bluuebunny/crossref_metadata_2025 huggingface dataset:
Climate change, chytridiomycosis or condition: an experimental test of amphibian survival
Authors: GARNER TRENTON W. J. , ROWCLIFFE J. MARCUS , FISHER MATTHEW C.
Published: June 2010

Variation in climatic temperature is frequently cited as playing a powerful role in driving host/pathogen dynamics, and as a consequence future patterns of disease are predicted to change owing to global warming. However, the preponderance of data is correlative and few quantitative tests exist that dissect the relative effects of changing temperature on host and pathogen responses. In this study, we experimentally tested the effect overwintering temperature had on the susceptibility of a vertebrate ectothermic host, the common toad Bufo bufo, to infection and mortality caused by Batrachochytrium dendrobatidis (Bd). We show that a ‘poor’ (warmer) overwintering regime increases the probability of infection, supporting the concept that the dynamics of infection are altered by changing environmental temperature profiles. We also show once established, the proliferation of Bd in the host was better in toadlets that experienced a ‘good’ (colder) winter. We find no relationship between survival and the different overwintering regimes or any consistent deleterious influence of disease on survival. Instead, survival is overwhelmingly predicted by measurements of condition (mass, mass lost over winter) that were not determined by overwintering temperature. We conclude that the survival of a cohort of overwintering toadlets is primarily driven by patterns of growth during the larval period rather than winter temperature or infectious disease.
ERG rearrangement in small cell prostatic and lung cancer
Authors: Scheble Veit J , Braun Martin , Wilbertz Theresia , Stiedl Ann‐Cathrin , Petersen Karen , Schilling David , Reischl Markus , Seitz Gerhard , Fend Falko , Kristiansen Glen , Perner Sven
Published: June 2010

Scheble V J, Braun M, Wilbertz T, Stiedl A‐C, Petersen K, Schilling D, Reischl M, Seitz G, Fend F, Kristiansen G & Perner S (2010) Histopathology 56, 937–943 ERG rearrangement in small cell prostatic and lung cancer

Aims:  Small cell prostatic cancer is a rare but aggressive disease. Currently, its histogenetic origin is unclear and its distinction from metastatic small cell lung cancer is challenging. The aim of our study was to determine whether the ERG rearrangement commonly observed in acinar prostatic cancer can distinguish small cell prostatic cancer from small cell lung cancer samples.

Methods and results:  We assessed 15 small cell prostatic cancers and 22 small cell lung cancers for ERG rearrangement using fluorescence in situ hybridization. Commonly used and novel immunohistochemical markers (i.e. androgen receptor, calcium activated nucleotidase 1, Golgi phosphoprotein 2, prostate‐specific antigen, prostate‐specific membrane antigen, CD56, epithelial membrane antigen, thyroid transcription factor 1, chromogranin A, synaptophysin and Ki67) were further studied. ERG rearrangement occured in 86% of small cell prostatic cancers but in none of the small cell lung cancers and was the best marker to differentiate between both tumours (P < 0.0001).

Conclusions:  The ERG rearrangement is commonly observed in small cell prostatic cancer, supporting the hypothesis that ERG rearrangement occurs in aggressive prostatic cancers. Furthermore, the ERG rearrangement is the most significant marker to differentiate between small cell prostatic cancer and small cell lung cancer. Moreover, our data suggest that small cell prostatic cancer is not a tumour entity on its own, but a dedifferentiated variant of common acinar prostatic cancer.
A study of indefinite for dysplasia in Barrett’s oesophagus: reproducibility of diagnosis, clinical outcomes and predicting progression with AMACR (α‐methylacyl‐CoA‐racemase)
Authors: Sonwalkar Sunil A , Rotimi Olorunda , Scott Nigel , Verghese Eldo , Dixon Mike , Axon Anthony T R A , Everett Simon M
Published: June 2010

Sonwalkar S A, Rotimi O, Scott N, Verghese E, Dixon M, Axon A T R A & Everett S M (2010) Histopathology 56, 900–907 A study of indefinite for dysplasia in Barrett’s oesophagus: reproducibility of diagnosis, clinical outcomes and predicting progression with AMACR (α‐methylacyl‐CoA‐racemase)

Aims:  To assess interobserver variation in the diagnosis of dysplasia in Barrett’s oesophagus, especially indefinite dysplasia (IND) using the revised Vienna classification. A secondary aim was to study clinical outcome of IND cases and to evaluate expression of alpha‐methyl‐CoA racemase (AMACR) as a marker predictive of progression.

Methods and results:  Cases of Barrett’s oesophagus and dysplasia over a 20 year period were assessed. Three experienced histopathologists reviewed 101 cases on set criteria in a blinded fashion. Slides were immunostained for AMACR and evaluated for the presence, extent and location of AMACR expression. Clinical and progression data were collected. Overall agreement for the diagnosis of dysplasia was fair (k = 0.35) but that for IND was poor (k = 0.18). 6 IND cases progressed after a median follow‐up of 31.4 months to a higher grade. The sensitivity of AMACR for the detection of abnormality was 22% for IND and specificity 100%. The positive predictive value of AMACR for progression was 0.44 and the negative predictive value was 0.92.

