bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural and Enzymatic Characterization of Os3BGlu6, a Rice β-Glucosidase Hydrolyzing Hydrophobic Glycosides and (1→3)- and (1→2)-Linked Disaccharides
Authors: Seshadri Supriya , Akiyama Takashi , Opassiri Rodjana , Kuaprasert Buabarn , Cairns James Ketudat 
Published: July 2009

Abstract
Glycoside hydrolase family 1 (GH1) β-glucosidases play roles in many processes in plants, such as chemical defense, alkaloid metabolism, hydrolysis of cell wall-derived oligosaccharides, phytohormone regulation, and lignification. However, the functions of most of the 34 GH1 gene products in rice (Oryza sativa) are unknown. Os3BGlu6, a rice β-glucosidase representing a previously uncharacterized phylogenetic cluster of GH1, was produced in recombinant Escherichia coli. Os3BGlu6 hydrolyzed p-nitrophenyl (pNP)-β-d-fucoside (k cat /K m = 67 mm −1 s−1), pNP-β-d-glucoside (k cat /K m = 6.2 mm −1 s−1), and pNP-β-d-galactoside (k cat /K m = 1.6 mm −1s−1) efficiently but had little activity toward other pNP glycosides. It also had high activity toward n-octyl-β-d-glucoside and β-(1→3)- and β-(1→2)-linked disaccharides and was able to hydrolyze apigenin β-glucoside and several other natural glycosides. Crystal structures of Os3BGlu6 and its complexes with a covalent intermediate, 2-deoxy-2-fluoroglucoside, and a nonhydrolyzable substrate analog, n-octyl-β-d-thioglucopyranoside, were solved at 1.83, 1.81, and 1.80 Å resolution, respectively. The position of the covalently trapped 2-F-glucosyl residue in the enzyme was similar to that in a 2-F-glucosyl intermediate complex of Os3BGlu7 (rice BGlu1). The side chain of methionine-251 in the mouth of the active site appeared to block the binding of extended β-(1→4)-linked oligosaccharides and interact with the hydrophobic aglycone of n-octyl-β-d-thioglucopyranoside. This correlates with the preference of Os3BGlu6 for short oligosaccharides and hydrophobic glycosides.
A Phosphofructokinase B-Type Carbohydrate Kinase Family Protein, NARA5, for Massive Expressions of Plastid-Encoded Photosynthetic Genes in Arabidopsis
Authors: Ogawa Taro , Nishimura Kenji , Aoki Takehiko , Takase Hisabumi , Tomizawa Ken-Ichi , Ashida Hiroki , Yokota Akiho 
Published: July 2009

Abstract
To date, there have been no reports on screening for mutants defective in the massive accumulation of Rubisco in higher plants. Here, we describe a screening method based on the toxic accumulation of ammonia in the presence of methionine sulfoximine, a specific inhibitor of glutamine synthetase, during photorespiration initiated by the oxygenase reaction of Rubisco in Arabidopsis (Arabidopsis thaliana). Five recessive mutants with decreased amounts of Rubisco were identified and designated as nara mutants, as they contained a mutation in genes necessary for the achievement of Rubisco accumulation. The nara5-1 mutant showed markedly lower levels of plastid-encoded photosynthetic proteins, including Rubisco. Map-based cloning revealed that NARA5 encoded a chloroplast phosphofructokinase B-type carbohydrate kinase family protein of unknown function. The NARA5 protein fused to green fluorescent protein localized in chloroplasts. We conducted expression analyses of photosynthetic genes during light-induced greening of etiolated seedlings of nara5-1 and the T-DNA insertion mutant, nara5-2. Our results strongly suggest that NARA5 is indispensable for hyperexpression of photosynthetic genes encoded in the plastid genome, particularly rbcL.
Extracellular NM23 protein promotes the growth and survival of primary cultured human acute myelogenous leukemia cells
Authors: Okabe‐Kado Junko , Kasukabe Takashi , Honma Yoshio , Kobayashi Hirofumi , Maseki Nobuo , Kaneko Yasuhiko 
Published: July 2009

