bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Variations in the Amylose Content of Grains Located at Different Positions in the Rice Panicle and the Effect of Milling
Authors: Zhang Xiao‐Ming , Shi Chun‐Hai , Wu Jian‐Guo , Hisamitus Horiuchi , Katsura Tomita , Feng Shui‐Ying , Bao Gen‐Liang , Ye Sheng‐Hai 
Published: June 2003

AbstractThe results of the presented study indicated that the amylose content (AC) of individual rice kernels varied among grains in the same panicle and even in outer and inner layers of the same grain. The differences among the grains in panicles were 19.1% in the Indica variety “Zhenong 5” and 22.8% in the Japonica variety “Zhehu 9827”. The amylose content of the grains in the upper position or the first branches of panicles and of the early flowering grains or heavier grains was higher than that of others. Amylose content increases by 1.36% in Japonica rice variety “Zhehu 9827” and 1.92% in Indica rice variety “Zhenong 5”, when the degree of milling increases by 10%. The tendency of changes in AC was the same for grains of the Japonica and the Indica rice variety. Furthermore, for analyzing AC the grains in a panicle should carefully be selected. Errors coming from sampling could be decreased by using a multiplicity of grains rather than single‐grain samples.
Isolation and characterization of the TRP1 gene from the yeast Yarrowia lipolytica and multiple gene disruption using a TRP blaster
Authors: Cheon Seon Ah , Han Eun Jung , Kang Hyun Ah , Ogrydziak David M. , Kim Jeong‐Yoon 
Published: June 2003

AbstractThe TRP1 gene encoding N‐(5′‐phosphoribosyl)‐anthranilate isomerase was isolated from the yeast Yarrowia lipolytica, in which only a few genetic marker genes are available. The Y. lipolytica TRP1 gene (YlTRP1) cloned by complementation of Y. lipolytica trp1 mutation was found to be a functional homologue of Saccharomyces cerevisiae TRP1. Since YlTRP1 could be used for counterselection in medium containing 5‐fluoroanthranilic acid (5‐FAA), we constructed TRP blasters that contained YlTRP1 flanked by a direct repeat of a sequence and allowed the recycling of the YlTRP1 marker. Using the TRP blasters the sequential disruption of target genes could be carried out within the same strain of Y. lipolytica. The nucleotide sequence of the YlTRP1 gene has been deposited at GenBank under Accession No. AF420590. Copyright © 2003 John Wiley & Sons, Ltd.
Relationship between G+C content, ORF‐length and mRNA concentration in Saccharomyces cerevisiae
Authors: Marín Antonio , Gallardo Mercedes , Kato Yuki , Shirahige Katsuhiko , Gutiérrez Gabriel , Ohta Kunihiro , Aguilera Andrés 
Published: June 2003

AbstractRNA biogenesis is a tightly‐regulated process. The levels and timing of expression of a gene depends on its particular function. However, gene expression levels may also depend on structural features. Here we describe the analysis of gene expression of 4977 ORFs using DNA microarrays covering the whole genome of three different S. cerevisiae strains, wild‐type and tho2 and thp1 mutants with a general effect on mRNA biogenesis. We show that transcripts from G+C‐rich ORFs accumulate at higher concentrations than those from G+C‐poor ones, in different ORF‐length categories in all strains tested. In addition, we found a negative correlation between ORF length and G+C content. Our results indicate that length and G+C content of a gene have a clear effect on its levels of expression. We discuss the biological relevance of these results, as well as different ways that these structural features could modulate mRNA biogenesis. Copyright © 2003 John Wiley & Sons, Ltd.
Candida albicans TDH3 gene promotes secretion of internal invertase when expressed in Saccharomyces cerevisiae as a glyceraldehyde‐3‐phosphate dehydrogenase‐invertase fusion protein
Authors: Delgado M. Luisa , Gil M. Luisa , Gozalbo Daniel 
Published: June 2003

