bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of gallium focused ion beam milling on preparation of aluminium thin foils
Authors: UNOCIC K.A. , MILLS M.J. , DAEHN G.S.
Published: May 2010

Focussed ion beam milling has greatly extended the utility of the atom probe and transmission electron microscope because it enables sample preparation with a level of dimensional control never before possible. Using focussed ion beam it is possible to extract the samples from desired and very specific locations. The artefacts associated with this sample preparation method must also be fully understood. In this work, issues specifically relevant to the focussed ion beam milling of aluminium alloys are presented. After using the focussed ion beam as a sample preparation technique it is evident that gallium will concentrate in three areas of the sample: on the surface, on grain boundaries and at interphase boundaries. This work also shows that low‐energy Ar ion nanomilling is potentially quite effective for removing gallium implantation layers and gallium from the internal surfaces of aluminium thin foils.
The changing faces of Streptococcus antigen I/II polypeptide family adhesins
Authors: Brady L. Jeannine , Maddocks Sarah E. , Larson Matthew R. , Forsgren Nina , Persson Karina , Deivanayagam Champion C. , Jenkinson Howard F.
Published: May 2010

Streptococcus mutans antigen I/II (AgI/II) protein was one of the first cell wall‐anchored adhesins identified in Gram‐positive bacteria. It mediates attachment of S. mutans to tooth surfaces and has been a focus for immunization studies against dental caries. The AgI/II family polypeptides recognize salivary glycoproteins, and are also involved in biofilm formation, platelet aggregation, tissue invasion and immune modulation. The genes encoding AgI/II family polypeptides are found among Streptococcus species indigenous to the human mouth, as well as in Streptococcus pyogenes, S. agalactiae and S. suis. Evidence of functionalities for different regions of the AgI/II proteins has emerged. A sequence motif within the C‐terminal portion of Streptococcus gordonii SspB (AgI/II) is bound by Porphyromonas gingivalis, thus promoting oral colonization by this anaerobic pathogen. The significance of other epitopes is now clearer following resolution of regional crystal structures. A new picture emerges of the central V (variable) region, predicted to contain a carbohydrate‐binding trench, being projected from the cell surface by a stalk formed by an unusual association between an N‐terminal α‐helix and a C‐terminal polyproline helix. This presentation mode might be important in determining functional conformations of other Gram‐positive surface proteins that have adhesin domains flanked by α‐helical and proline‐rich regions.
A novel nucleotide kinase encoded by gene 1.7 of bacteriophage T7
Authors: Tran Ngoc Q. , Lee Seung‐Joo , Richardson Charles C. , Tabor Stanley
Published: May 2010

Gene 1.7 of bacteriophage T7 confers sensitivity of both phage T7 and its host Escherichia coli to dideoxythymidine (ddT). We have purified the product of gene 1.7, gp1.7. It exists in two forms of molecular weight 22 181 and 17 782. Only the C‐terminal half of the protein is required to confer ddT sensitivity. We show that gp1.7 catalyses the phosphorylation of dGMP and dTMP to dGDP and dTDP, respectively, by using either GTP, dGTP or dTTP as the phosphate donor. Either form of gp1.7 exhibit identical kinase activity as compared with wild‐type gp1.7 that contains a mixture of both forms. The Km of 70 µM and Kcat of 4.3 s−1 for dTMP are similar to those found for E. coli thymidylate kinase. However, unlike the host enzyme, gp1.7 efficiently catalyses the conversion of the chain‐terminating dideoxythymidylate (ddTMP) to ddTDP. This finding explains the sensitivity of phage T7 but not E. coli to exogenous ddT. Gp1.7 is unusual in that it has no sequence homology to any known nucleotide kinase, it has no identifiable nucleotide‐binding motif and its activity is independent of added metal ions. When coupled with nucleoside diphosphate kinase, gp1.7 exponentially converts dTMP to dTTP.
DipM links peptidoglycan remodelling to outer membrane organization in Caulobacter
Authors: Goley Erin D. , Comolli Luis R. , Fero Katherine E. , Downing Kenneth H. , Shapiro Lucy
Published: May 2010

Cell division in Gram‐negative organisms requires coordinated invagination of the multilayered cell envelope such that each daughter receives an intact inner membrane, peptidoglycan (PG) layer and outer membrane (OM). Here, we identify DipM, a putative LytM endopeptidase in Caulobacter crescentus, and show that it plays a critical role in maintaining cell envelope architecture during growth and division. DipM localized to the division site in an FtsZ‐dependent manner via its PG‐binding LysM domains. Although not essential for viability, ΔdipM cells exhibited gross morphological defects, including cell widening and filamentation, indicating a role in cell shape maintenance and division that we show requires its LytM domain. Strikingly, cells lacking DipM also showed OM blebbing at the division site, at cell poles and along the cell body. Cryo electron tomography of sacculi isolated from cells depleted of DipM revealed marked thickening of the PG as compared to wild type, which we hypothesize leads to loss of trans‐envelope contacts between components of the Tol–Pal complex. We conclude that DipM is required for normal envelope invagination during division and to maintain a sacculus of constant thickness that allows for maintenance of OM connections throughout the cell envelope.
A protein critical for cell constriction in the Gram‐negative bacterium Caulobacter crescentus localizes at the division site through its peptidoglycan‐binding LysM domains
Authors: Poggio Sebastian , Takacs Constantin N. , Vollmer Waldemar , Jacobs‐Wagner Christine
Published: May 2010

