bluuebunny/crossref_metadata_2025 huggingface dataset:
FlAsH-FALI Inactivation of a Protein at the Third-Instar Neuromuscular Junction: Figure 1.
Authors: Habets Ron L.P. , Verstreken Patrik
Published: April 2011

INTRODUCTION

Fluorescein-assisted light inactivation (FALI) is a powerful method for studying acute loss of protein function, even if the corresponding mutations lead to early lethality. In this protocol, FALI is mediated by the membrane-permeable FlAsH (4′,5′-bis(1,3,2-dithioarsolan-2-yl)fluorescein) compound that binds with high specificity to the genetically encoded tetracysteine tag and thus allows the inactivation of protein function in vivo with exquisite spatial (<40 Å) and temporal (<30 sec) resolution. It also enables the analysis of kinetically distinct processes such as synaptic vesicle exocytosis and endocytosis. This protocol describes efficient inactivation of a protein using FlAsH-FALI at the neuromuscular junction (NMJ) of third-instar larvae. Note that FlAsH-FALI in other tissues is also theoretically possible with minor adaptations to the protocol described here. We explain controls for positional effects, for unspecific FlAsH binding to endogenous proteins, and for phototoxicity. Following FlAsH-FALI, protein function can be studied using a number of secondary assays, including electrophysiology, immunohistochemistry, and electron microscopy or FM1-43 labeling of synaptic vesicle pools.
Megaselia abdita: Cuticle Preparation from Injected Embryos: Figure 1.
Authors: Rafiqi Ab. Matteen , Lemke Steffen , Schmidt-Ott Urs
Published: April 2011

INTRODUCTION

This protocol describes how to prepare cuticles of Megaselia abdita embryos that have been injected through the chorion and developed under oil.
Making Yeast Cell Extracts for Purifying TAP-tagged Protein Complexes
Authors: Link Andrew J. , Weaver Connie , Farley Adam
Published: April 2011

INTRODUCTION

One approach to identifying protein–protein interactions is the biochemical purification of a target protein from cells or tissues under nondenaturing conditions followed by the mass spectrometric identification of the components of the purified protein complex. The combination of highly specific protein purification strategies and mass spectrometry has proven to be a very successful approach for identifying protein–protein interactions. The tandem affinity purification (TAP) affinity tag and purification method allows efficient recovery of proteins present at low cellular concentrations under native conditions. Expressing the target protein at its natural levels avoids the assembly of overexpressed proteins into nonphysiological complexes. This protocol describes the preparation of extract from TAP-tagged yeast cells for use in purifying TAP-tagged protein complexes.
Maintaining Live Cells and Tissue Slices in the Imaging Setup
Authors: Dailey Michael E. , Marrs Glen S. , Kurpius Dana
Published: April 2011

INTRODUCTION

Live imaging provides exciting opportunities to study dynamic cellular events as they occur in real time. However, such experimental procedures present several challenges. This article discusses some of the major considerations relating to the maintenance of live biological samples during ex vivo imaging and presents some relatively simple low-cost solutions for keeping samples healthy.
Fast computation of scattering maps of nanostructures using graphical processing units
Authors: Favre-Nicolin Vincent , Coraux Johann , Richard Marie-Ingrid , Renevier Hubert
Published: April 2011

Scattering maps from strained or disordered nanostructures around a Bragg reflection can be either computed quickly using approximations and a (fast) Fourier transform or obtained using individual atomic positions. In this article, it is shown that it is possible to compute up to 4 × 1010 reflections atoms s−1using a single graphics card, and the manner in which this speed depends on the number of atoms and points in reciprocal space is evaluated. An open-source software library (PyNX) allowing easy scattering computations (including grazing-incidence conditions) in the Python language is described, with examples of scattering from non-ideal nanostructures.
Mothers’ perceptions of factors affecting their abilities to care for infants with allergy
Authors: Alanne Soili , Laitinen Kirsi , Söderlund Riitta , Paavilainen Eija
Published: April 2011

Aim.  To identify mothers’ perceptions of the supporting or straining factors involved in motherhood generally and in relation to child’s atopic dermatitis and food allergy at the age of 1–2 years.

Background.  The main person who cares for a child in the early years of life is usually the mother. Information on mothers’ perceptions of factors affecting their resources to care for their healthy or allergic children is incomplete and needs to be identified.

Design.  A descriptive qualitative study with semi‐structured interviews.

Method.  Altogether 32 interviews with mothers having a one‐year‐old healthy or allergic child and repeat interviews with mothers of allergic children aged two years were conducted. Deductive content analysis was applied, based on the concept of empowerment and an empowerment model for family nursing.

Results.  A child’s food allergy at the age of one year caused considerable strain for the mother, chiefly because the adoption of new foods into the child’s diet was delayed, and waking at night because of itching related to atopic dermatitis. This delay was still causing the mother additional strain, when the child was two years old. Specific allergy‐related strengthening factors were the mother’s own knowledge, hope for an improvement, social support and work.

Conclusions.  This qualitative study succeeded in identifying strain situations for mothers with one‐ and two‐year‐old allergic children. Such mothers should be recognised and provided with individual support by nurses and healthcare professionals to focus on the supporting factors.

