bluuebunny/crossref_metadata_2025 huggingface dataset:
Retroviral Vector Production by Transient Transfection
Authors: Cornetta Kenneth , Pollok Karen E. , Miller A. Dusty
Published: April 2008

INTRODUCTION

This protocol describes vector production by transient transfection. The production of retroviral vectors requires a full-length copy of the vector RNA to be incorporated into virions. This is accomplished by coexpressing vector RNA and the viral proteins required for virion formation from expression plasmids. To avoid generation of replication-competent virus, the viral genes are carried by separate plasmids. Generally, the gag and pol genes are on one plasmid, and the viral envelope gene is on a second plasmid. The viral protein-coding regions can be expressed using various promoters to decrease homology and thereby decrease recombination. Because these plasmids do not contain the packaging (ψ) sequence, the viral genes are unlikely to be incorporated into virions.
Generation of Stable Vector-Producing Cells for Retroviral Vectors: Figure 1.
Authors: Cornetta Kenneth , Pollok Karen E. , Miller A. Dusty
Published: April 2008

INTRODUCTION

This procedure describes the generation of clonal vector-producing cells that will provide an unlimited amount of unrearranged retroviral vector. The procedure involves transfection of one packaging cell line to generate a vector that is used to transduce a second packaging cell line. The resultant vector-producing clones generally contain a single integrated copy of the retroviral vector, and virus produced from this integrated vector is as genetically homogeneous as possible. Although the vector produced by a given packaging cell line can sometimes be used to transduce the same cell line, the transduction rate is typically low because of receptor blockage by the Env protein made by the target packaging cells. Indeed, this procedure will select for target cells that express low Env protein levels and thus are less resistant to transduction, but at the same time will ultimately produce less vector because of low Env production. Therefore, to obtain the highest vector titers, it is important to use pairs of packaging cells such that receptor blockage is not an issue. In this example, we use PE501 ecotropic packaging cells for transfection and broad-host-range PT67 packaging cells to make stable vector-producing cells.
Transduction of Cell Lines by Retroviral Vectors
Authors: Cornetta Kenneth , Pollok Karen E. , Miller A. Dusty
Published: April 2008

INTRODUCTION

This protocol is suitable for transduction of many adherent cell lines. The number of target cells transduced can be varied as needed by maintaining the ratio of surface area to volume and using plates/flasks of various sizes. The protocol can also easily be adapted for non-adherent cells using similar vector-to-cell ratios.
Labeling the Nucleus with Fluorescent Dyes for Imaging: Figure 1.
Authors: Chazotte Brad
Published: April 2008

INTRODUCTION

The eukaryotic cell has evolved to compartmentalize its functions and transport various metabolites among cellular compartments. Therefore, in cell biology, the study of organization and structure/function relationships is of great importance. The nucleus contains almost all of the cell’s DNA and is bounded by a double membrane. Inside and adjacent to the inner membrane of the nuclear envelope is the nuclear lamina. It is composed of a fibrous meshwork comprising one or more of three major intermediate filament-like polypeptides: lamins A, B, and C. The outer nuclear membrane is contiguous with the endoplasmic reticulum. Several fluorescent stains are available that label DNA and allow easy visualization of the nucleus in interphase cells and chromosomes in mitotic cells. These stains include Hoechst and 4′,6-diamidino-2-phenylindole (DAPI), which are used in this article. Although not as bright as the vital Hoechst stains for DNA, DAPI has greater photostability. One advantage of Hoechst 33342 over DAPI is that the former is membrane-permeant and is, therefore, useful for imaging living cells, because it does not require cell fixation or permeabilization.
Desorption Electrospray Ionization (DESI) Analysis of Tryptic Digests/Peptides
Authors: Takats Zoltan , Wiseman Justin M. , Ifa Demian R. , Cooks R. Graham
Published: April 2008

INTRODUCTION

The analytical utility of desorption electrospray ionization (DESI) is such that it can be applied to qualitative proteomics research in the same way as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) methods, although little work has yet been reported in this regard. Because DESI is a surface analysis technique and easily automated, it can be implemented for high-throughput applications, which include the analysis of chromatographic fractions of digested proteins. The analysis of tryptic peptides follows the same protocols as in typical MALDI or ESI methods, except that the mixture is spotted directly onto an insulating surface, allowed to dry, and analyzed directly without adding matrix compounds (as in the case of MALDI methods). The spectral characteristics are similar to those of ESI in that both singly and multiply charged analyte ions are detected. Spectra are highly similar to electrospray spectra of tryptic digests with regard to the overwhelming presence of multiply charged ions of peptides. DESI-mass spectrometry (DESI-MS) is an emerging technique with great promise, but its application range is still being investigated. Therefore, the protocol for DESI-MS analysis of tryptic digests/peptides presented here provides general procedures used for the applications that have been investigated so far. Optimal ion source parameters and surface types may vary, depending on the application.
Maximum Parsimony Method for Phylogenetic Prediction
Authors: Mount David W.
Published: April 2008

INTRODUCTION

Maximum parsimony predicts the evolutionary tree or trees that minimize the number of steps required to generate the observed variation in the sequences from common ancestral sequences. For this reason, the method is also sometimes referred to as the minimum evolution method. A multiple sequence alignment (msa) is required to predict which sequence positions are likely to correspond. These positions will appear in vertical columns in the msa. For each aligned position, phylogenetic trees that require the smallest number of evolutionary changes to produce the observed sequence changes from ancestral sequences are identified. This analysis is continued for every position in the sequence alignment. Finally, those trees that produce the smallest number of changes overall for all sequence positions are identified. This method is best suited for sequences that are quite similar and is limited to small numbers of sequences.
Distance Methods for Phylogenetic Prediction
Authors: Mount David W.
Published: April 2008