During division of Gram‐negative bacteria, invagination of the cytoplasmic membrane and inward growth of the peptidoglycan (PG) are followed by the cleavage of connective septal PG to allow cell separation. This PG splitting process requires temporal and spatial regulation of cell wall hydrolases. In Escherichia coli, LytM factors play an important role in PG splitting. Here we identify and characterize a member of this family (DipM) in Caulobacter crescentus. Unlike its E. coli counterparts, DipM is essential for viability under fast‐growth conditions. Under slow‐growth conditions, the ΔdipM mutant displays severe defects in cell division and FtsZ constriction. Consistent with its function in division, DipM colocalizes with the FtsZ ring during the cell cycle. Mutagenesis suggests that the LytM domain of DipM is essential for protein function, despite being non‐canonical. DipM also carries two tandems of the PG‐binding LysM domain that are sufficient for FtsZ ring localization. Localization and fluorescence recovery after photobleaching microscopy experiments suggest that DipM localization is mediated, at least in part, by the ability of the LysM tandems to distinguish septal, multilayered PG from non‐septal, monolayered PG.
DipM, a new factor required for peptidoglycan remodelling during cell division in Caulobacter crescentus
Authors: Möll Andrea , Schlimpert Susan , Briegel Ariane , Jensen Grant J. , Thanbichler Martin
Published: May 2010

In bacteria, cytokinesis is dependent on lytic enzymes that facilitate remodelling of the cell wall during constriction. In this work, we identify a thus far uncharacterized periplasmic protein, DipM, that is required for cell division and polarity in Caulobacter crescentus. DipM is composed of four peptidoglycan binding (LysM) domains and a C‐terminal lysostaphin‐like (LytM) peptidase domain. It binds to isolated murein sacculi in vitro, and is recruited to the site of constriction through interaction with the cell division protein FtsN. Mutational analyses showed that the LysM domains are necessary and sufficient for localization of DipM, while its peptidase domain is essential for function. Consistent with a role in cell wall hydrolysis, DipM was found to interact with purified murein sacculi in vitro and to induce cell lysis upon overproduction. Its inactivation causes severe defects in outer membrane invagination, resulting in a significant delay between cytoplasmic compartmentalization and final separation of the daughter cells. Overall, these findings indicate that DipM is a periplasmic component of the C. crescentus divisome that facilitates remodelling of the peptidoglycan layer and, thus, coordinated constriction of the cell envelope during the division process.
Bacillus subtilis BY‐kinase PtkA controls enzyme activity and localization of its protein substrates
Authors: Jers Carsten , Pedersen Malene Mejer , Paspaliari Dafni Katerina , Schütz Wolfgang , Johnsson Christina , Soufi Boumediene , Macek Boris , Jensen Peter Ruhdal , Mijakovic Ivan
Published: May 2010

Bacillus subtilis BY‐kinase PtkA was previously shown to phosphorylate, and thereby regulate the activity of two classes of protein substrates: UDP‐glucose dehydrogenases and single‐stranded DNA‐binding proteins. Our recent phosphoproteome study identified nine new tyrosine‐phosphorylated proteins in B. subtilis. We found that the majority of these proteins could be phosphorylated by PtkA in vitro. Among these new substrates, single‐stranded DNA exonuclease YorK, and aspartate semialdehyde dehydrogenase Asd were activated by PtkA‐dependent phosphorylation. Because enzyme activity was not affected in other cases, we used fluorescent protein tags to study the impact of PtkA on localization of these proteins in vivo. For several substrates colocalization with PtkA was observed, and more importantly, the localization pattern of the proteins enolase, YjoA, YnfE, YvyG, Ugd and SsbA was dramatically altered in ΔptkA background. Our results confirm that PtkA can control enzyme activity of its substrates in some cases, but also reveal a new mode of action for PtkA, namely ensuring correct cellular localization of its targets.
More than just activity control: phosphorylation may control all aspects of a protein's properties
Authors: Stülke Jörg
Published: May 2010

Protein phosphorylation is a major post‐translational modification of proteins. Due to the introduction of a very large, strongly charged group, phosphorylation often has a dramatic effect on the characteristics of the protein, including alterations in activity or interaction properties. In this issue of Molecular Microbiology, Jers et al. have addressed the effect of protein tyrosine phosphorylation in Bacillus subtilis. They demonstrate that tyrosine phosphorylation stimulates the activity of several but not all targets. In addition, the subcellular localization of several proteins was shown to depend on tyrosine phosphorylation that is catalysed by the BY kinase PtkA. This is the first report showing that phosphorylation controls protein localization in bacteria and adds another important function to this post‐translational modification.
Gastroesophageal reflux and altered motility in lung transplant rejection
Authors: Castor J. M. , Wood R. K. , Muir A. J. , Palmer S. M. , Shimpi R. A.
Published: May 2010

Background Lung transplantation has become an effective therapeutic option for selected patients with end stage lung disease. Long‐term survival is limited by chronic rejection manifest as bronchiolitis obliterans syndrome (BOS). The aspiration of gastric contents has been implicated as a causative or additive factor leading to BOS. Gastroesophageal reflux (GER) and altered foregut motility are common both before and after lung transplantation. Further, the normal defense mechanisms against reflux are impaired in the allograft. Recent studies using biomarkers of aspiration have added to previous association studies to provide a growing body of evidence supporting the link between rejection and GER. Further, the addition of high‐resolution manometry (HRM) and impedance technology to characterize bolus transit and the presence and extent of reflux regardless of pH might better identify at‐risk patients. Although additional prospective studies are needed, fundoplication appears useful in the prevention or treatment of post‐transplant BOS.