An elevated serum level of NM23‐H1 protein is found in acute myelogenous leukemia (AML), and predicts a poor treatment outcome in AML patients. To investigate the potential pathological link between the elevated serum level of this protein and poor prognosis, we examined the extracellular effects of recombinant NM23‐H1 protein on the in vitro growth and survival of primary cultured AML cells at concentrations equivalent to the levels found in the serum of AML patients. Extracellular NM23‐H1 protein promoted the in vitro growth and survival of AML cells and this activity was associated with the cytokine production and activation of the MAPK and signal transducers and activators of transcription signaling pathways. Inhibitors specific to MAPK signaling pathways inhibited the growth‐ and survival‐promoting activity of NM23‐H1. These findings indicate the novel biological action of extracellular NM23‐H1 and its association with poor prognosis, and suggest an important role for extracellular NM23‐H1 in the malignant progression of leukemia and a potential therapeutic target for these malignancies. (Cancer Sci 2009; 100: 1885–1894)
Impact of a Bereavement and Donation Service incorporating mandatory ‘required referral’ on organ donation rates: a model for the implementation of the Organ Donation Taskforce’s recommendations
Authors: Murphy F. , Cochran D. , Thornton S. 
Published: July 2009

SummaryIn 2008 the Organ Donation Taskforce published its recommendations for increasing organ donation in the UK by 50% over 5 years. Bolton NHS Trust has addressed the problem of low rates of organ donation by amalgamating Bereavement and Donation Services and introducing a trigger to refer automatically all potential organ donors to the regional transplant donor co‐ordinators. We audited the ability of the new service to deliver the aims and recommendations of the Organ Donation Taskforce. Following the changes in service provision the number of tissue donors rose from six in 2002 to 246 in 2007. In the same period solid organ donation rates remained unchanged. The introduction of an automatic trigger for referral of potential donors in 2007 resulted in 31 referrals and 11 successful multi‐organ donors. The current service exceeds the aims of the Taskforce and offers the potential to meet UK organ donation targets without resorting to an ‘opt out’ system of presumed consent.
Effects of different doses of remifentanil on the end‐tidal concentration of sevoflurane required for tracheal intubation in children
Authors: He L. , Wang X. , Zhang X‐F. , Tang S‐R. 
Published: July 2009

SummaryWe investigated the effects of different doses of remifentanil on the end‐tidal concentration of sevoflurane required for tracheal intubation in children without the use of neuromuscular blocking drugs. One hundred and thirty paediatric patients, aged 3–8 years, were randomly allocated to receive no remifentanil (group control) or remifentanil 0.1 μg.kg−1.min−1 (group remi0.1), 0.2 μg.kg−1.min−1 (group remi0.2), 0.3 μg.kg−1.min−1 (group remi0.3). All patients were anaesthetised using 5% sevoflurane. After loss of eyelash reflex, remifentanil 1 μg.kg−1 was injected over 1 min followed by an appropriate group‐dependent infusion and the end‐tidal sevoflurane concentration was changed. Predetermined end‐tidal sevoflurane concentrations for each group were determined using the Dixon up‐and‐down method. After the target concentration of sevoflurane was maintained for 5 min, the child’s trachea was intubated. Successful intubation was defined as excellent or good intubating conditions. The end‐tidal concentration (SD) of sevoflurane for successful tracheal intubation in 50% of children (ED50) were 5.16 (0.22)% in control, 3.27 (0.18)%, 1.81 (0.20)% and 1.01 (0.11)%, in remi0.1, remi0.2, and remi0.3 groups, respectively. Using probit analysis, the 95% effective dose (ED95) of sevoflurane were 5.60% (95% CI 5.35–7.66), 3.77% (95% CI 3.45–7.74), 2.18% (95% CI 1.96–3.86), 1.19% (95% CI 1.06–1.82) in control, remi0.1, remi0.2, and remi0.3 groups, respectively.
The effect of heart rate variability on request for labour epidural analgesia
Authors: Lao H.‐C. , Hseu S.‐S. , Huang C.‐J. , Chan K.‐H. , Kuo C.‐D. 
Published: July 2009