INTRODUCTION

Phylogenetic analysis of a multiple sequence alignment (msa) can be performed using distance methods, which are based on genetic distances between sequence pairs in an msa. The genetic distance between two sequences is the fraction of aligned positions in which the sequence has been changed. In contrast, sequence identity is the fraction of the aligned positions that are identical. Gaps may be ignored in distance calculations or treated like substitutions. A scoring or substitution matrix may also be used, making the calculation slightly more complicated, although the principle is the same. Sequence pairs that have the smallest distances are “neighbors.” On a tree, these sequences share a node or common ancestor position and are each joined to that node by a branch.
Desorption Electrospray Ionization: Proteomics Studies by a Method That Bridges ESI and MALDI
Authors: Takats Zoltan , Wiseman Justin M. , Ifa Demian R. , Cooks R. Graham
Published: April 2008

INTRODUCTION

Desorption electrospray ionization (DESI) is a desorption ionization (DI) method by nature, and, like matrix-assisted laser desorption/ionization (MALDI), it is used for the analysis of material present on a surface. DESI includes features reminiscent of electrospray ionization (ESI) in respect to both its instrumental and mechanistic aspects. However, the analyte in the DESI experiment is not in solution as in ESI. Instead, a microelectrospray ion source is used to produce charged droplets, ionic clusters, and/or gas-phase ions (depending on chosen experimental conditions), and these are directed at the sample surface. The sample is present in the ambient environment. An electrical potential of several kilovolts (kV) is applied to the spray solution, and pneumatic nebulization is used to assist in desolvation. Ionization of molecules present on the sample surface occurs upon the impact of the ESI-originated, charged particles with the surface. Surfaces include deposited samples on sample holder targets as well as surfaces of natural objects such as biological tissues or minerals.
Improvement of the breaking strength of wound by combined treatment with recombinant human G‐CSF, recombinant human M‐CSF, and a TGF‐β1 receptor kinase inhibitor in rat skin
Authors: Sugiyama Kenji , Ishii Genichiro , Ochiai Atsushi , Esumi Hiroyasu
Published: April 2008

Effective doses of ionizing radiation during preoperative radiotherapy occasionally cause wound complications after subsequent surgery. The authors attempted to accelerate radiation‐impaired wound healing in animal models. Recombinant human granulocyte colony‐stimulating factor (rhG‐CSF), recombinant human macrophage colony‐stimulating factor (rhM‐CSF), and an inhibitor of transforming growth factor (TGF)‐β1 receptor kinase, SB431542, were injected s.c. into a full‐thickness incisional wound site in the dorsal skin of rats after local irradiation of X‐ray (30 Gy). Wound healing of irradiated skin was assessed using the breaking strength of the wound and histological analyses. The impaired wound healing in irradiated skin was found to be associated with impaired mobilization of bone marrow‐derived cells and enhanced expression of TGF‐β1 mRNA. The breaking strength of the wound in the irradiated skin was approximately one‐eighth of that in the non‐irradiated skin; however, following combined treatment with the above three compounds the breaking strength increased to approximately one‐half of that in the non‐irradiated skin. Histological analysis of the wounded skin revealed an increase in formation of collagen fibers and the panniculus carnosus following the combined treatment. Moreover, the increased breaking strength was associated with an increase in a subpopulation of fibrocytes (collagen I/ED1 double positive cells). These findings suggested that a combined treatment with rhG‐CSF, rhM‐CSF, and SB431542 is promising as a means of improving radiation‐impaired wound healing. (Cancer Sci 2008; 99: 1021–1028)
CYP2A6and the plasma level of 5‐chloro‐2, 4‐dihydroxypyridine are determinants of the pharmacokinetic variability of tegafur and 5‐fluorouracil, respectively, in Japanese patients with cancer given S‐1
Authors: Fujita Ken‐ichi , Yamamoto Wataru , Endo Shoji , Endo Hisashi , Nagashima Fumio , Ichikawa Wataru , Tanaka Ryuhei , Miya Toshimichi , Araki Kazuhiro , Kodama Keiji , Sunakawa Yu , Narabayashi Masaru , Miwa Keisuke , Ando Yuichi , Akiyama Yuko , Kawara Kaori , Kamataki Tetsuya , Sasaki Yasutsuna
Published: April 2008