Conclusion:  Fair agreement was achieved for the diagnosis of dysplasia but poor agreement for IND. A proportion of IND cases progress. Re‐diagnosis or consensus diagnosis did not predict progression. AMACR shows promise as a marker to indicate IND patients in need of more intensive surveillance.
Smoothelin immunohistochemistry is a useful adjunct for assessing muscularis propria invasion in bladder carcinoma
Authors: Bovio Ian M , Al‐Quran Samer Z , Rosser Charles J , Algood Chester B , Drew Peter A , Allan Robert W
Published: June 2010

Bovio I M, Al‐Quran S Z, Rosser C J, Algood C B, Drew P A & Allan R W (2010) Histopathology 56, 951–956  Smoothelin immunohistochemistry is a useful adjunct for assessing muscularis propria invasion in bladder carcinoma

Aims:  To prospectively evaluate the utility of smoothelin immunohistochemical expression for the evaluation of muscularis propria (MP) in diagnostic transurethral resection of bladder tumour (TURBT) specimens and cystectomies.

Methods and results:  Smoothelin immunohistochemistry was performed on a total of 26 TURBT and cystectomy specimens. All but two cases (24/26) demonstrated strong (3+) or moderate (2+) immunoreactivity of the MP with smoothelin. Muscularis mucosae (MM) never displayed strong (3+) reactivity, and in only one case did the MM have moderate (2+) reactivity; in this case the MP had strong (3+) reactivity. MM intensity mirrored the intensity of reactivity of blood vessels in all cases (26/26). Using moderate or strong immunoreactivity as a cut‐off, smoothelin had a sensitivity of 92% for detecting MP and a specificity of 97% for distinguishing between MP and MM. In all unequivocal MP‐invasive and lamina proporia‐invasive cases by haematoxylin and eosin (H&E), smoothelin immunohistochemistry confirmed the original light microscopic diagnosis. In four cases in which there was equivocal MP involvement by H&E, smoothelin helped establish MP invasion.

Conclusions:  Smoothelin immunohistochemistry has diagnostic utility in the evaluation of MP invasion in urothelial carcinoma. Smoothelin could be used as an adjunct to traditional H&E‐stained light microscopy and may help reduce the number of equivocal diagnoses.
p16INK4A overexpression is frequently detected in tumour‐free tonsil tissue without association with HPV
Authors: Klingenberg Boris , Hafkamp Harriët C , Haesevoets Annick , Manni Johannes J , Slootweg Pieter J , Weissenborn Soenke J , Klussmann Jens P , Speel Ernst‐Jan M
Published: June 2010

Klingenberg B, Hafkamp H C, Haesevoets A, Manni J J, Slootweg P J, Weissenborn S J, Klussmann J P & Speel E‐J M (2010) Histopathology 56, 957–967 p16INK4Aoverexpression is frequently detected in tumour‐free tonsil tissue without association with HPV

Aims:  Oncogenic human papillomavirus (HPV) type 16 has been strongly associated with tonsillar squamous cell carcinoma (TSCC) and appears to be of prognostic significance. Because HPV+ TSCC also accumulates p16INK4A, this cyclin‐dependent kinase inhibitor has been proposed as a potential biomarker for HPV in clinical diagnosis. The aim of this study was to determine the prevalence of HPV in tumour‐free tonsillar tissue and the value of p16INK4A overexpression in predicting its presence.

Methods and results:  p16INK4A overexpression was detected by immunohistochemistry in tissue sections of tumour‐free tonsils of 262 patients. They were treated for non‐oncological reasons (snoring or chronic/recurrent tonsillitis) consisting of tonsillectomy. Genomic DNA isolated from these tissues was subjected to HPV‐specific polymerase chain reaction (PCR) analysis. p16INK4A immunoreactivity was detected in 28% of samples in both crypt epithelium (49/177) and lymphoid germinal centres (52/187), which correlated with each other (P < 0.0001). No reactivity was observed in superficial squamous cell epithelium. HPV16 and 18 were detected by PCR analysis in 2/195 cases (1%), which, however, were negative on fluorescence in situ hybridization analysis and discrepant on p16INK4A immunostaining.

Conclusions:  No proof was found for the presence of HPV in tumour‐free tonsil tissue, despite increased p16INK4A expression in a quarter of tonsil cases. Other mechanisms than HPV infection are therefore implicated in p16INK4A up‐regulation.
Transgenic analysis of the BmBLOS2 gene that governs the translucency of the larval integument of the silkworm, Bombyx mori
Authors: Fujii T. , Daimon T. , Uchino K. , Banno Y. , Katsuma S. , Sezutsu H. , Tamura T. , Shimada T.
Published: June 2010

The larval integument of the silkworm, Bombyx mori, is opaque because urate granules accumulate in the epidermis. Although the biosynthetic pathway of uric acid is well studied, little is known about how uric acid accumulates as urate granules in epidermal cells. In the distinct oily (od) mutant silkworm, the larval integument is translucent because of the inability to construct urate granules. Recently, we have found that the od mutant has a genomic deletion in the B. mori homologue of the human biogenesis of lysosome‐related organelles complex1, subunit 2 (BLOS2) gene (BmBLOS2). Here, we performed a molecular and functional characterization of BmBLOS2. Northern blot analysis showed that BmBLOS2 was ubiquitously expressed in various tissues. We analysed the structure of a newly isolated mutant (odB) allelic to od and found a premature stop codon in the coding sequence of BmBLOS2 in this new mutation. Moreover, the translucent phenotype was rescued by the germ‐line transformation of the wild‐type BmBLOS2 allele into the od mutant. Our results suggest that BmBLOS2 is responsible for the od mutant phenotype and plays a crucial role in biogenesis of urate granules in the larval epidermis of the silkworm. The relationships amongst Hermansky–Pudlak syndrome (HPS) genes in mammals, granule group genes in Drosophila and translucent mutant genes in B. mori are discussed.
Pollination services provided to small and large highbush blueberry fields by wild and managed bees
Authors: Isaacs Rufus , Kirk Anna K.
Published: June 2010

1. Plantings of pollinator‐dependent crops vary from large, intensively managed fields to small fields that are managed less intensively, yet there is relatively little information on how pollinator populations and their contribution to crop productivity vary across this gradient.

