bind DNA. We show that HPV‐16 E7 is indeed a potent inhibitor of TBP DNA binding activity. Further, this activity of E7 is increased following CK2 phosphorylation of E7, consistent with it having an increased affinity for TBP. Finally, a mutant E7 protein defective in its ability to bind TBP, has no effect upon TBP binding to DNA. These results demonstrate that one consequence of the E7–TBP interaction is abolition of TBP DNA binding activity, and may provide an explanation for the transcriptional inhibitory effects of E7. J. Cell. Biochem. 85: 663–669, 2002. © 2002 Wiley‐Liss, Inc.
Characterization of the upstream mouse Cbfa1/Runx2 promoter*
Authors: Xiao Z. S. , Liu Shi‐Guang , Hinson T. K. , Quarles L. D. 
Published: August 2002

AbstractCbfa1 (or Runx2/AML‐3/PEPB2α) is a transcriptional activator of osteoblastic differentiation. To investigate the regulation of Cbfa1 expression, we isolated and characterized a portion of the 5′‐flanking region of the Cbfa1 gene containing its “bone‐related” or P1 promoter and exon 1. We identified additional coding sequence in exon 1 and splice donor sites that potentially give rise to a novel Cbfa1 isoform containing an 18 amino acid insert. In addition, primer extension mapping identified in the Cbfa1 promoter a minor mRNA start site located ∼0.8 kb 5′ upstream of the ATG encoding the MASN/p57 isoform and ∼0.4 kb upstream of the previously reported start site. A luciferase reporter construct containing 1.4 kb of the mouse Cbfa1 promoter was analyzed in Ros 17/2.8 and MC3T3‐E1 osteoblast cell lines that express high levels of Cbfa1 transcripts. The activity of this construct was also examined in non‐osteoblastic Cos‐7 and NIH3T3 cells that do not express Cbfa1 and mesenchymal‐derived cell lines, including CH3T101/2, C2C12, and L929 cells, that express low levels of mature Cbfa1 transcripts. The 1.4 kb 5′ flanking sequence of the Cbfa1 gene directed high levels of transcriptional activity in Ros 17/2.8 and MC3T3‐E1 osteoblasts compared to non‐osteoblasts Cos‐7 cells, but this construct also exhibited high levels of expression in C310T1/2, L929, and C2C12 cells as well as NIH3T3 cells. In addition, Cbfa1 mRNA expression, but not the activity of the Cbfa1 promoter, was upregulated in a dose‐dependent manner in pluripotent mesenchymal C2C12 by bone morphogenetic protein‐2 (BMP‐2). These data indicate that Cbfa1 is expressed in osteogenic as well as non‐osteogenic cells and that the regulation of Cbfa1 expression is complex, possibly involving both transcriptional and post‐transcriptional mechanisms. Additional studies are needed to further characterize important regulatory elements and to identify additional regions of the promoter and/or post‐transcriptional events responsible for the cell‐type restricted regulation of Cbfa1 expression. J. Cell. Biochem. 82: 647–659, 2001. © 2001 Wiley‐Liss, Inc.
Characterisation of the temporal sequence of osteoblast gene expression during estrogen‐induced osteogenesis in female mice
Authors: Plant Andrea , Tobias Jonathan H. 
Published: August 2002