S‐1 is an oral anticancer agent composed of tegafur (FT), 5‐chloro‐2,4‐dihydroxypyridine (CDHP), and potassium oxonate. CDHP is added to prevent degradation of 5‐fluorouracil (5‐FU) by inhibiting dihydropyrimidine dehydrogenase. CYP2A6 is involved in the biotransformation of FT to 5‐FU. Thus, we prospectively analyzed the effects of theCYP2A6genotype, plasma level of CDHP, and patient characteristics on the pharmacokinetic (PK) variability of FT and 5‐FU. Fifty‐four Japanese patients with metastatic or recurrent cancers who received S‐1 were enrolled. TheCYP2A6polymorphisms (*4A, *7, and *9) with deficient or reduced activity were analyzed. All subjects were classified into three groups according to theirCYP2A6genotype: wild type (*1/*1), one‐variant allele (*1/any), or two‐variant alleles (combination other than *1). The PK of FT, 5‐FU, and CDHP were measured on day 1 of treatment. Multivariate regression analysis revealed that oral clearance of FT was associated with theCYP2A6genotype (analysis of variance [ANOVA],P = 0.000838). The oral clearance of FT seen in patients with the two‐variant alleles was significantly lower than those in wild type and the one‐variant allele (95% confidence intervals 0.75–2.41 and 0.41–1.82, respectively; Tukey‐Kramer test). The area under the time–concentration curve (AUC) of 5‐FU was significantly correlated with the AUC of CDHP (ANOVA,P = 0.00126). The AUC of 5‐FU and CDHP were inversely correlated with creatinine clearance (ANOVA,P = 0.0164 andP = 0.000762, respectively). Although theCYP2A6variants are the cause of the PK variability of FT, the AUC of CDHP affected by renal function is the key determinant of the variability in the PK of 5‐FU. (Cancer Sci2008; 99: 1049–1054)
Nuclear factor‐κB dependency of doxorubicin sensitivity in gastric cancer cells is determined by manganese superoxide dismutase expression
Authors: Cho Sung Jin , Park Jong‐Wan , Kang Jae Seung , Kim Woo Ho , Juhnn Yong‐Sung , Lee Jae‐Seon , Kim Young‐Hoon , Ko Young San , Nam Seon Young , Lee Byung Lan
Published: April 2008

The role of nuclear factor‐κB (NF‐κB) activation in cancer cell apoptosis appears to be tailored specifically for each cell type and the type of NF‐κB inducer. The present study aimed to determine whether or not NF‐κB activation is associated with chemosensitivity to doxorubicin (DOX) using the DOX‐sensitive SNU‐601 and DOX‐resistant SNU‐216 gastric cancer cell lines. The effect of NF‐κB activation on DOX (1 µg/mL) sensitivity was analyzed after the suppression of NF‐κB activation using transfection of the super‐suppressive mutant form of IκBα (mIκBα) or pretreatment with pyrrolidine dithiocarbamate. In addition, the association between NF‐κB and manganese superoxide dismutase (MnSOD) in relation to DOX sensitivity was analyzed after the modulation of MnSOD expression. The NF‐κB activity was much higher in DOX‐resistant SNU‐216 cells than in DOX‐sensitive SNU‐601 cells before and after DOX treatment. Overexpression of mIκBα or pyrrolidine dithiocarbamate pretreatment decreased the DOX resistance in SNU‐601 cells with low MnSOD expression, but not in SNU‐216 cells with high MnSOD expression. In comparison, the overexpression of MnSOD, which also suppressed NF‐κB activation in both cell lines, increased DOX resistance in SNU‐601 cells. Blocking of MnSOD expression using RNA interference techniques increased DOX sensitivity in SNU‐216 cells, which was further augmented by the additional inhibition of NF‐κB activity. Our results showed that whether NF‐κB contributes to DOX sensitivity in gastric cancer cells is determined by the level of MnSOD expression. Thus, targeting both MnSOD and NF‐κB may be helpful for increasing the efficacy of DOX treatment of DOX‐resistant SNU gastric cancer cells. (Cancer Sci 2008; 99: 1117–1124)
Evidence of the in vivo esterification of budesonide in human airways
Authors: Van Den Brink Karen I. Maassen , Boorsma Martin , Staal‐van den Brekel A. Jeske , Edsbäcker Staffam , Wouters Emiel F. , Thorsson Lars
Published: April 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• In vitro studies with human bronchial epithelial cells have shown that budesonide undergoes rapid, extensive and reversible intracellular esterification, a finding that is believed to contribute to retention and prolonged duration of action.

• A case report has suggested retention of budesonide in the lungs of patients undergoing surgical resection.

• This study evaluated, for the first time, the esterification (and distribution) of inhaled budesonide and fluticasone propionate in vivo in human lung.

WHAT THIS STUDY ADDS

• This study has unequivocally shown that budesonide undergoes esterification in human lungs in vivo.

• Budesonide was detectable in central and peripheral lung tissue for 10 h, and budesonide oleate up to 43 h after inhalation.

• The sustained retention of budesonide esters in the lungs probably contributes to the prolonged duration of action and once‐daily efficacy of budesonide.

AIMS

Budesonide, unlike fluticasone propionate, undergoes fatty acid esterification in the lungs, and there is a need to characterize fully the distribution and fate of the two drugs after inhalation in humans.

METHODS

This open‐label, randomized study was performed in adults undergoing whole lung or lobar resection resulting from lung cancer. Patients were given single 1000‐μg doses of both budesonide and fluticasone propionate via dry powder inhalers before surgery. Tissue samples from peripheral and central lung, an ex vivo bronchial brush sample and intercostal muscle, together with plasma samples, were taken during surgery and analysed by liquid chromatography plus tandem mass spectrometry.

RESULTS

Lung tissue samples were obtained from 22 patients at surgery, 1–43 h after drug dosing. Budesonide was detectable from earliest sampling in central and peripheral lung tissue up to 10 h (in six of 22 samples), fluticasone propionate up to 22 h after inhalation (in 16 of 22 samples), and budesonide oleate up to 43 h after inhalation (in 21 of 22 samples). Budesonide, but not fluticasone propionate, was detected in intercostal muscle for up to 10 h after inhalation. Bronchial brush samples showed the presence of fluticasone propionate for up to 18 h, suggesting the presence of undissolved drug powder particles in the airway lumen.