Purpose This review will highlight the existing literature on the relationship of gastroesophageal reflux and altered motility to lung transplant rejection, particularly BOS. The article will conclude with a discussion of the evaluation and management of patients undergoing lung transplantation at our center.
Substance P and vasoactive intestinal peptide are reduced in right transverse colon in pediatric slow‐transit constipation
Authors: King S. K. , Sutcliffe J. R. , Ong S.‐y. , Lee M. , Koh T. L. , Wong S. Q. , Farmer P. J. , Peck C. J. , Stanton M. P. , Keck J. , Cook D. J. , Chow C. W. , Hutson J. M. , Southwell B. R.
Published: May 2010

Background Slow‐transit constipation (STC) is recognized in children but the etiology is unknown. Abnormalities in substance P (SP), vasoactive intestinal peptide (VIP) and nitric oxide (NO) have been implicated. The density of nerve fibers in circular muscle containing these transmitters was examined in colon from children with STC and compared to other pediatric and adult samples.

Methods Fluorescence immunohistochemistry using antibodies to NO synthase (NOS), VIP and SP was performed on colonic biopsies (transverse and sigmoid colon) from 33 adults with colorectal cancer, 11 children with normal colonic transit and anorectal retention (NAR) and 51 with chronic constipation and slow motility in the proximal colon (STC). The percentage area of nerve fibers in circular muscle containing each transmitter was quantified in confocal images.

Key Results In colon circular muscle, the percentage area of nerve fibers containing NOS > VIP > SP (6 : 2 : 1). Pediatric groups had a higher density of nerve fibers than adults. In pediatric samples, there were no regional differences in NOS and VIP, while SP nerve fiber density was higher in sigmoid than proximal colon. STC children had lower SP and VIP nerve fiber density in the proximal colon than NAR children. Twenty‐three percent of STC children had low SP nerve fiber density.

Conclusions & Inferences There are age‐related reductions in nerve fiber density in human colon circular muscle. NOS and VIP do not show regional variations, while SP nerve fiber density is higher in distal colon. 1/3 of pediatric STC patients have low SP or VIP nerve fiber density in proximal colon.
Ameliorating effects of mirtazapine on visceral hypersensitivity in rats with neonatal colon sensitivity
Authors: Yin J. , Wang W. , Winston J. H. , Zhang R. , Chen J. D. Z.
Published: May 2010

Background The aim was to investigate the effects of mirtazapine on visceral hypersensitivity and gastric emptying in an established rodent model of colonic sensitization.

Methods Twenty colonic sensitized rats and 20 matched controls were used. Visceral sensitivity during colorectal distension (CRD) was assessed by the measurement of abdominal electromyogram (EMG) with the pressures of 20, 40, and 60 mmHg. Mirtazapine with doses of 1, 5, and 10 mg kg−1were administered orally. Gastric emptying and small intestinal transit were performed in a separated experiment after gavage of 1.5 mL of phenol red solution.

Key Results (i) Visceral hypersensitivity after neonatal colonic sensitization was confirmed. (ii) Mirtazapine dose‐dependently reduced visceral hypersensitivity in the colonic sensitized rats. The increases in EMG during CRD at 40, 60 mmHg were, 17.59 ± 6.49 and 26.04 ± 8.30, respectively, with saline session, and substantially reduced to 10.0 ± 5.95 (P = 0.02vscorresponding saline) and 12.58 ± 7.43 (P < 0.001vssaline) with mirtazapine at 10 mg kg−1. Similar findings were noted at doses of 5 and 1 mg kg−1at a lesser degree. In the control rats, mirtazapine‐reduced visceral sensitivity only during CRD at 60 mmHg. (iii) Mirtazapine 10 mg kg−1significantly accelerated gastric emptying (P = 0.045) but slightly and marginally delayed intestinal transit (P = 0.058) the colonic sensitized rats.

Conclusions & Inferences Mirtazapine dose‐dependently ameliorates visceral hypersensitivity in colonic sensitized rats. Mirtazapine at a high dose improves delayed gastric emptying in colonic sensitized rats but slightly and marginally delays small intestinal transit. Its roles in altering gastrointestinal motility need further investigation.
Xylem phenology and wood production: resolving the chicken‐or‐egg dilemma
Authors: LUPI CARLO , MORIN HUBERT , DESLAURIERS ANNIE , ROSSI SERGIO
Published: May 2010

Delays in the start of the growing season reduce the period available for growth and the amount of xylem production. However, a higher number of developing tracheids could prolong cell differentiation and, consequently, lengthen the growing season. The relationship between the amount and duration of cell production in the xylem remains an unresolved issue. The aim of this study was to resolve the chicken‐or‐egg causality dilemma about duration of growth and cell production through simple‐ and double‐cause models. This was achieved by (1) analysing the intra‐annual growth dynamics of the xylem in Picea mariana (Mill.) BSP during 2006–2009 in two contrasting sites of the boreal forest of Quebec, Canada, and (2) extracting the dates of onset and ending of xylem formation and the number of radial cells along the tree ring.

A higher number of cells was linked to an earlier onset (r = 0.74) and later ending (r = 0.61) of cell differentiation. The absence of a relationship between the residuals of the onset and ending of xylogenesis (rp = −0.06) indicated that cell production influenced the correlation between the two phenophases of the xylem.

