bluuebunny/crossref_metadata_2025 huggingface dataset:
Perchlorate in the environment: the emerging emphasis on natural occurrence
Authors: Parker David R.
Published: March 2009

Environmental context. Perchlorate is an emerging environmental contaminant that has a unique ability to interfere with normal iodine uptake by the human thyroid gland, and thus has the potential to adversely affect normal growth and development of infants and children. In the last decade, perchlorate’s environmental behaviour has been intensely studied in the United States, but has received little attention elsewhere. Recent evidence strongly suggests that perchlorate occurs at low levels naturally, and is ubiquitously present in the human diet. An atmospheric source for this natural occurrence is strongly implicated, and the naturally occurring isotopes of oxygen and chlorine offer considerable promise for unravelling the chemical mechanisms responsible. Abstract. Salts of perchlorate (ClO4–) are widely used in solid rocket propellants, and in a variety of munitions, explosives, and pyrotechnics; it is an emerging environmental pollutant that has caused widespread water contamination in the United States and probably other locales worldwide. Perchlorate interferes with normal iodine uptake by the human thyroid, and may thus lead to a lowered production of key hormones that are needed for proper growth and development. Debate about ‘safe’ levels of perchlorate is being fuelled by considerable evidence of declining iodine intake in many western nations. With the advent of more sensitive analytical methods, perchlorate is being found as a nearly ubiquitous contaminant in water, beverages, fresh produce, and other sources of human exposure. Recent evidence, including isotopic forensics, makes a strong case for more widespread natural occurrence of perchlorate, outside of the long-established occurrence in caliches of the Atacama Desert in Chile. Many questions about this low-level occurrence remain, including the role of microbial metabolism in attenuating the concentrations typically found in surface- and groundwaters.
Fractionation of stable isotopes in perchlorate and nitrate during in situ biodegradation in a sandy aquifer
Authors: Hatzinger Paul B. , Böhlke John Karl , Sturchio Neil C. , Gu Baohua , Heraty Linnea J. , Borden Robert C.
Published: March 2009

Environmental context. Perchlorate (ClO4–) and nitrate (NO3–) are common co-contaminants in groundwater, with both natural and anthropogenic sources. Each of these compounds is biodegradable, so in situ enhanced bioremediation is one alternative for treating them in groundwater. Because bacteria typically fractionate isotopes during biodegradation, stable isotope analysis is increasingly used to distinguish this process from transport or mixing-related decreases in contaminant concentrations. However, for this technique to be useful in the field to monitor bioremediation progress, isotope fractionation must be quantified under relevant environmental conditions. In the present study, we quantify the apparent in situ fractionation effects for stable isotopes in ClO4– (Cl and O) and NO3– (N and O) resulting from biodegradation in an aquifer. Abstract. An in situ experiment was performed in a shallow alluvial aquifer in Maryland to quantify the fractionation of stable isotopes in perchlorate (Cl and O) and nitrate (N and O) during biodegradation. An emulsified soybean oil substrate that was previously injected into this aquifer provided the electron donor necessary for biological perchlorate reduction and denitrification. During the field experiment, groundwater extracted from an upgradient well was pumped into an injection well located within the in situ oil barrier, and then groundwater samples were withdrawn for the next 30 h. After correction for dilution (using Br– as a conservative tracer of the injectate), perchlorate concentrations decreased by 78% and nitrate concentrations decreased by 82% during the initial 8.6 h after the injection. The observed ratio of fractionation effects of O and Cl isotopes in perchlorate (ϵ18O/ϵ37Cl) was 2.6, which is similar to that observed in the laboratory using pure cultures (2.5). Denitrification by indigenous bacteria fractionated O and N isotopes in nitrate at a ratio of ~0.8 (ϵ18O/ϵ15N), which is within the range of values reported previously for denitrification. However, the magnitudes of the individual apparent in situ isotope fractionation effects for perchlorate and nitrate were appreciably smaller than those reported in homogeneous closed systems (0.2 to 0.6 times), even after adjustment for dilution. These results indicate that (1) isotope fractionation factor ratios (ϵ18O/ϵ37Cl, ϵ18O/ϵ15N) derived from homogeneous laboratory systems (e.g. pure culture studies) can be used qualitatively to confirm the occurrence of in situ biodegradation of both perchlorate and nitrate, but (2) the magnitudes of the individual apparent ϵ values cannot be used quantitatively to estimate the in situ extent of biodegradation of either anion.
Why are non-photosynthetic tissues generally 13C enriched compared with leaves in C3 plants? Review and synthesis of current hypotheses
Authors: Cernusak Lucas A. , Tcherkez Guillaume , Keitel Claudia , Cornwell William K. , Santiago Louis S. , Knohl Alexander , Barbour Margaret M. , Williams David G. , Reich Peter B. , Ellsworth David S. , Dawson Todd E. , Griffiths Howard G. , Farquhar Graham D. , Wright Ian J.
Published: March 2009