CONCLUSION

Sustained retention of esterified budesonide in the lungs supports the prolonged duration of action of budesonide and suitability for once‐daily administration.
Eosinophils: Biological Properties and Role in Health and Disease
Authors: HOGAN SIMON P. , ROSENBERG HELENE F. , MOQBEL REDWAN , PHIPPS SIMON , FOSTER PAUL S. , LACY PAIGE , KAY A. BARRY , ROTHENBERG MARC E.
Published: April 2008

Eosinophils are pleiotropic multifunctional leukocytes involved in initiation and propagation of diverse inflammatory responses, as well as modulators of innate and adaptive immunity. In this review, the biology of eosinophils is summarized, focusing on transcriptional regulation of eosinophil differentiation, characterization of the growing properties of eosinophil granule proteins, surface proteins and pleiotropic mediators, and molecular mechanisms of eosinophil degranulation. New views on the role of eosinophils in homeostatic function are examined, including developmental biology and innate and adaptive immunity (as well as their interaction with mast cells and T cells) and their proposed role in disease processes including infections, asthma, and gastrointestinal disorders. Finally, strategies for targeted therapeutic intervention in eosinophil‐mediated mucosal diseases are conceptualized.
Zinc speciation and isotopic exchangeability in soils polluted with heavy metals
Authors: Diesing W. E. , Sinaj S. , Sarret G. , Manceau A. , Flura T. , Demaria P. , Siegenthaler A. , Sappin‐Didier V. , Frossard E.
Published: April 2008

Correct characterization of heavy metal availability is a prerequisite for the management of polluted soils. Our objective was to describe zinc (Zn) availability in polluted soils by measuring the isotopic exchangeability of Zn in soil/solution (E value) and in soil/plant systems (L value), by assessing the transfer of Zn and 65Zn in the fractions of a six‐step selective sequential extraction (SSE) in incubated soils and by identifying Zn forms in soils by means of extended X‐ray absorption fine structure (EXAFS) spectroscopy. We distinguished three pools of exchangeable Zn: the pool of Zn exchangeable within 1 minute, which is observed in all soils, Zn exchangeable in the medium term, and the slowly and not exchangeable Zn. The amount of Zn present in the first two pools was similar to the L value measured with Thlaspi caerulescens. The first three steps of the SSE solubilized the first pool and a fraction of the second pool. Most of the second pool and a fraction of the third pool were extracted in the fourth step of the SSE, while the rest of the third pool was extracted in the final steps of the SSE. The EXAFS study conducted on two soils showed that more than half of the Zn was present in species weakly bound to organic compounds and/or outer sphere inorganic and organic complexes. Other species included strongly sorbed Zn species and Zn species in crystalline minerals. The EXAFS study of selected SSE residues showed that the specificity and the efficiency of the extractions depended on the properties of the soil studied.
Recombinant factor VIIa to prevent surgical bleeding in factor XI deficiency
Authors: O'CONNELL N. M. , RIDDELL A. F. , PASCOE G. , PERRY D. J. , LEE C. A.
Published: April 2008

Summary.  Factor XI (FXI) deficiency is associated with bleeding after invasive procedures. Risks of human plasma‐derived FXI replacement products include transfusion transmitted infection, thrombosis and fluid overload. This study was designed to test the hypothesis that recombinant factor VIIa (rFVIIa) is an effective haemostatic agent in patients with FXI deficiency undergoing surgery. Fourteen FXI deficient patients [five severely deficient (FXI:C <20 U dL−1) and nine partially deficient (FXI:C 20–70 U dL−1] received rFVIIa to prevent surgical bleeding during five major, four minor and six dental procedures. Minor surgical and dental procedures were covered with two doses of rFVIIa (90 μg kg−1 i.v.), the first pre‐operatively and the second 4 h postoperatively. Major surgery was covered with 90 μg kg−1 i.v. two hourly for the first 24 h and four hourly for the second 24 h. Oral tranexamic acid was given for 7 days postoperatively. Effective haemostasis was observed in all cases and no alternative haemostatic agents or blood transfusions were required. Three adverse events were recorded; an acute cerebrovascular accident in a patient with a history of cardiovascular disease, an allergic reaction and local phlebitis. In this study, rFVIIa was an effective alternative to plasma‐derived FXI replacement for the prevention of surgical bleeding in FXI deficient patients but rFVIIa may not be suitable for patients with pre‐existing risk factors for thrombosis.
The impact of severe haemophilia on the social status and quality of life among Austrian haemophiliacs
Authors: HARTL H. K. , REITTER S. , EIDHER U. , RAMSCHAK H. , AY C. , PABINGER I.
Published: April 2008

