bluuebunny/crossref_metadata_2025 huggingface dataset:
First Report of Diplodia seriata Causing Loquat Fruit Rot in Spain
Authors: Palou L. , Taberner V. , Montesinos-Herrero C.
Published: February 2013

Spain is the second largest loquat (Eriobotrya japonica Lindl.) producer in the world, with about 40,000 t per year. ‘Algerie’ is the main cultivar planted in Alicante province (SE of Spain; Lat. 38.40° N, Long. 0.08° W), where more than 80% of Spanish commercial loquat plantations are located. In a survey of fruit losses at harvest, irregular brownish superficial dry spots (5 to 15 mm) located mainly near the stem end were observed on fruits from different orchards. After incubation at 20°C for 14 days, the spots on fruit expanded rapidly and turned to dark brown or black, producing black, unilocular, ostiolate, and thick-walled pycnidia. Isolation was performed by disinfecting the surface of symptomatic fruits with alcohol and aseptically cutting pieces of infected peel tissue and plating them in potato dextrose agar (PDA) dishes. The potential causal agent (isolate IVIA GCA-5) was identified in the Spanish Type Culture Collection (CECT, University of Valencia, Valencia, Spain). The fungus grew rapidly on both PDA and malt extract agar (MEA) at 26°C, covering the entire plate surface with dark gray mycelium within 4 days. The plate reverse was dark gray to black. The conidia were brown and aseptate, with the apex broadly rounded and the base rounded or truncate, and 23 × 11 μm (n = 50). The identification of Diplodia seriata De Not. was molecularly confirmed with the amplification with the primers ITS1 and ITS4 and subsequent sequencing of the internal transcribed spacer ITS1-5.8S-ITS2 region of the rDNA extracted from the isolate IVIA GCA-5 (GenBank Accession No. JX987099). Furthermore, the region D1/D2 in the 5′ end of the 28S rDNA gene was amplified with the primers NL1 and NL4 and sequenced (JX997743). A nucleotide BLAST analysis showed in both cases 100% identity with D. seriata [EF127892 (3) and AY928050, respectively]. To fulfill Koch's postulates, 5-mm diameter mycelial plugs from 7-day-old colonies of isolate IVIA GCA-5 grown on PDA at 25°C were aseptically transferred to skin wounds on superficially disinfected ‘Algerie’ loquats (one plug per fruit; n = 9). Wounded but not inoculated fruit were used as controls. The experiment was repeated three times. Inoculated fruit developed lesions of 18 to 100 mm after 7 to 21 days of incubation at 20°C. No lesion was observed on controls. The fungus was consistently reisolated from inoculated fruit. D. seriata is a broadly spread pathogen causing cankers, blight, dieback, and fruit rots in vines and many fruit trees. In Spain, it has been reported to cause fruit rot of olive (1) and branch dieback in olive (2) and grapevine (4). To our knowledge, this is the first report worldwide of D. seriata causing loquat fruit rot.

References: (1) J. Moral et al. Plant Dis. 92:311, 2008. (2) J. Moral et al. Phytopathology 100:1340, 2010. (3) A. J. L. Phillips et al. Fungal Divers. 25:141, 2007. (4) J. R. Úrbez-Torres et al. Plant Dis. 90:835, 2006.
Bioinformatics analysis reveals biophysical and evolutionary insights into the 3-nitrotyrosine post-translational modification in the human proteome
Authors: Ng John Y. , Boelen Lies , Wong Jason W. H.
Published: February 2013

Protein 3-nitrotyrosine is a post-translational modification that commonly arises from the nitration of tyrosine residues. This modification has been detected under a wide range of pathological conditions and has been shown to alter protein function. Whether 3-nitrotyrosine is important in normal cellular processes or is likely to affect specific biological pathways remains unclear. Using GPS-YNO2, a recently described 3-nitrotyrosine prediction algorithm, a set of predictions for nitrated residues in the human proteome was generated. In total, 9.27 per cent of the proteome was predicted to be nitratable (27 922/301 091). By matching the predictions against a set of curated and experimentally validated 3-nitrotyrosine sites in human proteins, it was found that GPS-YNO2 is able to predict 73.1 per cent (404/553) of these sites. Furthermore, of these sites, 42 have been shown to be nitrated endogenously, with 85.7 per cent (36/42) of these predicted to be nitrated. This demonstrates the feasibility of using the predicted dataset for a whole proteome analysis. A comprehensive bioinformatics analysis was subsequently performed on predicted and all experimentally validated nitrated tyrosine. This found mild but specific biophysical constraints that affect the susceptibility of tyrosine to nitration, and these may play a role in increasing the likelihood of 3-nitrotyrosine to affect processes, including phosphorylation and DNA binding. Furthermore, examining the evolutionary conservation of predicted 3-nitrotyrosine showed that, relative to non-nitrated tyrosine residues, 3-nitrotyrosine residues are generally less conserved. This suggests that, at least in the majority of cases, 3-nitrotyrosine is likely to have a deleterious effect on protein function and less likely to be important in normal cellular function.
Identification and characterization of complex dual nuclear localization signals in human bocavirus NP1
Authors: Li Qian , Zhang Zhenfeng , Zheng Zhenhua , Ke Xianliang , Luo Huanle , Hu Qinxue , Wang Hanzhong
Published: February 2013