AbstractWe have checked the ability of the Candida albicans GAPDH polypeptide, which lacks a conventional N‐terminal signal peptide, to reach the cell wall in Saccharomyces cerevisiae by using an intracellular form of the yeast invertase as a reporter protein. A hybrid TDH3–SUC2 gene containing the C. albicans TDH3 promoter sequences and a coding region encoding a fusion protein formed by the C. albicans GAPDH polypeptide, fused at its C‐terminus with the yeast internal invertase, was constructed in a centromer derivative plasmid and transformed into a Suc− S. cerevisiae strain. Transformants displayed invertase activity measured in intact whole cells, and were able to grow on sucrose as the sole fermentable carbon source. Northern blot analysis with both TDH3 and SUC2 probes detected a single mRNA species of the expected size (about 2.7 kb), and Western immunoblot analysis of cell‐free extracts, using a monoclonal antibody (mAb49) against a C. albicans GAPDH epitope, showed the presence of a 90 kDa polypeptide corresponding to the GAPDH‐invertase fusion protein. This indicates that the TDH3 gene is able to direct part of the encoded gene product to the cell wall, and that any putative motifs for this targeting should be within the GAPDH amino acid sequence. Further analysis, using the same approach, of a panel of seven N‐ and C‐terminal GAPDH truncates revealed that the region required for the cell wall targeting is located within the N‐terminal half of the protein. Copyright © 2003 John Wiley & Sons, Ltd.
Development of a reporter system for the yeast Schwanniomyces occidentalis: influence of DNA composition and codon usage
Authors: Janatova Ivana , Costaglioli Patricia , Wesche Jorgen , Masson Jean‐Michel , Meilhoc Eliane 
Published: June 2003

AbstractIn this paper we report on searching for suitable reporters to monitor gene expression and protein secretion in the amylolytic yeast Schwanniomyces occidentalis. Several potential reporter and marker genes, formerly shown to be functional in other yeasts, were cloned downstream from the homologous invertase gene (INV) promoter and their activity was followed in conditions of repression and derepression of the INV promoter. However, neither β‐glucuronidase nor β‐lactamase nor phleomycin resistance‐conferring gene, all originating from E. coli, were expressed in S. occidentalis cells to such a level to allow for monitoring of their activity. All the reporter genes tested have a higher percentage of GC (47–62%) in their DNA compared to the DNA composition of S. occidentalis genes that are more AT‐rich (36% GC). The codon usage of all the reporter genes also varies from that of 16 so far sequenced S. occidentalis genes. This suggests that an appropriate composition of DNA and a codon usage similar to S. occidentalis genes might be very important parameters for an efficient expression of a heterologous gene in Schwanniomyces occidentalis. Indeed, two genes originating from Staphylococcus aureus, with an AT‐content in their DNA similar to that of S. occidentalis, were functionally expressed in S. occidentalis cells. Both a phleomycin resistance‐conferring gene and a chloramphenicol acetyltransferase‐encoding gene thus represent suitable reporters of gene expression and protein secretion in S. occidentalis. Additionally, we show in this work that the transcription‐regulating region and the signal peptide sequence of the S. occidentalis invertase gene were efficient to direct gene expression and subsequent protein secretion in Saccharomyces cerevisiae. Copyright © 2003 John Wiley & Sons, Ltd.
Genes involved in β‐oxidation, energy metabolism and glyoxylate cycle are induced by Candida albicans during macrophage infection
Authors: Prigneau Odile , Porta Amalia , Poudrier Jacques A. , Colonna‐Romano Sergio , Noël Thierry , Maresca Bruno 
Published: June 2003

AbstractThe ability of intracellular pathogens to cause infection is related to their capacity to survive and grow inside macrophages or in other cell types. Candida albicans latent virulence is likely to be related to a similar mechanism of avoiding killing by specialized cells and to the resulting ability to grow in such hostile environments. Using a differential display reverse transcription polymerase chain reaction technique, we have identified seven genes induced in C. albicans during macrophage phagocytosis. Sequence analyses and database searches revealed that these cDNAs coded for proteins homologous to yeast metabolic proteins. Interestingly, four of them are putative peroxisomal proteins, and two are involved in environmental signal sensing and transduction. Among the seven genes induced by C. albicans, six represent new information that were not described in other infection models. Copyright © 2003 John Wiley & Sons, Ltd.
Construction of a set of Saccharomyces cerevisiae vectors designed for recombinational cloning
Authors: Van Mullem Vincent , Wery Maxime , De Bolle Xavier , Vandenhaute Jean 
Published: June 2003