Summary.  Although many studies of the impact of haemophilia on the quality of life were conducted, there is hardly any data on the social status of haemophiliacs. It was the aim of our study to obtain data on the social status of Austrian haemophiliacs and to compare these with an age‐ and sex‐matched reference population. Furthermore, we collected data on the quality of life of haemophilia patients. We conducted a case–control study in two Austrian haemophilia centres with 53 patients (mean age 36.7 ± 10.6 years) and 104 male controls (mean age 36.7 ± 11.1 years). Socio‐demographic data were collected using a standardized questionnaire and quality‐of‐life data using the SF‐36. More patients (56.6%) than controls (37.5%) were married (P = 0.023), whereas more controls (17.3%) than patients (3.8%) had a partner with whom they were not married (P = 0.016). The percentage having children was equivalent in both groups (47% and 41% respectively), but controls had more children (mean number 1.5 in patients and 2.1 in controls, P < 0.007). A greater number of patients was unemployed (34% of patients, 9% of controls, P < 0.001) as well as retired (23% and 4% respectively; P < 0.001). Patients had worse scores regarding physical functioning, role‐physical, bodily pain and general health (P < 0.001), whereas vitality, social functioning, role‐emotional and mental health were similar in both groups. Despite their disability, most of the Austrian haemophiliacs share a sound family environment. This suggests that they are highly capable of coping with their chronic disease and is indicated by good scores for role‐emotional and mental health.
Application of intron 9 and intron 25 dinucleotide repeats of the factor VIII gene for carrier diagnosis in haemophilia A
Authors: VENCESLÁ A. , BAENA M. , FARES TAIE L. , CORNET M. , BAIGET M. , TIZZANO E. F.
Published: April 2008

Summary.  We describe the usefulness of two dinucleotide repeats located in intron 9 and in intron 25 of the factor VIII gene for carrier diagnosis of haemophilia A. We analyzed 100 unrelated Spanish women and 34 women from haemophilia A (HA) families in whom known intragenic markers were unhelpful in determining their carrier status. The heterozygosity rate of intron 9 and intron 25 markers in the 100 control women was lower (0.28 and 0.38, respectively) than the values obtained with common markers routinely used in our laboratory. However, the application of intron 9 and intron 25 markers was effective in identifying the at‐risk X chromosome in 11 of 34 (32%) of the uninformative women from HA families. The combined use of these repeats with current markers may facilitate the identification of the X chromosome in HA families for application in carrier, prenatal and pre‐implantation diagnoses.
Molar incisor hypomineralization: a study of aetiological factors in a group of UK children*
Authors: WHATLING ROSEMARY , FEARNE JANICE M
Published: April 2008

Objectives.  The objectives were to investigate the aetiology of molar incisor hypomineralization (MIH), and to discuss previously published studies.

Subjects and methods.  One hundred and nine children attending the Department of Paediatric Dentistry at the Royal London Hospital were included in the study: 57 children with MIH and 52 controls. Their mothers completed a medical history interview.

Results.  No significant associations were found with MIH and delivery and birth complications, breastfeeding, immunization history, other illnesses and allergies, general anaesthetics, fluoride history, and trauma or abscesses affecting the primary predecessors. A family history of enamel defects was more commonly reported for MIH children, but the association was not statistically significant. However, MIH was significantly more common among those whose mothers had experienced problems during pregnancy (P = 0.025), those who had chickenpox between the ages of 3 and 3.99 (P = 0.047), and those for whom amoxycillin was the only antibiotic they had received (P = 0.028).

Conclusions.  The aetiology of MIH remains unclear, and this study questions whether it is because of a lone aetiological insult. This study recommends further research looking at the links with chickenpox occurring around the third year of life and amoxycillin.
Clinical performance of a diode laser fluorescence device for the detection of occlusal caries in primary teeth
Authors: KAVVADIA KATERINA , LAGOUVARDOS PANAGIOTIS
Published: April 2008

Objectives.  To correlate the DIAGNOdentTM readings (LF) with those of direct visual (DV) examination, indirect visual (IDV) examination, bitewing radiography (BWR), and pit and fissure opening (PFO) for the detection of occlusal caries in primary teeth; to determine the validity of this device using PFO as reference; and to evaluate its reliability.

Methods.  Two calibrated operators examined 130 teeth with 405 examination sites using DV, IDV and radiographic examinations, and LF. Of the 405 sites, 155 were visually scored for caries after pit and fissure opening.

Results.  Pit and fissure opening was found to significantly correlate with all methods (rhoLF = 0.48, rhoBWX = 0.48, rhoDV = 0.44, rhoIDV = 0.41). For enamel lesions, higher sensitivity (0.76) was found with DV, while higher specificity (0.88) with the LF. For lesions into dentin, however, higher sensitivity (0.78) was found with the LF, while higher specificity (0.98) with the BWR. The device's accuracy was found to be 0.61 for enamel lesions, while for lesions into dentin 0.70, and its reliability was excellent (ICC = 0.97).

Conclusion.  The LF device presented high reliability in the detection of occlusal caries in primary teeth and its performance was similar to DV and radiographic examinations.
Historical biogeography and post‐glacial recolonization of South American temperate rain forest by the relictual marsupial Dromiciops gliroides
Authors: Himes Christopher M. T. , Gallardo Milton H. , Kenagy G. J.
Published: April 2008

Aim  Long‐term climatic variation has generated historical expansions and contractions of species ranges, with accompanying fragmentation and population bottlenecks, which are evidenced by spatial variation in genetic structure of populations. We examine here hypotheses concerning dispersal and vicariance in response to historical geoclimatic change and potential isolation produced by mountains and water barriers.

Location  The temperate rain forest of southern South America, which is distributed from coastal Chile, including the large continental island of Chiloé, across the Andes into Argentina.