2. We determined the relative importance of wild bees and managed honey bees Apis mellifera L. for crop pollination in the blueberry Vaccinium corymbosum L. system of Michigan, USA, by comparing bee communities in small, isolated blueberry fields with those in large blueberry fields (stocked with managed honey bee hives) and measuring the difference in yields of open‐pollinated and pollinator‐excluded clusters of flowers. We combined these assessments to calculate the contribution of bees to crop production in this system.

3. Wild bees were the dominant pollinators in small fields, comprising 58% of flower‐visiting bees, whereas 97% of bees in large fields were honey bees. Large fields had four times as many bees visiting flowers than small fields, but only one‐tenth as many bumble bees as small fields.

4. Levels of fruit set exceeded 85% in all fields and were similar between field sizes. Berry weight increased with flower exposure to pollinators and was positively correlated with bee abundance. Berry weights from open‐pollinated flowers were twice as high in large fields compared with small fields.

5. By combining berry weight increases attributable to honey bees and wild bees in blueberry fields with measurements of the bee community adjusted for pollinator efficiency, we calculated the relative contributions of honey bees and wild bees to pollination of Michigan blueberries. We estimate that wild bees provide 82% of the pollination in small fields but only 12% of the total pollination services across this system, mostly through their secondary role in large fields.

6. Synthesis and applications. Wild bees are the primary pollinators of small blueberry fields, but these insects are at low abundance in large fields, perhaps due to a lack of nesting resources or competition for resources with honey bees. Our findings highlight the dependence of commercial fruit producers on honey bees and suggest that increasing the pollination contribution of other bees, particularly bumble bees, in large fields will require that growers adopt wild bee conservation strategies or stock their fields with managed colonies. Quantifying the contributions of managed and wild pollinating bees across the range of crop production scenarios will help to direct development of integrated crop pollination strategies to minimize the risk of pollination deficits affecting food production.
Contrasting landscape epidemiology of two sympatric rabies virus strains
Authors: BARTON HEATHER D. , GREGORY ANDREW J. , DAVIS ROLAN , HANLON CATHLEEN A. , WISELY SAMANTHA M.
Published: June 2010

Viral strain evolution and disease emergence are influenced by anthropogenic change to the environment. We investigated viral characteristics, host ecology, and landscape features in the rabies‐striped skunk disease system of the central Great Plains to determine how these factors interact to influence disease emergence. We amplified portions of the N and G genes of rabies viral RNA from 269 samples extracted from striped skunk brains throughout the distribution of two different rabies strains for which striped skunks were the reservoir. Because the distribution of these two strains overlapped on the landscape and were present in the same host population, we could evaluate how viral properties influenced epidemiological patterns in the area of sympatry. We found that South Central Skunk rabies (SCSK) exhibited intense purifying selection and high infectivity, which are both characteristics of an epizootic virus. Conversely, North Central Skunk rabies (NCSK) exhibited relaxed purifying selection and comparatively lower infectivity, suggesting the presence of an enzootic virus. The host population in the area of sympatry was highly admixed, and skunks among allopatric and sympatric areas had similar effective population sizes. Spatial analysis indicated that landscape features had minimal influence on NCSK movement across the landscape, but those same features were partial barriers to the spread of SCSK. We conclude that NCSK and SCSK have different epidemiological properties that interact differently with both host and landscape features to influence rabies spread in the central Great Plains. We suggest a holistic approach for future studies of emerging infectious diseases that includes studies of viral properties, host characteristics, and spatial features.
Septal and lateral wall localization of PBP5, the major D,D‐carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment
Authors: Potluri Lakshmiprasad , Karczmarek Aneta , Verheul Jolanda , Piette Andre , Wilkin Jean‐Marc , Werth Nadine , Banzhaf Manuel , Vollmer Waldemar , Young Kevin D. , Nguyen‐Distèche Martine , Den Blaauwen Tanneke
Published: June 2010

The distribution of PBP5, the major D,D‐carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild‐type cells and in mutants lacking one or more D,D‐carboxypeptidases. In addition to being scattered around the lateral envelope, PBP5 was also concentrated at nascent division sites prior to visible constriction. Inhibiting PBP2 activity (which eliminates wall elongation) shifted PBP5 to midcell, whereas inhibiting PBP3 (which aborts divisome invagination) led to the creation of PBP5 rings at positions of preseptal wall formation, implying that PBP5 localizes to areas of ongoing peptidoglycan synthesis. A PBP5(S44G) active site mutant was more evenly dispersed, indicating that localization required enzyme activity and the availability of pentapeptide substrates. Both the membrane bound and soluble forms of PBP5 converted pentapeptides to tetrapeptides in vitro and in vivo, and the enzymes accepted the same range of substrates, including sacculi, Lipid II, muropeptides and artificial substrates. However, only the membrane‐bound form localized to the developing septum and restored wild‐type rod morphology to shape defective mutants, suggesting that the two events are related. The results indicate that PBP5 localization to sites of ongoing peptidoglycan synthesis is substrate dependent and requires membrane attachment.
Propagation of giant migrating contractions between the small intestine, cecum and colon during radiation
Authors: Otterson M. F. , Leming S. C. , Fox C. J. , Moulder J. E.
Published: June 2010

Background  Radiation increases the frequency of small intestinal and colonic giant migrating contractions (GMCs). These contractions contribute to the diarrhea and cramping after radiation therapy and are coordinated with one another across the ileocolonic (IC) junction.