Human bocavirus (HBoV), closely related to canine minute virus (MVC) and bovine parvovirus (BPV), is a new member of the Bocavirus genus within the Parvoviridae family. The non-structural protein NP1 of HBoV is a nuclear localized protein and plays an important role in DNA replication as well as in the evasion of host innate immunity. In the current study, we provide the first evidence that NP1 possesses a non-classical nuclear localization signal (ncNLS) (amino acids 7–50). Embedded within this ncNLS is a classical bipartite nuclear localization signal (cNLS) (amino acids 14–28), capable of transporting a heterologous cytoplasmic protein β-galactosidase fusion protein (β-gal-EGFP) to the nucleus via the classical importin α/β1-mediated pathway. Amino acids 7–50 containing the cNLS and the ncNLS of NP1 or full-length NP1 interact with importin α1, importin β1 and importin β1Δ, which lacks the importin α binding domain, indicating that the nuclear import of NP1 is through both conventional importin α/β1 heterodimer- and non-classical importinß1-mediated pathways. Given that the arrangement of a cNLS embedded within an ncNLS is unusual in viral proteins, our data together reveal a novel molecular mechanism underlying the nuclear import of HBoV NP1, providing a basis for further understanding its biological function.
Adenoviral E2 IVa2 protein interacts with L4 33K protein and E2 DNA-binding protein
Authors: Ahi Yadvinder S. , Vemula Sai V. , Mittal Suresh K.
Published: February 2013

Adenovirus (AdV) is thought to follow a sequential assembly pathway similar to that observed in dsDNA bacteriophages and herpesviruses. First, empty capsids are assembled, and then the genome is packaged through a ring-like structure, referred to as a portal, located at a unique vertex. In human AdV serotype 5 (HAdV5), the IVa2 protein initiates specific recognition of viral genome by associating with the viral packaging domain located between nucleotides 220 and 400 of the genome. IVa2 is located at a unique vertex on mature capsids and plays an essential role during genome packaging, most likely by acting as a DNA packaging ATPase. In this study, we demonstrated interactions among IVa2, 33K and DNA-binding protein (DBP) in virus-infected cells by in vivo cross-linking of HAdV5-infected cells followed by Western blot, and co-immunoprecipitation of IVa2, 33K and DBP from nuclear extracts of HAdV5-infected cells. Confocal microscopy demonstrated co-localization of IVa2, 33K and DBP in virus-infected cells and also in cells transfected with IVa2, 33K and DBP genes. Immunogold electron microscopy of purified HAdV5 showed the presence of IVa2, 33K or DBP at a single site on the virus particles. Our results provide indirect evidence that IVa2, 33K and DBP may form a complex at a unique vertex on viral capsids and cooperate in genome packaging.
Targeted knock-down of cellular prion protein expression in myelinating Schwann cells does not alter mouse prion pathogenesis
Authors: Halliez Sophie , Chesnais Nathalie , Mallucci Giovanna , Vilotte Marthe , Langevin Christelle , Jaumain Emilie , Laude Hubert , Vilotte Jean-Luc , Béringue Vincent
Published: February 2013

In naturally acquired transmissible spongiform encephalopathies, the pathogenic agents or prions spread from the sites of initial peripheral uptake or replication to the brain where they cause progressive and fatal neurodegeneration. Routing via the peripheral nervous system is considered to be one of the main pathways to the central nervous system. Replication of prions in Schwann cells is viewed as a potentially important mechanism for efficient prion spread along nerves. Here we used a Cre-loxP mouse transgenetic approach to disrupt host-encoded prion protein (PrPC) specifically in myelinating Schwann cells. Despite the use of infection routes targeting highly myelinated nerves, there was no alteration in mouse prion pathogenesis, suggesting that conversion-dependent, centripetal spread of prions does not crucially rely on PrPC expressed by myelinating Schwann cells.
The Epiphytic Fungus Pseudozyma aphidis Induces Jasmonic Acid- and Salicylic Acid/Nonexpressor of PR1-Independent Local and Systemic Resistance
Authors: Buxdorf Kobi , Rahat Ido , Gafni Aviva , Levy Maggie
Published: February 2013

Pseudozyma spp. are yeast-like fungi, classified in the Ustilaginales, which are mostly epiphytic or saprophytic and are not pathogenic to plants. Several Pseudozyma species have been reported to exhibit biological activity against powdery mildews. However, previous studies have reported that Pseudozyma aphidis, which can colonize plant surfaces, is not associated with the ‎‎collapse of powdery ‎mildew colonies. In this report, we describe a novel P. aphidis strain and study its interactions with its plant host and the plant pathogen Botrytis cinerea. This isolate was found to secrete extracellular metabolites that inhibit various fungal pathogens in vitro and significantly reduce B. cinerea infection in vivo. Moreover, P. aphidis sensitized Arabidopsis (Arabidopsis thaliana) plants’ defense machinery via local and systemic induction of PATHOGENESIS-RELATED1 (PR1) and PLANT DEFENSIN1.2 (PDF1.2) expression. P. aphidis also reduced B. cinerea infection, locally and systemically, in Arabidopsis mutants impaired in jasmonic acid (JA) or salicylic acid (SA) signaling. Thus, in addition to direct inhibition, P. aphidis may inhibit B. cinerea infection via induced resistance in a manner independent of SA, JA, and Nonexpressor of PR1 (NPR1). P. aphidis primed the plant defense machinery and induced stronger activation of PDF1.2 after B. cinerea infection. Finally, P. aphidis fully or partially reconstituted PR1 and PDF1.2 expression in npr1-1 mutant and in plants with the SA hydroxylase NahG transgene, but not in a jasmonate resistant1-1 mutant, after B. cinerea infection, suggesting that P. aphidis can bypass the SA/NPR1, but not JA, pathway to activate PR genes. Thus, either partial gene activation is sufficient to induce resistance, or the resistance is not directed solely through PR1 and PDF1.2 but probably through other pathogen-resistance genes or pathways as well.
Time‐lag in extinction dynamics in experimental populations: evidence for a genetic Allee effect?
Authors: Vercken Elodie , Vincent Flora , Mailleret Ludovic , Ris Nicolas , Tabone Elisabeth , Fauvergue Xavier
Published: February 2013