Methods  We investigated our hypotheses in the phylogenetically and biogeographically relictual marsupial Dromiciops gliroides. We examined 56 specimens, which resulted from field samples and museum study skins from 21 localities. We evaluated the influence of two major barriers, the Andean cordillera and the waterway between the mainland and the large island of Chiloé, by performing Bayesian and maximum‐likelihood phylogenetic analyses on sequences of 877 base pairs of mitochondrial DNA. We further tested the contribution of the proposed geographical barriers using analysis of molecular variance (amova). We also evaluated the responses of populations to historical north–south shifts of habitat associated with glacial history and sea‐level change.

Results  Our analyses revealed a phylogeny with three clades, two of which are widespread and contain nearly all the haplotypes: a northern clade (36–39° S) and a southern clade (40–43° S). These two clades contain forms from both sides of the Andes. Within the southern clade, island and mainland forms were not significantly differentiated. Tests of recent demographic change revealed that southern populations have experienced recent expansion, whereas northern populations exhibit long‐term stability. The direction of recent gene flow and range expansion is predominantly from Chile to Argentina, with a modest reciprocal exchange across the Andes. Recent gene flow from the island of Chiloé to the mainland is also supported.

Main conclusions  The genetic structure of contemporary D. gliroides populations suggests recent gene flow across the Andes and between the mainland and the island of Chiloé. Differences in demographic history that we detected between northern and southern populations have resulted from historical southward shifts of habitat associated with glacial recession in South America. Our results add to a growing literature that demonstrates the value of genetic data to illuminate how environmental history shapes species range and population structure.
Mapping the evolutionary twilight zone: molecular markers, populations and geography
Authors: Diniz‐Filho José Alexandre Felizola , De Campos Telles Mariana Pires , Bonatto Sandro L. , Eizirik Eduardo , De Freitas Thales Renato O. , De Marco Paulo , Santos Fabrício R. , Sole‐Cava Antônio , Soares Thannya Nascimento
Published: April 2008

Since evolutionary processes, such as dispersal, adaptation and drift, occur in a geographical context, at multiple hierarchical levels, biogeography provides a central and important unifying framework for understanding the patterns of distribution of life on Earth. However, the advent of molecular markers has allowed a clearer evaluation of the relationships between microevolutionary processes and patterns of genetic divergence among populations in geographical space, triggering the rapid development of many research programmes. Here we provide an overview of the interpretation of patterns of genetic diversity in geographical and ecological space, using both implicit and explicit spatial approaches. We discuss the actual or potential interaction ofphylogeography,molecular ecology,ecological genetics,geographical genetics,landscape geneticsandconservation geneticswith biogeography, identifying their respective roles and their ability to deal with ecological and evolutionary processes at different levels of the biological hierarchy. We also discuss how each of these research programmes can improve strategies for biodiversity conservation. A unification of these research programmes is needed to better achieve their goals, and to do this it is important to develop cross‐disciplinary communication and collaborations among geneticists, ecologists, biogeographers and spatial statisticians.
A guar‐enriched wholemeal bread reduces postprandial glucose and insulin responses
Authors: Ellis Peter R. , Burley Victoria J. , Leeds Anthony R. , Peterson David B.
Published: April 2008

We have developed a guar wholemeal bread and tested its physiological efficacy and sensory qualities. The objective of the study was to estimate the optimum concentration of guar in bread required to reduce postprandial glycaemia and insulinaemia without adversely affecting the quality of the bread. Following overnight fasts, 15 normal weight, non‐diabetic subjects consumed meals of wholemeal bread alone (controls) and supplemented with guar at three different concentrations (5, 10 and 15% replacement of wheat flour). Blood samples for glucose and insulin analysis were taken preprandially, and at 30 and 60 min after the start of each meal. The acceptability of the wholemeal control and guar breads was assessed using a hedonic scaling method. A significantly lower blood glucose rise at 30 min was found after the 15% guar bread meal compared to the control. Plasma insulin responses at 30 and 60 min were significantly lower after the 10 and 15% guar bread meals compared to the controls. Hedonic scores indicated that the subjects found the 5 and 10% guar breads acceptable and in the case of the 5% variety the mean score was identical to the control. We suggest that a guar concentration of between 5 and 10% replacement of wheat flour (wholemeal or white) should be used for the baking of guar wholemeal bread but further work is needed to define the precise quantity of guar required.
Dietary assessment in children with juvenile chronic arthritis
Authors: Portinsson S. , Åkesson A. , Svantesson H. , Akesson B.
Published: April 2008

Dietary habits and nutrient intake were studied in girls with juvenile chronic arthritis (JCA) and in healthy controls. The intake of iron and vitamin D was lower than the recommended dietary allowances in both groups, while the intake of the other nutrients studied were close to or exceeded the recommendations. Only minor differences in nutrient intake between patients and controls were observed.