SummaryMaternal heart rate variability of 62 parturients were compared based on their choice of using (epidural group, 44 women) or not using (control, 18 women) epidural analgesia. Baseline heart rate variability and visual analogue scores were recorded when the cervix of the parturient dilated to 2–4 cm, and paired data were collected 1 h later. We found that parturients in the epidural group had greater heart rate variability at the beginning of labour. Multiple logistic regression analysis identified percentage of absolute difference in successive RR intervals exceeding 20 ms as the best indicator of choosing epidural analgesia. Almost all heart rate variability measures were unchanged 1 h later in both groups. We concluded that parturients who chose epidural analgesia had greater heart rate variability, and that percentage of absolute difference in successive RR intervals exceeding 20 ms could reflect their likelihood of requesting epidural analgesia at the beginning of labour.
Anti-epidermal growth factor receptor monoclonal antibodies in cancer therapy
Authors: Martinelli E , De Palma R , Orditura M , De Vita F , Ciardiello F 
Published: July 2009

Summary
The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor involved in the proliferation and survival of cancer cells. EGFR is the first molecular target against which monoclonal antibodies (mAb) have been developed for cancer therapy. Here we review the mechanisms underlying the effects of EGFR-specific mAb in cancer therapy. The efficacy of EGFR-specific mAb in cancer occurs thanks to inhibition of EGFR-generated signalling; furthermore, the effects of antibodies on the immune system seem to play an important role in determining the overall anti-tumour response. In this review, attention is focused on cetuximab and panitumumab, two mAb introduced recently into clinical practice for treatment of metastatic colorectal and head and neck cancer which target the external part of EGFR.
Phylogenetic analysis identifies many uncharacterized actin‐like proteins (Alps) in bacteria: regulated polymerization, dynamic instability and treadmilling in Alp7A
Authors: Derman Alan I. , Becker Eric C. , Truong Bao D. , Fujioka Akina , Tucey Timothy M. , Erb Marcella L. , Patterson Paula C. , Pogliano Joe 
Published: July 2009

SummaryActin, one of the most abundant proteins in the eukaryotic cell, also has an abundance of relatives in the eukaryotic proteome. To date though, only five families of actins have been characterized in bacteria. We have conducted a phylogenetic search and uncovered more than 35 highly divergent families of actin‐like proteins (Alps) in bacteria. Their genes are found primarily on phage genomes, on plasmids and on integrating conjugative elements, and are likely to be involved in a variety of functions. We characterize three Alps and find that all form filaments in the cell. The filaments of Alp7A, a plasmid partitioning protein and one of the most divergent of the Alps, display dynamic instability and also treadmill. Alp7A requires other elements from the plasmid to assemble into dynamic polymers in the cell. Our findings suggest that most if not all of the Alps are indeed actin relatives, and that actin is very well represented in bacteria.
Sinorhizobium meliloti CpdR1 is critical for co‐ordinating cell cycle progression and the symbiotic chronic infection
Authors: Kobayashi Hajime , De Nisco Nicole J. , Chien Peter , Simmons Lyle A. , Walker Graham C. 
Published: July 2009