These results demonstrated that a higher number of cells produced delay the ending of xylem maturation, so extending the duration of wood formation.
Intraspecific variation in the Populus balsamifera drought transcriptome
Authors: HAMANISHI ERIN T. , RAJ SHEROSHA , WILKINS OLIVIA , THOMAS BARB R. , MANSFIELD SHAWN D. , PLANT AINE L. , CAMPBELL MALCOLM M.
Published: May 2010

Drought is a major limitation to the growth and productivity of trees in the ecologically and economically important genus Populus. The ability of Populus trees to contend with drought is a function of genome responsiveness to this environmental insult, involving reconfiguration of the transcriptome to appropriately remodel growth, development and metabolism. Here we test hypotheses aimed at examining the extent of intraspecific variation in the drought transcriptome using six different Populus balsamifera L. genotypes and Affymetrix GeneChip technology. Within a given genotype there was a positive correlation between the magnitude of water‐deficit induced changes in transcript abundance across the transcriptome, and the capacity of that genotype to maintain growth following water deficit. Genotypes that had more similar drought‐responsive transcriptomes also had fewer genotypic differences, as determined by microarray‐derived single feature polymorphism (SFP) analysis, suggesting that responses may be conserved across individuals that share a greater degree of genotypic similarity. This work highlights the fact that a core species‐level response can be defined; however, the underpinning genotype‐derived complexities of the drought response in Populus must be taken into consideration when defining both species‐ and genus‐level responses.
Origin and evolution of polygonal cracks in hydrous sulphate sands, White Sands National Monument, New Mexico
Authors: CHAVDARIAN GREGORY V. , SUMNER DAWN Y.
Published: May 2010

The nucleation and propagation of polygonal cracks in hydrous sulphate dunes at White Sands National Monument, New Mexico, are affected by water availability, transport through the sand and exchange with the atmosphere. These gypsum sands are cohesive enough to crack due to capillary forces and gypsum cements formed during evaporation. Surface cracks form five‐sided polygons with a variety of triple‐junction angles. Cracks extend down to variable depths and polygons increase in size with depth, showing a maturation similar to experimentally produced polygonal columns in drying corn starch. Results from two years of monitoring crack geometries, temperature and humidity demonstrate that cracks form when water is lost to the atmosphere through the transport of water vapour. Subsurface relative humidity below 5 to 10 cm is almost always maintained at 100% by the evaporation and condensation of water in thin films on grains. The amplitude of daily temperature and thus absolute humidity changes decreases with depth, consistent with lower evaporation and condensation rates with increasing depth. Changes in absolute humidity and the contrast between humidity in pore spaces versus the overlying atmosphere result in significant water loss from the dunes except during times of precipitation and frost/dew condensation. This water loss allows cracks to nucleate and propagate into the dunes. This study hypothesizes that crack tips propagate into sand to the depth at which thin films of water on grains are drying, and that this depth varies from the surface of the dune during precipitation events to depths greater than 45 cm when dunes are drier.
Evolutionary conservation, diversity and specificity of LTR‐retrotransposons in flowering plants: insights from genome‐wide analysis and multi‐specific comparison
Authors: Du Jianchang , Tian Zhixi , Hans Christian S. , Laten Howard M. , Cannon Steven B. , Jackson Scott A. , Shoemaker Randy C. , Ma Jianxin
Published: May 2010

The availability of complete or nearly complete genome sequences from several plant species permits detailed discovery and cross‐species comparison of transposable elements (TEs) at the whole genome level. We initially investigated 510 long terminal repeat‐retrotransposon (LTR‐RT) families comprising 32 370 elements in soybean (Glycine max (L.) Merr.). Approximately 87% of these elements were located in recombination‐suppressed pericentromeric regions, where the ratio (1.26) of solo LTRs to intact elements (S/I) is significantly lower than that of chromosome arms (1.62). Further analysis revealed a significant positive correlation between S/I and LTR sizes, indicating that larger LTRs facilitate solo LTR formation. Phylogenetic analysis revealed seven Copia and five Gypsy evolutionary lineages that were present before the divergence of eudicot and monocot species, but the scales and timeframes within which they proliferated vary dramatically across families, lineages and species, and notably, a Copia lineage has been lost in soybean. Analysis of the physical association of LTR‐RTs with centromere satellite repeats identified two putative centromere retrotransposon (CR) families of soybean, which were grouped into the CR (e.g. CRR and CRM) lineage found in grasses, indicating that the ‘functional specification’ of CR pre‐dates the bifurcation of eudicots and monocots. However, a number of families of the CR lineage are not concentrated in centromeres, suggesting that their CR roles may now be defunct. Our data also suggest that the envelope‐like genes in the putative Copia retrovirus‐like family are probably derived from the Gypsy retrovirus‐like lineage, and thus we propose the hypothesis of a single ancient origin of envelope‐like genes in flowering plants.
A rice fungal MAMP‐responsive MAPK cascade regulates metabolic flow to antimicrobial metabolite synthesis
Authors: Kishi‐Kaboshi Mitsuko , Okada Kazunori , Kurimoto Leona , Murakami Shinya , Umezawa Toshiaki , Shibuya Naoto , Yamane Hisakazu , Miyao Akio , Takatsuji Hiroshi , Takahashi Akira , Hirochika Hirohiko
Published: May 2010

Plants recognize potential microbial pathogens through microbial‐associated molecular patterns (MAMPs) and activate a series of defense responses, including cell death and the production of reactive oxygen species (ROS) and diverse anti‐microbial secondary metabolites. Mitogen‐activated protein kinase (MAPK) cascades are known to play a pivotal role in mediating MAMP signals; however, the signaling pathway from a MAPK cascade to the activation of defense responses is poorly understood. Here, we found in rice that the chitin elicitor, a fungal MAMP, activates two rice MAPKs (OsMPK3 and OsMPK6) and one MAPK kinase (OsMKK4). OsMPK6 was essential for the chitin elicitor‐induced biosynthesis of diterpenoid phytoalexins. Conditional expression of the active form of OsMKK4 (OsMKK4DD) induced extensive alterations in gene expression, which implied dynamic changes of metabolic flow from glycolysis to secondary metabolite biosynthesis while suppressing basic cellular activities such as translation and cell division. OsMKK4DD also induced various defense responses, such as cell death, biosynthesis of diterpenoid phytoalexins and lignin but not generation of extracellular ROS. OsMKK4DD‐induced cell death and expression of diterpenoid phytoalexin pathway genes, but not that of phenylpropanoid pathway genes, were dependent on OsMPK6. Collectively, the OsMKK4–OsMPK6 cascade plays a crucial role in reprogramming plant metabolism during MAMP‐triggered defense responses.
Impact of hookworm infection and deworming on anaemia in non‐pregnant populations: a systematic review
Authors: Smith Jennifer L. , Brooker Simon
Published: May 2010