Relevance to clinical practice.  Mothers of allergic children should be recognised. An inquiry or questionnaire based on personal abilities and the family’s internal and external capabilities may contribute to identifying these mothers. Mothers should be provided with individual encouragement and support from mother and child health clinics by focusing on factors felt to be strengthening ones.
SEC14 is a specific requirement for secretion of phospholipase B1 and pathogenicity of Cryptococcus neoformans
Authors: Chayakulkeeree Methee , Johnston Simon Andrew , Oei Johanes Bijosono , Lev Sophie , Williamson Peter Richard , Wilson Christabel Frewen , Zuo Xiaoming , Leal Ana Lusia , Vainstein Marilene Henning , Meyer Wieland , Sorrell Tania Christine , May Robin Charles , Djordjevic Julianne Teresa
Published: April 2011

Secreted phospholipase B1 (CnPlb1) is essential for dissemination of Cryptococcus neoformans to the central nervous system (CNS) yet essential components of its secretion machinery remain to be elucidated. Using gene deletion analysis we demonstrate that CnPlb1 secretion is dependent on the CnSEC14 product, CnSec14‐1p. CnSec14‐1p is a homologue of the phosphatidylinositol transfer protein ScSec14p, which is essential for secretion and viability in Saccharomyces cerevisiae. In contrast to CnPlb1, neither laccase 1‐induced melanization within the cell wall nor capsule induction were negatively impacted in CnSEC14‐1 deletion mutants (CnΔsec14‐1 and CnΔsec14‐1CnΔsfh5). Similar to the CnPLB1 deletion mutant (CnΔplb1), CnΔsec14‐1 was hypovirulent in mice and did not disseminate to the CNS by day 14 post infection. Furthermore, macrophage expulsion of live CnΔsec14‐1 and CnΔplb1 (vomocytosis) was reduced. Individual deletion of CnSEC14‐2, a closely related CnSEC14‐1 homologue, and CnSFH5, a distantly related SEC fourteen like homologue, did not abrogate CnPlb1 secretion or virulence. However, reconstitution of CnΔsec14‐1 with CnSEC14‐1 or CnSEC14‐2 restored both phenotypes, consistent with functional genetic redundancy. We conclude that CnPlb1 secretion is SEC14‐dependent and that C. neoformans preferentially exports virulence determinants to the cell periphery via distinct pathways. We also demonstrate that CnPlb1 secretion is essential for vomocytosis.
Cannabinoid receptor expression in HIV encephalitis and HIV‐associated neuropathologic comorbidities
Authors: Cosenza‐Nashat M. A. , Bauman A. , Zhao M.‐L. , Morgello S. , Suh H.‐S. , Lee S. C.
Published: March 2011

M. A. Cosenza‐Nashat, A. Bauman, M.‐L. Zhao, S. Morgello, H.‐S. Suh and S. C. Lee (2011) Neuropathology and Applied Neurobiology37, 464–483

Cannabinoid receptor expression in HIV encephalitis and HIV‐associated neuropathologic comorbidities

Aims: Cannabinoids have been proposed for treating various neurodegenerative disorders and as adjunct therapy for HIV+ patients with neurologic sequelae. The expression of cannabinoid receptors (CB1 and CB2) has been reported in neurodegenerative diseases and in simian immunodeficiency virus encephalitis, yet the receptor expression in the central nervous system of HIV+ individuals is not known. Methods: An anti‐CB1 antibody and two anti‐CB2 antibodies were employed for immunohistochemistry in the cerebral cortex and white matter of HIV encephalitis (HIVE) and HIV‐associated comorbidities, as well as control brains (HIV− and HIV+). Results: By quantitative image analysis, we observed that CB1 was increased in HIVE brains and those with comorbidities, while CB2 was significantly increased in the white matter of HIVE. Morphologically, CB1 was present in neurones, and both CB1 and CB2 were present in meningeal macrophages and subpial glia in all brains. In HIVE, CB1 was found in white matter microglia and perivascular cells, while CB2 was increased in microglia, astrocytes and perivascular macrophages. Double immunofluorescence with cell‐specific markers and immunoblots on primary cultured microglia and astrocytes substantiated the glial localization of the cannabinoid receptors and specificity of the antibodies. Conclusions: Our study indicates that cannabinoid receptor expression occurs in glia in HIVE brains, and this may have ramifications for the potential use of cannabinoid ligands in HIV‐infected patients.
The microRNA miR393 re‐directs secondary metabolite biosynthesis away from camalexin and towards glucosinolates
Authors: Robert‐Seilaniantz Alexandre , MacLean Dan , Jikumaru Yusuke , Hill Lionel , Yamaguchi Shinjiro , Kamiya Yuji , Jones Jonathan D. G.
Published: April 2011

flg22 treatment increases levels of miR393, a microRNA that targets auxin receptors. Over‐expression of miR393 renders plants more resistant to biotroph pathogens and more susceptible to necrotroph pathogens. In contrast, over‐expression of AFB1, an auxin receptor whose mRNA is partially resistant to miR393 degradation, renders the plant more susceptible to biotroph pathogens. Here we investigate the mechanism by which auxin signalling and miR393 influence plant defence. We show that auxin signalling represses SA levels and signalling. We also show that miR393 represses auxin signalling, preventing it from antagonizing SA signalling. In addition, over‐expression of miR393 increases glucosinolate levels and decreases the levels of camalexin. Further studies on pathogen interactions in auxin signalling mutants revealed that ARF1 and ARF9 negatively regulate glucosinolate accumulation, and that ARF9 positively regulates camalexin accumulation. We propose that the action of miR393 on auxin signalling triggers two complementary responses. First, it prevents suppression of SA levels by auxin. Second, it stabilizes ARF1 and ARF9 in inactive complexes. As a result, the plant is able to mount a full SA response and to re‐direct metabolic flow toward the most effective anti‐microbial compounds for biotroph resistance. We propose that miR393 levels can fine‐tune plant defences and prioritize resources.
Members of the RKD transcription factor family induce an egg cell‐like gene expression program
Authors: Kőszegi Dávid , Johnston Amal J. , Rutten Twan , Czihal Andreas , Altschmied Lothar , Kumlehn Jochen , Wüst Samuel E. J. , Kirioukhova Olga , Gheyselinck Jacqueline , Grossniklaus Ueli , Bäumlein Helmut
Published: April 2011