SummaryATP‐driven proteolysis plays a major role in regulating the bacterial cell cycle, development and stress responses. In the nitro ‐fixing symbiosis with host plants, Sinorhizobium meliloti undergoes a profound cellular differentiation, including endoreduplication of the
ome. The regulatory mechanisms governing the alterations of the S. meliloti cell cycle in planta are largely unknown. Here, we report the characterization of two cpdR homologues, cpdR1 and cpdR2, of S. meliloti that encode single‐domain response regulators. In Caulobacter crescentus, CpdR controls the polar localization of the ClpXP protease, thereby mediating the regulated proteolysis of key protein(s), such as CtrA, involved in cell cycle progression. The S. meliloti cpdR1‐null mutant can invade the host cytoplasm, however, the intracellular bacteria are unable to differentiate into bacteroids. We show that S. meliloti CpdR1 has a polar localization pattern and a role in ClpX positioning similar to C. crescentus CpdR, suggesting a conserved function of CpdR proteins among α‐proteobacteria. However, in S. meliloti, free‐living cells of the cpdR1‐null mutant show a striking morphology of irregular coccoids and aberrant DNA replication. Thus, we demonstrate that CpdR1 mediates the co‐ordination of cell cycle events, which are critical for both the free‐living cell division and the differentiation required for the chronic intracellular infection.
Functional Role of Feline Zona Pellucida Protein 4 Trefoil Domain: A Sperm Receptor or Structural Component of the Domestic Cat Zona Pellucida?
Authors: Braun BC , Ringleb J , Waurich R , Viertel D , Jewgenow K 
Published: July 2009

ContentsThe trefoil domain (TFD) is a protein structure characterized by six cysteines, which form a typical three‐loop structure by three disulphide bridges. It is assumed that two of these loops generate a hydrophobic groove, which could be a binding site for carbohydrate residues or proteins. The zona pellucida (ZP) protein, ZP4, contains such a TFD. The carbohydrate‐/protein‐binding property of TFD allows us to assume a potential sperm receptor function of this domain. Additionally, gastrointestinal trefoil peptides are stable against proteases; therefore, a structural role of TFD within the ZP might also be possible. We were able to show that the synthesized and natural folded feline TFD (fTFD) expresses the typical protease resistance that vanished under reducing conditions and after substitution of cysteine residues within the peptide. Furthermore, an antibody directed against the first loop of fTFD was almost unable to bind to intact in vitro mature cat oocytes. Pre‐incubation of oocytes in the reducing agent (DDT), however, improved antibody binding substantially. Therefore, we suggest structural masking of the fTFD domain within the intact ZP. An interaction between fTFD and feline sperm cells was examined using several methods, including immunocytochemistry, immunoelectron microscopy, co‐immunoprecipitation and far western blot, but we found no indication for an involvement of TFD in the primary sperm binding to the ZP. To summarize, there is increasing evidence that the TFD of fZP4 has a structural rather than a sperm‐binding function.
In Vitro Compaction of Germinal Vesicle Chromatin is Beneficial to Survival of Vitrified Cat Oocytes
Authors: Comizzoli P , Wildt DE , Pukazhenthi BS 
Published: July 2009

ContentsThe immature cat oocyte contains a large‐sized germinal vesicle (GV) with decondensed chromatin that is highly susceptible to cryo‐damage. The aim of the study was to explore an alternative to conventional cryopreservation by examining the influence of GV chromatin compaction using resveratrol (Res) exposure (a histone deacetylase enhancer) on oocyte survival during vitrification. In Experiment 1, denuded oocytes were exposed to 0, 0.5, 1.0 or 1.5 mmol/l Res for 1.5 h and then evaluated for chromatin structure or cultured to assess oocyte meiotic and developmental competence in vitro. Exposure to 1.0 or 1.5 mmol/l Res induced complete GV chromatin deacetylation and the most significant compaction. Compared to other treatments, the 1.5 mmol/l Res concentration compromised the oocyte ability to achieve metaphase II (MII) or to form a blastocyst. In Experiment 2, denuded oocytes were exposed to Res as in Experiment 1 and cultured in vitro either directly (fresh) or after vitrification. Both oocyte types then were assessed for meiotic competence, fertilizability and ability to form embryos. Vitrification exerted an overall negative influence on oocyte meiotic and developmental competence. However, ability to reach MII, achieve early first cleavage, and develop to an advanced embryo stage (8–16 cells) was improved in vitrified oocytes previously exposed to 1.0 mmol/l Res compared to all counterpart treatments. In summary, results reveal that transient epigenetic modifications associated with GV chromatin compaction induced by Res is fully reversible and beneficial to oocyte survival during vitrification. This approach has allowed the production of the first cat embryos from vitrified immature oocytes.
Diagnosis of Endometritis in the Bitch: A New Approach
Authors: Fontaine E , Levy X , Grellet A , Luc A , Bernex F , Boulouis HJ , Fontbonne A 
Published: July 2009

