bluuebunny/crossref_metadata_2025 huggingface dataset:
Phosphorylation-mediated inactivation of coactivator-associated arginine methyltransferase 1
Authors: Higashimoto Ken , Kuhn Peter , Desai Dhaval , Cheng Xiaodong , Xu Wei
Published: July 2007

Multiple protein arginine methyltransferases are involved in transcriptional activation of nuclear receptors. Coactivator-associated arginine methyltransferase 1 (CARM1)-mediated histone methylation has been shown to activate nuclear receptor-dependent transcription; however, little is known about the regulation of its enzymatic activity. Here, we report that the methyltransferase activity of CARM1 is negatively regulated through phosphorylation at a conserved serine residue. When the serine residue is mutated to glutamic acid, which mimics the phosphorylated serine residue, the mutant CARM1 exhibits diminished ability to bind the methyl donor adenosylmethionine and diminished histone methylation activity. Moreover, such mutation leads to the inhibition of CARM1 transactivation of estrogen receptor-dependent transcription. Our results provide an example for the regulation of protein arginine methyltransferase activity by phosphorylation. As CARM1 is a potent transcriptional coactivator of estrogen receptor, our results suggest that phosphorylation of CARM1 serves as a unique mechanism for inactivating CARM1-regulated estrogen-dependent gene expression.
Interaction analysis of prenylated Rab GTPase with Rab escort protein and GDP dissociation inhibitor explains the need for both regulators
Authors: Wu Yao-Wen , Tan Kui-Thong , Waldmann Herbert , Goody Roger S. , Alexandrov Kirill
Published: July 2007

Prenylated Rab GTPases regulate intracellular vesicle trafficking in eukaryotic cells by associating with specific membranes and recruiting a multitude of Rab-specific effector proteins. Prenylation, membrane delivery, and recycling of all 60 members of the Rab GTPase family are regulated by two related molecules, Rab escort protein (REP) and GDP dissociation inhibitor (GDI). Biophysical analysis of the interaction of prenylated proteins is complicated by their low solubility in aqueous solutions. Here, we used expressed protein ligation to construct a semisynthetic fluorescent analogue of prenylated Rab7, Rab7-NBD-farnesyl. This molecule is soluble in the absence of detergent but is otherwise similar in its behavior to naturally prenylated Rab7 GTPase. To obtain information on the interaction of natively mono- and diprenylated Rab7 GTPases with REP and GDI molecules, we stabilized the former molecules in solution by using the β-subunit of Rab geranylgeranyl transferase, which we demonstrate to function as an unspecific chaperone of prenylated proteins. Using competitive titrations of mixtures of natively prenylated and fluorescent Rab, we demonstrate that monogeranylgeranylated Rab7 binds to the REP protein with a
K
d
value of ≈70 pM. The affinity of doubly prenylated Rab7 is ≈20-fold weaker. In contrast, GDI binds both prenylated forms of Rab7 with comparable affinities (
K
d
= 1–5 nM) but has extremely low affinity to unprenylated Rab molecules. The obtained data allow us to formulate a thermodynamic model for the interaction of RabGTPases with their regulators and membranes and to explain the need for both REP and GDI in Rab function.
Human endometriosis is associated with plasma cells and overexpression of B lymphocyte stimulator
Authors: Hever Aniko , Roth Richard B. , Hevezi Peter , Marin Maria E. , Acosta Jose A. , Acosta Hector , Rojas Jose , Herrera Rosa , Grigoriadis Dimitri , White Evan , Conlon Paul J. , Maki Richard A. , Zlotnik Albert
Published: July 2007

Endometriosis affects 10–20% of women of reproductive age and is associated with pelvic pain and infertility, and its pathogenesis is not well understood. We used genomewide transcriptional profiling to characterize endometriosis and found that it exhibits a gene expression signature consistent with an underlying autoimmune mechanism. Endometriosis lesions are characterized by the presence of abundant plasma cells, many of which produce IgM, and macrophages that produce BLyS/BAFF/TNFSF13B, a member of the TNF superfamily implicated in other autoimmune diseases. B lymphocyte stimulator (BLyS) protein was found elevated in the serum of endometriosis patients. These observations suggest a model for the pathology of endometriosis where BLyS-responsive plasma cells interact with retrograde menstrual tissues to give rise to endometriosis lesions.
Reaction coordinate of an enzymatic reaction revealed by transition path sampling
Authors: Quaytman Sara L. , Schwartz Steven D.
Published: July 2007

The transition path sampling method previously applied in our group to the reaction catalyzed by lactate dehydrogenase was used to generate a transition path ensemble for this reaction. Based on analysis of the reactive trajectories generated, important residues behind the active site were implicated in a compressional motion that brought the donor–acceptor atoms of the hydride closer together. In addition, residues behind the active site were implicated in a relaxational motion, locking the substrate in product formation. Although this suggested that the compression-relaxation motions of these residues were important to catalysis, it remained unproven. In this work, we used committor distribution analysis to show that these motions are integral components of the reaction coordinate.
Comparative Maps of Motion and Assembly of Filamentous Actin and Myosin II in Migrating Cells
Authors: Schaub Sébastien , Bohnet Sophie , Laurent Valérie M. , Meister Jean-Jacques , Verkhovsky Alexander B.
Published: July 2007