Methods  We investigated the coordination of contractile activity between the small intestine, cecum and colon in five canines following circumferential myotomy on the ileum at the IC junction and compared it to intact animals. Studies were performed before and during a radiation schedule.

Key Results  Myotomy increased the frequency of small intestinal GMCs prior to irradiation. In intact animals, the duration and amplitude of cecal GMCs decreased when multiple contractions occurred in rapid succession. This is in contrast to small intestinal and colonic GMCs and suggests a different mechanism of propagation for GMCs within the cecum. Ileal myotomy dramatically decreased the frequency of propagating radiation‐induced colonic GMCs. The total number of colonic GMCs was not altered. Colonic contractile activity was disrupted in intact animals during irradiation. However, after ileal myotomy, irradiation did not affect the pattern of colonic contractile states. Diarrhea in irradiated animals with myotomy started earlier than intact animals. This may be related to the frequency of small intestinal GMCs.

Conclusions & Inferences  Our findings suggest importance of the enteric neural connections at the IC region to contractile disorders of both the small and large intestine. The anatomic relationship between the canine IC junction is similar to the human ileo‐appendiceal‐colonic region and surgical manipulations of this area may likewise affect human contractile activity.
Molecular analysis of ureide accumulation under drought stress in Phaseolus vulgaris L.
Authors: ALAMILLO JOSEFA M. , DÍAZ‐LEAL JUAN LUÍS , SÁNCHEZ‐MORAN MA. VICTORIA , PINEDA MANUEL
Published: June 2010

Under water deficit, ureidic legumes accumulate ureides in plant tissues, and this accumulation has been correlated with the inhibition of nitrogen fixation. In this work we used a molecular approach to characterize ureide accumulation under drought stress in Phaseolus vulgaris. Accumulation of ureides, mainly allantoate, was found in roots, shoots and leaves, but only a limited transient increase was observed in nodules from drought‐stressed plants. We show that ureide accumulation is regulated at the transcriptional level mainly through induction of allantoinase (ALN), whereas allantoate amidohydrolase (AAH), involved in allantoate degradation, was slightly reduced, indicating that inhibition of this enzyme, key in ureide breakdown in aerial tissues, is not the main cause of allantoate accumulation. Expression of the ureide metabolism genes analysed in this study was induced by abscisic acid (ABA), suggesting the involvement of this plant hormone in ureide accumulation. Moreover, we observed that increases of ureide levels in P. vulgaris drought‐stressed tissues were similar in non‐nodulated, nitrate‐fed plants, and in plants cultured under nitrogen‐fixation conditions. Our results indicate that ureide accumulation in response to water deficit is independent from de novo synthesis of ureides in nodules, and therefore uncoupled from nitrogen fixation.
Mechanisms independent of abscisic acid (ABA) or proline feedback have a predominant role in transcriptional regulation of proline metabolism during low water potential and stress recovery
Authors: SHARMA SANDEEP , VERSLUES PAUL E.
Published: June 2010

Proline accumulation in response to abiotic stress is controlled partially by transcriptional regulation of key enzymes including Δ1‐pyrroline‐carboxylate synthetase1 (P5CS1), proline dehydrogenase (ProDH), ornithine amino transferase (OAT) and Δ1‐pyrroline‐carboxylate dehydrogenase (P5CDH). For these genes, the role of abscisic acid (ABA), role of feedback regulation by high proline and the mechanisms of gene regulation upon stress release remain unclear. An ABA‐deficient (aba2‐1) mutant, mutants deficient in proline accumulation (p5cs1), as well as double mutants deficient in both, were used to determine the importance of these factors in transcriptional regulation of proline metabolism. Upregulation of P5CS1 by low water potential was less dependent on ABA than that of stress‐marker genes used for comparison. ProDH downregulation by low water potential and upregulation by stress release was not impaired in aba2‐1, p5cs1 or p5cs1/aba2‐1 compared with wild type despite differing ABA and proline levels in these mutants. Thus, ProDH is a model for characterization of novel regulatory mechanisms associated with low water potential and stress recovery. Both OAT and P5CDH were upregulated during low water potential. This contrasts with previous salt stress experiments and raises questions about the flux of metabolites through proline metabolism under low water potential when high levels of proline accumulate.
Ammonium‐dependent respiratory increase is dependent on the cytochrome pathway in Arabidopsis thaliana shoots
Authors: HACHIYA TAKUSHI , WATANABE CHIHIRO K. , BOOM CAROLINA , THOLEN DANNY , TAKAHARA KENTARO , KAWAI‐YAMADA MAKI , UCHIMIYA HIROFUMI , UESONO YUKIFUMI , TERASHIMA ICHIRO , NOGUCHI KO
Published: June 2010

Oxygen uptake rates are increased when concentrated ammonium instead of nitrate is used as sole N source. Several explanations for this increased respiration have been suggested, but the underlying mechanisms are still unclear. To investigate possible factors responsible for this respiratory increase, we measured the O2 uptake rate, activity and transcript level of respiratory components, and concentration of adenylates using Arabidopsis thaliana shoots grown in media containing various N sources. The O2 uptake rate was correlated with concentrations of ammonium and ATP in shoots, but not related to the ammonium assimilation. The capacity of the ATP‐coupling cytochrome pathway (CP) and its related genes were up‐regulated when concentrated ammonium was sole N source, whereas the ATP‐uncoupling alternative oxidase did not influence the extent of the respiratory increase. Our results suggest that the ammonium‐dependent increase of the O2 uptake rate can be explained by the up‐regulation of the CP, which may be related to the ATP consumption by the plasma‐membrane H+‐ATPase.
Keep on growing under drought: genetic and developmental bases of the response of rosette area using a recombinant inbred line population
Authors: TISNÉ SÉBASTIEN , SCHMALENBACH INGA , REYMOND MATTHIEU , DAUZAT MYRIAM , PERVENT MARJORIE , VILE DENIS , GRANIER CHRISTINE
Published: June 2010