ContentsThere are a few investigations into endometritis in the bitch and its relationship with failure to conceive remains unclear. This may be because of the difficulty in collecting uterine samples for further investigations. Recently, transcervical catheterization by vaginal endoscopy has been introduced allowing the evaluation of the endometrium. In this study, uterine cytology and bacteriology were evaluated in 26 infertile bitches. Endometritis was bacterial in origin in most cases (70% of affected bitches), but these results may be underestimated, as some other pathogens (anaerobic bacteria, mycoplasms and fungi) were not investigated. Endometritis, in our opinion, should be investigated in each case of unexplained infertility in bitches. The method used here seems reliable although defining more accurate classification criteria will improve the efficiency of this non‐invasive technique.
Post‐Partum Involution of the Canine Uterus – Gross Anatomical and Histological Features
Authors: Orfanou DC , Ververidis HN , Pourlis A , Fragkou IA , Kokoli AN , Boscos CM , Taitzoglou IA , Tzora A , Nerou CM , Athanasiou L , Fthenakis GC 
Published: July 2009

ContentsWe aimed to study the normal puerperium in the bitch. Ovariohysterectomy was performed in nine bitches, each at a different day after normal whelping; their genital tract was subject to gross anatomical examination, as well as to histological examination and electron microscopy scanning. Corpora albicans were evenly distributed in the left and right ovaries and placental sites were evenly distributed among left and right uterine horns. Placental sites were initially of dark green to grey colour, later becoming dark brown; their length and height progressively decreased. Height of the myometrium and diameter of the uterine glands progressively decreased. Trophoblast‐like cells were consistently observed at the placental sites and on the surface of the interplacental areas, at all time points where hysterectomy had been performed. It is suggested that involution of the canine genital tract can last up to 3 months and is slow. Continuous (up to D84 post‐partum) presence of prominent placental sites should be considered a normal feature of canine uterine post‐partum involution.
Lactoferrin in Canine Sera: A Pyometra Study
Authors: Bartoskova A , Adlerova L , Kudlackova H , Leva L , Vitasek R , Faldyna M 
Published: July 2009

ContentsThe concentration of lactoferrin was measured in canine sera from groups of healthy male dogs as well as pregnant and non‐pregnant female dogs and was compared with that of bitches with pyometra. Lactoferrin concentrations were higher in bitches with pyometra. The role of elevated lactoferrin concentrations in the suppression of lymphocyte activity was examined in sera from bitches with pyometra in a series of investigations. Although the sera from bitches with pyometra were capable of suppressing lymphocyte activity, lactoferrin was not found to be involved in this action.
Effects of the Anti‐Progestin Aglepristone on the Uterine Tissue of Cats Administered Medroxyprogesterone Acetate
Authors: Muphung W , Rungsipipat A , Chatdarong K 
Published: July 2009