To understand the mechanism of cell migration, one needs to know how the parts of the motile machinery of the cell are assembled and how they move with respect to each other. Actin and myosin II are thought to be the major structural and force-generating components of this machinery ( Mitchison and Cramer, 1996 ; Parent, 2004 ). The movement of myosin II along actin filaments is thought to generate contractile force contributing to cell translocation, but the relative motion of the two proteins has not been investigated. We use fluorescence speckle and conventional fluorescence microscopy, image analysis, and computer tracking techniques to generate comparative velocity and assembly maps of actin and myosin II over the entire cell in a simple model system of persistently migrating fish epidermal keratocytes. The results demonstrate contrasting polarized assembly patterns of the two components, indicate force generation at the lamellipodium–cell body transition zone, and suggest a mechanism of anisotropic network contraction via sliding of myosin II assemblies along divergent actin filaments.
European guidelines for quality assurance in cervical cancer screening: recommendations for cervical cytology terminology
Authors: Herbert A. , Bergeron C. , Wiener H. , Schenck U. , Klinkhamer P. , Bulten J. , Arbyn M.
Published: July 2007

There are many different systems of cytology classification used in the member states of the European Union (EU) and many different languages. The following short annexe to Chapter 3 of the European Guidelines for Quality Assurance in Cervical Cancer Screening provides a framework that will allow different terminologies and languages to be translated into standard terminology based on the Bethesda system (TBS) for cytology while retaining the cervical intraepithelial neoplasia (CIN) classification for histology. This approach has followed extensive consultation with representatives of many countries and professional groups as well as a discussion forum published in Cytopathology (2005;16:113).

This article will describe the reporting of specimen adequacy, which is dealt with in more detail elsewhere in Chapter 3 of the guidelines, the optional general categorization recommended in TBS, the interpretation/cytology result and other comments that may be made on reports such as concurrent human papillomavirus testing and the use of automation review and recommendations for management. The main categories in TBS will be described in the context of CIN, dyskaryosis and dysplasia terminologies so that all may be translated into the same framework. These guidelines should allow European countries to adapt their terminology in such a way as to make their screening programmes comparable with each other as well as with programmes elsewhere in the world.
Landscape, habitat characteristics and the genetic population structure of two caddisflies
Authors: WILCOCK HELEN R. , BRUFORD MICHAEL W. , NICHOLS RICHARD A. , HILDREW ALAN G.
Published: July 2007

1. In a region of south‐eastern England, we investigated the hierarchical genetic structure of populations of two stream‐dwelling caddisflies (Trichoptera: Polycentropodidae) with contrasting distributions:Plectrocnemia conspersainhabits numerous small, patchily distributed seeps and streams, while the confamilialPolycentropus flavomaculatusis found in fewer but larger streams and rivers. We also contrasted the genetic structure ofP. conspersain the lowland south‐east with that in an upland region in the north west.

2. Microsatellite genotypes were obtained from samples of both species taken from a ‘core area’ and at sites 15, 40 and 100 km from this core (two regions forP. conspersa, totalling 45 sites and 1405 larvae; one region forP. flavomaculatus, totalling 10 sites and 269 larvae).

3. The genetic structure ofP. conspersadiffered in the two regions. In the upland north‐west, significant genetic differentiation was observed at a spatial scale of around 40 km from the core, while there was no structure in the lowland south‐east up to around 100 km. Areas of high altitude did not appear directly to reduce gene flow, whereas other potential landscape barriers, including particular geological formations, large urban areas and the sea had a pronounced effect.

4. Weak genetic differentiation inP. conspersaacross large distances, particularly in the lowland south‐east, suggests that it disperses strongly, facilitating gene flow within and between catchments. Conversely, forP. flavomaculatuswe found strong genetic differentiation between almost all sites, suggesting that dispersal is much more limited.

5. Greater dispersal in the patchily distributedP. conspersathan inP. flavomaculatus, which occupies larger and presumably more persistent habitats, could be a general feature of other similarly distributed aquatic insects. While higher relief is potentially a partial barrier to dispersal,P. conspersamust have effective gene flow through such apparently inhospitable terrain, perhaps attributable to dispersal between neighbouring small and ephemeral populations. Indeed, its exploitation of headwaters and seeps requires the ability to disperse between such sites. Apparently it cannot, however, overcome more continuous barriers, consisting of large tracts of landscape with few habitable larval sites. Such landscapes, including those created by humans, may have a stronger effect on population connectivity and colonization in the longer term.
Faecal pellets in streams: their binding, breakdown and utilization
Authors: JOYCE PAUL , WARREN LUKE L. , WOTTON ROGER S.
Published: July 2007

1. Faecal pellets of Gammarus (shredders) and Simulium larvae (suspension feeders) are bound by exopolymers. Immediately after egestion, Gammarus pellets are covered by a peritrophic membrane that breaks up within hours, although pellets remain intact because of internal binding materials.

2. Although they expand soon after egestion, the faecal pellets of Gammarus and Simulium remain intact for more than 30 days. Their internal structure is altered and the main agents of this change are bacteria that have survived passage through the gut (and become bound within pellets).

3. When disrupted physically, freshly egested (1‐ to 2‐day old) Simulium faecal pellets break up into relatively large pieces whereas freshly egested Gammarus faecal pellets break apart into much smaller pieces. Disruption of 30‐day old Simulium faecal pellets results in similar sized pieces to those from freshly egested pellets, but disruption of 30‐day old Gammarus pellets produces pieces that are two orders of magnitude larger than those resulting from disruption of freshly egested pellets.

4. Faecal pellets of Gammarus and Simulium are eaten by stream invertebrates and are sites of microbial breakdown. Faecal pellets are a source of organic matter for benthic invertebrates, bacteria and, indirectly, for plants.
Predictable changes in trophic community structure along a spatial disturbance gradient in streams
Authors: LEPORI FABIO , MALMQVIST BJÖRN
Published: July 2007

1. We assessed insect and resource standing stocks along a spatial gradient of flood disturbance in 19 sub‐alpine Swedish streams to test the prediction that change in trophic structure arises from the joint action of disturbance, which affect basal resources, and resource‐control, which ties the response of the consumers to the response of the resources.