Propagule pressure, i.e. the number of individuals introduced, is thought to be a major predictor of the establishment success of introduced populations in the field. Its influence in laboratory experimental systems has however been questioned. In fact, other factors involved in long‐term population persistence, like habitat size, were usually found to explain most of the dynamics of experimental populations.

To better understand the respective influence of short‐ and long‐term factors and their potential interaction on extinction dynamics in experimental systems, we investigated the influence of propagule pressure, habitat size and genetic background on the early dynamics of laboratory‐based populations of a hymenopteran parasitoid.

The amount of demographic variance differed between establishment and persistence phase and was influenced by habitat size and genetic background (geographic strain), but independent of propagule pressure. In contrast, the probability of extinction within five generations depended on the genetic background and on the interaction between propagule pressure and habitat size. Vulnerability to extinction in small size habitats was increased when populations were founded with a small number of individuals, but this effect was delayed until the third to fifth generations.

These results indicate that demographic stochasticity is influential during population establishment, but is not affected by the genetic variability of propagules. On the other hand, extinction might be influenced by a genetic Allee effect triggered by the combination of low propagule pressure and genetic drift. Finally, we documented consistent differences between genetic backgrounds in both deterministic and stochastic population dynamics patterns, with major consequences on extinction risk and ultimately population establishment.
Routine early versus deferred provisional tirofiban treatment in patients with acute coronary syndrome undergoing percutaneous coronary intervention
Authors: Zhang You , Gao Chuanyu , Liu Hongzhi , Wang Xianpei , Yang Honghui , Li Muwei , Wang Xianqing , Zhu Zhongyu , Hu Dayi
Published: February 2013

The present study examined the optimal timing of tirofiban administration in moderate‐ or high‐risk non‐ST segment elevated acute coronary syndrome (NSTE‐ACS) patients undergoing percutaneous coronary intervention (PCI).

Eligible patients were randomized into two groups. Tirofiban was administered routinely at ≥ 4 h before angiography (routine early group; n = 141 patients) or provisionally only for bailout after angiography (deferred provisional group; n = 145 patients). The parameters analysed were: creatine kinase MB isoenzyme (CK‐MB), thrombolysis in myocardial infarction (TIMI) flow, thrombotic complications during PCI, efficacy end‐points (death, myocardial infarction or target vessel revascularization) at 7, 30 and 180 days and safety end‐points (bleeding or thrombocytopenia).

In the deferred provisional group, 48 patients (33.1%) required bailout tirofiban. Tirofiban was administered 5.8 h earlier in the routine early compared with the deferred provisional group. The routine early group showed a lower percentage increase in CK‐MB (in U/L) 12–24 h after PCI compared with the deferred provisional group (0 (−4.0, 3.0) vs 0.4 (−3.0, 5.0), respectively; P = 0.045), as well as higher pre‐PCI TIMI 3 (i.e. normal) flow (78.7% vs 64.8%, respectively; P = 0.042) and a lower incidence of thrombotic events (5.0% vs 33.1%, respectively; P < 0.0001). There were no significant differences in efficacy and safety end‐points.

In patients with moderate‐ or high‐risk NSTE‐ACS, early tirofiban combined with dual antiplatelet therapy was associated with better patency before PCI, attenuated minor myocardial damage and a lower prevalence of thrombotic complications during PCI, but had no significant benefit on the post‐PCI TIMI 3 flow or short‐term prognosis.
A short note on extractive foraging behavior in gray snub‐nosed monkeys
Authors: XIANG Zuofu , LIANG Wenbin , NIE Shuaiguo , LI Ming
Published: February 2013

Extractive foraging (EF) involves the exploitation of hidden or embedded foods, generally any food that is not visible to the naked eye. Therefore, EF is particularly important for survival in marginal habitats as it provides seasonal fallback foods in low food availability seasons. Although many studies consider primates’ EF behavior and category, colobine species are usually categorized as non‐extractive foragers and few studies quantitatively examine their EF behavior. In this study, we examined the EF behavior of one colobine species, the gray snub‐nosed monkey (Rhinopithecus brelichi), at Yangaoping in Fanjingshan National Nature Reserve, Guizhou. We recorded 6 categories of EF behaviors. The most frequently sought‐out foods were seeds, young bamboo and invertebrates. Extracted foods accounted for an average of 26.02% of feeding records. As the monkey engages in little EF behavior in the winter when the food availability is low, these results seem to do not support the hypothesis that EF serves to secure additional resources during lean times in marginal or seasonal habitats. According to these findings, we suggestR. brelichshould be considered as an extractive forager. Our study also highlights the need for increased representation of colobines in the EF literature to better inform the discussion concerning its link to primate brain evolution.
Helminth parasites of freshwater fishes from Cuatro Ciénegas, Coahuila, in the Chihuahuan Desert of Mexico: inventory and biogeographical implications
Authors: AGUILAR‐AGUILAR Rogelio , MARTÍNEZ‐AQUINO Andrés , ESPINOSA‐PÉREZ Héctor , PÉREZ‐PONCE DE LEÓN Gerardo
Published: February 2013