Variation in leaf development caused by water deficit was analysed in 120 recombinant inbred lines derived from two Arabidopsis thaliana accessions, Ler and An‐1. Main effect quantitative trait loci (QTLs) and QTLs in epistatic interactions were mapped for the responses of rosette area, leaf number and leaf 6 area to water deficit. An epistatic interaction between two QTLs affected the response of whole rosette area and individual leaf area but only with effects in well‐watered condition. A second epistatic interaction between two QTLs controlled the response of rosette area and leaf number with specific effects in the water deficit condition. These effects were validated by generating and phenotyping new appropriate lines. Accordingly, a low reduction of rosette area was observed for lines with a specific allelic combination at the two interacting QTLs. This low reduction was accompanied by an increase in leaf number with a lengthening of the vegetative phase and a low reduction in individual leaf area with low reductions in epidermal cell area and number. Statistical analyses suggested that responses of epidermal cell area and number to water deficit in individual leaves were partly caused by delay in flowering time and reduction in leaf emergence rate, respectively.
Characterization of Immune Responses Following Intranasal Immunization with the Mycobacterium bovis CFP‐10 Protein Expressed by Attenuated Salmonella typhimurium
Authors: Zhang H. , Wen K. , Shen J. , Geng S. , Huang J. , Pan Z. , Jiao X.
Published: June 2010

Culture filtrate protein 10 (CFP‐10) from Mycobacterium bovis or Mycobacterium tuberculosis (MTB) is an immunodominant T‐cell antigen expressed during the early stages of infection. Because lungs are most commonly associated with primary M. bovis infections, specific immunity at this site is desirable for protection. Therefore, in this study, immune responses generated in mouse lung, spleen and Peyer’s patches were examined following intranasal (i.n.) immunization with Salmonella typhimurium– expressing CFP‐10. Cells harvested from the lungs and Peyer’s patches of immunized mice and then stimulated with CFP‐10 produced significant levels of IFN‐γ and these mice developed elevated serum IgG and lung IgA anti‐CFP‐10 responses, suggesting that this approach induced potent anti‐CFP‐10 mucosal immunity. Our study demonstrates that i.n. administration of CFP‐10 expressed by S. typhimurium represents an effective way to induce efficient immune response to M. bovis antigen.
Identification of protein stability determinants in chloroplasts
Authors: Apel Wiebke , Schulze Waltraud X. , Bock Ralph
Published: June 2010

Although chloroplast protein stability has long been recognised as a major level of post‐translational regulation in photosynthesis and gene expression, the factors determining protein stability in plastids are largely unknown. Here, we have identified stability determinants in vivo by producing plants with transgenic chloroplasts that express a reporter protein whose N‐ and C‐termini were systematically modified. We found that major stability determinants are located in the N‐terminus. Moreover, testing of all 20 amino acids in the position after the initiator methionine revealed strong differences in protein stability and indicated an important role of the penultimate N‐terminal amino acid residue in determining the protein half life. We propose that the stability of plastid proteins is largely determined by three factors: (i) the action of methionine aminopeptidase (the enzyme that removes the initiator methionine and exposes the penultimate N‐terminal amino acid residue), (ii) an N‐end rule‐like protein degradation pathway, and (iii) additional sequence determinants in the N‐terminal region.
The hydroxyproline‐rich glycoprotein domain of the Arabidopsis LRX1 requires Tyr for function but not for insolubilization in the cell wall
Authors: Ringli Christoph
Published: June 2010

Extensins, hydroxyproline‐rich repetitive glycoproteins with Ser–Hyp4 motifs, are structural proteins in plant cell walls. The leucine‐rich repeat extensin 1 (LRX1) of Arabidopsis thaliana is an extracellular protein with both a leucine‐rich repeat and an extensin domain, and has been demonstrated to be important for cell‐wall formation in root hairs. lrx1 mutants develop defective cell walls, resulting in a strong root hair phenotype. The extensin domain is essential for protein function and is thought to confer insolubilization of LRX1 in the cell wall. Here, in vivo characterization of the LRX1 extensin domain is described. First, a series of LRX1 extensin deletion constructs was produced that led to identification of a much shorter, functional extensin domain. Tyr residues can induce intra‐ and inter‐molecular cross‐links in extensins, and substitution of Tyr in the extensin domain by Phe led to reduced activity of the corresponding LRX1 protein. An additional function of Tyr (or Phe) is provided by the aromatic nature of the side chain. This suggests that these residues might be involved in hydrophobic stacking, possibly as a mechanism of protein assembly. Finally, modified LRX1 proteins lacking Tyr in the extensin domain are still insolubilized in the cell wall, indicating strong interactions of extensins within the cell wall in addition to the well‐described Tyr cross‐links.
Exome sequencing: locating causative genes in rare disorders
Authors: Pussegoda KA
Published: June 2010

Exome sequencing identifies the cause of a mendelian disorder

Ng et al. (2009)

Nature Genetics. Advanced online publication.
Genetic variants associated with cisplatin‐induced hearing loss
Authors: Pussegoda KA
Published: June 2010

Genetic variants in TPMT and COMT are associated with hearing loss in children receiving cisplatin chemotherapy

Ross et al. (2009)

Nature Genetics 41:1345–1349
Prolongation of renal allograft survival by anergic cells: advantages and limitations
Authors: Bashuda Hisashi , Shimizu Akira , Uchiyama Masateru , Okumura Ko
Published: June 2010

Bashuda H, Shimizu A, Uchiyama M, Okumura K. Prolongation of renal allograft survival by anergic cells: advantages and limitations. Clin Transplant 2010: 24 (Suppl. 22): 6–10. © 2010 John Wiley & Sons A/S.