Objectives To summarise age‐ and intensity‐stratified associations between human hookworm infection and anaemia and to quantify the impact of treatment with the benzimidazoles, albendazole and mebendazole, on haemoglobin and anaemia in non‐pregnant populations.

Methods Electronic databases (MEDLINE, EMBASE, PubMed) were searched for relevant studies published between 1980 and 2009, regardless of language, and researchers contacted about potential data. Haemoglobin concentration (Hb) was compared between uninfected individuals and individuals harbouring hookworm infections of different intensities, expressed as standardised mean differences (SMD) and 95% confidence intervals (CI). Meta‐analysis of randomised control trials (RCTs) investigated the impact of treatment on Hb and anaemia.

Results Twenty‐three cross‐sectional studies, six pre‐ and post‐intervention studies and 14 trials were included. Among cross‐sectional studies, moderate‐ and heavy‐intensity hookworm infections were associated with lower Hb in school‐aged children, while all levels of infection intensity were associated with lower Hb in adults. Among RCTs using albendazole, impact of treatment corresponded to a 1.89 g/l increase (95%CI: 0.13–3.63) in mean Hb while mebendazole had no impact. There was a positive impact of 2.37 g/l (95%CI: 1.33–3.50) on mean Hb when albendazole was co‐administered with praziquantel, but no apparent additional benefit of treatment with benzimidazoles combined with iron supplementation. The mean impact of treatment with benzimidazoles alone on moderate anaemia was small (relative risk (RR) 0.87) with a larger effect when combined with praziquantel (RR 0.61).

Conclusions Anaemia is most strongly associated with moderate and heavy hookworm infection. The impact of anthelmintic treatment is greatest when albendazole is co‐administered with praziquantel.
Successful treatment of canine necrolytic migratory erythema (superficial necrolytic dermatitis) due to metastatic glucagonoma with octreotide
Authors: Oberkirchner Ursula , Linder Keith E. , Zadrozny Leah , Olivry Thierry
Published: May 2010

Necrolytic migratory erythema (NME; also known as superficial necrolytic dermatitis) is a syndrome most often associated with certain chronic liver diseases or pancreatic glucagonomas. In humans with glucaconoma‐associated NME, skin lesions usually respond to octreotide, a somatostatin analogue that inhibits glucagon release. In this report an 11‐year‐old golden retriever dog with pancreatic glucononoma and metastasis to the regional lymph nodes, spleen and liver was diagnosed with NME. The dog exhibited erosions, ulcers and crusts on the paws, pressure points, muzzle, periocular area and prepuce. The dog was also anorexic and had difficulty walking. Because metastasis precluded surgery, treatment was initiated with subcutaneous octreotide (2 μg/kg twice daily). Skin lesions and systemic clinical signs improved markedly within 5 days. The dosage was increased to nearly 3 μg/kg twice daily and signs almost completely resolved within 10 days. Anorexia was the major adverse effect observed. During the following month, both dosage (1–3.7 μg/kg) and frequency (two to four times daily) of the octreotide injections were adjusted to permit control of clinical signs while maintaining adequate appetite. Temporary cessation of octreotide administration resulted in the rapid recurrence of skin lesions. Resuming injections led to improvement of clinical signs within 48 h. The dog was later euthanized because of progressive metastatic disease. In conclusion, subcutaneous octreotide injections were beneficial in this dog with glucagonoma‐associated NME. This somatostatin analogue could be a valuable option to treat canine patients with non‐resectable or relapsing pancreatic glucagonoma‐associated NME.
The efficacy of imiquimod 5% cream (Aldara®) in the treatment of aural plaque in horses: a pilot open‐label clinical trial
Authors: Torres Sheila M. F. , Malone Erin D. , White Stephen D. , Koch Sandra N. , Watson Johanna L.
Published: May 2010

Aural plaques affect at least 22% of horses and can be asymptomatic or cause ear sensitivity. Immunohistochemical and electron microscopy studies have shown a strong association between aural plaques and papilloma virus. The purpose of this study was to investigate the efficacy of imiquimod 5% cream, an immune response modifier with potent antiviral activity, in the treatment of equine aural plaques. Twenty‐one horses were enrolled and 16 completed the study. Imiquimod 5% cream was applied three times a week, every other week. When both ears were affected only the worst affected ear was treated. Adverse effects in all horses included marked local inflammation, exudation and thick crust formation at the site of treatment and the adjacent skin. Removal of the crust before treatment was painful and required sedation in most horses. Complete resolution of lesions was noted in all horses immediately post‐treatment and the long‐term resolution rate was 87.5%. Duration of therapy ranged from 1.5 to 8 months (median: 2.9 mean: 3.5). All horses were followed‐up for 12–22 months after treatment was discontinued and only two horses had a recurrence of lesions. Clinical signs related to the aural plaques prior to treatment were reported in 11 of 16 (68.8%) horses and included resistance to touching the ears and bridling. Complete resolution of these signs was reported by the owners in all of the horses followed‐up for at least 12 months. In conclusion, the topical application of imiquimod 5% cream is an efficacious treatment for aural plaques in horses.
PREPARATION AND PROPERTIES OF ACTIVE WATER INSOLUBLE DERIVATIVES OF ADENOSINE DEAMINASE
Authors: Lucacchini A. , Ronca G. , Montali U. , Rossi C. A.
Published: May 2010

Adenosine deaminase (adenosine aminohydrolase E.C.3.5.4.4.) purified from calf intestinal mucosa, has been bound to polyacrylamide derivatives by reaction with their diazo or acyl azide groups.