AbstractThe Gateway™ technology is becoming an increasingly popular method for cloning ORFs by recombination. It allows the transfer of any ORF flanked by specific recombination sites into any vectors harbouring the corresponding sites. Here we describe the construction of a set of 20 Saccharomyces cerevisiae Gateway™ compatible vectors. These plasmids bear an URA3 or TRP1 selection marker. They are designed for expression without tag sequence or for C‐ or N‐terminal protein tagging with 3HA (haemagglutinin), 13MYC, 4TAP (tandem affinity purification) or GST (glutathione S‐transferase) epitopes. The centromeric vectors allow expression of DNA sequence in yeast under tetracycline‐regulatable promoters, while expression from the high copy vectors is driven by PGK promoter. To test their applicability, the genes encoding the RNA polymerase I subunit Rpa12p or the TFIIS transcription factor were cloned in these vectors. Their expression was demonstrated using Western blotting or complementation assays. Copyright © 2003 John Wiley & Sons, Ltd.
Bone loss following tendon laceration, repair and passive mobilization
Authors: Ditsios Konstantinos , Boyer Martin I. , Kusano Nozomu , Gelberman Richard H. , Silva Matthew J. 
Published: June 2003

AbstractLittle is known about the localized changes in bone mass that occur following tendon or ligament injury. Interruption of normal load transfer at the insertion site will presumably lead to a localized loss of bone, although few data exist to support this claim. To test this hypothesis, we transected the canine flexor digitorum profundus (FDP) tendon from its insertion, and either repaired it using a trans‐osseous suture technique or left it unrepaired (laceration only). Post‐operatively, forelimbs in the repair group were cast immobilized except for 10 min of daily passive mobilization rehabilitation, whereas in the laceration only group dogs were allowed full weight bearing. At 5–42 days post‐injury, we assessed bone mineral density (BMD) using pQCT and osteoclast surface by histomorphometry. We measured significant bone loss in the distal phalanx after combined FDP tendon laceration, repair, and postoperative passive mobilization, with BMD decreases of 20%, 40%, and 41% at 10, 21, and 42 days (p < 0.01). Moreover, we observed that passive mobilization and tendon laceration each contributed independently to the observed bone loss. At 42 days, BMD was reduced by 21% in bones that were not injured but were subjected to the post‐operative passive mobilization protocol, while BMD was reduced by 28% in bones subjected to tendon laceration and full weight bearing (p < 0.01). In both the passive mobilization and laceration specimens, we counted significantly increased osteoclasts after only 7–10 days, and these increases persisted through 42 days (p < 0.05). We conclude that rapid and sustained bone resorption leads to significant bone loss in the 6‐week period following flexor tendon injury and repair. This bone loss may impact healing by impeding the restoration of a strong tendon‐bone interface. © 2003 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
VEGF in 105 pheochromocytomas: enhanced expression correlates with malignant outcome
Authors: SALMENKIVI KAISA , HEIKKIL PÄIVI , LIU JIANQI , HAGLUND CAJ , AROLA JOHANNA 
Published: May 2003

Pheochromocytomas are rare sympathoadrenal tumors that are highly vascular. Their malignancy is extremely difficult to estimate on the basis of histopathological features. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors involved in both tumor growth and metastasis. In our search for new prognostic markers, we investigated the expression of VEGF in normal adrenal gland, in 105 primary pheochromocytomas, and in 6 metastases by using immunohistochemistry and Northern blot analysis. We also calculated the microvessel density of these tumors by staining the endothelial cells with monoclonal CD34 antibody.VEGF messenger ribonucleic acid was found in all pheochromocytomas studied. Immunohistochemically, VEGF was not found in normal adrenal medullary cells. Interestingly, all malignant pheochromocytomas (n=8), regardless of their primary location, had strong or moderate VEGF immunoreactivity, while most benign adrenal pheochromocytomas (26 of 37, 70.3%) were either negative or only weakly positive. The staining was heterogenous in extraadrenal pheochromocytomas as well as in a group of tumors that had histologically suspicious features but had not metastasized, here called borderline tumors (n=29). The microvessel density varied greatly in all of the tumor groups, and no statistical difference was found between these groups.Here we report moderate to strong VEGF expression in malignant pheochromocytomas, and negative or weak expression in benign adrenal pheochromocytomas. Normal medullary cells are immunohistochemically negative. Thus, low VEGF expression in pheochromocytomas favors a benign diagnosis.
Two‐year follow‐up of Helicobacter pylori infection in C57BL/6 and Balb/cA mice
Authors: WANG XIN , WILLÉN ROGER , SVENSSON MARIE , LJUNGH ÅSA , WADSTRÖM TORKEL 
Published: May 2003