Non-photosynthetic, or heterotrophic, tissues in C3 plants tend to be enriched in 13C compared with the leaves that supply them with photosynthate. This isotopic pattern has been observed for woody stems, roots, seeds and fruits, emerging leaves, and parasitic plants incapable of net CO2 fixation. Unlike in C3 plants, roots of herbaceous C4 plants are generally not 13C-enriched compared with leaves. We review six hypotheses aimed at explaining this isotopic pattern in C3 plants: (1) variation in biochemical composition of heterotrophic tissues compared with leaves; (2) seasonal separation of growth of leaves and heterotrophic tissues, with corresponding variation in photosynthetic discrimination against 13C; (3) differential use of day v. night sucrose between leaves and sink tissues, with day sucrose being relatively 13C-depleted and night sucrose 13C-enriched; (4) isotopic fractionation during dark respiration; (5) carbon fixation by PEP carboxylase; and (6) developmental variation in photosynthetic discrimination against 13C during leaf expansion. Although hypotheses (1) and (2) may contribute to the general pattern, they cannot explain all observations. Some evidence exists in support of hypotheses (3) through to (6), although for hypothesis (6) it is largely circumstantial. Hypothesis (3) provides a promising avenue for future research. Direct tests of these hypotheses should be carried out to provide insight into the mechanisms causing within-plant variation in carbon isotope composition.
A non-cell-autonomous mechanism for the control of plant architecture and epidermal differentiation involves intercellular trafficking of BREVIPEDICELLUS protein
Authors: Rim Yeonggil , Jung Jin-Hee , Chu Hyosub , Cho Won Kyong , Kim Seon-Won , Hong Jong Chan , Jackson David , Datla Raju , Kim Jae-Yean
Published: March 2009

Intercellular trafficking of maize KNOTTED1 and its homologous KNOTTED1-related homeobox (KNOX) proteins has been reported; however, little is known about the functional significance of KNOX trafficking in plant development. In this study, we showed that intercellular movement of BREVIPEDICELLUS (BP or KNAT1), the closest Arabidopsis homologue of KNOTTED1, is tissue-specific and takes place through a selective pathway. When BP was fused to a red fluorescent mCherry construct, it could move from the mesophyll to epidermal cells of leaves, although it could not move out from the cortex/endodermis of roots. Using a trichome rescue-trafficking assay, we also showed that BP fusion could confer gain-of-trafficking function to the cell-autonomous GLABROUS1 (GL1) protein. In the wild type, BP transcripts are expressed in the sub-epidermal cortical cell layers of the inflorescence stem and pedicel. However, bp mutant phenotypes include defects in epidermal cell differentiation suggesting a non-cell-autonomous function. Expression of a GFP:BP fusion under the control of a BP promoter specific to the stem cortex layers resulted in epidermal GFP fluorescence suggesting its movement from subepidermis to epidermis. Here, we provide evidence from complementation analyses using cell autonomous or non-cell-autonomous BP fusions that the intercellular trafficking of BP protein is important for plant architecture and epidermal differentiation.
Heat tolerance and expression of protein synthesis elongation factors, EF-Tu and EF-1α, in spring wheat
Authors: Bukovnik Urška , Fu Jianming , Bennett Miranda , Prasad P. V. Vara , Ristic Zoran
Published: March 2009

Protein elongation factors, EF-Tu and EF-1α, have been implicated in cell response to heat stress. We investigated the expression (accumulation) of EF-Tu and EF-1α in mature plants of spring wheat cultivars Kukri and Excalibur, and tested the hypothesis that cultivars with contrasting tolerance to heat stress differ in the accumulation of these elongation factors under prolonged exposure to high temperature (16 days at 36/30°C). In addition, we investigated the expression of EF-Tu and EF-1α in young plants experiencing a 24-h heat shock (43°C). Excalibur showed better tolerance to heat stress than Kukri. Heat stress induced accumulation of EF-Tu and EF-1α in mature plants of both cultivars, but to a greater extent in Excalibur. Young plants did not show appreciable accumulation of EF-Tu in response to heat shock. However, these plants showed increased accumulation of EF-1α and the accumulation appeared greater in Excalibur than in Kukri. The results support the hypothesis that EF-Tu plays a role in heat tolerance in spring wheat. The results also suggest that EF-1α may be of importance to wheat response to heat stress.
Proteomic analysis of 'hybrid necrosis' in wheat (Triticum aestivum) leaves
Authors: Pan Xing Lai , Jiang Qi Yan , Pan Qian Ying , Wen Xue Fei , Shi Yin Hong , Wang Yong Jie , Pan Tian Yuan , Xie San Gang , Zhang Gui Yun , Wu Shen Jie , Chai Yong Feng , Zhang Chang Sheng , Wu Zong Xin , Shen Shi Hua
Published: March 2009

Wheat hybrid necrosis has been genetically characterised for many years, but the specific gene(s) and the protein products involved in the processes remains unknown. In this study, protein expression in the base (B), mid (M) and tip (T) segments of the FL-2 leaves of a necrotic hybrid, PZF1 and its parents, Pan555 and Zheng891, was analysed and compared using a high throughput proteomic approach. Twenty-three protein spots, with significant variations in intensity across the necrotic leaf segments, were analysed by MALDI-TOF-MS, of which, 18 were matched to protein accessions in the NCBI database. Several of these proteins are enzymes involved in the methylation cycle, including AdoHcy hydrolase, AdoMet synthase 3 and methionine synthase 1; AdoHcy hydrolase was downregulated sharply in M and T, and AdoMet synthase 3 and methionine synthase 1 were upregulated gradually from M to T. This result suggests that methylation-associated processes, including epigenetic mechanisms, may play a role in the initiation and development of hybrid necrosis. Several energy cycle-associated proteins and cytoprotective proteins were also differentially expressed across the leaf segments, suggesting their direct association with or possible involvement in the necrotic processes. The significant imbalance of a heat-shock protein, a transposon protein and a RNA- and ssDNA-binding protein also makes these proteins potential molecular components in the necrotic processes.
Desiccation protects two Antarctic mosses from ultraviolet-B induced DNA damage
Authors: Turnbull Johanna D. , Leslie Simon J. , Robinson Sharon A.
Published: March 2009