In contrast to animals, the life cycle of higher plants alternates between a gamete‐producing (gametophyte) and a spore‐producing (sporophyte) generation. The female gametophyte of angiosperms consists of four distinct cell types, including two gametes, the egg and the central cell, which give rise to embryo and endosperm, respectively. Based on a combined subtractive hybridization and virtual subtraction approach in wheat (Triticum aestivum L.), we have isolated a class of transcription factors not found in animal genomes, the RKD (RWP‐RK domain‐containing) factors, which share a highly conserved RWP‐RK domain. Single‐cell RT‐PCR revealed that the genes TaRKD1 and TaRKD2 are preferentially expressed in the egg cell of wheat. The Arabidopsis genome contains five RKD genes, at least two of them, AtRKD1 and AtRKD2, are preferentially expressed in the egg cell of Arabidopsis. Ectopic expression of the AtRKD1 and AtRKD2 genes induces cell proliferation and the expression of an egg cell marker. Analyses of RKD‐induced proliferating cells exhibit a shift of gene expression towards an egg cell‐like transcriptome. Promoters of selected RKD‐induced genes were shown to be predominantly active in the egg cell and can be activated by RKD in a transient protoplast expression assay. The data show that egg cell‐specific RKD factors control a transcriptional program, which is characteristic for plant egg cells.
Severe α‐1 antitrypsin deficiency caused by Q0Ourém allele: clinical features, haplotype characterization and history
Authors: Vaz Rodrigues L , Costa F , Marques P , Mendonça C , Rocha J , Seixas S
Published: April 2011

Vaz Rodrigues L, Costa F, Marques P, Mendonça C, Rocha J, Seixas S. Severe α‐1 antitrypsin deficiency caused by Q0Ourém allele: clinical features, haplotype characterization and history.

α‐1 Antitrypsin deficiency (AATD) caused by null alleles is associated with the total lack of protein and generally it translates into more severe clinical features of pulmonary disease. This is the case of Q0Ourém, a rare variant found in several families of Central Portugal caused by the L353fsX376 mutation. A total of 41 patients carrying at least one copy of Q0Ourém were evaluated for SERPINA1 levels, respiratory function values and lung parenchyma status (chest X‐ray and computerized tomography scan). Q0Ourém haplotype background was characterized using seven microsatellites flanking SERPINA1 and Q0Ourém age was estimated by a statistical method relying on the decay of haplotype sharing at linked markers (DHSMAP).

Homozygous patients showed a compromised lung function and extensive emphysema. SQ0Ourém, although having serum levels below the 11 µM threshold, did not necessarily result in signs of disease. MQ0Ourém were found to be a heterogeneous group, mainly composed of normal individuals. Eight Q0Ourém haplotypes were identified and the allele was estimated to have arisen 650 years ago. Q0Ourém was associated with mild to severe AATD and has a single origin, probably linked to the major Ourém settlements where the occurrence of severe AATD may not be explained by recent consanguinity.
Avoiding high relative air humidity during critical stages of leaf ontogeny is decisive for stomatal functioning
Authors: Fanourakis Dimitrios , Carvalho Susana M. P. , Almeida Domingos P. F. , Heuvelink Ep
Published: April 2011

Plants of several species, if grown at high relative air humidity (RH ≥85%), develop stomata that fail to close fully in case of low leaf water potential. We studied the effect of a reciprocal change in RH, at different stages of leaf expansion of Rosa hybrida grown at moderate (60%) or high (95%) RH, on the stomatal closing ability. This was assessed by measuring the leaf transpiration rate in response to desiccation once the leaves had fully expanded. For leaves that started expanding at high RH but completed their expansion after transfer to moderate RH, the earlier this switch took place the better the stomatal functioning. Leaves initially expanding at moderate RH and transferred to high RH exhibited poor stomatal functioning, even when this transfer occurred very late during leaf expansion. Applying a daily abscisic acid (ABA) solution to the leaves of plants grown at continuous high RH was effective in inducing stomatal closure at low water potential, if done before full leaf expansion (FLE). After FLE, stomatal functioning was no longer affected either by the RH or ABA level. The results indicate that the degree of stomatal adaptation depends on both the timing and duration of exposure to high RH. It is concluded that stomatal functionality is strongly dependent on the humidity at which the leaf completed its expansion. The data also show that the effect of ambient RH and the alleviating role of ABA are restricted to the period of leaf expansion.
Chromatin remodelling in plant light signalling
Authors: Fisher Alex J. , Franklin Keara A.
Published: April 2011

