bluuebunny/crossref_metadata_2025_split huggingface dataset:
Whole-Mount Fluorescence Immunocytochemistry on Xenopus Embryos
Authors: Lee Chanjae , Kieserman Esther , Gray Ryan S. , Park Tae Joo , Wallingford John 
Published: February 2008

INTRODUCTIONImmunocytochemistry (ICC) is widely exploited in studying mammalian systems, but is underutilized among Xenopus developmental biologists. This stems, in part, from the relatively small number of Xenopus antibodies available for use in research. Common misconceptions about ICC in Xenopus embryos also prevail, discouraging researchers from trying the procedure. However, ICC with Xenopus is simple and effective. This article describes methods for whole-mount ICC in Xenopus embryos. Also included are simple procedures to quench autofluorescence of Xenopus and to remove surface pigment from embryos which may interfere with fluorescence imaging. The methods described here are useful for detecting tissue-specific probes (e.g., 12/101 to detect somites). They are also effective for imaging the cytoskeleton (e.g., α-tubulin to detect microtubules) or localizing specific proteins at the subcellular level (e.g., ZO-1 to detect tight junctions). In addition, combining ICC with in situ hybridization is simple and highly effective.
Rab-A2 and Rab-A3 GTPases Define atrans-Golgi Endosomal Membrane Domain inArabidopsisThat Contributes Substantially to the Cell Plate
Authors: Chow Cheung-Ming , Neto Hélia , Foucart Camille , Moore Ian 
Published: February 2008

AbstractThe Ypt3/Rab11/Rab25 subfamily of Rab GTPases has expanded greatly in Arabidopsis thaliana, comprising 26 members in six provisional subclasses, Rab-A1 to Rab-A6. We show that the Rab-A2 and Rab-A3 subclasses define a novel post-Golgi membrane domain in Arabidopsis root tips. The Rab-A2/A3 compartment was distinct from but often close to Golgi stacks and prevacuolar compartments and partly overlapped the VHA-a1 trans-Golgi compartment. It was also sensitive to brefeldin A and accumulated FM4-64 before prevacuolar compartments did. Mutations in RAB-A2a that were predicted to stabilize the GDP- or GTP-bound state shifted the location of the protein to the Golgi or plasma membrane, respectively. In mitosis, KNOLLE accumulated principally in the Rab-A2/A3 compartment. During cytokinesis, Rab-A2 and Rab-A3 proteins localized precisely to the growing margins of the cell plate, but VHA-a1, GNOM, and prevacuolar markers were excluded. Inducible expression of dominant-inhibitory mutants of RAB-A2a resulted in enlarged, polynucleate, meristematic cells with cell wall stubs. The Rab-A2/A3 compartment, therefore, is a trans-Golgi compartment that communicates with the plasma membrane and early endosomal system and contributes substantially to the cell plate. Despite the unique features of plant cytokinesis, membrane traffic to the division plane exhibits surprising molecular similarity across eukaryotic kingdoms in its reliance on Ypt3/Rab11/Rab-A GTPases.
Epigenetic Modification of Centromeric Chromatin: Hypomethylation of DNA Sequences in the CENH3-Associated Chromatin inArabidopsis thalianaand Maize
Authors: Zhang Wenli , Lee Hye-Ran , Koo Dal-Hoe , Jiang Jiming 
Published: February 2008

AbstractThe centromere in eukaryotes is defined by the presence of a special histone H3 variant, CENH3. Centromeric chromatin consists of blocks of CENH3-containing nucleosomes interspersed with blocks of canonical H3-containing nucleosomes. However, it is not known how CENH3 is precisely deposited in the centromeres. It has been suggested that epigenetic modifications of the centromeric chromatin may play a role in centromere identity. The centromeres of Arabidopsis thaliana are composed of megabase-sized arrays of a 178-bp satellite repeat. Here, we report that the 178-bp repeats associated with the CENH3-containing chromatin (CEN chromatin) are hypomethylated compared with the same repeats located in the flanking pericentromeric regions. A similar hypomethylation of DNA in CEN chromatin was also revealed in maize (Zea mays). Hypomethylation of the DNA in CEN chromatin is correlated with a significantly reduced level of H3K9me2 in Arabidopsis. We demonstrate that the 178-bp repeats from CEN chromatin display a distinct distribution pattern of the CG and CNG sites, which may provide a foundation for the differential methylation of these repeats. Our results suggest that DNA methylation plays an important role in epigenetic demarcation of the CEN chromatin.
Expressive vocabulary in 18‐month‐old children in relation to demographic factors, mother and child characteristics, communication style and shared reading
Authors: Westerlund M. , Lagerberg D. 
Published: February 2008