As part of an ongoing inventory of the helminth parasites of freshwater fishes in Mexico, 570 individual fish were collected between Apr 2008 and Oct 2011 in 26 localities along the Cuatro Ciénegas region in Coahuila State, northern Mexico. Seventeen species of hosts, mostly corresponding to Nearctic freshwater elements, were studied. A total of 8324 individual worms were collected during this survey, representing 25 species of helminths, of which 9 were digeneans, 3 monogeneans, 3 acanthocephalans, 9 nematodes and 1 cestode. Most of the records in this checklist represent new host or locality records. The information provided in this checklist may be helpful for our understanding of the biodiversity and historical biogeography of this host–parasite system, because in the Cuatro Ciénegas region occur a Nearctic freshwater fish fauna, along with Neotropical and endemic elements, and from a biogeographical point of view, this may represent a transitional area.
Analysis of differently expressed proteins involved in metastatic niche of lung
Authors: Zheng Yan , Lin Yongbin , Situ Dongrong , Jiang Long , Su Xiaodong , Long Hao
Published: February 2013

Background

The “seed and soil” hypothesis for metastasis was a pivotal milestone in the study of malignant disease. Recently, growing studies have focused on the tumor secretory factors that may mediate preparation of the “metastatic soil.” A suitable environment for the metastatic lesions was created by many inflammatory cytokines in the lung, whichwas verified by in vivo experimental models. In 2005, a pre‐metastatic niche and metastatic niche modelwere suggested by David Lyden and Bethan Psaila, to delineate the interactions between malignant cells and their microenvironment at the metastatic site, whichsoon became the most importanthypothesis. However, the evidence is limited to animal models. More clinical evidence is needed to support this hypothesis.

Methods

Human lung specimens were taken from different regions within metastatic lung tissue and normal lung tissue. Differently expressed proteins were analyzed by using the two dimensional fluorescent difference gel electrophoresis (2‐D DIGE) technology: about 0.5–1 cm (Tissue 1) of lung tissue was taken adjacent to the metastatic tumor; about 1 cm (Tissue 2) of lung tissue was taken far away from the metastatic tumor; and normal lung tissue of the inflammatory pseudotumor (Tissue 3) was taken at least 3 cm away from the pseudotumor. We usedmatrix‐assisted laser desorption/ ionization time of flight mass spectrometry (MALDI‐TOF/TOFMS) analysis to identify differently expressed proteins in T1, T2, andT3 samples.

Results

T1 samples were different from T2 samples in the expression of 27 proteins. T2 samples had different expressions in 24 proteins, compared to T3 samples.Nine proteins were expressed differently between T1 and T3 samples. These proteins are mainly involved inenergy metabolism, protect the tumor cell from immunologic engraftment of metastatic tumor cells, and migration. Some of thesehave been reported to be related to the tumor metastatic niche hypothesis: Type VI collagen, heat shock protein 90, and Fibrinogen.

Conclusion

The type VI collagen, heat shock protein 90, and Fibrinogen were selected aspotential niche proteins. These findings support the metastatic niche hypothesis and encourage further studies.
Transversus abdominis plane block or subcutaneous wound infiltration after open radical prostatectomy: a randomized study
Authors: SKJELSAGER A. , RUHNAU B. , KISTORP T. K. , KRIDINA I. , HVARNESS H. , MATHIESEN O. , DAHL J. B.
Published: February 2013

Background

Open radical retropubic prostatectomy (ORRP) is associated with moderate pain. We hypothesized that a transversus abdominis plane (TAP) block would reduce post‐operative pain, morphine consumption and opioid‐related side effects compared with wound infiltration and placebo in this population.

Methods

This was a randomized, double‐blind and placebo‐controlled study. The operations were performed with patients in general anaesthesia. Patients were allocated to receive either bilateral TAP block (n = 23), wound infiltration (n = 25) or placebo (n = 25). Treatment was 40 ml ropivacaine 0.75% and placebo was 40 ml saline 0.9%. Pre‐operatively, all patients received oral gabapentin, ibuprofen and paracetamol, followed by oral paracetamol and ibuprofen at regular doses and intervals, and patient‐controlled analgesia with IV morphine from 0 h to 24 h post‐operatively.

Results

Visual analogue scale pain score during mobilization 4 h post‐operatively (primary outcome) did not differ significantly between the TAP block and placebo group (TAP 28 ± 22 mm vs. placebo 33 ± 18 mm, P = 0.64). Pain scores (as area under the curve) during the first 24 h were not significantly different among any of the three groups, neither at rest nor during mobilization. Morphine consumption (0–24 h) did not differ significantly between groups {TAP block = 15 [8–23] mg, infiltration 15 [8–36] mg, placebo 15 [3–30] mg, [median (interquartile range)]}. Levels of nausea, sedation and number of vomits were not different among the groups.