0.2–1.6 mg of protein were bound to 100 mg of insoluble product and retained 2–11% of free enzyme activity. The most active preparations were obtained in presence of purine riboside, a competitive inhibitor of adenosine deaminase.

Km, pH‐activity curve, activation energy for the breakdown of the enzyme substrate complex are the same for insoluble derivatives and native enzyme.

Insoluble adenosine deaminase is more resistent to heat denaturation, pH extremes and lyophilization than the native enzyme.
STEREOCHEMICAL CRITERIA FOR POLYPEPTIDE AND PROTEIN CHAIN CONFORMATIONS.
Authors: Balasubramanian R. , Ramakrishnan C.
Published: May 2010

Conformational study on a pair of alanyl peptide units has been made, considering the various contribution to the total potential energy, and the (φ, φ) energy maps corresponding to 3 different values of τ and ω around their ideal values (of 110° and 180° respectively) are presented. The global minima has been found to occur close to the right‐handed 3 → 1 hydrogen‐bonded region. The effect of distortions on the energy map as well as the effect of the various individual contributions to the total energy has been discussed. Also the barrier heights between the different regions of low energy in the (φ, φ) maps have been analyzed. Finally a comparison of the results with similar studies of other workers has been made
AMINO ACIDS AND PEPTIDES.
Authors: Ali Akhtar , Cook Ronald M. , Weinstein Boris
Published: May 2010

A protected undecapeptide that corresponds to the amino acids found at the end of the protein chain in rubredoxin has been prepared by classical peptide methods.
CONFORMATIONAL ENERGY CALCULATIONS OF ENZYME‐SUBSTRATE INTERACTIONS. I. Computation of Preferred Conformations of Some Substrates of α‐Chymotrypsin
Authors: Platzer Karen E. B. , Momany Frank A. , Scheraga Harold A.
Published: May 2010

Empirical energy calculations are used to determine all low‐energy conformations of the isolated α‐chymotrypsin substrates N‐acetyl‐L‐phenylalanine amide, N‐acetyl‐L‐tyrosine amide, N‐formyl‐L‐tyrosine amide, and N‐acetyl‐L‐tryptophan amide. The computed conformations are in agreement with the available experimental data for L‐phenylalanyl, L‐tyrosyl and L‐tryptophanyl side chains in proteins, and with NMR and X‐ray data for these side chains in small peptides. In the following papers, certain of these low‐energy substrate conformations will be shown to bind selectively to the enzyme chymotrypsin to form stable noncovalent enzyme‐substrate complexes.
SPECIFIC FLUORESCENT DERIVATIVES OF MACROMOLECULES.
Authors: Schoellmann Guenther
Published: May 2010

1‐Dimethylamino‐5‐naphthalene sulfonyl (dansyl)‐derivatives of phenylalanine and lysine were synthesized and employed to introduce highly fluorescent groups into the active centers of chymotrypsin and trypsin utilizing the active‐site directed labeling technique. N‐alpha‐dansyl‐L‐phenylalanyl chloromethane reacted irreversibly with chymotrypsin. A second order rate constant of 8.8 M‐1 sec‐1 was found. Trypsin was specifically inactivated by N‐alpha‐dansyl‐L‐lysyl bromomethane with a second order rate constant of 3.5 M‐1 sec‐1.

Both enzymes were completely and stoichiometrically inactivated by reaction of the inhibitors with a single histidine residue as determined by amino acid analysis. From previous work with structurally similar inhibitors, histidine residue‐57 of chymotrypsin and histidine residue‐46 of trypsin are inferred to have been modified in the reaction. After performic acid rearrangement and acid hydrolysis, 3‐carboxymethyl‐histidine could be identified showing that the alkylation had occurred at nitrogen‐3 of the histidine residue involved.

Because of their fluorescent properties the new inhibitors can be considered as probes into the microenvironment of the active site of these enzymes.
FRACTIONATION AND PROPERTIES OF AMINOPEPTIDASE B DURING PURIFICATION AND STORAGE
Authors: Mäkinen Pirkko‐Liisa , Mäkinen Kauko K.
Published: May 2010

Purified preparations of rat liver aminopeptidase B were found to display heterogeneity which was demonstrated in isoelectric focusing and in electrophoresis on polyacrylamide columns. Most of the heterogeneity was generated during the purification of the enzyme. Acid treatment at pH 5.1 and ammonium sulphate fractionations caused considerable heterogeneity and the presence of thiols reduced it. Ampholine ampholytes themselves did not cause the generation of the heterogeneity but isoelectric focusing rendered its demonstration possible. A special type of heterogeneity was observed when aged preparations of the enzyme were found to undergo aggregation. The aggregate forms moved slowly or not at all in electrophoresis on polyacrylamide. The pIs of the main forms appearing in isoelectric focusing were around 5 and 7. The aggregate forms had a pI below 4. The enzymic properties of the different forms, whether appearing in isoelectric focusing, during ageing, or in purification were essentially similar, indicating that the heterogeneity involved changes only in those amino acid residues which were not responsible for substrate binding or catalysis.
REVISED STRUCTURE OF SHEEP BETA‐LIPOTROPIC HORMONE
Authors: Chrétien M. , Gilardeau C. , Li Choh Hao
Published: May 2010