Helicobacter pylori infection is associated with chronic gastritis, peptic ulcer disease, gastric adenocarcinoma and MALT lymphoma. We previously found high‐grade lymphoma after 13 months' H. pylori infection in C57BL/6 mice. In this study we followed H. pylori infection by three different isolates in C57BL/6 and Balb/cA mice for 23 months. Six‐week‐old C57BL/6 and Balb/cA mice were infected with H. pylori strains 119p (CagA+, VacA+), SS1 (CagA+, VacA+) and G50 (CagA‐, VacA‐). Mice were followed at 2 weeks, 10 weeks and 23 months post‐inoculation (p.i.) by culture, histopathology and serology. Strain G50 was only reisolated from mice 2 weeks p.i. There was no difference in colonization between strain 119p and SS1 at 10 weeks p.i., whereas SS1 gave 100% colonization versus 119p gave 50% 23 months p.i.. Interestingly, the inflammation score was higher in mice infected with strain 119p than with SS1 10‐week p.i., and there were lymphoepithelial lesions in mice infected with strain 119p and G50 but not with SS1 at 23 months post‐infection. Eight mice infected with strains 119p and G50 developed gastric lymphoma (grade 5 and 4). One C57BL/6 mouse infected with strain 119p developed hepatocellular carcinoma after 23 months. Immunoblot showed specific bands of 26–33 kDa against H. pylori in infected mice, and two mice infected with strain SSI reacted with antibodies to the 120 kDa CagA toxin.Conclusion: A reproducible animal model for H. pylori‐induced lymphoma and possibly hepatocellular carcinoma is described. Strain diversity may lead to different outcomes of H. pylori infection.
Photodynamic therapy (PDT) using topically applied δ‐aminolevulinic acid (ALA) for the treatment of oral leukoplakia
Authors: Sieroń Aleksander , Adamek Mariusz , Kawczyk‐Krupka Aleksandra , Mazur Sebastian , Ilewicz Leszek 
Published: June 2003

AbstractBackground:  Photodynamic therapy (PDT) is a non‐invasive method for topical and selective treatment of pre‐malignant lesions of oral cavity. The aim of our study was to determine therapeutic response to PDT in patients with oral leukoplakia.Methods:  Twelve patients participated in our study. Lesions affected a variety of intraoral sites. The most common location was buccal, gingival and mandibular mucosa. Patients were treated with topically applied 10% δ‐aminolevulinic acid (ALA) and light from an argon‐pumped dye laser. Irradiation was performed in several (6–8) sessions using light at 635 nm wavelength, delivering a total dose of 100 J/cm2 per session.Results:  A complete response was obtained in 10 out of 12 treated patients. One recurrence was observed during 6 months.Conclusion:  Photodynamic therapy appears to be a feasible alternative to conventional therapy of pre‐malignant lesions of oral cavity.
Id proteins are overexpressed in human oral squamous cell carcinomas
Authors: Nishimine Masayoshi , Nakamura Mitsutoshi , Mishima Kenji , Kishi Munehiro , Kirita Tadaaki , Sugimura Masahito , Konishi Noboru 
Published: June 2003

AbstractBackground:  The helix‐loop‐helix (HLH) proteins Id‐1, Id‐2 and Id‐3 have been demonstrated to inhibit the activity of transcription factors and play an important role in regulating cell growth and tissue‐specific differentiation.Methods:  To elucidate the involvement of Id in human oral squamous cell carcinoma (OSCC), we examined 83 surgical specimens and eight normal gingival mucosae for the expression of Id proteins by immunohistochemistry; in addition, some specimens of the OSCC and the normal gingivae were also examined for the expression of Id‐1 mRNA by in situ hybridization (ISH), while Western blots were performed on six of the tumours and on cell lysates of five OSCC cell lines. We also explored the correlation between Id expressions and cellular proliferation indicating Ki‐67 or clinical parameters.Results:  We discovered a higher and more frequent expression of Id‐1 protein (27.7%) in human OSCC compared to that of Id‐2 and Id‐3 proteins (6.0 and 7.2%, respectively). Western blot analysis detected Id‐1 protein in four of six tumour samples and in all cell lines. ISH demonstrated strong cytoplasmic localization of Id‐1 mRNA in tumour samples at significantly higher levels compared to those of normal gingival mucosae.Conclusion:  Id‐1 protein expression significantly correlates with lymph node status, indicating that increased Id expression may relate to the metastatic behaviour in human OSCC.
Shifts in cellular localization of moesin in normal oral epithelium, oral epithelial dysplasia, verrucous carcinoma and oral squamous cell carcinoma
Authors: Kobayashi Hiroichi , Sagara Junji , Masumoto Junya , Kurita Hiroshi , Kurashina Kenji , Taniguchi Shuníichiro 
Published: June 2003