Conclusion

Neither TAP block nor wound infiltration with ropivacaine improved a basic multimodal analgesic regimen with paracetamol, ibuprofen and gabapentin after ORRP.
The relationship between minimum alcohol prices, outlet densities and alcohol‐attributable deaths inBritishColumbia, 2002–09
Authors: Zhao Jinhui , Stockwell Tim , Martin Gina , Macdonald Scott , Vallance Kate , Treno Andrew , Ponicki William R. , Tu Andrew , Buxton Jane
Published: February 2013

Aim

To investigate relationships between periodic increases in minimum alcohol prices, changing densities of liquor stores and alcohol‐attributable (AA) deaths inBritishColumbia,Canada.

Design

Cross‐section (16 geographic areas) versus time–series (32 annual quarters) panel analyses were conducted withAAdeaths as dependent variables and price, outlet densities and socio‐demographic characteristics as independent variables.

Setting and participants

Populations of 16HealthServiceDeliveryAreas inBritishColumbia,Canada.

Measurements

Age–sex‐standardized rates of acute, chronic and whollyAAmortality; population densities of restaurants, bars, government and private liquor stores; minimum prices of alcohol in dollars per standard drink.

Findings

A 10% increase in average minimum price for all alcoholic beverages was associated with a 31.72% [95% confidence interval (CI): ± 25.73%,P < 0.05] reduction in whollyAAdeaths. Significantly negative lagged associations were also detected up to 12 months after minimum price increases for wholly but not for acute or chronicAAdeaths. Significant reductions in chronic and totalAAdeaths were detected between 2 and 3 years after minimum price increases. Significant but inconsistent lagged associations were detected for acuteAAdeaths. A 10% increase in private liquor stores was associated with a 2.45% (95%CI: ± 2.39%,P < 0.05), 2.36% (95%CI: ± 1.57%,P < 0.05) and 1.99% (95%CI: ± 1.76%,P < 0.05) increase in acute, chronic and totalAAmortality rates.

Conclusion

Increases in the minimum price of alcohol inBritishColumbia,Canada, between 2002 and 2009 were associated with immediate and delayed decreases in alcohol‐attributable mortality. By contrast, increases in the density of private liquor stores were associated with increases in alcohol‐attributable mortality.
Outcomes of Descemet’s stripping endothelial keratoplasty in eyes with failed therapeutic penetrating keratoplasty
Authors: Chaurasia Sunita , Murthy Somasheila , Ramappa Muralidhar , Mohamed Ashik , Garg Prashant
Published: February 2013

Purpose:  To report the results of Descemet’s stripping endothelial keratoplasty (DSEK) for failed therapeutic penetrating keratoplasty (PK).

Methods:  Twenty‐seven eyes of 27 patients undergoing DSEK for failed therapeutic PK were analysed.

Results:  The mean age of the patients was 36 ± 13.9 years (range: 14–70 years). The median size of the therapeutic graft was 10 mm (inter‐quartile range; IQR 9.5–11 mm). Descemet’s membrane stripping was performed in all eyes. Graft clarity was achieved in 20/27 eyes. Six eyes had primary graft failure, and one had interface keratitis in the early postoperative period. In all eyes with primary graft failure, there was progressive stromal vascularization, which led to haziness in the graft. Late postoperative complications were rejection in four eyes, infection in two eyes, secondary graft failure in one eye and vortex keratopathy in one eye. Graft size was found to be a significant risk factor for graft rejection in this series. At the last follow‐up visit, the best‐corrected visual acuity was ≥20/40 in 5/27 eyes (18.5%), 20/60–20/40 in 12/27 eyes (44.4%), 20/100–20/60 in 3/27 eyes (1.5%) and ≤20/200 in 7/27 eyes (25.9%).

Conclusions:  Considering the limited success of repeat PK in failed large therapeutic keratoplasty, DSEK is a viable option for visual rehabilitation in these eyes, however; visual acuity may be limited due to sub‐epithelial/stromal or interface scarring.
Expression of thrombospondin‐2 as a marker in proliferative diabetic retinopathy
Authors: Abu El‐Asrar Ahmed M. , Nawaz Mohd I. , Ola Mohammad S. , De Hertogh Gert , Opdenakker Ghislain , Geboes Karel
Published: February 2013

Purpose:  To determine the expression of the endogenous anti‐angiogenic and pro‐fibrotic matricellular protein thrombospondin (TSP)‐2 and its receptors CD36 and CD47 in proliferative diabetic retinopathy (PDR). In addition, we examined the expression of TSP‐2 in the retinas of diabetic rats.

Methods:  Epiretinal membranes from 14 patients with PDR and nine patients with proliferative vitreoretinopathy were studied by immunohistochemistry. Vitreous samples from 30 PDR and 25 nondiabetic patients were studied by enzyme‐linked immunosorbent assay. Vitreous samples and retinas of rats were examined by Western blotting.

Results:  In epiretinal membranes, vascular endothelial cells and myofibroblasts expressed TSP‐2, CD36 and CD47. In PDR membranes, significant correlations were observed between numbers of blood vessels expressing the panendothelial cell marker CD34 and numbers of blood vessels and stromal cells expressing TSP‐2, CD36 and CD47. The numbers of blood vessels and stromal cells expressing CD34, TSP‐2, CD36 and CD47 were significantly higher in membranes with active neovascularization when compared with those with quiescent disease. Thrombospondin‐2 levels in vitreous samples from PDR patients were significantly higher than those in control patients without diabetes (p < 0.001). Western blot analysis revealed a significant increase in the expression of intact and cleaved TSP‐2 in vitreous samples from PDR patients and in the retinas of diabetic rats compared to nondiabetic controls.