The structure and compaction of chromatin exerts a major regulatory influence on eukaryotic transcription. Changes in both histone composition and post‐translational modifications of individual histone proteins can lead to remodelling of higher order chromatin structure. Chromatin remodelling regulates transcriptional activity through modifying gene accessibility, via DNA/histone interactions and the recruitment of non‐histone proteins to DNA. Plant growth and development is regulated by the integration of multiple environmental signals. Of these, light is one of the most important. Chromatin remodelling processes have been identified in plants following a variety of different light treatments. These include the initiation of seedling de‐etiolation, changes in photon irradiance and ultraviolet‐B radiation exposure. In this review, we will summarize the roles of chromatin remodelling in plant photomorphogenesis and discuss these in the wider context of plant environmental adaptation.
60Co‐γ radiation induces differential acetylation and phosphorylation of histones H3 and H4 in wheat
Authors: Raut V. V. , Sainis J. K.
Published: April 2011

Histone modifications occur during DNA damage and repair in eukaryotes. These modifications were analysed in wheat seedlings exposed to 60Co‐γ radiation. Seedling height was not significantly affected in the first 2 days after irradiation up to 150 Gy. Subsequently, in the next 2 weeks, there was 30–40% reduction in seedling height, indicating that there were late effects of irradiation. The histones isolated from irradiated seedlings were analysed in the initial stages for modifications of H3 and H4 using antibodies. Global acetylation of H3 decreased and H4 increased in a dose‐dependent manner till 100 Gy. The time course of individual modifications showed that for H3K4 and H3K9, acetylation decreased, whereas for H3S10 phosphorylation increased. There were fluctuations in acetylation of H4K5, H4K12 and H4K16, whereas H4K8 showed hyper‐acetylation. The results indicate that γ radiation induced DNA damage and repair in wheat seedlings and initiated differential acetylation of H3 and H4. This is the first report in plants on site‐specific H3 and H4 modifications in response to exposure to ionizing radiation.
Initial Lung Lesions in Two Calves Experimentally Infected with Haemophilus somnus
Authors: Tegtmeier C. , Bloch B. , Jensen N. E. , Jensen H. E.
Published: March 2011

The initial lung lesions in two calves intrabronchially inoculated with Haemophilus somnus are described. The animals were euthanized within 7 h after challenge. The in situ location of H. somnus and accompanying lesions were examined by light microscopy, immunohistochemistry and transmission electron microscopy (TEM).

Inoculation with H. somnus resulted in the development of acute pulmonary lesions within 3.5 h. H. somnus antigen was demonstrated only within the luminal spaces of the airways and in one area of bronchio‐associated lymphoid tissue (BALT). As observed by TEM, the bacteria were phagocytized by both neutrophils and alveolar macrophages. Antigen was never demonstrated in the pulmonary intravascular macrophages.
Patterns of Seroconversion to Aujeszky's Disease Virus in Unvaccinated Chronically Infected Swedish Weaner Pig‐Producing Herds
Authors: Engel M. , Wierup M. , Novén B.
Published: March 2011

This study was carried out in nine unvaccinated Swedish weaner pig‐producing herds, ranging in size from 20 to 134 breeding animals, which had experienced a previous outbreak of Aujeszky's disease (AD) and in which seropositive sows were still present. The objective was to quantify the seroconversion rate to Aujeszky's disease virus (ADV) in breeding animals and to find out whether unvaccinated herds could become free from ADV by using only seronegative animals for replacement. Serum samples were initially obtained from all adult pigs in the herds. Animals testing negative, as well as replacement animals, were subsequently tested every second or third month and the herds were followed for 10–28 months. The herd owners were advised to keep seropositive and seronegative animals separate whenever possible and to implement sanitary measures against the reintroduction of the virus into the herds.

All herds experienced long consecutive periods (≥ 4 months, median 9 months) without seroconversion. Three herds became free from ADV during the serological study; in two of these herds no seroconversion was observed. In those seven herds where seroconversion occurred, between 9 and 86% of the susceptible pigs became infected. The reinfection was indicative of the reactivation of latent virus in all herds but two, where the reintroduction of the virus was most likely. The pattern of seroconversion was either sporadic, involving not more than three animals at a time, or epidemic, involving a large proportion of the susceptible animals in the herd. Severe clinical outbreaks hit the two largest herds and these only became ADV‐free after the conclusion of the study when vaccine was used, which reduced the incidence of seroconversion to zero. The results from the present study show that it is possible for smaller herds to achieve freedom from ADV without any radical control methods, provided that only ADV‐free replacement animals are used and the virus is not reintroduced. Moreover, the finding that long periods can elapse without transmission suggests that when infection is discovered in a herd, the risk of massive spread is not necessarily imminent.
Comparison of Agar Gel Immunodiffusion Test, Enzyme‐linked Immunosorbent Assay and Western Blotting for the Detection of BLV Antibodies
Authors: Dolz G. , Moreno E.
Published: March 2011

An indirect enzyme‐linked immunosorbent assay (ELISA) for the diagnosis of bovine leukaemia virus (BLV) infection was developed and compared with the agar gel immunodiffusion test (AGIDT). Western blotting (WB) was used as confirmatory test. ELISA and AGIDT had specificities that were comparable with that of WB, however, ELISA showed a higher sensitivity than AGIDT. The ELISA was useful for screening a large number of samples, whereas WB was important for detecting the antibody response against the individual BLV‐proteins. Different types of positive serological reactions were discerned in WB, that correlated with reactions of sera in AGIDT and ELISA. The most important antigen in WB and ELISA was the BLV protein p24, whereas the BLV glycoproteins gp51 and gp30 were of special importance in AGIDT. The relevance of repeatedly testing the antibody response in BLV‐infected herds for control and eradication programmes using assays with higher sensitivity than AGIDT was demonstrated.
Longitudinal Study on the Susceptibility to Bacteriophages of Staphylococcus aureus Strains Isolated from Dairy Farms in Trinidad
Authors: Adesiyun A. A. , Romain H. T.
Published: March 2011