The interindividual variation in dietary habits was considerable in both groups. The girls with JCA consumed less fruit, vegetables and juices and also ate these foods less often in composite meals. The intake of ascorbic acid was positively correlated with plasma ascorbic acid concentration, but no correlation of plasma concentration of 25‐hydroxyvitamin D and α‐tocopherol with the consumption of the respective vitamin was found.
The use of a visual analogue scale in measuring nutritional status
Authors: George Andrew , Gilmour Elizabeth R. , Crichton Sheila , Eastwood Martin
Published: April 2008

A 100 mm visual analogue scale (VAS) has been used to measure nutritional status in 94 patients receiving supplementary nutrition or evaluation in a Gastrointestinal Unit. The patients described normal distribution with a median value of 50 mm (range 30 mm‐77 mm). There is good correlation between VAS and objective anthropometric measures. VAS is a useful simple measure of recording nutritional status.
Radiation‐induced glioblastoma in a medulloblastoma patient: A case report with molecular features
Authors: Gessi Marco , Maderna Emanuela , Guzzetti Sara , Cefalo Graziella , Massimino Maura , Solero Carlo L. , Finocchiaro Gaetano , Pollo Bianca
Published: April 2008

We report a case of glioblastoma (GBM) occurring 8 years after radiation therapy for a medulloblastoma. A 15‐year‐old boy underwent surgery and radiotherapy for a medulloblastoma and 8 years later he developed a second tumor at the same site. The second lesion showed different histological and molecular features, was diagnosed as a glioblastoma and fulfilled the criteria of radiation‐induced neoplasm. Mutational analysis of the p53 gene showed a C to G transition at codon 176 in tumor DNA. LOH was detected at 17p and 19q. The tumor also showed O6‐methylguanine‐DNA methyl‐transferase (MGMT) promoter methylation and no amplification of EGF receptor. In conclusion, the radiation‐induced MGMT hyper‐methylation and p53 mutations may have a role in the development of a subgroup of radio‐induced glioma (RIG), suggesting that these molecular alterations directly cooperate in the genesis of the post‐irradiation GBM. Moreover RIGs seem to be a heterogeneous group of tumors that may resemble either primary or secondary GBM.
Characterization of Numb expression in astrocytomas
Authors: Yan Benedict , Omar Feroz Mohd , Das Kakoli , Ng Wai Hoe , Lim Chinghway , Shiuan Koh , Yap Celestial T. , Salto‐Tellez Manuel
Published: April 2008

During early neurodevelopment, asymmetric segregation of Numb in mitotic progenitor cells influences the fate of daughter cells, whereby one daughter retains the progenitor phenotype while the other proceeds along a differentiation pathway. Numb has also been reported to function as a tumor suppressor in breast cancers and medulloblastomas. Given its role in maintaining neural progenitor pools in animal models and its reported role as a tumor suppressor, Numb could potentially contribute to astrocytoma oncogenesis. We characterized Numb expression in both human astrocytoma tissue samples and glioblastoma cell lines. We found that Numb is expressed in all grades of astrocytomas, being predominantly cytoplasmic in higher‐grade astrocytomas but nuclear in pilocytic astrocytomas. Numb is also present in normal neurons, but not in normal astrocytes. In cultured glioblastoma cells, Numb concentrates in the perinuclear region and process tips. Numb expression in astrocytomas recapitulates that of progenitor cells during neurodevelopment, and suggests a role for Numb in astrocytoma oncogenesis.
Neuronal toxicity of expanded polyglutamine depends on intracellular distribution in addition to the expression level
Authors: Satoh Mamoru , Shimada Atsuyoshi , Kawamura Noriko , Chiba Yoichi , Yoshikawa Keisuke , Ishii Sanae , Furukawa Ayako , Kumagai Naoko , Hosokawa Masanori
Published: April 2008

The proteins that accumulate in pathologic lesions of neurodegenerative disorders are thought to be closely associated with neuronal cell damage. However, whether or not the formation of cytoplasmic or nuclear inclusions by expanded polyglutamine (polyQ) is directly toxic to neurons has been controversial to date. We prepared a culture model system in which polyQ tracts were transfected into Neuro2a, cells of neuronal origin, to study novel factors involved in cell toxicity of polyQ tracts to neuronal cells. Pathogenic polyQ tracts of 79 repeats (Q79C) when expressed in cytoplasm of Neuro2a cells changed in their intracellular distribution patterns from homogeneous, via punctate aggregates, to massive aggregates with incubation time. Some polyQ tracts formed nuclear inclusions. Cytoplasmic massive aggregates of Q79C tended to be associated with apoptotic fate of Neuro2a cells. Cells exhibiting cytoplasmic massive inclusions had the highest expression level of polyQ tracts among cells with four patterns of intracellular distribution. The elevation in the expression levels of polyQ tracts was not due to the difference in the initial transfection efficiency. When compared among cells expressing polyQ tracts at similar levels, damages were most remarkable in cells with cytoplasmic massive aggregate in terms of shrunken cellular and nuclear sizes. Cells with the other patterns of polyQ tract distribution such as cytoplasmic homogeneous, cytoplasmic punctate and nuclear inclusions were relatively spared. These data suggest that the severity of cell damages depends on the type of intracellular distribution of polyQ tracts, in addition to the expression level of polyQ tracts.
‘It'll never happen to me’: understanding public awareness of local flood risk
Authors: Burningham Kate , Fielding Jane , Thrush Diana
Published: April 2008

Following the severe flood events of 1998 and 2000, the United Kingdom's Environment Agency prioritised the need to increase public flood risk awareness. Drawing on data collected during research undertaken for the Environment Agency, this paper contributes to understanding of one aspect of flood awareness: people's recognition that their property is in an area that is potentially at risk of flooding. Quantitative analyses indicate that class is the most influential factor in predicting flood risk awareness, followed by flood experience and length of time in residence. There are also significant area differences. Our qualitative work explores how those defined as ‘at risk’ account for their lack of awareness or concern about their risk status. We conclude that the problem is often not simply a lack of awareness, but rather, assessments of local risk based on experience that underestimate the impact of rare or extreme events. We underline the importance of engaging with local perspectives on risk and making local people part of ‘awareness‐raising’ processes.
A TAXONOMIC REVISION OF BIARUM
Authors: Boyce Peter C.
Published: April 2008