Conclusions:  Upregulation of TSP‐2 may be a protective mechanism against inflammation and angiogenesis associated with PDR.
Eye laterality: a comprehensive analysis in refractive surgery candidates
Authors: Linke Stephan J. , Druchkiv Vasyl , Steinberg Johannes , Richard Gisbert , Katz Toam
Published: February 2013

Purpose:  To explore eye laterality (higher refractive error in one eye) and its association with refractive state, spherical/astigmatic anisometropia, age and sex in refractive surgery candidates.

Methods:  Medical records of 12 493 consecutive refractive surgery candidates were filtered. Refractive error (subjective and cycloplegic) was measured in each subject and correlated with eye laterality. Only subjects with corrected distance visual acuity (CDVA) of >20/22 in each eye were enrolled to exclude amblyopia. Associations between eye laterality and refractive state were analysed by means of t‐test, chi‐squared test, Spearman’s correlation and multivariate logistic regression analysis, respectively.

Results:  There was no statistically significant difference in spherical equivalent between right (−3.47 ± 2.76 D) and left eyes (−3.47 ± 2.76 D, p = 0.510; Pearson’s r = 0.948, p < 0.001). Subgroup analysis revealed (I) right eye laterality for anisometropia >2.5 D in myopic (−5.64 ± 2.5 D versus −4.92 ± 2.6 D; p = 0.001) and in hyperopic (4.44 ± 1.69 D versus 3.04 ± 1.79 D; p = 0.025) subjects, (II) a tendency for left eye cylindrical laterality in myopic subjects, and (III) myopic male subjects had a higher prevalence of left eye laterality. (IV) Age did not show any significant impact on laterality.

Conclusions:  Over the full refractive spectrum, this study confirmed previously described strong interocular refractive correlation but revealed a statistically significant higher rate of right eye laterality for anisometropia >2.5 D. In general, our results support the use of data from one eye only in studies of ocular refraction.
Acute–onset diplopia
Authors: O’Colmain Una , Gilmour Claire , MacEwen Caroline J.
Published: February 2013

Purpose:  To investigate the clinical features of all patients with acute–onset diplopia presenting to the ophthalmology department.

Methods:  The notes of every patient who presented with acute onset (<4‐week duration) diplopia to the ophthalmology clinic over a 2‐year period were reviewed. Data regarding clinical features, underlying aetiology, past medical history, investigations and outcomes were extracted.

Results:  One hundred and forty‐nine patients presented with 53.7% having an isolated third, fourth or sixth nerve palsy, 10.7% a mechanical cause, 10.1% a dysfunction of higher control, 8.1% decompensation of a pre‐existing heterophoria, 6.7% idiopathic, 5.4% causes of monocular diplopia and 5.3% another diagnosis. Neuroradiological investigation identified that <5% of patients had a serious underlying pathology, which required immediate management; 80.5% had a diagnosis and underlying aetiology, which were obvious at presentation based only on clinical information and evaluation.

Conclusions:  Acute onset diplopia is an uncommon and challenging presentation for the ophthalmologist to manage. These results demonstrate that the aetiology is commonly identifiable at the first presentation based on clinical evaluation, only a small percentage require urgent radiological investigation and a small minority of cases are likely to have serious emergent pathology.
Positive pressure during penetrating keratoplasty can be solved with a modified graft‐over‐host technique
Authors: Dekaris Iva , Gabrić Nikica , Pauk Maja , Drača Nataša
Published: February 2013

Purpose:  To describe a modified graft‐over‐host (MGOH) surgical technique which was used to prevent serious complications in cases of positive vitreal pressure during penetrating keratoplasty (PKP) and to compare the endothelial cell density (ECD) loss and clinical outcome between eyes undergoing MGOH or standard PKP.

Methods:  In the proposed MGOH technique, the donor graft is sutured over the recipient cornea as soon as signs of positive pressure are noticed, even before cutting the recipient cornea, and the surgery is continued by consecutive cutting of the recipient and continuous suturing of the donor cornea, quadrant by quadrant. Donor grafts were prospectively followed up for the graft clarity, ECD loss and the amount of astigmatism.

Results:  Of 220 cases scheduled for standard PKP in years 2009–2011, eight were operated by MGOH technique. Despite the fact that the donor cornea is situated over the recipient almost throughout the whole MGOH procedure, the ECD loss is equal between eyes operated on by MGOH technique and standard PKP, and none of the grafts had primary graft failure. The only disadvantage was an increased amount of astigmatism that was not as significant once the sutures were taken out.

Conclusion:  Modified graft‐over‐host technique seems to be a safe and effective method to prevent complications during PKP complicated by positive vitreal pressure.
Angiopoietin‐like protein 4 (ANGPTL4) is induced by high glucose in retinal pigment epithelial cells and exhibits potent angiogenic activity on retinal endothelial cells
Authors: Yokouchi Hirotaka , Eto Koki , Nishimura Wataru , Takeda Norio , Kaburagi Yasushi , Yamamoto Shuichi , Yasuda Kazuki
Published: February 2013

Purpose:  Hyperglycaemia has been identified as major risk factor for diabetic retinopathy (DR). It is widely accepted that the progression of DR is mainly due to a local imbalance of pro‐ versus anti‐angiogenic factors in the retina. In this study, we investigated whether retinal pigment epithelial (RPE) cells produced pro‐angiogenic factors under high glucose (HG) conditions in vitro.