Abstract:  Even in the era of pharmacological calcineurin inhibitors, a current major challenge in organ transplantation remains the development of immunosuppressive regimens that protect against rejection. One potentially effective procedure is the use of donor‐specific anergic T cells generated ex vivo and adoptively transferred back into the recipient after transplantation. In our own work, we first investigated the effect of anergic cells on the prolongation of graft survival in non‐human primates. In six animals, half of the recipients survived for over one yr (all animals died or were killed within eight yr). The cause of death was acute renal failure because of cellular rejection (one), uncontrolled bleeding after renal biopsy (two), hydronephrosis probably because of ureteral stenosis (one), and chronic rejection (one). The remaining animal was killed at the end of the study. No infection, malignancy, or signs of graft versus host disease (GVHD) was observed in any of these monkeys. Except for the one animal that died of acute cellular rejection, there was no evidence of tubular infiltration by mononuclear cells, glomerular damage, or parenchymal necrosis. In all animals surviving for more than one yr, a mild grade of interstitial fibrosis, an increase in mesangial matrix, or glomerulopathy was noted. In two of three monkeys, no vascular narrowing of the luminal area caused by fibrointimal thickening of arteries was noted, and arteriosclerotic change was dominant. In this chapter, we summarize the efficacy and limitations of our strategy.
A case of pregnancy‐induced thrombotic thrombocytopenic purpura with a kidney allograft recipient
Authors: Iwami Daiki , Harada Hiroshi , Hotta Kiyohiko , Miura Masayoshi , Seki Toshimori , Togashi Masaki , Hirano Tetsuo
Published: June 2010

Iwami D, Harada H, Hotta K, Miura M, Seki T, Togashi M, Hirano T. A case of pregnancy‐induced thrombotic thrombocytopenic purpura with a kidney allograft recipient. Clin Transplant 2010: 24 (Suppl. 22): 66–69. © 2010 John Wiley & Sons A/S.

Abstract:  A 32‐yr‐old female patient, who had been suffering from diffuse crescentic glomerulonephritis and a consequent end‐stage renal disease, successfully underwent living‐related ABO‐incompatible kidney transplantation after a desensitization therapy including anti‐CD20 monoclonal antibody. Forty‐six months after the transplantation, the recipient became pregnant. At the 17th gestational week, the patient was admitted for the management of pregnancy‐induced hypertension and aggressive deterioration of kidney graft function. At the 21st gestational week, the patient lost her kidney graft and was re‐induced into regular hemodialysis. The patient was also suffering from progressive hemolytic anemia, thrombocytopenia, and neurologic symptoms with decreased activity of von Willebrand factor‐cleaving protease, a disintegrin‐like and metalloprotease with thrombospondin type 1 motifs 13 (ADAMTS13). From these findings and a kidney allograft biopsy, the patient was diagnosed as thrombotic thrombocytopenic purpura concurrent with acute T‐cell‐mediated rejection. The patient immediately underwent plasma exchange as well as steroid pulse therapy. Despite these treatments, thrombocytopenia and intrauterine growth retardation progressed. The patient underwent a caesarian section at the 24th gestational week. Consequently, her platelet count recovered drastically. However, the patient lost her neonate five d after giving a birth, and the patient’s graft function had never recovered.
ABO‐incompatible renal transplantation in Epstein syndrome
Authors: Ogura Masao , Kikuchi Eriko , Kaito Hiroshi , Kamei Koichi , Matsuoka Kentaro , Tanaka Hideaki , Kuroda Tatsuo , Sekine Takashi , Ito Shuichi
Published: June 2010

Ogura M, Kikuchi E, Kaito H, Kamei K, Matsuoka K, Tanaka H, Kuroda T, Sekine T, Ito S. ABO‐incompatible renal transplantation in Epstein syndrome. Clin Transplant 2010: 24 (Suppl. 22): 31–34. © 2010 John Wiley & Sons A/S.