2. Trophic structure, quantified as scores of non‐metric multidimensional scaling based on the biomass of insect trophic groups, changed predictably along the disturbance gradient. In early summer, predators and algae feeders decreased relative to suspension feeders with increasing disturbance; in autumn, algae feeders decreased relative to leaf feeders with increasing disturbance.

3. Across the disturbance gradient, the biomass of algae‐, deposit‐ and leaf‐feeders was principally controlled by the availability of the respective resource (algae, fine detritus and coarse detritus), while disturbance only had subsidiary effects on algae feeders in early summer.

4. Overall, patterns in trophic‐group biomass along the disturbance gradient were more likely to reflect indirect effects of disturbance via impact on the resources, which reverberated to the consumers because of resource‐control, rather than direct effects. In contrast with the view that stream communities are the result of stochastic colonization following disturbance events, in the study streams the trophic structure of insect assemblages is predictable and partly organized by resource‐control across a broad range of disturbance conditions.
Kinetics of NO3‐ net fluxes in Pinus pinaster, Rhizopogon roseolus and their ectomycorrhizal association, as affected by the presence of NO3‐ and NH4+
Authors: GOBERT ANTHONY , PLASSARD CLAUDE
Published: July 2007

The impact of mineral N supply, N‐free or NO3‐ with or without NH4+, on the subsequent uptake of NO3‐ by maritime pine seedlings associated with the ectomycorrhizal fungus Rhizopogon roseolus was studied using ion‐selective microelectrodes. NO3‐ net fluxes into N‐starved non‐mycorrhizal short roots (NMSRs) were low and measurable only over the NO3‐ concentration range of 0–70 µM. The simple kinetics observed in those roots may reflect the dominant operation of a high‐affinity NO3‐ transport system (HATS) which is constitutive. NO3‐ pretreatment increased the NO3‐ net fluxes and led to a complex kinetics that may reflect the operation of other HATS. A simple kinetics was observed in plants pre‐incubated at high NH4+ concentration. In contrast, NO3‐ uptake kinetics presented only one saturation phase in the fungus, whether associated with the plant or not. NO3‐ uptake was greater after a pretreatment in N‐free or NO3‐ solution, but NH4+ pretreatment led to a threefold reduction in NO3‐ uptake. These results suggest that the regulation of NO3‐ transport systems varies between the host and the fungal partner. This variation is likely to contribute to the positive effect of mycorrhizal association on N uptake in plants when the N supply is low and fluctuating.
Systematic status of the genus Formosaphis Takahashi and the evolution of galls based on the molecular phylogeny of Pemphigini (Hemiptera: Aphididae: Eriosomatinae)
Authors: ZHANG HECAI , QIAO GEXIA
Published: July 2007

Abstract Aphids of the tribe Pemphigini (Hemiptera: Aphididae: Eriosomatinae) can induce pseudo‐galls or galls on their primary host plants. Those belonging to subtribe Prociphilina often produce pseudo‐galls, and those of the other subtribe, Pemphigina, can form not only pseudo‐galls but also true galls. Pseudo‐galls are all formed on the leaves, whereas true galls, although all similar in shape, can be located on the joint of the leaf blade and the petiole, the middle of the petiole and the branches of the host plant. From a first phylogenetic analysis of Pemphigini based on nuclear elongation factor‐1α (EF‐1α) sequences, no support was found for the monophyly of Pemphigini, but subtribe Pemphigina was monophyletic with robust support. Formosaphis obviously clusters with Pemphigina which has Populus as a primary host. Formosaphis belongs to Pemphigini and its unknown primary host is probably Populus. The evolution of galls in Pemphigina is discussed based on the phylogenetic tree drawn from EF‐1α sequences and mitochondrial cytochrome oxidase subunit I (COI) sequences. The results suggest that, in this subtribe, the closed gall is preceded by an open pseudo‐gall, and the galls move their locations from the joint of the leaf blade and the petiole to the middle of the petiole and, eventually, to the branch of the host plant. Such an evolutionary tendency may provide aphids with more protection and nutrition.
MicroRNAs in loblolly pine (Pinus taedaL.) and their association with fusiform rust gall development
Authors: Lu Shanfa , Sun Ying‐Hsuan , Amerson Henry , Chiang Vincent L.
Published: July 2007

MicroRNAs (miRNAs) are endogenous small RNAs that can have large‐scale regulatory effects on development and on stress responses in plants. The endemic rust fungusCronartium quercuumf. sp.fusiformecauses fusiform rust disease in pines, resulting in the development of spindle‐shaped galls (cankers) on branches or stems. This disease is the most destructive disease of pines in the southern USA. To test whether miRNAs play roles in fusiform rust gall development, we cloned and identified 26 miRNAs from stem xylem of loblolly pine (Pinus taeda), which belong to four conserved and seven loblolly pine‐specific miRNA families. Forty‐three targets for nine of these 11 families were experimentally validatedin vivo. Sequence analysis suggested that the target cleavage site may be determined not only by the miRNA sequence but also by the target sequence. Members of three loblolly pine‐specific miRNA families target a large number of non‐protein coding transcripts, and one of these families could also initiate secondary phased production from its target of a putative trans‐acting short interfering RNA (ta‐siRNA). Expression of 10 of these 11 miRNA families was significantly repressed in the galled stem. PCR‐based transcript quantification showed complex expression patterns of these miRNAs and their targets in the galled tissues and in tissues surrounding the gall. We further predict 82 plant disease‐related transcripts that may also response to miRNA regulation in pine. These results reveal a new genetic basis for host–pathogen interactions in the development of fusiform rust gall.
A comparative study of nutrients utilization, alkaline phosphatase activity and creatinine concentration in the serum of sheep and goats fed diets based on olive leaves
Authors: Yáñez‐Ruiz D. R. , Molina‐Alcaide E.
Published: July 2007