AbstractOsteoblast differentiation under in vitro conditions is associated with increased expression of non‐collagenous bone proteins including osteocalcin, osteopontin, and osteonectin, the exact function of which remain poorly understood. To determine whether these proteins play an important role in the formation of mineralised bone matrix by osteoblasts in vivo, we analysed the time‐course of their expression during estrogen‐induced osteogenesis in female mice, and compared this with the formation of new cancellous bone. Female mice were sacrificed prior to or following treatment with 17β‐estradiol for up to 32 days (500 μg/animal/week). Total RNA was extracted from femurs, and changes in expression of genes for a range of osteoblast‐derived proteins assessed by Northern blot analysis. In parallel experiments, the time course of cancellous bone formation was determined by measuring bone mineral density (BMD) of the distal femur. Estrogen led to a rapid increase in BMD, which reached significance by Day 16. This was preceded by three‐fold increases in expression of alkaline phosphatase (ALP) and type I collagen (COL I) at Days 8 and 12 respectively. In contrast, osteocalcin, osteopontin, and osteonectin expression showed no change during this initial period, although modest increases were observed at later times (i.e., Days 20 and 24). Our results suggest that osteocalcin, osteopontin, and osteonectin are not involved in the initial phase of the osteogenic response to estrogen, suggesting that these non‐collagenous bone proteins do not play a direct role in the formation of mineralised bone matrix by osteoblasts in vivo. J. Cell. Biochem. 82: 683–691, 2001. © 2001 Wiley‐Liss, Inc.
Calculation of electronic g‐tensors for transition metal complexes using hybrid density functionals and atomic meanfield spin‐orbit operators
Authors: Kaupp Martin , Reviakine Roman , Malkina Olga L. , Arbuznikov Alexei , Schimmelpfennig Bernd , Malkin Vladimir G. 
Published: August 2002

AbstractWe report the first implementation of the calculation of electronic g‐tensors by density functional methods with hybrid functionals. Spin‐orbit coupling is treated by the atomic meanfield approximation. g‐Tensors for a set of small main group radicals and for a series of ten 3d and two 4d transition metal complexes have been compared using the local density approximation (VWN functional), the generalized gradient approximation (BP86 functional), as well as B3‐type (B3PW91) and BH‐type (BHPW91) hybrid functionals. For main group radicals, the effect of exact‐exchange mixing is small. In contrast, significant differences between the various functionals arise for transition metal complexes. As has been shown previously, local and in particular gradient‐corrected functionals tend to underestimate the “paramagnetic” contributions to the g‐tensors in these cases and thereby recover only about 40–50% of the range of experimental g‐tensor components. This is improved to ca. 60% by the B3PW91 functional, which also gives slightly reduced standard deviations. The range increases to almost 100% using the half‐and‐half functional BHPW91. However, the quality of the correlation with experimental data worsens due to a significant overestimate of some intermediate g‐tensor values. The worse performance of the BHPW91 functional in these cases is accompanied by spin contamination. Although none of the functionals tested thus appears to be ideal for the treatment of electronic g‐tensors in transition metal complexes, the B3PW91 hybrid functional exhibited the overall most satisfactory performance. Apart from the validation of hybrid functionals, some aspects in the treatment of spin‐orbit contributions to the g‐tensor are discussed. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 794–803, 2002
Parameterization of OPLS–AA force field for the conformational analysis of macrocyclic polyketides
Authors: Kahn Kalju , Bruice Thomas C. 
Published: August 2002

AbstractThe parameters for the OPLS–AA potential energy function have been extended to include some functional groups that are present in macrocyclic polyketides. Existing OPLS–AA torsional parameters for alkanes, alcohols, ethers, hemiacetals, esters, and ketoamides were improved based on MP2/aug‐cc‐pVTZ and MP2/aug‐cc‐pVDZ calculations. Nonbonded parameters for the sp3 carbon and oxygen atoms were refined using Monte Carlo simulations of bulk liquids. The resulting force field predicts conformer energies and torsional barriers of alkanes, alcohols, ethers, and hemiacetals with an overall RMS deviation of 0.40 kcal/mol as compared to reference data. Densities of 19 bulk liquids are predicted with an average error of 1.1%, and heats of vaporization are reproduced within 2.4% of experimental values. The force field was used to perform conformational analysis of smaller analogs of the macrocyclic polyketide drug FK506. Structures that adopted low‐energy conformations similar to that of bound FK506 were identified. The results show that a linker of four ketide units constitutes the shortest effector domain that allows binding of the ketide drugs to FKBP proteins. It is proposed that the exact chemical makeup of the effector domain has little influence on the conformational preference of tetraketides. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 977–996, 2002
Necessary existence criteria for mandatory representation designs MRD (4, k; ν)
Authors: Grüttmüller Martin , Rees R. S. 
Published: August 2002