Methods:  Cultured human retinal endothelial (RE) cells were exposed to conditioned medium from retinal pigment epithelium cells (ARPE‐19) grown in HG medium and assessed for tube formation. Based on the expression profiles of ARPE‐19, we investigated whether ANGPTL4 was a major angiogenic factor released from ARPE‐19 under HG conditions using cultured human RE cells as the test system for experiments with recombinant protein, conditioned medium from ARPE‐19 and RNA interference (RNAi).

Results:  The conditioned medium from ARPE‐19 cultured under HG conditions promoted tube formation of cultured human RE cells. GeneChip analysis showed that ANGPTL4 was one of the highest upregulated genes under HG conditions. In addition, recombinant ANGPTL4 promoted all of the elements of angiogenesis in human RE cells in vitro. The results of experiments using conditioned medium from ARPE‐19 combined with RNAi demonstrated that ANGPTL4 was a major angiogenic factor released from ARPE‐19 under HG conditions.

Conclusions:  ANGPTL4 was induced by high glucose in RPE cells and exhibited potent angiogenic activity on RE cells. Our results are unique and may potentially add a new candidate to the long list of molecules involved in diabetic retinopathy.
Predictive value of neonatal brain MRI on the neurodevelopmental outcome of preterm infants by 5 years of age
Authors: Setänen Sirkku , Haataja Leena , Parkkola Riitta , Lind Annika , Lehtonen Liisa
Published: February 2013

Aim

To study the prognostic value of MRI in preterm infants at term equivalent age for cognitive development at 5 years of age.

Methods

A total of 217 very low birth weight/very low gestational age infants who all received brain MRI at term equivalent age were categorized into 4 groups based on the brain MRI findings. Cognitive development was assessed at 5 years of chronological age by using a short form of Wechsler Preschool and Primary Scale of Intelligence – Revised. This information was combined with neurosensory diagnoses by 2 years of corrected age.

Results

Of all infants 31 (17.0%) had Full Scale Intelligence Quotient (FSIQ) <85, 14 (6.5%) had cerebral palsy and 4 (1.8%) had severe hearing impairment. A total of 41 (22.0%) infants had some neurodevelopmental impairment at 5 years of age. Considering cognitive outcome (FSIQ <85), the positive predictive value of several major MRI pathologies was 43.8%, and the negative predictive value of normal finding or minor pathologies was 92.0% and 85.7%, respectively.

Conclusion

The MRI of the brain at term equivalent age may be valuable in predicting neurodevelopmental outcome in preterm infants by 5 years of age. The findings should always be interpreted alongside the clinical information of the infant. Furthermore, MRI should not replace a long‐term clinical follow‐up for very preterm infants.
Humerus development in moles (Talpidae, Mammalia)
Authors: Bickelmann Constanze , Jiménez Rafael , Richardson Michael K. , Sánchez‐Villagra Marcelo R.
Published: February 2013

The humerus of fossorial moles has a highly derived anatomy, reflecting the ecological specialization of these animals for digging. It is short and broad, with enlarged muscle attachment sites and pronounced articulations compared to non‐fossorial sister taxa and other mammals. Both condyles are rotated in opposite directions, resulting in a torsion which is unique among eutherian mammals. The development of this exceptional bone was studied in embryonic stages of the fossorial Iberian mole (Talpa occidentalis) from mesenchymal condensation to incipient ossification based on histological serial sections using 3D reconstruction methods. For comparison, embryonic stages of the semi‐fossorial Japanese shrew mole (Urotrichus talpoides) as well as a sister taxon of moles, the terrestrial North American least shrew (Cryptotis parva), were studied. Results show that the humerus of Talpa already shows its derived anatomy with broadened muscle attachment sites and distinct articulations at early cartilaginous stages, when ossification has just started in the mid‐diaphyseal region. The torsion takes place simultaneously with the medial rotation of the forelimbs. The supracondylar foramen is closed in all studied Talpa embryos, but patent in Cryptotis and Urotrichus. This is an example of developmental penetrance, suggesting that variation of adult elements can be found at early stages as well.
Comparison of γδ T cell responses and farnesyl diphosphate synthase inhibition in tumor cells pretreated with zoledronic acid
Authors: Idrees Atif S. M. , Sugie Tomoharu , Inoue Chiyomi , Murata‐Hirai Kaoru , Okamura Haruki , Morita Craig T. , Minato Nagahiro , Toi Masakazu , Tanaka Yoshimasa
Published: February 2013