Antarctic mosses live in a frozen desert, and are characterised by the ability to survive desiccation. They can tolerate multiple desiccation-rehydration events over the summer growing season. As a result of recent ozone depletion, such mosses may also be exposed to ultraviolet-B radiation while desiccated. The ultraviolet-B susceptibility of Antarctic moss species was examined in a laboratory experiment that tested whether desiccated or hydrated mosses accumulated more DNA damage under enhanced ultraviolet-B radiation. Accumulation of cyclobutane pyrimidine dimers and pyrimidine (6–4) pyrimidone dimers was measured in moss samples collected from the field and then exposed to ultraviolet-B radiation in either a desiccated or hydrated state. Two cosmopolitan species, Ceratodon purpureus (Hedw.) Brid. and Bryum pseudotriquetrum (Hedw.) Gaertn., B.Mey. & Scherb, were protected from DNA damage when desiccated, with accumulation of cyclobutane pyrimidine dimers reduced by at least 60% relative to hydrated moss. The endemic Schistidium antarctici (Cardot) L.I. Savicz & Smirnova accumulated more DNA damage than the other species and desiccation was not protective in this species. The cosmopolitan species remarkable ability to tolerate high ultraviolet-B exposure, especially in the desiccated state, suggests they may be better able to tolerate continued elevated ultraviolet-B radiation than the endemic species.
Is body shape of varanid lizards linked with retreat choice?
Authors: Thompson Graham G. , Clemente Christofer J. , Withers Philip C. , Fry Bryan G. , Norman Janette A.
Published: March 2009

In our earlier analysis of Varanus body shape, size was a dominating factor with some qualitative phylogenetic patterns and grouping of species into ecological categories. With a phylogeny and an improved capacity to account for the effects of size, we have reanalysed our morphometric data for male Australian goannas (Varanus spp.) using an increased number of specimens and species to examine whether variations in body shape can be accounted for by retreat choice, as it can for Western Australian Ctenophorus dragon lizards. After accounting for body size in the current analysis, four ecotypes based on retreat choice (i.e. those that retreat to oblique crevices between large rocks or rock faces, those that retreat to burrows dug into the ground, those that retreat to spaces under rocks or in tree hollows, and those that retreat to trees but not tree hollows) accounted for much of the variation in body shape. There is a phylogenetic pattern to the ecotypes, but accounting for phylogenetic effects did not weaken the link between body shape and ecotype based on retreat choice. This suggests that there are large differences in body shape among ecotypes, and shape is relatively independent of phylogeny. The strong link between shape and choice of retreat site in Varanus spp. is consistent with that for Ctenophorus spp. We speculate on why there might be a strong link between retreat choice and body shape for both Varanus and Ctenophorus.
“Vanishing Penis” and Urinary Retention due to Locally Destructive Penile Cancer
Authors: Sountoulides Petros , Bantis Athanasios , Zachos Ioannis , Asouhidou Irene , Pantazakos Athanasios
Published: March 2009

Penile carcinomas are relatively rare. They usually arise from precancerous lesions and present in the form of ulcerative or exophytic tumors. They rarely give rise to urinary symptoms and complications, and are usually easy to diagnose. We present a case of an 82-year-old man with chronic urinary retention due to urethral dissemination by a locally destructive penile lesion. The penis was literally “vanished” by the lesion down to the level of the pubic bone without, interestingly, having spread to the local lymph nodes or given rise to distant metastases. A temporary suprapubic catheter was placed, followed by a perineal urethrostomy in order to reverse the established renal failure.
Localization of Liv2 as an Immature Hepatocyte Marker in EB Outgrowth
Authors: Takashimizu Ikkei , Tanaka Yoshiki , Yoshie Susumu , Kano Yoshiya , Ichikawa Hinako , Cui Li , Ogiwara Naoko , Johkura Kohei , Sasaki Katsunori
Published: March 2009

The objective of this study was to establish Liv2, a surface marker of mouse immature hepatocytes (hepatoblasts), as a selection tool for embryonic stem (ES) cell–derived immature hepatocytes by acquiring basic data on Liv2 in normal mouse embryos and by confirming Liv2 expression in mouse ES-derived cells. The estimated molecular weight of Liv2 was 4045 kDa, and immunoreactivity was definitively detected in the cell membrane of fetal hepatocytes on embryonic day (E) 9.5, declined gradually until E12.5, and subsequently became undetectable. Liv2 was localized on and close to the cell membrane. Embryoid bodies (EB) were formed from mouse ES cells whose undifferentiated state was confirmed with immunostaining of Nanog by the hanging drop method. A few Liv2-positive cells occurred as a cluster in EB outgrowth on day 7, but only some of these were albumin (ALB)-positive on day 13. These cells had the same pattern of immunoreactivity, i.e., localization on the cell membrane, as immature hepatocytes in the developing liver, although there were other types of cells with a different pattern of immunoreactivity that were seen only as a granular pattern in the cytoplasm and without ALB or the neuronal marker nestin. These results suggest that Liv2 may be useful as a surface marker for immature hepatocytes derived from ES cells. This application would allow for the sole selection of immature hepatocytes and provide a useful tool for regenerative medicine.
Generating Timed Pregnancies in Fruit Bats (Carollia perspicillata): Figure 1.
Authors: Rasweiler John J. , Cretekos Chris J. , Behringer Richard R.
Published: March 2009