Abstract:  Epstein syndrome (ES) is an autosomal dominant hereditary disease characterized by hereditary nephritis, sensory deafness, and thrombocytopenia. We herein report the case of a 20‐yr‐old man with ES who underwent ABO blood type‐incompatible living‐donor kidney transplantation from his mother. He was given platelet transfusion, and his pre‐operative number of platelets were 108 × 103/μL. After transplantation, urine output and the decrease in serum creatinine (sCr) were within the acceptable ranges. On the seventh post‐operative day (POD), sCr had risen and urine output decreased. Anti‐type A antibody rapidly elevated from <2 times (×2) just before transplantation to 64 times (×64), and the patient required hemodialysis again. Resistance index (RI) by ultrasound increased from an average of 0.5 ∼ 0.6 on POD 1 to an average of 0.7 ∼ 0.8 on POD 7. However, several biopsies (POD 4, 7, and 10) showed no obvious findings of acute rejection except for intense C4d deposition. Because acute antibody‐mediated rejection was not completely ruled out, he was treated with methyl‐prednisolone pulse therapy, plasma exchange, cyclophosphamide, and immunoglobulin. Regardless, his titer of anti‐type A antibody was still high, and he still presented oliguria. We performed an emergent splenectomy. Consequently, the levels of anti‐type A antibody decreased, the RI also dropped to an average of 0.6. However, on POD 19 and 25 (platelets were 27 × 103/μL and 36 × 103/μL), he developed a massive intraperitoneal hematoma around the graft and region of the removed spleen, which pushed the graft out and caused acute tubular necrosis, resulting in anuria. The RI rose to an average of 0.8 ∼ 1.0 after these episodes. He also experienced bleeding from a duodenal ulcer on POD 21. However, his renal function has fully recovered after acute hemodialysis for 35 d. The latest sCr was 1.5 mg/dL with a recovery in RI to 0.6. Although his platelet count was maintained at a minimum of 50 × 103/μL, he had several severe bleeding episodes, concluding that sufficient platelets are necessary after transplantation in ES.
Clinicopathological analysis of acute vascular rejection cases after renal transplantation
Authors: Shimizu Tomokazu , Ishida Hideki , Shirakawa Hiroki , Omoto Kazuya , Tsunoyama Kuniko , Tokumoto Tadahiko , Tanabe Kazunari
Published: June 2010

Shimizu T, Ishida H, Shirakawa H, Omoto K, Tsunoyama K, Tokumoto T, Tanabe K. Clinicopathological analysis of acute vascular rejection cases after renal transplantation. Clin Transplant 2010: 24 (Suppl. 22): 22–26. © 2010 John Wiley & Sons A/S.

Abstract:  Histopathological change of acute vascular rejection (AVR) is characterized by intimal arteritis and transmural arteritis. In this report, we discuss the clinicopathological analysis of AVR cases after renal transplantation (RTX).

Patients:  AVR was diagnosed in 17 patients from 17 renal transplant patients followed in our institute between January 2003 and September 2008. We retrospectively reviewed these 17 patients.

Results:  Among 17 cases of AVR, 10 cases were mild (v1 in Banff 07 classification), five were moderate (v2), and two were severe (v3). Interstitial inflammation (i1–i3) was present in all 17 biopsies. Moderate to severe tubulitis (t2–t3) was present in seven biopsies, and transplant glomerulitis (g1–g3) was present in 11, peritubular capillaritis (ptc1–ptc3) was in 15 of 17 biopsies. C4d deposition in peritubular capillary (PTC) was observed in 6 of 17 cases. By assaying with plastic beads coated with anti‐human leukocyte antigen (HLA) antigen performed in 17 cases, the circulating ant‐HLA alloantibody was detected in 10 patients, of which 5/10 were donor‐specific antibodies (DSA). Acute antibody‐mediated rejection (AAMR) was diagnosed in three cases. Many of v1 cases, steroid pulse therapy (SP) were effective. In v2 and v3 cases, six of seven were steroid‐resistant rejection and were need more anti‐rejection therapy (ART), such as muromonab CD3 (OKT3) injection, gusperimus (DSG) injection, plasmapheresis, intravenous immune globulin, and injection of rituximab. Ten of 17 patients recovered their renal allograft functions by ART, and 16 of 17 patients’ grafts are functioning. Deterioration of renal allografts’ function after biopsies was seen in seven patients with one of them lost their graft.

Conclusions:  In some cases, AVR might be provoked by anti‐donor antibodies. The prognosis of the graft exhibiting AVR was relatively good in present immunosuppression and ART.
Protocol biopsies for focal segmental glomerulosclerosis treated with plasma exchange and rituximab in a renal transplant patient
Authors: Sakai Ken , Takasu Jirou , Nihei Hiroshi , Yonekura Takashi , Aoki Yujirou , Kawamura Takeshi , Mizuiri Sonoo , Aikawa Atsushi
Published: June 2010

Sakai K, Takasu J, Nihei H, Yonekura T, Aoki Y, Kawamura T, Mizuiri S, Aikawa A. Protocol biopsies for focal segmental glomerulosclerosis treated with plasma exchange and rituximab in a renal transplant patient. Clin Transplant 2010: 24 (Suppl. 22): 60–65. © 2010 John Wiley & Sons A/S.

Abstract:  We discuss a renal transplant patient with focal segmental glomerulosclerosis (FSGS) treated with plasma exchange and rituximab. A 45‐yr‐old woman underwent cadaveric renal transplantation in May 2008. She had started hemodialysis support in 1991. Immediately after transplantation, massive proteinuria (1–5 g/d) appeared. Graft biopsy at one h showed minor glomerular abnormalities with partial foot process effacement on electric microscopy. Protocol biopsy at three months after transplantation for persistent proteinuria showed obvious FSGS under light microscopy. Plasma exchange and rituximab administration were subsequently initiated in August 2008, and proteinuria disappeared within a month after starting these treatments. Protocol graft biopsy one yr after transplantation (2009) showed increased global sclerosis and a decrease in segmental sclerosis. In addition, foot process effacement had recovered by one yr after transplantation. Plasma exchange and subsequent rituximab administration led to clinical remission of post‐transplant FSGS with improvement in podocyte structure. Rituximab should be considered soon after several sessions of plasmapheresis in transplant patients with recurrent FSGS.
A case of second renal transplantation with acute antibody‐mediated rejection complicated with BK virus nephropathy
Authors: Atsumi Hirokatsu , Asaka Mitsuhiro , Kimura Shogo , Imura Junko , Fujimoto Keiji , Chikazawa Yoshihiro , Nakagawa Masaru , Okuyama Hiroshi , Yamaya Hideki , Moriyama Manabu , Tanaka Tatsuro , Suzuki Koji , Yokoyama Hitoshi
Published: June 2010

Atsumi H, Asaka M, Kimura S, Imura J, Fujimoto K, Chikazawa Y, Nakagawa M, Okuyama H, Yamaya H, Moriyama M, Tanaka T, Suzuki K, Yokoyama H. A case of second renal transplantation with acute antibody‐mediated rejection complicated with BK virus nephropathy. Clin Transplant 2010: 24 (Suppl. 22): 35–38. © 2010 John Wiley & Sons A/S.