AbstractA mandatory representation design MRD (K; ν) is a pairwise balanced design on ν points with block sizes from the set K in which for each k ∈ K there is at least one block in the design of size k. A survey of known results on MRDs has been given in [J Comb Math Comb Comput, to appear] by Rees. MRDs with K = {3, k} have been extensively studied by Mendelsohn and Rees [J Comb Theory Ser A 49 ( 1988), 349–362], Rees [J Stat Plann Inference 86 (2000), 567–594], Rees and Grüttmüller [Congr Numerantium, 147 (2000), 41–51] and Grüttmüller [J Comb Des 8 (2000) 122–131; On the existence of pairwise balanced designs, Ph.D. Thesis, University of Rostock, 1999]. In this paper, we investigate necessary conditions for the existence of MRDs with K = {4, k}. © 2001 John Wiley & Sons, Inc. J Combin Designs 9: 309–333, 2001
Quantitative PCR determination of human cytomegalovirus in blood cells
Authors: Satou J. , Funato T. , Satoh N. , Abe Y. , Ishii K.K. , Sasaki T. , Kaku M. 
Published: August 2002

AbstractWe evaluated a rapid and sensitive method to determine human cytomegalovirus (CMV) DNA levels in blood cells using a quantitative polymerase chain reaction (PCR) technique. This method is based on real‐time detection of PCR using a dual fluorescence‐labeled probe and a sequence detector. Ten copies of CMV DNA were detected, when 1 μg of DNA from blood samples was used with this method, and a good correlation was obtained between increased concentrations of copy numbers calculated and measured copy numbers of CMV DNA (r = 0.999). Forty normal subjects exhibited no copies of CMV DNA. On the other hand, a 6‐month‐old girl tested positive for increased levels 4 weeks after liver transplant. This method is simple, accurate, and sensitive for the quantitative detection of CMV DNA in vivo, indicating possible applications for the diagnosis and monitoring of CMV infection. J. Clin. Lab. Anal. 15:122–126, 2001. © 2001 Wiley‐Liss, Inc.
A cognitive perspective on resistance in psychotherapy
Authors: Newman Cory F. 
Published: August 2002

AbstractThe phenomena that comprise resistance in therapy are described and discussed from a cognitive viewpoint. The cases of Brian, Julie, and Victoria are reviewed to illustrate similar and contrasting manifestations of resistance, and suggestions for further individualized conceptualization and intervention are presented. Emphasis is placed on gaining an accurate, empathic understanding of the client's difficulties in changing. This is coupled with methods that allow the client some autonomy in charting the course of therapy, at the same time reinforcing the importance of the therapist's active involvement in educating and stimulating the client toward greater awareness and adaptability. In order to be maximally effective in encouraging such changes, therapists must be aware of their own tendencies to react adversely to client resistance. It is vital that therapists maintain their professionalism and confidence in persevering toward a better understanding of what will be helpful to their clients. © 2002 John Wiley & Sons, Inc. J Clin Psychol/In Session 58: 165–174, 2002.
Comparing adults in Los Angeles County who have and have not been homeless
Authors: Cousineau Michael R. 
Published: August 2002

AbstractThis study compares the formerly homeless with those who have not been homeless on several characteristics, based on a telephone survey of the general adulate population. The study was conducted in Los Angeles County. Researchers estimate how many and what percentage of adults (aged 18 or older) have been homeless in the past 5 years and the types of places people stayed while they were homeless. An estimated 370,000 adults have experienced homelessness within the past 5 years, 5.7% of the adult population (95% confidence interval [CI] 5.2–6.2). A third were literally homeless (in a shelter, street, or car). Just over half (56%) stayed with a friend or relative while homeless. Nine percent had a mixed experience. Compared to those who were not homeless, the formerly homeless are disproportionately poor, African American, not in the job market, on public assistance, and in poor health. There are few differences when comparing place of birth, citizenship status, or length of residence in Los Angeles County. Yet many homeless have been able to achieve some economic stability. Implications for the development of intervention and prevention programs are discussed. © 2001 John Wiley & Sons, Inc.
Transforming growth factor‐β isoforms differently stimulate proα2 (I) collagen gene expression during wound healing process in transgenic mice
Authors: Kinbara Takuro , Shirasaki Fumiaki , Kawara Shigeru , Inagaki Yutaka , de Crombrugghe Benoit , Takehara Kazuhiko 
Published: August 2002