INTRODUCTION

When female short-tailed fruit bats (Carollia perspicillata) are bred, they are first kept sexually segregated for some months to adapt to captivity (in the case of recently captured animals) and to ensure that none are already pregnant. In the case of captive-reared females that had previously been housed with males, 4 mo should be sufficient. In the case of wild-caught females, 7 or 8 mo may not be too long, because introduction into captivity can substantially prolong existing early pregnancies (i.e., at or earlier than the primitive streak stage). Wild-caught females are also much more prone than captive-reared females to take pregnancies into delay after captive breeding. It is important to note that females should not be housed in sexual isolation for prolonged periods, because this can eventually lead to the development of markedly hyperplastic uteri and reduced fertility. Although this is a very serious problem for zoos maintaining all-female colonies of Carollia, it is not the case with our captive colony because the adult females are regularly bred, both to replenish the colony and to inhibit the development of hyperplastic uteri. The generation of timed pregnancies is described in this protocol. For breeding purposes, a stud male with prominent testes is introduced into each cage of females. Vaginal aspirates are then checked each morning thereafter for the presence of spermatozoa.
Collection of Embryos from Short-Tailed Fruit Bats (Carollia perspicillata): Figure 1.
Authors: Rasweiler John J. , Cretekos Chris J. , Behringer Richard R.
Published: March 2009

INTRODUCTION

Embryos from the short-tailed fruit bat (Carollia perspicillata) can be collected from females that are pregnant as a result of matings established in a laboratory colony or during the breeding season in the wild. The entire reproductive tract containing the single conceptus is dissected from the female, and the uterus is measured. If the uteri contain embryos that can be dissected out, measured, and staged, this information can be compared to published data for normal pregnancies. The results can then be used to predict when conceptuses at other stages of development might be collected. Such data, by themselves, do not establish whether those pregnancies may have previously included a period of delay at the primitive streak stage. As described in this protocol, the embryo is usually dissected from the reproductive tract free of decidual tissue and its extraembryonic membranes, including the yolk sac and amnion. Once isolated, the embryo can then be processed for subsequent analysis.
Fixation and Storage of Embryos from Short-Tailed Fruit Bat (Carollia perspicillata)
Authors: Rasweiler John J. , Cretekos Chris J. , Behringer Richard R.
Published: March 2009

INTRODUCTION

This protocol describes fixation of embryos and tissues of the short-tailed fruit bat C. perspicillata. The fixed embryos can be stored at −20°C for prolonged periods and can be used for whole-mount in situ hybridization, whole-mount immunohistochemistry, or for embedding and sectioning in paraffin or frozen blocks for a variety of histological procedures.
Alcian Blue/Alizarin Red Staining of Cartilage and Bone of Short-Tailed Fruit Bat (Carollia perspicillata)
Authors: Rasweiler John J. , Cretekos Chris J. , Behringer Richard R.
Published: March 2009

INTRODUCTION

This protocol is used to stain embryo skeleton with alcian blue and alizarin red at later stages of development, when there is significant replacement of the cartilaginous early skeleton with ossified bone. It has been used with good results on the short-tailed fruit bat Carollia perspicillata and other bat species from CS 20 through neonatal stages.
Alcian Blue/Alizarin Red Staining of Cartilage and Bone in Mouse: Figure 1.
Authors: Ovchinnikov Dmitry
Published: March 2009

INTRODUCTION

The vertebrate skeleton forms by endochondral and intramembranous bone formation. During endochondral bone formation, mesenchyme condensations give rise to cartilages that are eventually replaced by bone. However, there are some permanent cartilages that do not ossify, such as the cartilage of the trachea and articular cartilage of the joints, and intramembranous bone formation occurs directly without a cartilage template. This article describes a method for simultaneously visualizing cartilage and mineralized tissues (notably bone) in the developing mouse. It makes use of alcian blue and alizarin red stains, and it is best used for later fetal, newborn, and early post-natal stages of development. Neonatal skeletons are especially well suited for this technique.
Collection of Embryos from Darwin’s Finches (Thraupidae, Passeriformes)
Authors: Abzhanov Arhat
Published: March 2009

INTRODUCTION

There are no breeding colonies of Darwin’s finches anywhere in the world. Thus, all of the embryonic material is collected in the wild. This protocol describes how, in a field setting, fertilized eggs are collected and incubated at a precise temperature and how the resulting embryos are harvested and processed for in situ hybridization, antibody staining, and microarray analyses. In addition, the protocol includes steps for preparing the heads of older embryos for histological staining of bone and cartilage with alcian blue and alizarin red. It is likely that the same or similar methods can be used to obtain embryonic tissue from other species of songbirds. The main limitation of this protocol is that, when used in the field without external constant electricity sources, it requires power generators that need to run on a more or less constant basis, as well as stockpiles of supplies (fuel, oil, and fresh water).
Cetyltrimethyl Ammonium Bromide (CTAB) DNA Miniprep for Plant DNA Isolation
Authors: Clarke Joseph D.
Published: March 2009

INTRODUCTION

The presence of polysaccharides in a DNA preparation can inhibit use of techniques such as polymerase chain reaction (PCR). Cetyltrimethyl ammonium bromide (CTAB) is a surfactant useful for isolation of DNA from tissues containing high amounts of polysaccharides. Under the high-salt conditions used in this protocol, the CTAB binds the polysaccharides, removing them from the solution. When used with Arabidopsis, this procedure yields quite pure DNA that is suitable for thermal asymmetric interlaced PCR (TAIL-PCR).
Dellaporta Miniprep for Plant DNA Isolation
Authors: Weigel Detlef , Glazebrook Jane
Published: March 2009