A 6‐month longitudinal study was conducted on 30 dairy cows in early lactation and their human handlers on six farms across Trinidad. Weekly samples of bulk milk, composite milk and anterior nares and hand swabs from human handlers were collected and cultured for Staphylococcus aureus on Baird‐Parker agar (BPA). The susceptibility of S. aureus strains to bacteriophages and the relatedness of strains isolated over the study period were determined. Sixty‐three (51.2%) of 123 strains of S. aureus from bulk milk were typable compared with 111 (57.3%) of 194 and 82 (61.7%) of 133 strains isolated from composite milk and human handlers, respectively. The differences were not statistically significant (P > 0.05; χ2). Bovine phage 42D lysed 3.3% (4 of 123), 16.5% (32 of 194) and 12.0% (16 of 133) of S. aureus strains isolated from bulk milk, composite milk and human handlers, respectively. The differences were statistically significant (P < 0.001; χ2). Amongst bulk milk isolates of S. aureus, 35 (31.8%) of 110 exhibited relatedness in 11 groups based on their phage patterns and groups. The mean maximum interval between the first and last detection of related S. aureus strains in a group was 11.5 ± 7.3 weeks. Amongst composite milk strains of S. aureus, 23 (46.0%) of 50, 25 (62.5%) of 40 and 22 (53.7%) of 41 exhibited relatedness on farms IB 2, IB 27 and IC 23, respectively, but the differences were not statistically significant (P > 0.05; χ2). On farm IB 2, five groups of related strains of S. aureus were detected with a mean maximum interval of detection of 18.2 ± 8.5 weeks compared to farm IB 27 where five groups of related strains were also observed but with an interval of 13.8 ± 8.2 weeks. On farm IC 23, a total of seven groups of related S. aureus strains were detected with a mean interval of 8.0 ± 5.5 weeks. For human strains of S. aureus from farm IB 2, nine (56.3%) of 16 strains isolated from anterior nares exhibited relatedness in three groups with a mean maximum interval of 13.3 ± 4.7 weeks compared to four (25.0%) of 16 hand swab isolates which exhibited relatedness in two groups with mean interval of detection of 11.0 ± 1.4 weeks. The differences were not statistically significant (P > 0.05; χ2). On farm IB 27, for anterior nares isolates, eight (72.7%) of 11 exhibited relatedness in two groups with a mean maximum interval of detection of 20.5 ± 2.1 weeks compared to hand swab isolates, with six (50.0%) of 12 showing relatedness in two groups and a mean interval of 10.5 ± 2.1 weeks.

It was concluded that dairy cows and their human handlers carried particular strains of S. aureus at various sites for extended periods, which served as continuous sources of contamination of milk and may play a significant role in the occurrence of subclinical mastitis, with an obvious economic impact.
Effects of recombinant human endostatin on peritoneal angiogenesis in peritoneal dialysis rats
Authors: ZHAO ZHAN‐ZHENG , CAO YING , LIU ZHANG‐SUO , XIAO JING , TANG LIN , WANG PEI , LIANG XIAN‐HUI
Published: April 2011

Aim:  To investigate the effects of recombinant human endostatin (Endostar) on peritoneum angiogenesis in a model of dialysate exposure in rats.

Methods:  Forty male Sprague–Dawley rats were randomized to five groups: normal (group 1); uraemia (group 2); 4.25% peritoneal dialysate (PD) uraemic (group 3); uraemia + PD + recombinant human endostatin 10 mg/kg PD (group 4); and uraemia + PD + recombinant human endostatin 40 mg/kg PD (group 5). The uraemic rats model was established by 5/6 nephrectomy. Endostatin was administrated by s.c. injection every other day, over 28 days. After 28 days of PD fluid exposure, immunohistochemistry and reverse transcript polymerase chain reaction were used to detect protein and mRNA expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in each group. Microvessel density (MVD) was measured by immunohistochemistry.

Results:  Compared with group 1, the mRNA and protein expressions of VEGF and bFGF were significantly upregulated in groups 2 and 3 (P < 0.05). Compared with group 3, the mRNA and protein expressions of VEGF and bFGF were significantly downregulated in groups 4 and 5 (P < 0.05). Compared with group 4, the mRNA and protein expressions of VEGF and bFGF were significantly downregulated in group 5 (P < 0.05). Compared with group 1, MVD was significantly upregulated in groups 2 and 3 (P < 0.05). Compared with group 3, MVD was significantly downregulated in groups 4 and 5 (P < 0.05).

Conclusion:  Endostar can effectively inhibit rat peritoneum neoangiogenesis and the effect was dose‐dependent.
Current and emerging antiglycaemic pharmacological therapies: The renal perspective
Authors: SHARIF ADNAN
Published: April 2011