AbstractBackground  Previous research has elucidated the associations between children's language development and reading habits, and maternal education, communication style, gender and birth order. Research including maternal age and child temperament is more scarce. We studied the associations of all these factors with children's expressive vocabulary and reading habits. We also analysed the relationships of reading with expressive vocabulary, and effect sizes associated with frequent reading.Methods  Questionnaires were completed by mothers of 1091 children aged 17–19 months visiting the Swedish Child Health Services. Expressive vocabulary was assessed by the Swedish Communication Screening at 18 months, a screening version of McArthur‐Bates Communicative Development Inventories. Mother's perception of ability to communicate was measured by a scale constructed ad hoc from the International Child Development Programmes, a parent education curriculum. Bates' ‘difficultness’ scale was used to assess temperament.Results  Good communication, low maternal age, female gender and frequent reading were significantly associated with expressive vocabulary. High maternal education, good communication, higher maternal age, female gender and being a first‐born child were significantly associated with frequent reading. Reading at least 6 times/week added more than 0.3 SD in vocabulary regardless of gender and communication.Conclusions  The findings support the importance of reading and communication quality to early language development. Knowledge of the relationship between children's vocabulary and book reading in a context of joint attention is both theoretically and practically valuable to speech and language pathologists, pre‐school teachers, child health workers and other professionals.
Objectively monitored patching regimens for treatment of amblyopia: randomised trial
Authors: Reading Richard 
Published: February 2008

.

Stewart C.E.,
Stephens D.A.,
Fielder A.R. &
Moseley M.J.
(
2007
)

,
335
,
707
–
711
.
DOI: 10.1136/bmj.39301.460150.55.
Objectives  To compare visual outcome in response to two prescribed rates of occlusion (6 h a day and 12 h a day).Design  Unmasked randomized trial.Setting  Research clinics in two London hospitals.Participants  In total, 97 children with a confirmed diagnosis of amblyopia associated with strabismus, anisometropia, or both.Interventions  An 18‐week period of wearing glasses (refractive adaptation) followed by occlusion prescribed (‘patching’) for 6 or 12 h a day.Main outcome measures  Visual acuity measured by logMAR letter recognition, objectively monitored rate of occlusion (hours a day).Results  The mean age of children at study entry was 5.6 (SD 1.5) years. Ninety were eligible for occlusion but 10 dropped out in this phase, leaving 80 children who were randomized to a prescribed dose rate of 6 (n = 40) or 12 (n = 40) h a day. The mean change in visual acuity of the amblyopic eye was not significantly different (P = 0.64) between the two groups [0.26 (95% confidence interval 0.21–0.31) log units in 6‐h group; 0.24 (0.19–0.29) log units in 12‐h group]. The mean dose rates (hours a day) actually received, however, were also not significantly different [4.2 (3.7–4.7) in 6‐h group vs. 6.2 (5.1–7.3) in 12‐h group; P = 0.06]. The visual outcome was similar for those children who received 3 to 6 h a day or more than 6 to 12 h a day, but significantly better than that in children who received less than 3 h a day. Children aged under 4 years required significantly less occlusion than older children. Visual outcome was not influenced by type of amblyopia.Conclusions  Substantial (6 h a day) and maximal (12 h a day) prescribed occlusion results in similar visual outcome. On average, the occlusion dose received in the maximal group was only 50% more than in the substantial group and in both groups was much less than that prescribed. Younger children required the least occlusion.
The serum levels of the EGF‐like homeotic protein dlk1 correlate with different metabolic parameters in two hormonally different children populations in Spain*
Authors: Hermida Carmen , Garcés Carmen , De Oya Manuel , Cano Beatriz , Martínez‐Costa Oscar H. , Rivero Samuel , García‐Ramírez José J. , Laborda Jorge , Aragón Juan J. 
Published: February 2008