ContentsThe effect of aglepristone, a progesterone receptor antagonist, on feline endometrial proliferation induced by medroxyprogesterone acetate (MPA), a contraceptive progestin, was investigated. Three groups each of six cats were studied: Group A, during inter‐oestrus; Group B, given 50 mg MPA; Group C, receiving 50 mg MPA followed by 10 mg/kg aglepristone twice at 24‐h interval 3 weeks later. Ovariohysterectomy was performed, and uterine tissue was collected from the mid part of the horn when the cats were confirmed as being in inter‐oestrus (Group A), 3 weeks after the MPA administration (Group B), or 2 weeks after aglepristone administration (Group C). Uterine weights, relative uterine weights and uterine diameters were determined. The endometrial and myometrial thicknesses and the endometrial/myometrial ratios were determined in histological sections. Histological sections were also immunostained for proliferative activity [anti‐proliferative cell nuclear antigen (PCNA) antibody], and progesterone receptor (anti‐hPR) and percentage of positive cells in luminal vs glandular epithelium were determined for both parameters. PCNA and all morphological parameters except endometrial thickness were increased (p < 0.05) by MPA, and progesterone receptor was decreased compared to untreated cats. Aglepristone treatment had no significant effect compared to MPA alone. However, glandular proliferative activity was usually but not significantly lower in Group C than in Group B, suggesting a potential effect of the anti‐progestin that might be clinically beneficial. Alternatively, this possible effect in the aglepristone group may reflect the longer time after MPA that these animals were examined. Whether lack of significant effect of the anti‐progestin was because of too high dose of MPA relative to the dose of aglepristone, the delay of 14 days in obtaining tissues after aglepristone treatment, or simply an insufficient duration of aglepristone treatment requires further study.
Effect of Catalase and Superoxide Dismutase on Motility, Viability and Acrosomal Integrity of Frozen‐Thawed Cat Spermatozoa
Authors: Thiangtum K , Pinyopummin A , Hori T , Kawakami E , Tsutsui T 
Published: July 2009

ContentsThe aim of this study was to evaluate the effect of antioxidant catalase (CAT) and superoxide dismutase (SOD) in semen extender on motility, viability and acrosomal integrity of frozen‐thawed cat spermatozoa. Semen was collected by using an artificial vagina from five domestic cats (two ejaculates/cat). Spermatozoa were diluted in egg yolk Ttris‐fructose citrate solution (EYT‐FC) without glycerol and cooled at 4°C for 1 h, then diluted further with EYT‐FC with glycerol (7% final concentration) and 400 IU/ml of CAT (treatment 1) or SOD (treatment 2) or without antioxidants (control). Before freezing using a styrofoam box, diluted spermatozoa filled in 0.25‐ml straws were equilibrated for 1 h at 4°C. After thawing, spermatozoa were assessed for motility, viability and acrosomal integrity. Cryopreservation significantly impaired sperm motility, viability and acrosomal integrity (p < 0.05). However, motility, viability and acrosomal integrity of frozen‐thawed cat spermatozoa in the EYT‐FC with CAT, SOD and without the antioxidants were not significantly different. The average percentages of spermatozoa motility after thawing compared between control, treatment 1 and treatment 2 group were 43.5 ± 3.2, 42 ± 4.1 and 38 ± 4.5; for viability: 44.8 ± 3.5, 50.6 ± 5.7 and 47.1 ± 4.1 and for acrosomal integrity: 45 ± 3.5, 44.9 ± 3.4 and 44.4 ± 3.3, respectively. In conclusion, adding CAT and SOD to EYT‐FC did not improve motility, viability and acrosomal integrity in cryopreserved cat spermatozoa.
Ultrasound as a Mechanical Method for Male Dog Contraception
Authors: Leoci R , Aiudi G , De Sandro Salvati A , Silvestre F , Binetti F , Lacalandra GM 
Published: July 2009

ContentsTherapeutic ultrasound (1.5 W/cm2) was investigated for its effect upon testicular size and consistency and semen quality in five dogs. Dogs were treated every other day with a total of three treatments and were examined up to 2 weeks after treatment. There was some tenderness after treatment and a significant decrease in testicular size and semen quality. This method may provide a suitable contraception technique in dogs and requires further investigation.