Diabetes mellitus and chronic kidney disease are two major global epidemics, with a significant overlap of patients with concomitant problems. Therapeutic guidelines for the treatment of diabetes mellitus are continuously updated to reflect the growing armamentarium of antiglycaemic agents at the disposal of clinicians. However, they rarely focus on the significant caveats and limitations associated with pharmacological delivery of glucose‐lowering treatment in the context of advancing kidney disease or in the presence of a renal allograft. Proposed consensus algorithms for the treatment of hyperglycaemia may not be appropriate for individuals with coexisting renal disease and it is imperative to ensure nephrologists maintain a thorough understanding of the limitations of antiglycaemic treatments in the presence of renal insufficiency or a renal allograft. The purpose of this review is to highlight the range of glucose‐lowering therapies at the disposal of the clinician, both currently available and in development, and discuss the advantages and disadvantages of these pharmacological agents from a renal perspective. A tailored and individualized approach to treatment of diabetes mellitus in the context of renal disease is essential to maintain optimum care and this article should act as a supplement to existing guidelines and treatment algorithms.
Inactivation of O6‐methyguanine‐DNA methyltransferase by promoter hypermethylation: Association of epithelial ovarian carcinogenesis in specific histological types
Authors: Roh Hyun‐Jin , Suh Dong‐Soo , Choi Kyung‐Un , Yoo Hang‐Jo , Joo Won‐Deok , Yoon Man‐Soo
Published: March 2011

Aim:  The aim of this study was to evaluate O6‐methyguanine‐DNA methyltransferase (MGMT) promoter hypermethylation, MGMT expression and microsatellite instability (MSI), as well as to elucidate their correlation with clinical and pathological parameters in epithelial ovarian cancer.

Methods:  Ovarian cancer tissue specimens (n = 86) were obtained after a staging operation. The MGMT gene was investigated by methylation‐specific polymerase chain reaction (MSP) and MGMT expression status was analyzed using immunohistochemistry. MSI status was examined by the fluorescence‐based PCR using five National Cancer Institute markers.

Results:  Negative MGMT expression was detected in 12 of 86 (14.0%) epithelial ovarian cancers. In 34 cases where MSP results were available, MGMT promoter hypermethylation was detected in five cases (14.7%) with mucinous or clear cell carcinomas, but not in any of other histological types (P = 0.031). Five out of six cases with negative MGMT expression showed MGMT promoter hypermethylation, whereas all of the 28 cases that retained expression of MGMT were unmethylated at the MGMT CpG island (P < 0.001). In 41 cases of MSI results available, seven (17.1%) cases showed MSI‐H‐phenotyped. Both MGMT promoter hypermethylation and negative MGMT expression were noted only in cases of mucinous or clear cell carcinoma in which MSI status were mostly MSS‐phenotyped; however, no significant correlation was found between MSI status and clinicopathological parameters.

Conclusions:  Negative MGMT expression was significantly correlated with MGMT promoter hypermethylation in MSS‐phenotyped tumors of mucinous or clear cell carcinoma. The results suggest that MGMT promoter hypermethylation might be associated with epithelial ovarian carcinogenesis in specific histological types.
Fetal therapy of severe symptomatic toxoplasmosis using azithromycin
Authors: Tamaru Shunsuke , Kikuchi Akihiko , Takagi Kimiyo , Wakamatsu Masao , Horikoshi Tsuguhiro , Ogiso Yoshifumi
Published: March 2011

Severe symptomatic fetal toxoplasmosis rarely occurs after the maternal primary infection of Toxoplasma gondii. We herein report our experience of fetal therapy of symptomatic toxoplasmosis using azithromycin. Ultrasound assessment at 23 weeks' gestation revealed fetal ascites, cardiac effusion, cardiomegaly, enlarged lateral ventricles and thickened placenta. Serum Toxoplasma gondii antibody titer was ×81 920. Toxoplasma immunoglobulin M was 2.4 index (normal, <0.8 index), and immunoglobulin G was ≥240 IU/mL (normal, <6 IU/mL). Maternal oral administration of azithromycin in addition to sulfadoxine, pyrimethamine and acetylspiramycin was conducted. Spontaneous vaginal delivery occurred at 32 weeks and a male infant weighing 2036 g was born. Hepatosplenomegaly, chorioretinitis, hydrocephalus, intracranial calcifications, ascites, and meningitis were confirmed after birth. The infant underwent therapy with pyrimethamine and sulfadiazine. It seems imperative to establish a new drug choice for fetal therapy of severe symptomatic toxoplasmosis in order to reduce the maternal and fetal risks of drug side‐effects.
Two cases of angular pregnancy complicated by preterm labor and placental abruption at mid‐pregnancy
Authors: Kwon Ji Young , Hwang Seong Jin , Shin Jae Eun , Yoon Won Sik , Shin Jong Chul , Park In Yang
Published: March 2011

Angular pregnancy is the rare condition in which the gestational sac is implanted in the lateral angle of the uterine cavity. There have been several reports on angular pregnancies; however, due to a lack of clinical understanding, angular pregnancy does not appear to be recognized as a clinical entity and many cases are likely to go undiagnosed. This report involves two women with angular pregnancies: one had placenta abruption and the other had preterm delivery at mid‐trimester and postpartum endometritis. The former was diagnosed early with Three‐dimensional sonography, but the other was not diagnosed until the time of cesarean section. This is the first report on the sonographic diagnosis of angular pregnancy. Through these cases, we emphasize the benefit of an accurate early diagnosis of angular pregnancy and careful prenatal care because of the possible risk of adverse perinatal outcome.
Case of acquired hemophilia with factor VIII inhibitor in a mother and newborn
Authors: Kotani Yasushi , Shiota Mitsuru , Umemoto Masahiko , Koike Eiji , Tsuritani Mitsuhiro , Hoshiai Hiroshi
Published: March 2011