The aim of this study was to compare, in goats and wethers, the nutritive utilization of diets including olive leaves (OL) and the possible detrimental effect of that by‐product. Three different diets were studied: OL, OL plus polyethylene glycol (PEG) (OLP) and OL supplemented with barley [164 g/kg dry matter (DM)] and faba beans (59 g/kg DM) (OLSUP). Apparent digestibility of nutrients and energy and nitrogen balances were determined along with creatinine concentrations and alkaline phosphatase (ALP) activity in the serum. The apparent digestibility of OL was low and similar in goats and wethers (54.4% and 53.5%, 22.2% and 21.6% and, 47.7% and 46.6% for DM, crude protein and neutral detergent fibre in goats and wethers, respectively). The addition of PEG did not improve (p > 0.05) digestibility of OL, although a slightly beneficial effect on the digestion of structural carbohydrates was observed (up to 8.4% and 7.10% in goats and wethers, respectively). The supplementation of OL with barley and faba beans increased (p < 0.001) the apparent digestibility of nutrients and the energy value. The consumption of the OL led to high ALP activity in the serum. Despite the higher sensitivity of wethers to Cu levels, our results show a similar digestive use of OL by wethers and goats and suggest the need of further comparative investigations focusing on the effect of the presence of high Cu levels in the by‐product on the animal health.
Dietary crude protein has minimal effect on the activity of selected enzymes of methionine catabolism in kittens fed diets near‐limiting in methionine*
Authors: Strieker M. J. , Morris J. G. , Avery E. H. , Freedland R. A. , Rogers Q. R.
Published: July 2007

Previous experiments have shown that increasing the dietary crude protein (CP) of cats does not increase urea cycle enzymes or alanine amino transferase as occurs in rats. Also when an essential amino acid (EAA) is limiting in a diet for growing kittens, the kittens do not exhibit an amino acid imbalance when other EAAs are added to the diet. To study the metabolic basis for these observations which are different from that found in omnivores and herbivores, the hypothesis that increased dietary CP decreases methionine catabolism, so more is spared for growth, was tested. Fifteen male kittens were randomly assigned to one of three dietary treatments. Each diet contained 2.5 g l‐methionine/kg diet and 200, 300 or 500 g CP/kg diet. The livers and kidneys were removed and assayed for methionine transaminase (MTA), cystathionase (CASE) and cystathionine synthase (CS). Free amino acid concentrations were determined in liver, kidney and plasma. The 300 and 500 g CP/kg groups had significantly greater kidney weights and body weight gains than the 200 g CP/kg group. Hepatic MTA activity was lower in the 300 than the 200 or 500 g CP/kg groups (p < 0.05). Renal MTA and CASE activities were 35% and 50% greater, respectively, for the 500 g CP/kg group than for the 200 g CP/kg diet group (p < 0.05). Renal CS activities for the 300 and 500 g CP/kg groups were 29% (p > 0.05) and 38% (p < 0.05) greater, respectively, than the 200 g CP/kg group. Cyst(e)ine concentrations were lower in the livers of the 500 g CP/kg group than the 200 g CP/kg group (p < 0.05). Cystathionine was lower in plasma and kidney from the 500 g CP/kg diet group than from the 200 g CP/kg diet group (p < 0.05). It was concluded that the metabolic basis for the increased growth of kittens fed diets marginally limiting in methionine, with increasing concentrations of dietary CP, was not mediated through decreased enzyme activity associated with the catabolism of methionine, but was the result of an increase in food (methionine) intake.
The absorption, tissue distribution and excretion of enteraly administered α‐ketoglutarate in rats
Authors: Filip R. , Pierzynowski S. G.
Published: July 2007

The absorption, tissue distribution and excretion of enteral alpha‐ketoglutarate (AKG) was studied in four experiments. Six male Sprague Dawley rats were used to investigate the excretion of AKG in urine and faeces. Thirty rats, randomly assigned to five groups, were used to investigate the distribution of AKG in body tissues. They were gavaged with AKG enriched with 3 μCi/kg BW of 14C uniformly marked AKG. Fourteen male Sprague Dawley rats were used to study the absorption of AKG (duodenum vs. ileum). Intestinal recovery of NaAKG vs. CaAKG was investigated in 36 rats. There was no significant excretion of non‐metabolized AKG in the urine and faeces. There was no significant difference in the systemic levels of AKG when comparing the proximal to distal small intestine infusion. Up to 50%, 30% and 20% of gastrically delivered AKG was recovered in the stomach, 0.5, 1 and 2 h after gavage; the jejunal recovery achieved a maximum of 3%, 30 min after gavage, and was not detectable 2 h later. There was a relatively high distribution of 14C‐AKG in the tissues (e.g. liver, brain, bones, skin, muscles), 3 h after gavage, up to 70% of the administered dose. In conclusion, the high rate of retention of the carbon from AKG allows the postulation that there is a non‐energetic mode of metabolism of intragastrically administered AKG. After conversion to final metabolites, AKG penetrates into all tissues and organs of rats, including the bone tissue. Intestinal absorption of AKG does not depend on the type of AKG salt administered.
Isolation of Sporothrix schenckii from the environmental sources of cutaneous sporotrichosis patients in Himachal Pradesh, India: results of a pilot study
Authors: Mehta Karan Inder Singh , Sharma Nand Lal , Kanga Anil K. , Mahajan Vikram K. , Ranjan Nitin
Published: July 2007