SummaryBackground  The Dlk1 gene encodes for dlk1, a transmembrane protein belonging to the EGF‐like repeat‐containing family. Dlk1 has been shown to act as a regulator of adipogenesis. Fc‐dlk1 transgenic mice show a decrease in adipose tissue and glucose tolerance, hypertriglyceridaemia and lower insulin sensitivity. Dlk1‐deficient mice show growth retardation, increased serum lipid metabolites and develop obesity. These data advocate for a role of dlk1 in the maintenance of lipid homeostasis, and suggest that dlk1 levels may influence the development of cardiovascular disease.Aim and methods  In this study, we analysed whether dlk1 serum levels could be indicative of the different hormonal or metabolic status shown by two Spanish children populations (6–8 years‐old), Orense and Murcia. We determined dlk1 serum levels by ELISA assay, using an antibody raised against the recombinant protein, and performed a correlation analysis against measurements of several hormonal and biochemical parameters in samples from 494 subjects.Results  We found a statistically significant positive correlation between serum levels of dlk1 and those of glucose (P < 0·05), total cholesterol (P < 0·01) and high‐density lipoprotein‐cholesterol (HDL‐C) (P < 0·01) in children from Murcia, but not from Orense's population, where dehydroepiandrosterone‐sulphate (DHEA‐S) levels were significantly higher (P < 0·01) and dlk1 correlated positively with insulin (P < 0·01), homeostasis model assessment (HOMA) (P < 0·01) and free fatty acids (FFA) (P < 0·05).Conclusions  dlk1 serum levels appear related to the anabolic status of the children in association with changes in the levels of DHEA‐S, which have been associated with hyperinsulinaemia and diabetes. Monitoring dlk1 levels may be important to evaluate the metabolic and hormonal stage of child development.
Control of IGF‐I levels with titrated dosing of lanreotide Autogel over 48 weeks in patients with acromegaly
Authors: Chanson Philippe , Borson‐Chazot Françoise , Kuhn Jean‐Marc , Blumberg Joëlle , Maisonobe Pascal , Delemer Brigitte ,   the Lanreotide Acromegaly Study Group
Published: February 2008

SummaryBackground  An essential criterion for control of acromegaly is normalization of IGF‐I levels. Somatostatin analogues act to suppress IGF‐I and GH levels.Objective  To assess the efficacy and safety of 48 weeks titrated dosing of lanreotide Autogel.Design  Open‐label, multicentre, phase III, 48‐week trial.Methods  Patients with active acromegaly (IGF‐I levels > 1·3 times upper limit of age‐adjusted normal range) were recruited. Twelve injections of lanreotide Autogel were given at 28‐day intervals: during the 16‐week fixed‐dose phase, patients received 90 mg; in the 32‐week dose‐titration phase, patients received 60, 90 or 120 mg according to GH and IGF‐I levels. Intention‐to‐treat analysis was performed to determine the proportion of patients with normalized age‐adjusted IGF‐I levels at study end. Secondary evaluations included GH levels, clinical acromegaly signs and safety.Results  Fifty‐seven of 63 patients completed the study. Lanreotide Autogel resulted in normalized age‐adjusted IGF‐I levels in 27 patients (43%, 95% CI 31–55). Mean GH levels decreased from 6·2 to 1·5 µg/l at study end, with 53 of 62 patients (85%) having GH levels ≤ 2·5 µg/l (95% CI 76·7–94·3) and 28 of 62 patients (45%) with levels < 1 µg/l (95% CI 32·8–57·6). Twenty‐four (38%) had both normal IGF‐I levels and GH levels ≤ 2·5 µg/l. Acromegaly symptoms reduced significantly in most patients throughout the study. The most common adverse events were gastrointestinal, as expected for somatostatin analogues.Conclusions  Using IGF‐I as primary end‐point, 48 weeks lanreotide Autogel treatment, titrated for optimal hormonal control, controlled IGF‐I and GH levels effectively, reduced acromegaly symptoms and was well tolerated.
Metabolic characteristics of subjects with normal glucose tolerance and 1‐h hyperglycaemia
Authors: Qian Lei , Fu Xuelian , Xu Lihong , Zheng Sheng , Zhou Weibin , Wang Xiao , Gu Yanyun , Lin Fan , Luo Min 
Published: February 2008