We report a mother and newborn in the puerperium with hemorrhage secondary to factor VIII inhibitor. A 31‐year‐old gravida 1 para 1 delivered at a local clinic with a massive postpartum hemorrhage. The activated partial thromboplastin time was prolonged and factor VIII inhibitor was detected. The persistent hemorrhage improved following treatment, including transfusion, steroid therapy, and bypass therapy with factor VII formulations. After hysteroscopic removal of the retained placenta, the hemorrhage decreased. The newborn developed significant swelling of the hands after routine blood sampling and factor VIII inhibitor was detected. The inhibitor disappeared without any special treatment in the 5th month postpartum in the mother and the 4th month postpartum in the newborn. Factor VIII inhibitor may be transferred via the placenta from the mother to the fetus. Therefore, the newborn should also be carefully observed in a case of massive hemorrhage after delivery.
Pivalic acid acts as a starter unit in a fatty acid and antibiotic biosynthetic pathway in Alicyclobacillus, Rhodococcus and Streptomyces
Authors: Řezanka Tomáš , Siristova Lucie , Schreiberová Olga , Řezanka Michal , Masák Jan , Melzoch Karel , Sigler Karel
Published: March 2011

A biosynthetic pathway using pivalic acid as a starter unit was found in three bacterial species, Alicyclobacillus acidoterrestris, Rhodococcus erythropolis and Streptomyces avermitilis. When deuterium‐labelled pivalic acid was added to A. acidoterrestris and R. erythropolis nutrient media it was incorporated into fatty acids to give rise to tert‐butyl fatty acids (t‐FAs). In addition, in R. erythropolis, pivalic acid was transformed into two starter units, i.e. isobutyric and 2‐methylbutyric acid, which served as precursors of corresponding iso‐even FAs and anteiso‐FAs. In S. avermitilis the biosynthesis also yielded all three branched FAs; apart from this pathway, both pivalic and 2‐methylbutyric acids were incorporated into the antibiotic avermectin.
Hydrothermal ecotones and streamer biofilm communities in the Lower Geyser Basin, Yellowstone National Park
Authors: Meyer‐Dombard D'Arcy R , Swingley Wesley , Raymond Jason , Havig Jeff , Shock Everett L , Summons Roger E
Published: April 2011

In Yellowstone National Park, a small percentage of thermal features support streamer biofilm communities (SBCs), but their growth criteria are poorly understood. This study investigates biofilms in two SBC hosting, and two non‐SBC springs. Sequencing of 16S rRNA clones indicates changing community structure as a function of downstream geochemistry, with many novel representatives particularly among the Crenarchaeota. While some taxonomic groups show little genetic variation, others show specialization by sample location. The transition fringe environment between the hotter chemosynthetic and cooler photosynthetic zones hosts a larger diversity of organisms in SBC bearing springs. This transition is proposed to represent an ecotone; this is the first description of an ecotone in a hydrothermal environment. The Aquificales are ubiquitous and dominate among the Bacteria in the hottest environments. However, there is no difference in species of Aquificales from SBC and non‐SBC locations, suggesting they are not responsible for the formation of SBCs, or that their role in SBC formation is competitively suppressed in non‐SBC sites. In addition, only SBC locations support Thermotogales‐like organisms, highlighting the potential importance these organisms may have in SBC formation. Here, we present a novel view of SBC formation and variability in hydrothermal ecosystems.
The modification of predicative bare nouns in French: a functional analysis1
Authors: Lauwers Peter
Published: April 2011

This paper offers a corpus‐based study of the (restricted) modification patterns of predicatively used bare nouns in French (e.g. Je suis bon catholique‘I am a good Catholic’). These are ‘truly’ bare in that they do not rely on a covert determiner. The aim of the paper is to determine which modification patterns are allowed and to provide a semantic‐functional explanation. Further, on the basis of this study, some conclusions can be drawn both about the categorial status of the bare predicative noun and about the internal architecture of type specification, the innermost functional layer within the NP. More specifically, the study of bare nouns leads to the identification of several lower‐level slots that are independent of the instantiation of a particular nominal entity in conceptual space. These are to be distinguished from higher‐level modifiers. To end, some additional syntactic contexts are provided in which bare nouns occur. These display identical restrictions with respect to modification, but differ in the array of lexical heads they allow for.
Proto‐Anatolian as a mora‐based language1
Authors: Yoshida Kazuhiko
Published: April 2011

It is not easy by any means to obtain prosodic information from documents written in ancient languages because it is not usually recorded therein. But the techniques of philology and linguistics sometimes enable us to derive prosodic evidence from written data. The two lenition rules in Proto‐Anatolian, which were considered to have operated either after an accented long vowel or between two short vowels, have been unified by Adiego into a single rule in moraic terms: lenition occurred after an unaccented mora. Furthermore, Hittite mediopassive present verbs of the nasal‐infix class, which cannot be adequately accounted for on the supposition that the basic units which carried accents in Proto‐Anatolian were syllables, come to receive a well‐motivated (both morphologically and typologically) and straightforward historical explanation from a moraic point of view. These two independent pieces of evidence inevitably lead us to argue that Proto‐Anatolian was a mora‐based language.
Importance of the multidrug and toxin extrusion MATE/SLC47A family to pharmacokinetics, pharmacodynamics/toxicodynamics and pharmacogenomics
Authors: Yonezawa Atsushi , Inui Ken‐ichi
Published: April 2011

The renal organic cation transport system mediates the tubular secretion of cationic compounds including drugs, toxins and endogenous metabolites into urine. It consists of a membrane potential‐dependent organic cation transporter at the basolateral membrane and an H+/organic cation antiporter at the brush‐border membrane. In 2005, human multidrug and toxin extrusion MATE1/SLC47A1 was identified as a mammalian homologue of bacterial NorM. Thereafter, human MATE2‐K/SLC47A2 and rodent MATE were found. Functional characterization revealed that MATE1 and MATE2‐K were H+/organic cation antiporter, mediating the renal tubular secretion of cationic drugs in cooperation with the basolateral organic cation transporter OCT2. Recently, substrate specificity, transcription mechanisms, structure, polymorphisms, in vivo contributions and clinical outcomes on MATE have been investigated intensively. In this review, we summarize recent findings on MATE1/SLC47A1 and MATE2‐K/SLC47A2 and discuss the importance of these transporters to the pharmacokinetics, pharmacodynamics/toxicodynamics and pharmacogenomics of cationic drugs.