Himachal Pradesh, India is a known endemic area for cutaneous sporotrichosis. No attempt has been made to isolate Sporothrix schenckii, the causative fungus, from environmental sources in this region or in India as such. This prospective study was carried out to isolate Sporothrix schenckii from different environmental samples collected from the vicinity of cutaneous sporotrichosis patients. All patients of cutaneous sporotrichosis diagnosed during March 2005–February 2006 were studied. Twenty‐one biopsy specimens and 62 environmental samples of soil, various thorns, corn‐stalk, grass‐blades and sphagnum moss were subjected to mycologic culture on Sabouraud’s glucose agar. Sporothrix schenckii was identified by colony characteristics, lacto‐phenol cotton blue mounts and demonstration of temperature dimorphism. These patients (F : M 15 : 6) were between 12 and 72 years of age and had cutaneous lesions for 45 days to 4 years. Lymphocutaneous and fixed cutaneous sporotrichosis was seen in 14 (66.6%) and 7 (33.3%) patients respectively. Extremities were involved in 16 (76.2%); and 5 (23.8%) patients had facial lesions. Ten (47.4%) biopsy specimens and six environmental (three soil, three corn‐stalk) samples were culture‐positive, which showed morphological characteristics suggesting Sporothrix schenckii. No variation in colony characteristics and mycelial morphology was observed in growth isolates from clinical or environmental samples. Temperature dimorphism was observed in all the 10 isolates obtained from the clinical specimens and in two isolates cultured from corn‐stalk. Corn‐stalks are evidently important sources of Sporothrix schenckii infection although subsequent contamination of wounds appears more important for development of clinical disease. Culture of Sporothrix schenckii from environmental sources may not be always possible to correlate with profile of injuries.
Amphotericin B‐induced cutaneous leucocytoclastic vasculitis: case report
Authors: Cagatay A. Atahan , Taranoglu Ozer , Alpay Nilufer , Tufan Fatih , Karadenız Asli , Kapmaz Mahir , Buyukbabanı Nesimi
Published: July 2007

A 37‐year‐old female patient was diagnosed with ulcerative colitis 8 months ago and medical treatment with oral azathioprine, low‐dose corticosteroids and 5‐ASA was started. Following 3 months without any symptoms, the patient had total colectomy and ileostomy. After this period, liposomal amphotericin B (3 mg kg−1 day−1) was given with the diagnosis of probable fungal infection. Palpable purpuric skin lesions on the anterior surface of both legs appeared on the 55th day of amphotericin B treatment. Histological examination of a skin biopsy was consistent with leucocytoclastic vasculitis. We present a case of cutaneous leucocytoclastic vasculitis in which amphotericin B might presumably be the aetiological factor.
Agrobacterium‐mediated transformation of Japanese lawngrass (Zoysia japonicaSteud.) containing a syntheticcryIA(b)gene fromBacillus thuringiensis
Authors: Zhang Lei , Wu D. , Zhang Li , Yang C.
Published: July 2007

Transgenic Japanese lawngrass plants containing a syntheticcryIA(b)gene fromBacillus thuringiensisunder the control of a maize ubiquitin promoter were developed byAgrobacterium‐mediated transformation. A total of 1540 embryogenic calluses derived from dormancy‐removal mature seeds were co‐cultured with the disarmed strain EHA105 harbouring the binary vector pKUB. Three days after co‐culture with EHA105 in the dark at 21°C, transientβ‐glucuronidase (GUS) expression frequency was 74.2%. After selection with 100 mg/l hygromycin B, a total of over 50 independent resistant cell clones and 25 regenerated plants were obtained. The integration and expression of thecryIA(b)gene into the genome was confirmed in 22 regenerated plants by the GUS histochemical assay, PCR amplification, Southern blotting and Western blotting analysis, with a transformation efficiency of 1.4%. The entire process from callus induction of mature seeds to production of transgenic plantlets was 80–100 days. T1progeny segregation analysis of these transgenic lines demonstrated that 59.1% of the transgenic events were inherited in a typical Mendelian fashion.
SPINAL α‐AMINO‐3‐HYDROXY‐5‐METHYL‐4‐ISOXAZOLEPROPIONIC ACID RECEPTORS MAY MEDIATE THE ANALGESIC EFFECTS OF EMULSIFIED HALOGENATED ANAESTHETICS
Authors: Hang Li‐Hua , Shao Dong‐Hua , Yang Ying‐Hong , Dai Ti‐Jun , Zeng Yin‐Ming
Published: July 2007

The present study was designed to investigate the relationship between spinal cord α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionic acid (AMPA) receptors and the analgesic effects of emulsified halogenated anaesthetics.

After having established the mouse model of analgesia by intraperitoneal or subcutaneous injections of appropriate doses of emulsified enflurane, isoflurane or sevoflurane, we injected different doses of AMPA intrathecally and observed effects on the pain threshold using the hot‐plate and acetic acid‐induced writhing tests.

The results showed that intrathecal injection of AMPA (0.25, 0.5 and 1.0 ng) did not affect the pain threshold on the hot‐plate test or the writhing times in conscious mice. In contrast, AMPA (0.25, 0.5 and 1.0 ng intrathecally) significantly and dose‐dependently decreased the pain threshold on the hot‐plate test and increased the writhing times in mice treated with emulsified anaesthetics.

These results suggest that spinal AMPA receptors may be important targets for the analgesic effects of emulsified enflurane, isoflurane and sevoflurane.
NITRIC OXIDE MODULATES AIR EMBOLISM‐INDUCED LUNG INJURY IN RATS WITH NORMOTENSION AND HYPERTENSION
Authors: Liu Yen Chin , Kao Shang Jyh , Chuang I Chun , Chen Hsing I
Published: July 2007

Air embolism the in lungs induces microvascular obstruction, mediator release and acute lung injury (ALI). Nitrite oxide (NO) plays protective and pathological roles in ALI produced by various causes, but its role in air embolism‐induced ALI has not been fully investigated.