SummaryObjective  Nondiabetic subjects with a 1‐h plasma glucose ≥ 11·1 mmol/l during an oral glucose tolerance test (OGTT) drew our attention to their somewhat confusing status and relative frequency among Chinese patients. The aim of this study was to clarify the metabolic characteristics of these subjects.Design and patients  A total of 2549 Chinese subjects were included in this study. Based on results of OGTT, these subjects were classified into three groups: normal glucose tolerance (NGT), impaired glucose regulation (IGR) and diabetes mellitus (DM). Then, according to the level of 1‐h plasma glucose, the NGT and IGR groups were subclassified, respectively, as: NGT without 1‐h hyperglycaemia (NGTN), NGT with hyperglycaemia at 1 h (NGT1H), IGR without 1‐h hyperglycaemia (IGRN), and IGR with hyperglycaemia at 1 h (IGR1H).Results  After adjustments for age and gender, the insulinogenic index (IGI) of NGT1H and IGR1H was found to be lower than for those with NGTN and of IGRN, respectively (P < 0·05). No statistical differences, however, were found in oral glucose insulin sensitivity (OGIS) between either of the 1‐h hyperglycaemic groups or of the corresponding NGTN or IGRN groups. Homeostasis model assessment for β‐cell function (HOMA‐B) of NGT1H was lower than that of NGTN (P < 0·05), while IGRN and IGR1H showed no difference. No differences in homeostasis model assessment for insulin resistance (HOMA‐IR) were found among NGTN, NGT1H, IGRN and IGR1H groups. The levels of triglycerides (TG) were not significantly different among NGT1H, IGRN and IGR1H, while TG in these groups were significantly higher than in NGTN (P < 0·05). LDL‐C was significantly higher and HDL significantly lower in NGT1H than in all other groups (P < 0·05).The IGR group was also subclassified as: isolated impaired fasting glucose (IFG), isolated impaired glucose tolerance (IGT) and combined glucose intolerance (CGI). The IGI of the NGT1H group was similar to the IGI that of combined glucose intolerance group but lower than those of IFG and IGT (P < 0·05).The OGIS of the NGT1H group was the highest among all groups (P < 0·05). HOMA‐B of IGT and NGT1H were higher than that of IFG (P < 0·05). There was no difference among all groups in HOMA‐IR. Plasma lipid levels were not significantly different between NGT1H and any other group.Conclusions  Chinese NGT subjects with a 1‐h plasma glucose ≥ 11·1 mmol/l are characterized by metabolic abnormalities, which may be caused by the impairment of early insulin release rather than aggravated insulin resistance.
Double‐stranded RNA mediates interferon regulatory factor 3 activation and interleukin‐6 production by engaging Toll‐like receptor 3 in human brain astrocytes
Authors: Kim Hyemi , Yang Eunjung , Lee Jeonggi , Kim Se‐Hoon , Shin Jeon‐Soo , Park Joo Young , Choi Sun Ju , Kim Se Jong , Choi In‐Hong 
Published: February 2008

SummaryToll‐like receptor 3 (TLR3) participates in the innate immune response by recognizing viral pathogens. In this study, human brain astrocytes were found to constitutively express TLR3, and this expression was increased by interferon‐γ (IFN‐γ) or double‐stranded RNA (dsRNA). Treatment employing dsRNA in astrocytes induced IFN regulatory factor 3 (IRF3) phosphorylation, dimer formation and nuclear translocation followed by STAT1 activation. This treatment also activated nuclear factor‐κB, p38 and c‐Jun N‐terminal kinase significantly, while activating extracellular signal‐regulated kinase to a lesser extent. Treatment with anti‐TLR3 antibody inhibited dsRNA‐mediated interleukin‐6 (IL‐6) production. In the presence of mitogen‐activated protein kinase inhibitors, astrocytes failed to secrete IL‐6 in response to dsRNA treatment. Therefore, dsRNA‐induced IL‐6 production is dependent on mitogen‐activated protein kinases and type I IFN production is dependent on IRF3 in brain astrocytes. These results suggest that brain inflammation, which produces inflammatory cytokines and type I IFNs, may enhance TLR3 expression in astrocytes. Additionally, upregulated TLR3 might modulate inflammatory processes by producing proinflammatory cytokines.
The periodontal pathogen Porphyromonas gingivalis cleaves apoB‐100 and increases the expression of apoM in LDL in whole blood leading to cell proliferation
Authors: Bengtsson T. , Karlsson H. , Gunnarsson P. , Skoglund C. , Elison C. , Leanderson P. , Lindahl M. 
Published: February 2008