Summary This paper contains a revision of the genus Biarum Schott. 21 species and 6 subspecies are recognised. All the taxa (except B. mendax Boyce) are illustrated by line drawings, and most of them are shown as paintings or as photographs in habitat or in cultivation. Distribution maps of all the species are given and a key to all taxa is provided.
Bayesian Semiparametric Cure Rate Model with an Unknown Threshold
Authors: NIETO‐BARAJAS LUIS E. , YIN GUOSHENG
Published: April 2008

Abstract.  We propose a Bayesian semiparametric model for survival data with a cure fraction. We explicitly consider a finite cure time in the model, which allows us to separate the cured and the uncured populations. We take a mixture prior of a Markov gamma process and a point mass at zero to model the baseline hazard rate function of the entire population. We focus on estimating the cure threshold after which subjects are considered cured. We can incorporate covariates through a structure similar to the proportional hazards model and allow the cure threshold also to depend on the covariates. For illustration, we undertake simulation studies and a full Bayesian analysis of a bone marrow transplant data set.
Nonlinear Censored Regression Using Synthetic Data
Authors: DELECROIX MICHEL , LOPEZ OLIVIER , PATILEA VALENTIN
Published: April 2008

Abstract.  The problem of estimating a nonlinear regression model, when the dependent variable is randomly censored, is considered. The parameter of the model is estimated by least squares using synthetic data. Consistency and asymptotic normality of the least squares estimators are derived. The proofs are based on a novel approach that uses i.i.d. representations of synthetic data through Kaplan–Meier integrals. The asymptotic results are supported by a small simulation study.
Alan Lyell and Lyell's syndrome
Authors: Revuz J
Published: April 2008

Alan Lyell has done a major contribution to the description of several diseases which could be Lyell's diseases if he had not been the most unpretentious gentelman in the field of dermatology.
Treatment of classic Kaposi sarcoma with a nicotine dermal patch: a phase II clinical trial
Authors: Goedert JJ , Scoppio BM , Pfeiffer R , Neve L , Federici AB , Long LR , Dolan BM , Brambati M , Bellinvia M , Lauria C , Preiss L , Boneschi V , Whitby D , Brambilla L
Published: April 2008

Background  Kaposi sarcoma (KS), a malignancy of dermal endothelial cells that is caused by human herpesvirus 8 (HHV8) infection, is sensitive to perturbations of immunity. Nicotine might be effective against KS because of its immunologic and vascular effects and because smoking is associated with a low risk of KS.

Objective and study design  We conducted a masked, randomized phase 2 clinical trial of transdermal nicotine and placebo patches to assess the safety and efficacy of nicotine against classic KS (cKS).

Subjects and methods  Three cKS lesions, predominantly nodules, in each of 24 non‐smoking patients were randomly assigned to 15 weeks continuous treatment with nicotine patch (escalated to 7 mg), identical masked placebo patch or no patch. Changes in lesion area and elevation from baseline through six follow‐up visits, by direct measurement and by two independent readers using digital photographs of the lesions, were compared using non‐parametric and regression methods. Changes in longitudinal levels of HHV8 antibodies and DNA in blood cells were similarly assessed.

Results  There were no systemic or serious adverse events, and compliance was good. One patient resumed smoking and discontinued patches, and two patients withdrew at week 12 for unrelated indications. Six (29%) of the remaining 21 suspended use of patches to relieve local skin irritation; four of these six completed the trial at reduced dose. Treatment assignment was not associated with significant or consistent changes in cKS lesion area or elevation, HHV8 viral load or antibodies.

Conclusion  Transdermal nicotine and placebo patches caused no serious toxicities but had no demonstrable effect on nodular cKS lesions or HHV8 levels.
The Impact of Checking the Health of Adults with Intellectual Disabilities on Primary Care Consultation Rates, Health Promotion and Contact with Specialists
Authors: Felce David , Baxter Helen , Lowe Kathy , Dunstan Frank , Houston Helen , Jones Glyn , Grey Jill , Felce Janet , Kerr Michael
Published: April 2008

Background  Studies have found that health checking in primary care led to the identification of previously unrecognized morbidity among adults with intellectual disabilities. The aim here was to evaluate whether health checking stimulated increased consultation with the general practitioner or another member of the primary care team, increased health promotion actions undertaken outside the health check or increased contact with specialists.

Method  Data on the above three categories of activity were abstracted from the medical records of 77 adult participants with intellectual disabilities for eight 6‐month periods before and seven 6 month periods after they had undergone a health check. Comparisons of access to care before and after the health check were made using non‐parametric statistics.

Results  On average, participants had 5.4 and 1.8 primary care and specialist consultations per year respectively. There were no significant differences in either rate before and after the health check. The frequency of health promotion actions increased significantly after the health check from a mean of 1.2 to 2.2/year.

Conclusions  Comparison of the primary care and specialist consultations rates of people with intellectual disabilities with those for the general population might suggest that the former have greater access to these services. However, comparison to the general practitioner consultation rates of patients with other chronic conditions would seem to indicate that contact with primary care may not be commensurate with need. Attention to health promotion is inadequate. Further research is required to substantiate whether health checking increases health promotion and how increased health promotion activity would affect the health of this population.