LINKED ARTICLES This article is part of a themed section on Transporters. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2011.164.issue‐7
Expression and gene polymorphisms of interleukin 28B and hepatitis B virus infection in a Chinese Han population
Authors: Li Wanyu , Jiang Yanfang , Jin Qinglong , Shi Xiaodong , Jin Jinglan , Gao Yanhang , Pan Yu , Zhang Hong , Jiang Jing , Niu Junqi
Published: March 2011

Background: Recent genome‐wide association studies found that genetic polymorphisms near the IL28B gene is strongly associated with sustained viral response and spontaneous viral clearance in chronically infected hepatitis C patients.

Aims: We aimed to evaluate the effects of IL28B variations on hepatitis B virus (HBV) infection in a Chinese Han population and to explore the association between IL28B polymorphisms and susceptibility to infection, viral clearance, disease progression, viral load and liver inflammation.

Methods: We determined three IL28B single gene polymorphisms (rs12979860, rs12980275 and rs8099917) in 203 individuals with chronic HBV infection, 203 individuals with self‐limited HBV infection and 203 individuals negative for all HBV seromarkers. Interleukin (IL)28B serum levels were evaluated in all subjects. Additionally, peripheral blood mononuclear cells from 42 chronically HBV‐infected individuals were subjected to whole‐genome expression studies.

Results: The association among genotype, allele and haplotype frequencies of IL28B with alanine aminotransferase levels and HBV DNA was established. However, no significant differences were observed in genotype or allele frequencies among chronically HBV‐infected, self‐limited and healthy subjects. The serum IL28B level was lower in patients with chronic HBV infection than in the self‐limited HBV‐infected or healthy subjects. The serum IL28B level was correlated with the subject's genotype. Gene expression micro‐array analysis showed enhanced IL28B expression in patients with low HBV viral load.

Conclusions: Variability at the IL28B locus is associated with HBV viral load and hepatic inflammation. Genetic variation of IL28B may prevent HBV progression by reducing viral load and liver inflammation, providing a valuable gene therapy tool.
Pediatric Dermatology Consultations: A Retrospective Analysis of Inpatient Consultations Referred to the Dermatology Service
Authors: PEÑATE YERAY , BORREGO LEOPOLDO , HERNÁNDEZ NOELIA , ISLAS DIANA
Published: March 2011

Abstract:  To analyze the care provided by consultant dermatologists to hospitalized pediatric patients, we retrospectively studied the records of inpatient pediatric consultation requests received by a hospital dermatology department between 2000 and 2009. The diagnoses were recorded according to the International Classification of Diseases, Ninth Revision (ICD‐9). In a high percentage of cases, the problem was addressed in a single visit, and few additional tests were ordered. The most common diagnostic groups were diseases of the skin and subcutaneous tissue (ICD‐9: 680–709). The most frequent specific diagnosis was atopic dermatitis (ICD‐9: 691.8). Our series shows that the role of dermatologists as consultants for pediatric inpatients is similar to their role in outpatient consultations.
Computer‐Assisted Numerical Analysis for Oxygen and Carbon Dioxide Mass Transfer in Blood Oxygenators
Authors: Turri Fabio , Yanagihara Jurandir Itizo
Published: April 2011

A two‐dimensional numeric simulator is developed to predict the nonlinear, convective‐reactive, oxygen mass exchange in a cross‐flow hollow fiber blood oxygenator. The numeric simulator also calculates the carbon dioxide mass exchange, as hemoglobin affinity to oxygen is affected by the local pH value, which depends mostly on the local carbon dioxide content in blood. Blood pH calculation inside the oxygenator is made by the simultaneous solution of an equation that takes into account the blood buffering capacity and the classical Henderson–Hasselbach equation. The modeling of the mass transfer conductance in the blood comprises a global factor, which is a function of the Reynolds number, and a local factor, which takes into account the amount of oxygen reacted to hemoglobin. The simulator is calibrated against experimental data for an in‐line fiber bundle. The results are: (i) the calibration process allows the precise determination of the mass transfer conductance for both oxygen and carbon dioxide; (ii) very alkaline pH values occur in the blood path at the gas inlet side of the fiber bundle; (iii) the parametric analysis of the effect of the blood base excess (BE) shows that is similar in the case of blood metabolic alkalosis, metabolic acidosis, or normal BE, for a similar blood inlet , although the condition of metabolic alkalosis is the worst case, as the pH in the vicinity of the gas inlet is the most alkaline; (iv) the parametric analysis of the effect of the gas flow to blood flow ratio (QG/QB) shows that variation with the gas flow is almost linear up to QG/QB = 2.0. is not affected by the gas flow as it was observed that by increasing the gas flow up to eight times, the grows only 1%. The mass exchange of carbon dioxide uses the full length of the hollow‐fiber only if QG/QB > 2.0, as it was observed that only in this condition does the local variation of pH and blood comprise the whole fiber bundle.