INTRODUCTION

This procedure provides a quick and inexpensive method for plant DNA isolation. When used with Arabidopsis, this procedure yields quite good-quality DNA that is suitable as a polymerase chain reaction (PCR) template for cleaved amplified polymorphic sequences (CAPS) or simple sequence length polymorphism (SSLP) markers. It yields enough DNA to test a large number of markers. It is recommended in cases when many markers will be tested on the same samples.
Quick Miniprep for Plant DNA Isolation
Authors: Weigel Detlef , Glazebrook Jane
Published: March 2009

INTRODUCTION

This procedure describes a quick miniprep for plant DNA isolation. It yields a small quantity of poorly purified DNA. However, many samples can be processed in a short time. When used with Arabidopsis, the method generally provides DNA of sufficient quality for polymerase chain reaction (PCR) amplification of cleaved amplified polymorphic sequences (CAPS) and simple sequence length polymorphism (SSLP) markers. Repeated freeze-thaw cycles should be avoided, because they result in samples that will no longer work as PCR templates.
Mass Spectrometry Characterization of Multiprotein Complexes: Figure 1.
Authors: Robinson Carol V.
Published: March 2009

INTRODUCTION

Although mass spectrometry measures species in the gas phase, it is becoming widely accepted that, under the appropriate conditions, noncovalent features of protein structure and interactions can be retained in mass spectra. Recent applications have shown that mass spectrometry can provide information not only about the stoichiometry of subunits within a complex, but also about subsets of interacting proteins within macromolecular complexes, the component oligomers in polydisperse assemblies, and changes in quaternary structure in response to phosphorylation of subunits. An important advance in this field has been the recent extension of established techniques to examine protein complexes isolated directly from cells, rather than assembled from recombinant subunits. This protocol provides guidance for setup and use of a mass spectrometer to analyze multiprotein complexes.
Transduction of Vertebrate Cells with Recombinant Baculovirus
Authors: Airenne Kari J. , Laitinen Olli H. , Mähönen Anssi J. , Ylä-Herttuala Seppo
Published: March 2009

INTRODUCTION

The baculoviral expression vector system (BEVS) has become one of the most popular systems to produce recombinant proteins. The finding that a modified baculovirus bearing a suitable promoter was able to efficiently transduce hepatocytes stimulated further interest in BEVS as a potential gene delivery vector for cells other than those of insect origin. Indeed, a wide variety of vertebrate cells can be transduced by recombinant baculoviruses. Such viruses can also mediate efficient gene delivery into several tissues of mammals in vivo. The popularity of BEVS over other expression systems is based on several unique features such as safety, ability to accommodate large transgenes, and low cytotoxicity in vertebrate cells even at very high virus load. This protocol describes a method for transformation of vertebrate cells with recombinant baculovirus.
Up‐regulated expression of ADAM17 in gastrointestinal stromal tumors: coexpression with EGFR and EGFR ligands
Authors: Nakagawa Motomichi , Nabeshima Kazuki , Asano Shigeyuki , Hamasaki Makoto , Uesugi Noriko , Tani Hiroki , Yamashita Yuichi , Iwasaki Hiroshi
Published: March 2009

Metalloproteinase activities of a disintegrin and metalloproteinases (ADAMs), matrix metalloproteinases (MMPs), and membrane type (MT‐)MMPs are involved in many aspects of tumor biology. ADAMs are transmembrane proteins that cleave membrane‐anchored proteins to release soluble factors, and thereby mediate important biological phenomena in tumors. The aim of this study was to analyze histopathology, expression and roles of metalloproteinases, especially ADAMs, in gastric gastrointestinal stromal tumor (GIST). Histopathology and immunohistochemical expression of ADAMs were examined in 89 gastric GISTs. In 11 GISTs, ADAM expression was examined at mRNA and protein levels by reverse transcription – polymerase chain reaction (RT‐PCR) and immunoblotting, respectively. RT‐PCR analysis showed frequent expression of ADAM9 (91%), ADAM10 (64%), ADAM17 (82%), MMP‐2 (82%), and MT1‐MMP (73%). However, ADAM17 and MMP‐2 were the only metalloproteinases that were up‐regulated in GISTs at the protein level compared with non‐neoplastic gastric tissues. ADAM17 was immunohistochemically expressed in 93% of GIST versus 16% of normal gastric tissues. Furthermore, CD117‐positive interstitial cells of Cajal in normal gastric tissues were all negative for ADAM17 with double immunostaining. Expressions of epidermal growth factor receptor (EGFR) and several EGFR ligands such as amphiregulin, heparin‐binding epidermal growth factor (HB‐EGF), betacellulin, and epiregulin were also demonstrated in GIST by RT‐PCR. Protein expression of EGFR, phosphorylated EGFR, amphiregulin, and HB‐EGF, both of which can be shed by ADAM17, was confirmed in tumors coexpressing ADAM17 by immunoblotting. Moreover, proteolytically cleaved soluble forms of amphiregulin were identified in tumor extracts. Considered together, the results suggest that ADAM17 may contribute to the progression and growth of GIST through shedding of EGFR ligands and consequent EGFR stimulation. ADAM17, as a major sheddase in GIST, could be potentially a suitable target in anticancer treatment of imatinib‐resistant GISTs. (Cancer Sci 2009; 100: 654–662)
Potent antitumor activity of double‐regulated oncolytic adenovirus‐mediated ST13 for colorectal cancer
Authors: Yu De Bin , Zhong Su Yang , Yang Min , Wang Yi Gang , Qian Qi Jun , Zheng Shu , Liu Xin Yuan
Published: March 2009