The amino‐acid sequence of the first 38 residues of sheep beta‐LPH has been determined with the automated Edman degradation method and found to be slightly different from the published structure. The sequence gly‐thr in positions 3 and 4, and ala‐glu in positions 16–17 are respectively thr‐gly and gln‐ala.
HAEMOLYTIC FACTOR FROM BULL SEMINAL VESICLE FLUID: CHEMICAL AND BIOCHEMICAL PROPERTIES, AMINO ACID COMPOSITION
Authors: Kysilka Čenék
Published: May 2010

Some chemical and biochemical properties of bull seminal vesicle haemolytic factor, the protein responsible for the haemolysis of red blood cells, are described. The molecule has only one N‐terminal amino acid, aspartic acid, although subunit arrangement was described previously. The haemolytic activities of the native and chemically modified protein were compared. The activities of the reduced, reduced and carboxamido‐methylated, and oxidized protein were all substantially decreased. N‐terminal analysis shows that the whole particle of haemolytic factor is probably composed of ten identical subunits. It is supposed that decrease of haemolytic activity results from a splitting of disulphide bonds.

No proteolytic activity was demonstrated; however, some evidence for phospholipolytic activity was observed. The isoelctric point was determined to be pH 4.5.

The amino acid composition of haemolytic factor is given.
INTERACTION OF A BASIC CHROMIUM‐ZIRCONIUM SULPHATE COMPLEX WITH TROPOCOLLAGEN
Authors: Davis Margaret H. , Scroggie J. G.
Published: May 2010

Rat‐tail tendon tropocollagen has been shown to react with a basic chromium‐zirconium sulphate complex (Zircryst: Na2Zr3Cr2(SO4)4(OH)12.6H2O) in pH 3 sulphuric acid. Additional crosslinks, both intra‐and inter‐molecular, are introduced into the collagen and the Zircryst moiety remains essentially intact throughout. This results in an increase in thermal stability of the protein so that ultimately denaturation does not occur until approximately 90°.
Periumbilical parasitic thumbprint purpura: Strongyloides hyperinfection syndrome acquired from a cadaveric renal transplant
Authors: Weiser J.A. , Scully B.E. , Bulman W.A. , Husain S. , Grossman M.E.
Published: May 2010

J.A. Weiser, B.E. Scully, W.A. Bulman, S. Husain, M.E. Grossman. Periumbilical parasitic thumbprint purpura: Strongyloides hyperinfection syndrome acquired from a cadaveric renal transplant. Transpl Infect Dis 2011: 13: 58–62. All rights reserved

Abstract: Periumbilical parasitic thumbprint purpura may be a presenting sign of hyperinfection strongyloidiasis in the immunocompromised host. We report a case of fatal hyperinfection strongyloidiasis acquired from a cadaveric renal allograft, diagnosed by the pathognomonic periumbilical thumbprint purpuric eruption, confirmed by skin biopsy and laboratory testing.
Diagnostic criteria in patients with complex regional pain syndrome assessed in an out‐patient clinic
Authors: VAN BODEGRAVEN HOF E. A. M. , GROENEWEG G. J. , WESSELDIJK F. , HUYGEN F. J. P. M. , ZIJLSTRA F. J.
Published: May 2010

Background: Specific criteria have been described and accepted worldwide for diagnosing patients with complex regional pain syndrome (CRPS). Nevertheless, a clear‐cut diagnosis cannot be confirmed in a number of cases.

Aim: The objective of this study was to investigate the effectiveness of the described diagnostic criteria used by several clinical disciplines.

Methods: We included 195 patients who were referred to our pain clinic within a period of 1 year. Data were collected on patient characteristics, signs, symptoms, disease‐related medication, and the background of the referring clinicians.

Results: The Harden and Bruehl criteria were confirmed in 95 patients (49%). These patients used a higher than average number of analgesics, opiates, and anti‐oxidants, and frequently received prescriptions for benzodiazepines instead of anti‐depressants. The mean disease duration was 29 ± 4.6 months and the mean visual analogue score for pain was 8.1 ± 0.19. A subgroup of patients had a colder temperature in the affected extremity compared with the unaffected extremity. This subgroup showed a longer disease duration and higher visual analogue scale pain.

Conclusion: The diagnostic criteria used to determine CRPS should be further improved. A large number of referred patients experienced substantial pain, without receiving adequate medication. Disease‐related medication is unrelated to CRPS‐specific disease activity. Knowledge of underlying mechanisms is warranted before an adequate pharmaceutical intervention can be considered.
Acute kidney injury with renal replacement therapy in trauma patients
Authors: BEITLAND S. , MOEN H. , OS I.
Published: May 2010

Background: Acute kidney injury (AKI) with renal replacement therapy (RRT) is rare in trauma patients. The primary aim of the study was to assess incidence, mortality and chronic RRT dependency in this patient group.

Methods: Adult trauma patients with AKI receiving RRT at a regional trauma referral center over a 12‐year period were retrospectively reviewed.