The purpose of the present investigation was to elucidate the involvement of NO and pro‐inflammatory cytokines in the pathogenesis of ALI following air infusion into isolated perfused lungs from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) rats.

The extent of ALI was evaluated by changes in lung weight, Evans blue dye leakage, the protein concentration in the bronchoalveolar lavage and pathological examination. We also measured nitrite/nitrate (NOx), tumour necrosis factor (TNF)‐α and interleukin (IL)‐1β concentrations in lung perfusate and determined cGMP in lung tissue.

The NO synthase (NOS) inhibitors NG‐nitro‐l‐arginine methyl ester (l‐NAME) and l‐N6‐(1‐iminoethyl)‐lysine (l‐Nil), as well as the NO donors sodium nitroprusside (SNP) and s‐nitroso‐N‐acetylpenicillamine (SNAP), were administered 30 min before air embolism at a concentration of 10−3 mol/L in the lung perfusate.

Air embolism‐induced ALI was enhanced by pretreatment with l‐NAME or l‐Nil, but was alleviated by SNP or SNAP pretreatement, in both SHR and WKY rats. In both SHR and WKY rats, AE elevated levels of NOx (2.6 and 28.7%, respectively), TNF‐α (52.7 and 158.6%, respectively) and IL‐1β (108.4 and 224.1%, respectively) in the lung perfusate and cGMP levels in lung tissues (35.8 and 111.2%, respectively). Pretreatment with l‐LAME or l‐Nil exacerbated, whereas SNP or SNAP abrogated, the increases in these factors, except in the case of NOx (levels were decreased by l‐LAME or l‐Nil pretreatment and increased by SNP or SNAP pretreatment).

Air embolism caused increases in the lung weight (LW)/bodyweight ratio, LW gain, protein concentration in bronchoalveolar lavage and Evans blue dye leakage. These AE‐induced changes were less in lungs isolated from SHR compared with normotensive WKY rats.

The results suggest that ALI and associated changes following air embolism in lungs isolated from SHR are less than those in WKY rats. Nitric oxide production through inducible NOS isoforms reduces air embolism‐induced lung injury and associated changes. Spontaneously hypertensive rats appear to be more resistant than WKY rats to air embolism challenge.
Miliary brain metastasis presenting with dementia: Progression pattern of cancer metastases in the cerebral cortex
Authors: Ogawa Masaya , Kurahashi Kozo , Ebina Akio , Kaimori Mitsuomi , Wakabayashi Koichi
Published: July 2007

We report an autopsy case of an 82‐year‐old woman with progressive dementia due to miliary brain metastasis from lung adenocarcinoma. The patient presented with dementia 5 months prior to death and suddenly died of pulmonary hemorrhage. Postmortem examination revealed normal appearance of the brain. However, there were numerous foci of cancer metastasis in all parts of the brain on light microscopic examination. The carcinoma cells were located in the perivascular (Virchow‐Robin) space and did not invade to the brain parenchyma. The carcinoma cells were also found in the subpial space. In the cerebral cortex, foci of metastasis appeared to spread in the following way: tiny foci of metastasis initially occur in the middle cortical layer, then spread to all layers through the perivascular space, and finally reach the subpial space and subcortical white matter. Although the junction between gray and white matter is a preferred site for usual brain metastasis, middle cortical layer was considered to be the initial site for metastasis in our patient. The perivascular pial sheath plays an important role for the development of miliary brain metastasis.
Neurotrophic and growth factor gene expression profiling of mouse bone marrow stromal cells induced by ischemic brain extracts
Authors: Qu Runjiang , Li Yi , Gao Qi , Shen Lihong , Zhang Jing , Liu Zhongwu , Chen Xiaoguang , Chopp Michael
Published: July 2007

Treatment of rodents after stroke with bone marrow stromal cells (BMSCs) improves functional outcome. However, the mechanisms underlying this benefit have not been ascertained. This study focused on the contribution of neurotrophic and growth factors produced by BMSCs to therapeutic benefit. Rats were subjected to middle cerebral artery occlusion and the ischemic brain extract supernatant was collected to prepare the conditioned medium. The counterpart normal brain extract from non‐ischemic rats was employed as the experimental control. Using microarray assay, we measured the changes of the neurotrophin associated gene expression profile in BMSCs cultured in different media. Furthermore, real‐time RT‐PCR and fluorescent immunocytochemistry were utilized to validate the gene changes. The morphology of BMSCs, cultured in the ischemic brain‐conditioned medium for 12 h, was dramatically altered from a polygonal and flat appearance to a fibroblast‐like long and thin cell appearance, compared to those in the normal brain‐conditioned medium and the serum replacement medium. Forty‐four neurotrophin‐associated genes in BMSCs were identified by microarray assay under all three culture media. Twelve out of the 44 genes (7 neurotrophic and growth factor genes, 5 receptor genes) increased in BMSCs cultured in the ischemic brain‐conditioned medium compared to the normal brain‐conditioned medium. Real time RT‐PCR and immunocytochemistry validated that the ischemic brain‐conditioned medium significantly increased 6/7 neurotrophic and growth factor genes, compared with the normal brain‐conditioned medium. These six genes consisted of fibroblast growth factor 2, insulin‐like growth factor 1, vascular endothelial growth factor A, nerve growth factor beta, brain‐derived neurotrophic factor and epidermal growth factor. Our results indicate that transplanted BMSCs may work as ‘small molecular factories’ by secreting neurotrophins, growth factors and other supportive substances after stroke, which may produce therapeutic benefits in the ischemic brain.
Decommissioning Western Australia's First Bauxite Mine: Co‐evolving vegetation restoration techniques and targets
Authors: Grant Carl , Koch John
Published: July 2007