AbstractThe role of many growth factors and cytokines in the process of wound healing has been intensively investigated in recent two decades. Among them, transforming growth factor‐βs (TGF‐βs) are well known to have a potent stimulatory effect on collagen synthesis as shown in various in vivo experimental systems. In the present study, we examined the effects of various growth factors on the promoter activity of the proα2 (I) collagen gene (COL1A2) during the wound healing process. For this purpose, we utilized transgenic mice harboring the −17 kb promoter sequence of the mouse COL1A2 linked to either a firefly luciferase or a bacterial β‐galactosidase gene. These mice exhibited normal phenotypic expression and the wound healing process was not impaired. Full thickness wounds were made by punch biopsy. We examined the effects of TGF‐β1, ‐β2, ‐β3, basic fibroblast growth factor, platelet‐derived growth factor, and connective tissue growth factor by applying them locally to the open wound every 2 days. Among the growth factors examined, all of the three isoforms of TGF‐β exhibited a more potent stimulatory effect on COL1A2 promoter activity than did other factors. In addition, while TGF‐β1 and ‐β2 significantly increased the number of fibroblasts which were positive for X‐Gal staining, TGF‐β3 treatment did not change the number of β‐galactosidase expressing cells. Accumulation of collagen fibers was observed to the same extent in the mice treated with TGF‐β1 and those with TGF‐β3. These findings suggest that TGF‐β1 and ‐β3 have similar but not identical regulatory mechanisms of COL1A2 expression, and that their pathophysiological roles in wound healing might be different from each other. J. Cell. Physiol. 190: 375–381, 2002. © 2002 Wiley‐Liss, Inc.
Profiling of retinoid mediated gene expression in synchronized human SCC cells using Atlas™ human cDNA expression arrays
Authors: Le Quan , Soprano Dianne Robert , Soprano Kenneth J. 
Published: August 2002

AbstractWhile retinoids have been demonstrated to inhibit growth of many tumor cells, including SCC cells, the molecular mechanism by which retinoids suppress growth has not been elucidated. We previously found that the growth of SCC cells was significantly inhibited by all‐trans‐retinoic acid (all‐trans‐RA) treatment, and this inhibition was dependent on the binding and activation of RARs. These nuclear receptors bind retinoids and alter the rate of transcription of specific genes. To identify targets of the activated RARs which mediate growth inhibition, we growth arrested SCC‐25 cells in G‐0 and examined the effect of all‐trans‐RA on synchronized SCC‐25 cells. All‐trans‐RA inhibited G‐1 progression in quiescent SCC‐25 cells stimulated by FBS. More specifically, we found that the all‐trans‐RA execution point maps to mid/late G‐1, 6 to 10 h after stimulation. Using this synchronized cell system, we examined the expression of cell cycle regulatory genes in quiescent SCC‐25 cells stimulated with FBS and treated with all‐trans‐RA. We found few changes in expression of these genes which could account for all‐trans‐RA inhibition of SCC‐25 cell growth. In order to compare the patterns of expression of a wider selection of genes in all‐trans‐RA treated and non‐treated SCC‐25 cells, we have used expression array technology. We successfully performed expression profiling experiments on the Atlas™ Human cDNA arrays which contain 1176 human genes. We have identified several up‐regulated and several down‐regulated gene expression changes mediated by all‐trans‐RA treatment in synchronized SCC‐25 cells. This novel information will be useful in defining the mechanism by which retinoids suppress the growth of SCC cells. J. Cell. Physiol. 190: 345–355, 2002. © 2002 Wiley‐Liss, Inc.