Following targeted gene virotherapy, the suppression of tumorigenicity 13 (ST13) gene was inserted into the double‐regulated oncolytic adenovirus SG500 to ensure more safety and potent antitumor activity against colorectal cancer in vitro and in vivo. We generated the ST13‐expressing oncolytic adenovirus SG500‐ST13, with which colorectal carcinoma cell lines SW620 and HCT116, and the lung fibroblast cell line WI38, were infected. Crystal violet staining was carried out to detect the cytopathic effect in cells, and the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide method was used to assay cell viability. The effect of apoptosis induced by SG500‐ST13 was confirmed by Hoechst staining and the TdT‐mediated dUTP‐biotin nick‐end labeling method. To further identify the antitumor effects of SG500‐ST13 on HCT116 xenografts in Balb/c nude mice, the induction of cell death was assessed by hematoxylin–eosin staining. Immunohistochemical study was also carried out. (Cancer Sci 2009; 100: 678–683)
Cost‐effective mammography screening in Korea: High incidence of breast cancer in young women
Authors: Lee Soon Young , Jeong Seong Hwa , Kim Youn Nam , Kim Jinheum , Kang Dae Ryong , Kim Hyeon‐Chang , Nam Chung Mo
Published: March 2009

The epidemiological characteristics of breast cancer in Korean women are different from the characteristics reported in Western women. The highest incidence rate occurs in Korean women in their 40s. The purpose of this study was to determine the most cost‐effective screening interval and target age range for Korean women from the perspective of the national healthcare system. A stochastic model was used to simulate breast cancer screenings by varying both the screening intervals and the age ranges. The effectiveness of mammography screening was defined as the probability of detecting breast cancer in the preclinical state and the cost was based on the direct cost of mammography screening and the confirmative tests. The age‐specific mean sojourn times and the sensitivity of the mammography were applied in the stochastic model. An optimal cost‐effectiveness was determined by the incremental cost‐effectiveness ratio and lifetime schedule sensitivity. Sensitivity analyses were undertaken to assess parameter uncertainty. The selected cost‐effective strategies were: (1) the current biennial mammography screenings for women who are at least 40 years old; (2) biennial screening for women between the ages of 35 and 75 years; and (3) a combination strategy consisting of biennial screening for women aged between 45 and 54 years, and 3‐year interval screening for women aged between 40 and 44 years and 55 and 65 years. Further studies should follow to investigate the effectiveness of mammography screening in women younger than 40 years in Asia as well as in Korea. (Cancer Sci 2009; 100: 1105–1111)
Risk of upper gastrointestinal bleeding with oral bisphosphonates and non steroidal anti‐inflammatory drugs: a case‐control study
Authors: ETMINAN M. , LÉVESQUE L. , FITZGERALD J. M. , BROPHY J. M.
Published: March 2009

Background  Gastrointestinal injuries including gastric ulcers have been reported with oral bisphosphonate therapy. However, the risk of the more serious upper gastrointestinal bleeding (UGB) especially in the community setting with these drugs remains unknown. Similarly, the risk of UGB among users of both bisphosphonates and non steroidal anti‐inflammatory drugs (NSAIDs) in the community is also unknown.

Aim  To explore the risk of more serious UGB among users of bisphosphonates and the risk of UGB among users of both bisphosphonates and NSAIDs in the community.

Methods  We conducted a case‐control study within a cohort of Quebec residents who had received a revascularization procedure from 1995 to 2004. Cohort members were followed up from the date of their first procedure until the earliest of: (1) study outcome, (2) date of death or (3) end of health care coverage. Cases were defined as those with the first diagnosis of a UGB. For each case, 20 controls were selected and matched to the cases by index date, age and cohort entry. Adjusted odds ratios for current use of bisphosphonates, NSAIDs and co‐therapy of both drugs were computed.

Results  Within the initial cohort, 3253 incident cases of UGBs and corresponding 65 060 matched controls were identified. The adjusted odds ratio (OR) for UGB by current users of bisphosphonates was 1.01 (95% CI, 0.72–1.43). Current NSAID use was associated with an increased risk of UGB OR = 1.75; 95% CI, 1.53–1.99. The OR for use of bisphosphonates and NSAIDs was elevated OR = 2.00; 95% CI, 1.12–3.57. This risk was still elevated for users of bisphosphonates and COX‐2 inhibitors [OR = 2.38 (95% CI, 1.26–4.50)].