AbstractBackground:  Moesin, a member of ERM (ezrin/radixin/moesin) family, links actin filaments of cell surface structure to the cell membrane. The purpose of the study is to assess the shifts in cellular distribution of moesin in normal oral epithelium, oral epithelial dysplasia (OED), verrucous carcinoma (VC), and oral squamous cell carcinoma (OSCC).Methods:  The expression of moesin was evaluated immuohistochemically in paraffin‐embedded tissues of 59 specimens of OSCC, 35 specimens of OED, 17 specimens of VC, and five specimens of normal oral epithelium.Results:  In the normal oral epithelia, all specimens showed a pattern of membranous expression against the anti‐moesin antibody in the basal layer cells. In the OED specimens, moesin was dominantly expressed in the cell membrane except for the cornified layer. In VC and OSCC specimens, almost the whole of the carcinoma cells were stained with anti‐moesin antibody. However, in OSCC samples, moesin was markedly expressed increasingly in the cytoplasm and decreasingly in the cell membrane, as compared with OED and VC. In addition, there was a significant correlation between the pattern of moesin expression and tumor differentiation in OSCC.Conclusions:  Our results suggest that it is useful to detect the moesin expression as adjunct to screening mucosal lesions in the oral cavity.
Auto‐fluorescence spectra of oral submucous fibrosis
Authors: Chen Hsin‐Ming , Wang Chih‐Yu , Chen Chin‐Tin , Yang Hsiang , Kuo Ying‐Shiung , Lan Wan‐Hong , Kuo Mark Yen‐Ping , Chiang Chun‐Pin 
Published: June 2003

AbstractBackground:  Oral submucous fibrosis (OSF) is a chronic oral mucosal disease characterized by progressive deposition of collagen in the subepithelial connective tissue and epithelial atrophy. Previous studies have shown that at 330‐nm excitation, the 380‐ and 460‐nm emission peaks of the auto‐fluorescence spectra for oral mucosal tissues reflect the collagen content in the subepithelial connective tissue and the nicotinamide adenine dinucleotide phosphate (NADH) content in the epithelial cells, respectively. Therefore, at 330‐nm excitation OSF mucosa may have a higher 380‐nm emission peak and a lower 460‐nm emission peak than the normal oral mucosa (NOM).Methods:  To test the above hypothesis, we measured the in vivo auto‐fluorescence spectra of 59 OSF mucosal sites and compared the measured spectra with auto‐fluorescence spectra obtained from 15 NOM samples from 15 healthy volunteers, five samples of friction hyperkeratosis (histologic diagnosis, hyperkeratosis and acanthosis) on OSF buccal mucosa (FHOSF), and 29 samples of oral leukoplakia (histologic diagnosis, hyperkeratosis and acanthosis) on OSF buccal mucosa (OLOSF).Results:  We found that the spectrum of the OSF mucosa had a significantly higher 380‐nm emission peak and a significantly lower 460‐nm emission peak than the spectra of NOM, FHOSF, and OLOSF samples. When the mean (±SD) fluorescence intensities at 380 ± 15 nm (I380 ± 15 nm) and 460 ± 15 nm (I460 ± 15 nm) emission peaks and the mean ratio of I460 ± 15 nm/I380 ± 15 nm were compared between groups, we found that OSF group had a significantly higher mean value of I380 ± 15 nm, a significantly lower mean value of I460 ± 15 nm, and a significantly lower mean ratio of I460 ± 15 nm/I380 ± 15 nm than the NOM, FHOSF, and OLOSF groups (P < 0.001). However, no significant differences in the mean values of I380 ± 15 nm, I460 ± 15 nm, and ratio of I460 ± 15 nm/I380 ± 15 nm were found between NOM and FHOSF or OLOSF samples as well as between FHOSF and OLOSF samples (P > 0.05).Conclusion:  Because OSF mucosa has a very unique pattern of auto‐fluorescence spectrum, we conclude that auto‐fluorescence spectroscopy is a good method for real‐time diagnosis of OSF.