Summary  The closure and decommissioning of Alcoa's first mine in Western Australia at Jarrahdale was a landmark for the company. This article describes the strategies and research contributing to successful mine rehabilitation in the Jarrah forest. Lessons learnt at Jarrahdale are being applied to other Western Australian mines. Over 30 years of research have resulted in a strong working knowledge of ecosystem function in this environment. The direct linkage of research to operational practices provides two‐way benefits for practitioners and researchers. The Jarrahdale mine is the first mine in Western Australia to have received a certificate of completion from the state government.
RISK MANAGEMENT OF ENDOSCOPIC SPHINCTEROTOMY FOR CHOLEDOCHOLITHIASIS
Authors: Ito Kei , Fujita Naotaka , Noda Yutaka , Kobayashi Go , Horaguchi Jun , Takasawa Osamu , Obana Takashi , Endo Takuro , Nakahara Kazunari
Published: July 2007

Endoscopic retrograde cholangiopancreatography (ERCP) is accepted as a safe, direct technique for evaluating pancreatobiliary diseases. Since the introduction of endoscopic sphincterotomy (EST), ERCP has evolved from a diagnostic procedure to a therapeutic one. ERCP with EST and stone removal is a valuable therapeutic modality in choledocholithiasis. Herein are reviewed the complications of EST in cases of choledocholithiasis along with a discussion of the risk management of this particular procedure. Pancreatitis is the most common ERCP‐related complication. Hemorrhage is primarily a complication related to sphincterotomy. Other complications include perforation, cholangitis, cholecystitis, and sedation‐related cardiopulmonary complications. Endoscopists who perform ERCP should have appropriate training and be knowledgeable of such complications.
COLONIC MICROPERFORATION AFTER ENDOSCOPIC PIECEMEAL MUCOSAL RESECTION
Authors: Uragami Naoyuki , Akiko Chino , Igarashi Masahiro , Takahashi Hiroshi , Fujita Rikiya
Published: July 2007

A 45‐year‐old woman had an Is + IIa laterally spreading tumor (granular type), 30 mm in size, located in the lower rectum. This lesion was removed by endoscopic piecemeal mucosal resection (EPMR) combined with argon plasma coagulation. The area of the defect after EPMR was completely closed with endoscopic clipping to avoid delayed bleeding and perforation. Perforation, such as tearing of the muscle layer, was not recognized during EPMR. However, abdominal radiography, which the authors perform routinely after EPMR, showed free air near the rectum. The patient did not complain of abdominal pain except for anal discomfort, and conservative treatment with antibiotics was selected. The surgeon inserted a tube from the anus to prevent expansion of the rectum and to keep unwanted digestive juices from leaking out from the damaged area. The maximum C‐reactive protein level was 17.2 mg/dL 2 days after EPMR, and on day 6 was <1.0 mg/dL. An oral diet was resumed, and the patient was discharged at 8 days after the EMR.
INFLAMMATORY PROTRUDING LESION IN THE INTRAHEPATIC BILIARY DUCT DIAGNOSED ON NARROW BAND IMAGING–VIDEOCHOLANGIOSCOPY
Authors: Yamauchi Junichi , Yazumi Shujiro , Kida Masaya , Shio Seiji , Tada Shinsuke , Ida Hiroshi , Shawkat Mohamed , Harada Daisuke , Chiba Tsutomu
Published: July 2007

A 66‐year‐old woman who had a repeated history of cholangitis since undergoing choledocho‐jejunostomy for post‐cholecystectomy common bile duct stricture, was referred to Kyoto University Hospital for further examination of the stricture of left intrahepatic biliary duct (IHBD). Endoscopic retrograde cholangiography (ERC) showed obstruction of the left IHBD with slightly protruding lesion. Peroral cholangioscopy (POCS) with narrow band imaging (NBI) demonstrated that a protruding lesion with papillary surface had neither tumor vessels on the surface of the tumor nor microvessel in each papillary projection. Biopsy presented only inflammation. Pathological analysis of the resected left lobe also showed only inflammatory change. NBI‐videocholangioscopy appears to be useful for diagnosing biliary disease by evaluating the tumor vessels including microvessel in papillary projection.
CONTACT ULTRA‐HIGH MAGNIFYING ENDOSCOPY CAN DIFFERENTIATE SQUAMOUS CELL CARCINOMA FROM NON‐CANCEROUS SQUAMOUS CELLS IN THE ESOPHAGUS: TWO CASES OF SUPERFICIAL ESOPHAGEAL CARCINOMA
Authors: Kaise Mitsuru , Goda Ken‐ich , Yoshida Yukinaga , Yonezawa Jin , Kato Masayuki , Tajiri Hisao
Published: July 2007

Endocytoscopy, a type of contact ultra‐high magnifying endoscopy, enables in vivo observation of cells in the gastrointestinal tract. To test its clinical relevance, endocytoscopy was conducted on ex vivo specimens from two cases of superficial esophageal carcinoma that were resected by endoscopic mucosal resection (EMR). Using a catheter‐type system with ×450 magnification, endocytoscopic observation was performed on small areas of cancerous and non‐cancerous squamous cells, which were subsequently retrieved for pathological examination on horizontal sections. In both cases, endocytoscopy identified non‐cancerous areas as cells with sparsely distributed round nuclei and a low nucleus‐cytoplasm (NC) ratio, which correlated well with histological sections. Similarly, the endocytoscopy correlated well with histological sections of cancerous areas and identified cells with densely distributed irregular nuclei with a high NC ratio. In conclusion, the high correlation between histological and endocytoscopic identification of cancerous and non‐cancerous lesions may enable endocytoscopic diagnosis that is of comparable accuracy to the current pathological methodology. A prospective in vivo study is required to confirm the evidence.
AUTOFLUORESCENCE IMAGING IS USEFUL TO ASSESS ACTIVITY OF ULCERATIVE COLITIS
Authors: Fujiya Mikihiro , Saitoh Yusuke , Watari Jiro , Moriichi Kentaro , Kohgo Yutaka
Published: July 2007