Results: Population‐based incidence of post‐traumatic AKI with RRT was 1.8 persons per million inhabitants per year (p.p.m./year) [95% confidence the interval (CI) 1.5–2.1 p.p.m./year]. In trauma patients admitted to hospital, incidence was 0.5‰ (95% CI 0.3–0.7‰) of those treated in intensive care unit (ICU), it was 8.3% (95% CI 5.9–10.8%). The median age was 46 years. Odds ratio (OR) for post‐traumatic AKI requiring RRT was higher in males than in females in general population (OR 5.6, 95% CI 2.2–14.0), and in trauma patients admitted to hospital (OR 4.4, 95% CI 1.9–10.3) and ICU (OR 4.5, 95% CI 1.9–10.7). The in‐hospital mortality rate was 24% (95% CI 11–37%), 3‐month mortality 36% (95% CI 21–51%) and 1‐year mortality 40% (95% CI 25–55%). Age was a risk factor for death after 1 year, with 57% (95% CI 7–109%) increased risk for each 10 years added. None of the survivors was dialysis‐dependent 3 months or 1 year after trauma.

Conclusion: AKI in trauma patients requiring RRT was rare in this single‐center study. More males than females were affected. Mortality was modest, and renal recovery was excellent as none of the survivors became dependent on chronic RRT.
Molecular evolutionary analysis of seminal receptacle sperm storage organ genes of Drosophila melanogaster
Authors: PROKUPEK A. M. , EYUN S.‐I. , KO L. , MORIYAMA E. N. , HARSHMAN L. G.
Published: May 2010

Sperm storage organs are common and broadly distributed among animal taxa. However, little is known about how these organs function at the molecular level. Additionally, there is a paucity of knowledge about the evolution of genes expressed in these organs. This investigation is an evolutionary expressed sequence tag (EST) study of genes expressed in the seminal receptacle, one of the sperm storage organs in Drosophila. The incidence of positive selection is higher for the seminal receptacle genes than Drosophila reproductive genes as a whole, but lower than genes associated with the spermatheca, a second type of Drosophila sperm storage organ. By identifying overrepresented classes of proteins and classes for which sperm storage function is suggested by the nature of the proteins, candidate genes were discovered. These candidates belong to protein classes such as muscle contraction, odorant binding and odorant receptor, protease inhibitor and immunity.
Bargaining in biology?
Authors: BINMORE K.
Published: May 2010

This paper surveys the economic theory of bargaining with a view to applications in biology, using Roughgarden’s recent Genial Gene as a case study in mistakes to be avoided.
Inheritance in tetraploid yeast revisited: segregation patterns and statistical power under different inheritance models
Authors: STIFT M. , REEVE R. , Van TIENDEREN P. H.
Published: May 2010

In their recent article, Albertin et al. (2009) suggest an autotetraploid origin of 10 tetraploid strains of baker’s yeast (Saccharomyces cerevisiae), supported by the frequent observation of double reduction meiospores. However, the presented inheritance results were puzzling and seemed to contradict the authors’ interpretation that segregation ratios support a tetrasomic model of inheritance. Here, we provide an overview of the expected segregation ratios at the tetrad and meiospore level given scenarios of strict disomic and tetrasomic inheritance, for cases with and without recombination between locus and centromere. We also use a power analysis to derive adequate sample sizes to distinguish alternative models. Closer inspection of the Albertin et al. data reveals that strict disomy can be rejected in most cases. However, disomic inheritance with strong but imperfect preferential pairing could not be excluded with the sample sizes used. The possibility of tetrad analysis in tetraploid yeast offers a valuable opportunity to improve our understanding of meiosis and inheritance of tetraploids.
Measuring sexual selection on females in sex‐role‐reversed Mormon crickets (Anabrus simplex, Orthoptera: Tettigoniidae)
Authors: ROBSON L. J. , GWYNNE D. T.
Published: May 2010

Although many studies examine the form of sexual selection in males, studies characterizing this selection in females remain sparse. Sexual selection on females is predicted for sex‐role‐reversed Mormon crickets, Anabrus simplex, where males are choosy of mates and nutrient‐deprived females compete for matings and nutritious nuptial gifts. We used selection analyses to describe the strength and form of sexual selection on female morphology. There was no positive linear sexual selection on the female body size traits predicted to be associated with male preferences and female competition. Instead, we detected selection for decreasing head width and mandible length, with stabilizing selection as the dominant form of nonlinear selection. Additionally, we tested the validity of a commonly used instantaneous measure of mating success by comparing selection results with those determined using cumulative mating rate. The two fitness measures yielded similar patterns of selection, supporting the common sampling method comparing mated and unmated fractions.
Trafficking, lateral mobility and segregation of the plant K+ channel KAT1*
Authors: Reuff M. , Mikosch M. , Homann U.
Published: May 2010

Functioning of ion channels depends not only on the control of their activity but also on their density and lateral organisation in the membrane. The two latter aspects have recently attracted increasing attention. Here, we summarize studies on trafficking and plasma membrane distribution of the plant K+ channel KAT1 from Arabidopsis thaliana. In guard cells, KAT1 was found to be subject to constitutive and pressure‐driven turnover and ABA‐stimulated endocytosis. These results point to a role of exo‐ and endocytosis in regulating KAT1 density and thus ion transport during guard cell functioning. Recent studies indicate that KAT1 density can also be adjusted at the site of ER export. Efficient ER export of KAT1 was shown to depend on an acidic motif that interacts with Sec24, a component of ER‐derived vesicles. Surface expression of ER export mutants of KAT1 can be rescued through heterotetrameric assembly with wild‐type KAT1, implying that not all subunits of the channel tetramer need to carry an ER export motif. Analysis of the distribution of KAT1 in the plasma membrane revealed segregation of the channel into microdomains, and low lateral mobility in both plant and mammalian cells. In plant cells, SNAREs have been shown to be involved in anchoring KAT1 in the plasma membrane. Studies on guard cells imply a role for the cell wall in organisation of KAT1 microdomains. Together, these findings underline the importance of investigating mechanisms of KAT1 trafficking and lateral organisation in order to fully understand channel functioning.