Abstract:  We reported a 40‐year‐old female case of second renal transplantation with antibody‐mediated rejection (AMR) complicated by BK virus nephropathy. She started hemodialysis (HD) at the age of 17 because of IgA nephropathy. At the age of 18, she underwent living‐donor kidney transplantation from her father, but two and a half years after transplantation, she developed chronic rejection. This time, she received cadaveric renal transplantation under the negative cross‐match (AHG‐LCT), and HLA‐AB 1 mismatch and ‐DR 1 mismatch. Immunosuppressive therapy was initiated using the following four immunosuppressants: methylprednisolone, mycophenolate mofetil, cyclosporine, and basiliximab. However, renal graft showed delayed function, the biopsy showed glomerulitis (g2), endarteritis (v1), and cellular infiltration (ptc3) consisting mainly of mononuclear cells in the peritubular capillary with diffusely positive C4d and anti‐SV 40 large T‐antigen‐positive renal tubular epithelial cells on post‐operative day 19. The donor‐specific antibody for HLA‐B46 was proven by the LAB screen method. We performed plasma exchange three times and administered immunoglobulin (15 g in total). Then, methylprednisolone pulse therapy was added, and the serum creatinine (SCr) levels gradually decreased. On post‐operative day 44, the patient was removed from HD and was discharged with SCr level of 3.3 mg/dL.
Perioperative anticoagulation and antiplatelet therapy in renal transplant: is there an increase in bleeding complication?
Authors: Eng Mary , Brock Guy , Li Xiaohong , Chen Yinlu , Ravindra Kadiyala V. , Buell Joseph F. , Marvin Michael R.
Published: June 2010

Eng M, Brock G, Li X, Chen Y, Ravindra KV, Buell JF, Marvin MR. Perioperative anticoagulation and antiplatelet therapy in renal transplant: is there an increase in bleeding complication?  Clin Transplant 2011: 25: 292–296. © 2010 John Wiley & Sons A/S.

Abstract: Background:  Renal transplant recipients may have comorbidities requiring anticoagulation or antiplatelet therapy. While the effects of warfarin may be neutralized with plasma infusion, those of aspirin and clopidogrel are not easily reversible and may be associated with an increased risk of bleeding. We conducted this study to evaluate the risk of bleeding complications in patients receiving perioperative anticoagulation or antiplatelet therapy.

Methods:  Medical records of patients who underwent renal transplantation from July 1, 2005 to April 30, 2009 were retrospectively reviewed. Patients receiving perioperative anticoagulation or antiplatelet therapy were identified. The incidence of reoperation, transfusion utilization and decrease in serum hemoglobin from pre‐operative value (ΔHgb) were compared to those on no therapy.

Results:  Of the 327 patients identified, 105 received pre‐operative anticoagulation or antiplatelet therapy, 28 received therapy post‐operatively, while 213 patients received no therapy. The incidence of reoperation, transfusion utilization and ΔHgb were not significantly increased with pre‐operative anticoagulation or antiplatelet therapy. With post‐operative heparin infusion, the incidence of reoperation and transfusion utilization were significantly increased (p values < 0.001). Patients with activated partial thromboplastin times (aPTT) >80 s experienced significant bleeding complications.

Conclusion:  A supratherapeutic aPTT with post‐operative heparin infusion was associated with the greatest risk of bleeding complication.
Anaphylaxis in Israel: Experience with 92 hospitalized children
Authors: Hoffer V. , Scheuerman O. , Marcus N. , Levy Y. , Segal N. , Lagovsky I. , Monselise Y. , Garty B. Z.
Published: June 2010

To cite this article: Hoffer V, Scheuerman O, Marcus N, Levy Y, Segal N, Lagovsky I, Monselise Y, Garty BZ. Anaphylaxis in Israel: Experience with 92 hospitalized children. Pediatr Allergy Immunol 2011; 22:172–177.

Little is known about the courses, causes, and clinical features of anaphylaxis in children outside the USA and Europe. Our objective was to evaluate the events of anaphylaxis in children admitted to the Schneider Children’s Medical Center of Israel, a major tertiary facility, over a 12‐year period. Ninety‐two children with anaphylaxis (50 boys, 42 girls) aged 14 days to 18 yr (mean, 7.4 yr) were hospitalized during the study period. The event occurred at home in 52 children (56%), in a medical institution in 24 (26%), outdoors in 13 (15%), at school in 2 (2%), and in an unspecified location in 1 (1%). The main causes were foods (43%), mainly milk and nuts, medications (22%), and hymenoptera venom (11%); in five children, anaphylaxis occurred during general anesthesia, and in 5, the causative agent could not be determined. Food‐induced anaphylaxis tended to occur in younger children. Forty‐eight children (52%) had a history of atopy (mainly asthma). Hospital treatment consisted of corticosteroids (85%), antihistamines (75%), epinephrine (72%), and β2 agonists (42%). Seven patients were admitted to intensive care units. There were no fatalities. EpiPen was used by only one of the 16 patients with more than one episode of anaphylaxis, indicating that patient and parent education in the application of the EpiPen needs to be improved.