The efficacy of autofluorescence imaging (AFI) for assessing ulcerative colitis (UC) activity was reviewed. AFI images of UC lesions could be classified into four categories: green; green with purple spots; purple with green spots; and purple. The strength of purple in AFI images of UC lesions is related to histologic severity, thus this classification is helpful to assess the activity of UC. The autofluorescence of AFI images was also quantified using color analysis software. Active lesions had significantly low autofluorescence/reflex ratio than inactive areas in UC patients. The ratio is inversely proportional to histologic severity. When the cut‐off is 0.9, the accuracy to predict histologic activity by AFI is 92%.
COMPLICATIONS OF ENDOSCOPIC SPHINCTEROTOMY FOR CHOLEDOCHOLITHIASIS: SAFETY AND BENEFITS OF PANCREATIC STENT PREPLACEMENT
Authors: Kutsumi Hiromu , Funatsu Eiji , Morita Yoshinori , Yoshida Masaru , Inokuchi Hideto , Azuma Takeshi
Published: July 2007

Endoscopic sphincterotomy (EST) is one of the most common procedures for treatment of choledocholithiasis. However, EST sometimes causes complications such as pancreatitis and bleeding. Especially, precut is more risky for pancreatitis. In contrast, bleeding after EST should be treated immediately. Clipping technique is the ideal method for bleeding but is very difficult technically. The authors have carried out precut with needle knife and treated bleeding with argon plasma coagulation after placement of pancreatic duct stent (PS) to prevent pancreatitis. This retrospective study showed the safety and benefit of stent preplacement for these two representative problems. Complications in 174 patients with choledocholithiasis treated by EST were collected and investigated retrospectively. In total, 92% were standard EST, 8% were precut over PS. Pancreatitis occurred in 13.1% of the standard EST group and in 7.1% of the precut over PS group. Bleeding after EST occurred in three cases (1.7%). Two patients were treated easily and safely by argon plasma coagulation after placement of PS.
TECHNIQUES AND PITFALLS OF ENDOSCOPIC SUBMUCOSAL DISSECTION FOR COLORECTAL TUMORS
Authors: Oka Shiro , Tanaka Shinji , Kaneko Iwao , Kanao Hiroyuki , Chayama Kazuaki
Published: July 2007

Endoscopic submucosal dissection (ESD) for colorectal tumors is steadily being developed. Safety and standardization of ESD for colorectal tumors have not been yet established because of the technical difficulties and the unsuitable anatomical characteristics of the colon and rectum. The authors mainly use a Flex knife for mucosal incision and a Hook knife for submucosal dissection to perform ESD safely. Skillful colonoscopic control, selection of scope, distal attachment tip hood, adequate high‐frequency generator and correct approach strategy should all be considered for safe performance of ESD. However, the incidence of indicative lesions is rare because the majority of colorectal tumors are adenomatous large laterally spreading tumors, which can be cured by intentional endoscopic piecemeal resection. At present, ESD for colorectal tumors should be performed only at central facilities that have expert colonoscopists. With the development of new devices and associated techniques, technical standardization of ESD for colorectal tumors is expected in the near future.
SUCCESSFUL ENDOSCOPIC CLOSURES OF COLONIC PERFORATIONS REQUIRING ABDOMINAL DECOMPRESSION AFTER ENDOSCOPIC MUCOSAL RESECTION AND ENDOSCOPIC SUBMUCOSAL DISSECTION FOR EARLY COLON CANCER
Authors: Saito Yutaka , Matsuda Takahisa , Kikuchi Tsuyoshi , Ikehara Hisatomo , Uraoka Toshio
Published: July 2007

Endoscopic submucosal dissection (ESD) for colorectal cancer is not widely accepted because of its technical difficulty and the risk of perforation. In addition, the risk of peritonitis cannot be completely eliminated even if a perforation is closed successfully. Reported here are two cases of early colon cancer in which the patients sustained iatrogenic perforations of the ascending colon during conventional endoscopic mucosal resection and of the sigmoid colon during ESD, respectively, requiring abdominal decompression with an 18 G Medicut needle. Both of these perforations were successfully treated by endoscopic clipping. In conclusion, conservative medical management may be possible in patients who have undergone successful closure of colonic perforations using endoscopic clipping. In order to perform immediate endoscopic closure, abdominal decompression has been useful to decrease patient discomfort and colonic lumen collapse. Now, CO2 insufflation is being used effectively for the prevention of pneumoperitoneum.
MULTICENTER STUDY DESIGN OF THE EX VIVO EVALUATION OF ENDOCYTOSCOPY IN ESOPHAGEAL SQUAMOUS CELL CARCINOMA
Authors: Muto Manabu , Fujishiro Mitsuhiro , Sato Yoshitaka , Niwa Yasumasa , Kaise Mitsuru , Kato Mototsugu , Takubo Kaiyo
Published: July 2007

Optical technological innovations enable us to visualize cellular nuclei endoscopically. Herein is described a protocol design for a multicenter study for the ex vivo evaluation of endocytoscopy. The present study was performed by the Endoscopy Forum Japan study group.