Conclusion  We found no evidence of an increase in the risk of UGB among current users of bisphosphonates. The risk of combined NSAID and bisphosphonate therapy was increased, but this risk was not higher than the risk for NSAID users alone.
Therapeutic opportunities for counteracting apoptosis resistance in childhood leukaemia
Authors: Fulda Simone
Published: March 2009

Evasion of apoptosis is a hallmark of human cancers, for example in haematological malignancies. Apoptosis is an intrinsic cell death program that is crucial in maintaining tissue homeostasis, for example in the haematopoietic system where there is a high turnover rate of cells. As a result, a decrease in the rate of apoptosis as well as an increase in proliferation favours tumorigenesis as well as tumour progression. Further, the anti‐leukaemic action of current treatment approaches, including chemo‐, radio‐ or immunotherapy, critically relies on intact cell death programs in cancer cells. Therefore, defects in apoptosis pathways are frequently associated with the resistance to anticancer therapies. In recent years, the identification and characterization of the molecules and pathways that are involved in the regulation and execution of cell death in leukaemia and lymphoma cells, for example tumour necrosis factor‐related apoptosis inducing ligand (TRAIL), ‘inhibitor of apoptosis’ (IAP) proteins and Bcl‐2, have set the ground for the development of novel diagnostic tools and molecular therapeutics targeting apoptosis pathways in haematological malignancies.
Notoriety to respectability: a short history of arsenic prior to its present day use in haematology
Authors: Doyle Derek
Published: March 2009

This paper looks at arsenic, and in particular the trioxide, from the days of the ancient Chinese, Egyptians, Greeks and Romans, through the 17th–20th centuries to its adoption by today’s haematologists. It looks at its commercial and medical uses, past and present, its notoriety as a poison, it’s reputation as a ‘tonic’ and therapeutic agent, many of the famous people associated with it including Thomas Fowler, William Withering and Robert Christison, and the promise an 18th century panacea now offers 21st century patients under the care of today’s haematologists and tomorrow’s oncologists.
A novel view on stem cell development: analysing the shape of cellular genealogies
Authors: Glauche I. , Lorenz R. , Hasenclever D. , Roeder I.
Published: March 2009

Objectives:  The analysis of individual cell fates within a population of stem and progenitor cells is still a major experimental challenge in stem cell biology. However, new monitoring techniques, such as high‐resolution time‐lapse video microscopy, facilitate tracking and quantitative analysis of single cells and their progeny. Information on cellular development, divisional history and differentiation are naturally comprised into a pedigree‐like structure, denoted as cellular genealogy. To extract reliable information concerning effecting variables and control mechanisms underlying cell fate decisions, it is necessary to analyse a large number of cellular genealogies.

Materials and Methods:  Here, we propose a set of statistical measures that are specifically tailored for the analysis of cellular genealogies. These measures address the degree and symmetry of cellular expansion, as well as occurrence and correlation of characteristic events such as cell death. Furthermore, we discuss two different methods for reconstruction of lineage fate decisions and show their impact on the interpretation of asymmetric developments. In order to illustrate these techniques, and to circumvent the present shortage of available experimental data, we obtain cellular genealogies from a single‐cell‐based mathematical model of haematopoietic stem cell organization.

Results and Conclusions:  Based on statistical analysis of cellular genealogies, we conclude that effects of external variables, such as growth conditions, are imprinted in their topology. Moreover, we demonstrate that it is essential to analyse timing of cell fate‐specific changes and of occurrence of cell death events in the divisional context in order to understand the mechanisms of lineage commitment.
Lability of organic carbon in lakes of different trophic status
Authors: OSTAPENIA A. P. , PARPAROV A. , BERMAN T.
Published: March 2009

1. We used first‐order kinetic parameters of biological oxygen demand (BOD), the constant of aerobic decomposition (k) and the asymptotic value of BOD (BODult), to characterise the lability of organic carbon pools in six lakes of different trophic state: L. Naroch, L. Miastro and L. Batorino (Belarus), L. Kinneret (Israel), L. Ladoga (Russia) and L. Mendota (U.S.A.). The relative contributions of labile and refractory organic carbon fractions to the pool of total organic carbon (TOC) in these lakes were quantified. We also determined the amounts of labile organic carbon within the dissolved and particulate TOC pools in the three Belarus lakes.

2. Mean annual chlorophyll concentrations (used as a proxy for lake trophic state) ranged from 2.3 to 50.6 μg L−1, labile organic carbon (OCL = 0.3BODult) from 0.75 to 2.95 mg C L−1 and k from 0.044 to 0.14 day−1.

3. Our data showed that there were greater concentrations of OCL but lower k values in more productive lakes.

4. In all cases, the DOC fraction dominated the TOC pool. OCL was a minor component of the TOC pool averaging about 20%, irrespective of lake trophic state.

5. In all the lakes, most (c. 85%) of the DOC pool was refractory, corresponding with published data based on measurements of bacterial production and DOC depletion. In contrast, a larger fraction (27–55%) of the particulate organic carbon (POC) pool was labile. The relative amount of POC in the TOC pool tended to increase with increasing lake productivity.

6. Long‐term BOD incubations can be valuable in quantifying the rates of breakdown of the combined particulate and dissolved organic carbon pools and in characterising the relative proportions of the labile and recalcitrant fractions of these pools. If verified from a larger number of lakes our results could have important general implications.
Human mid‐gestation amniotic fluid contains interleukin‐16 bioactivity
Authors: Thornton Catherine A. , Holloway Judith A. , Shute Janis K. , Holloway John W. , Diaper Norma D. , Warner John O.
Published: March 2009