Abstract.Objective.  Several studies support an association between periodontal disease and atherosclerosis with a crucial role for the pathogen Porphyromonas gingivalis. This study aims at investigating the proteolytic and oxidative activity of P. gingivalis on LDL in a whole blood system using a proteomic approach and analysing the effects of P. gingivalis‐modified LDL on cell proliferation.Methods.  The cellular effects of P. gingivalis in human whole blood were assessed using lumi‐aggregometry analysing reactive oxygen species production and aggregation. Blood was incubated for 30 min with P. gingivalis, whereafter LDL was isolated and a proteomic approach was applied to examine protein expression. LDL‐oxidation was determined by analysing the formation of protein carbonyls. The effects of P. gingivalis‐modified LDL on fibroblast proliferation were studied using the MTS assay.Results.  Incubation of whole blood with P. gingivalis caused an extensive aggregation and ROS production, indicating platelet and leucocyte activation. LDL prepared from bacteria‐exposed blood showed an increased protein oxidation, elevated levels of apoM and formation of two apoB‐100 N‐terminal fragments. Porphyromonas gingivalis‐modified LDL markedly increased the growth of fibroblasts. Inhibition of gingipain R suppressed the modification of LDL by P. gingivalis.Conclusions.  The ability of P. gingivalis to change the protein expression and proliferative capacity of LDL may represent a crucial event in periodontitis‐associated atherosclerosis.
PYK10, a β‐glucosidase located in the endoplasmatic reticulum, is crucial for the beneficial interaction between Arabidopsis thaliana and the endophytic fungus Piriformospora indica
Authors: Sherameti Irena , Venus Yvonne , Drzewiecki Corinna , Tripathi Swati , Dan Vipin Mohan , Nitz Inke , Varma Ajit , Grundler Florian M. , Oelmüller Ralf 
Published: February 2008

Summary Piriformospora indica, an endophyte of the Sebacinaceae family, promotes growth and seed production of many plant species, including Arabidopsis. Growth of a T‐DNA insertion line in PYK10 is not promoted and the plants do not produce more seeds in the presence of P. indica, although their roots are more colonized by the fungus than wild‐type roots. Overexpression of PYK10 mRNA did not affect root colonization and the response to the fungus. PYK10 codes for a root‐ and hypocotyl‐specific β‐glucosidase/myrosinase, which is implicated to be involved in plant defences against herbivores and pathogens. Expression of PYK10 is activated by the basic helix‐loop‐helix domain containing transcription factor NAI1, and two Arabidopsis lines with mutations in the NAI1 gene show the same response to P. indica as the PYK10 insertion line. PYK10 transcript and PYK10 protein levels are severely reduced in a NAI1 mutant, indicating that PYK10 and not the transcription factor NAI1 is responsible for the response to the fungus. In wild‐type roots, the message level for a leucine‐rich repeat protein LRR1, but not for plant defensin 1.2 (PDF1.2), is upregulated in the presence of P. indica. In contrast, in lines with reduced PYK10 levels the PDF1.2, but not LRR1, message level is upregulated in the presence of the fungus. We propose that PYK10 restricts root colonization by P. indica, which results in the repression of defence responses and the upregulation of responses leading to a mutualistic interaction between the two symbiotic partners.
Sequential glucosylation of a furofuran lignan, (+)‐sesaminol, by Sesamum indicum UGT71A9 and UGT94D1 glucosyltransferases
Authors: Noguchi Akio , Fukui Yuko , Iuchi‐Okada Asako , Kakutani Saki , Satake Honoo , Iwashita Takashi , Nakao Masahiro , Umezawa Toshiaki , Ono Eiichiro 
Published: February 2008

Summary(+)‐Sesaminol 2‐O‐triglucoside is the most abundant water‐soluble furofuran lignan in sesame seeds (Sesamum indicum) and is considered to be a beneficial compound for human health. The biosyntheses and physiological roles of lignan glycosides, however, remain elusive. Here we report the molecular identification and biochemical characterization of two Sesamum uridine diphosphate (UDP) glucose:lignan glucosyltransferases. Sesamum indicum UGT71A9 preferentially glucosylated at the 2‐hydroxyl group of (+)‐sesaminol, resulting in (+)‐sesaminol 2‐O‐glucoside. Similarly, two UGT71A9 homologs from Sesamum radiatum (UGT71A10) and Sesamum alatum (UGT71A8) also showed (+)‐sesaminol glucosylating activity, evidencing the functional conservation of (+)‐sesaminol 2‐O‐glucosyltransferases in the Sesamum genus. In addition, S. indicum UGT94D1 specifically glucosylated at the 6′‐hydroxyl group of the sugar moiety of (+)‐sesaminol 2‐O‐glucoside but not at that of flavonoid glucosides. The gene expression patterns of UGT71A9 and UGT94D1 during seed development were correlated with the glucosylating activities toward (+)‐sesaminol in planta, suggesting that the two lignan UDP‐glycosyltransferases participate in the sequential glucosylation steps in the biosynthesis of (+)‐sesaminol 2‐O‐triglucoside.