CD4‐positive cells are detectable in the human fetal gastrointestinal tract from 11 weeks of gestation. Interleukin‐16 (IL‐16) is a chemoattractant for CD4+ cells and, via fetal swallowing of amniotic fluid, could mediate the influx of CD4+ cells into the fetal gut. We have shown that IL‐16 was detectable in human amniotic fluid at 16–18 weeks of gestation (mid‐pregnancy) but was not detectable at term (late pregnancy; > 37 weeks of gestation). Similarly, mid‐pregnancy, but not late pregnancy, amniotic fluid contained chemotactic activity for CD4+ T cells, this activity was reduced by 58% in the presence of a neutralizing anti‐IL‐16 antibody. The levels of IL‐16 in fetal plasma at 16–24 weeks of gestation were very high, and decreased significantly by 25–36 weeks but at > 37 weeks remained significantly higher than adult levels. IL‐16 transcripts were detectable in whole tissue extracts of fetal gut, skin and placenta but not in amniocytes, and IL‐16 immunoreactivity was detectable in cells within the lamina propria of the fetal gut and within the skin, where it was associated with the basement membrane. Neither IL‐16 levels nor chemotactic activity for CD4+ T cells in mid‐pregnancy amniotic fluid was related to atopic outcomes at 1 year of age. IL‐16 might have an important role in the early development of the human immune system and/or in regulating fetal and maternal immunological responsiveness during pregnancy.
New dimensions in endodontic imaging: part 1. Conventional and alternative radiographic systems
Authors: Patel S. , Dawood A. , Whaites E. , Pitt Ford T.
Published: March 2009

Conventional radiographs used for the management of endodontic problems yield limited information because of the two‐dimensional nature of images produced, geometric distortion and anatomical noise. These factors often act in combination. This review paper assesses the limitations of periapical radiographs and seeks to clarify three‐dimensional imaging techniques that have been suggested as adjuncts to conventional radiographs. These include tuned aperture computed tomography, magnetic resonance imaging, ultrasound, computed tomography and cone beam computed tomography (CBCT). Of these techniques, CBCT appears to be an effective and safe way to overcome some of the problems associated with conventional radiographs.
Detection of periapical bone defects in human jaws using cone beam computed tomography and intraoral radiography
Authors: Patel S. , Dawood A. , Mannocci F. , Wilson R. , Pitt Ford T.
Published: March 2009

Aim  To compare the diagnostic accuracy of intraoral digital periapical radiography with that of cone beam computed tomography (CBCT) for the detection of artificial periapical bone defects in dry human jaws.

Methodology  Small and large artificial periapical lesions were prepared in the periapical region of the distal root of six molar teeth in human mandibles. Scans and radiographs were taken with a charged couple device (CCD) digital radiography system and a CBCT scanner before and after each periapical lesion had been created. Sensitivity, specificity, positive predictive values, negative predictive values and Receiver Operator Characteristic (ROC) curves as well as the reproducibility of each technique were determined.

Results  The overall sensitivity was 0.248 and 1.0 for intraoral radiography and CBCT respectively, i.e. these techniques correctly identified periapical lesions in 24.8% and 100% of cases, respectively. Both imaging techniques had specificity values of 1.0. The ROC Az values were 0.791 and 1.000 for intraoral radiography and CBCT, respectively.

Conclusions  With intraoral radiography, external factors (i.e. anatomical noise and poor irradiation geometry), which are not in the clinician’s control, hinder the detection of periapical lesions. CBCT removes these external factors. In addition, it allows the clinician to select the most relevant views of the area of interest resulting in improved detection of the presence and absence of artificial periapical lesions.
Lactobacillus farciminis treatment attenuates stress‐induced overexpression of Fos protein in spinal and supraspinal sites after colorectal distension in rats
Authors: Ait‐belgnaoui A. , Eutamene H. , Houdeau E. , Bueno L. , Fioramonti J. , Theodorou V.
Published: March 2009

Abstract  Irritable bowel syndrome (IBS), frequently associated with psychological distress, is characterized by hypersensitivity to gut wall distension. Some probiotics are able to alleviate IBS symptoms and reduce visceromotor response to mechanical stimuli in animals. Moreover, we have previously shown that Lactobacillus farciminis treatment abolished the hyperalgesia to colorectal distension (CRD) induced by acute stress. The aims of the present study were to determine whether (i) stress‐induced visceral hyperalgesia modifies the expression of Fos, a marker of general neuronal activation, induced by CRD, (ii) this activation can be modulated by L. farciminis treatment. Female rats were treated by L. farciminis and CRD was performed after partial restraint stress (PRS) or sham‐PRS. The expression of Fos protein was measured by immunohistochemistry. After CRD or PRS, Fos expression was increased in spinal cord section (S1), nucleus tractus solitarius (NTS), paraventricular nucleus (PVN) of the hypothalamus, and in the medial nucleus of the amygdala (MeA). The combination of both stimuli, PRS and CRD, markedly increased this Fos overexpression in the sacral spinal cord section, PVN and MeA, but not in NTS. By contrast, a pretreatment with L. farciminis significantly reduced the number of Fos positive cells in these area. This study shows that PRS enhances Fos protein expression induced by CRD at the spinal and supraspinal levels in rats. Lactobacillus farciminis treatment inhibited this enhancing effect, suggesting that the antinociceptive effect of this probiotic strain results from a decrease of the stress‐induced activation/sensitization of sensory neurons at the spinal and supraspinal level.
Measurement of sand creep on a flat sand bed using a high‐speed digital camera
Authors: WANG YUAN , WANG DAWEI , WANG LI , ZHANG YANG
Published: March 2009

In this study, a novel method for measuring sand creep along a flat bed was developed. Specifically, a high‐speed digital camera was employed to record the sand creep movement. With an unsigned image subtraction, a new image queue could be formed from the original experimental images. The particle tracking was then calculated, after which the velocity and trajectory of the creep of sand grains was obtained. In addition, the creep fluxes of the different sand samples could also be calculated based on the velocities of the creep grains.