Exposing human tumor cells to nitrogen‐containing bisphosphonates, such as zoledronic acid (Zol), greatly increases their susceptibility to killing by γδ T cells. Based on this finding and other studies, cancer immunotherapy using γδ T cells and nitrogen‐containing bisphosphonates has been studied in pilot clinical trials and has shown benefits. Although Zol treatment can render a wide variety of human tumor cells susceptible to γδ T cell killing, there has not been a systematic investigation to determine which types of tumor cells are the most susceptible to γδ T cell‐mediated cytotoxicity. In this study, we determined the Zol concentrations required to stimulate half maximal tumor necrosis factor‐α production by γδ T cells cultured with various tumor cell lines pretreated with Zol and compared these concentrations with those required for half maximal inhibition of farnesyl diphosphate synthase (FPPS) in the same tumor cell lines. The inhibition of tumor cell growth by Zol was also assessed. We found that FPPS inhibition strongly correlated with γδ T cell activation, confirming that the mechanism underlying γδ T cell activation by Zol is isopentenyl diphosphate (IPP) accumulation due to FPPS blockade. In addition, we showed that γδ T‐cell receptor‐mediated signaling correlated with γδ T cell tumor necrosis factor‐α production and cytotoxicity. Some lymphoma, myeloid leukemia, and mammary carcinoma cell lines were relatively resistant to Zol treatment, suggesting that assessing tumor sensitivity to Zol may help select those patients most likely to benefit from immunotherapy with γδ T cells.
Generalist genes and cognitive abilities in Chinese twins
Authors: Chow Bonnie Wing‐Yin , Ho Connie Suk‐Han , Wong Simpson Wai‐Lap , Waye Mary M.Y. , Bishop Dorothy V.M.
Published: February 2013

This study considered how far nonverbal cognitive, language and reading abilities are affected by common genetic influences in a sample of 312 typically developing Chinese twin pairs aged from 3 to 11 years. Children were individually given tasks of Chinese word reading, receptive vocabulary, phonological memory, tone awareness, syllable and rhyme awareness, rapid automatized naming, morphological awareness and orthographic skills, and Raven's Colored Progressive Matrices. Factor analyses on the verbal tasks adjusted for age indicated two factors: Language as the first factor and Reading as the second factor. Univariate genetic analyses indicated that genetic influences were substantial for nonverbal cognitive ability and moderate for language and reading. Multivariate genetic analyses showed that nonverbal cognitive ability, language and reading were influenced by shared genetic origins, although there were specific genetic influences on verbal skills that were distinct from those on nonverbal cognitive ability. This study extends the Generalist Genes Hypothesis to Chinese language and reading skills, suggesting that the general effects of genes could be universal across languages.
Fluorescence resonance energy transfer imaging of cell signaling from in vitro to in vivo: Basis of biosensor construction, live imaging, and image processing
Authors: Aoki Kazuhiro , Kamioka Yuji , Matsuda Michiyuki
Published: February 2013

The progress in imaging technology with fluorescent proteins has uncovered a wide range of biological processes in developmental biology. In particular, genetically‐encoded biosensors based on the principle of fluorescence resonance energy transfer (FRET) have been used to visualize spatial and temporal dynamics of intracellular signaling in living cells. However, development of sensitive FRET biosensors and their application to developmental biology remain challenging tasks, which has prevented their widespread use in developmental biology. In this review, we first overview general procedures and tips of imaging with FRET biosensors. We then describe recent advances in FRET imaging – namely, the use of optimized backbones for intramolecular FRET biosensors and transposon‐mediated gene transfer to generate stable cell lines and transgenic mice expressing FRET biosensors. Finally, we discuss future perspectives of FRET imaging in developmental biology.
Chronic childhood ataxia: the cause depends on how you look
Authors: Subramony Sankarasubramoney H
Published: February 2013

This commentary is on the original article by Salman et al on pages 341–347 of this issue.
Assessing the Role of Tourism in Poverty Alleviation: A Research Agenda
Authors: Winters Paul , Corral Leonardo , Mora Adela Moreda
Published: February 2013

Understanding the tourism‐poverty link is critical if tourism is to be used as a mechanism for reducing poverty. Yet, the available empirical analysis is insufficient for this. This article proposes a research agenda for closing this gap in the literature. It argues that, while analysing the link poses peculiar challenges, models exist to do so. Second, it contends that the key question is not whether the link exists but under what conditions it is strongest. Finally, it maintains that the best way to analyse the link is to incorporate accurate diagnosis and evaluations into tourism projects, using the approaches and concepts of the literature on impact evaluation.
Plasticity and metaplasticity of adult rat hippocampal mossy fibers induced by neurotrophin‐3
Authors: Ramos‐Languren L. E. , Escobar M. L.
Published: February 2013

Changes in synaptic efficacy and morphology are considered as the downstream mechanisms of consolidation of memories and other adaptive behaviors. In the last decade, neurotrophin‐3 (NT‐3) has emerged as one potent mediator of synaptic plasticity. In the adult brain, expression of NT‐3 is largely confined to the hippocampal dentate gyrus (DG). Our previous studies show that application of high‐frequency stimulation (HFS) sufficient to elicit long‐term potentiation (LTP) at the DG‐CA3 pathway as well as acute intrahippocampal microinfusion of brain‐derived neurotrophin factor produce mossy fiber (MF) structural reorganization. Here, we show that intrahippocampal microinfusion of NT‐3 induces a long‐lasting potentiation of synaptic efficacy in the DG‐CA3 projection accompanied by an MF structural reorganization of adult rats in vivo. It is considered that the capacity of synapses to express plastic changes is itself subject to variation depending on previous experience; taking into consideration the effects of NT‐3 on MF synaptic plasticity, we thus used intrahippocampal microinfusion of NT‐3 to analyse its effects on functional and structural plasticity induced by subsequent MF‐HFS sufficient to induce LTP in adult rats, in vivo. Our results show that NT‐3 modifies the ability of the MF pathway to present subsequent LTP by HFS, and modifies the structural reorganization pattern. The modifications in synaptic efficacy and morphology elicited by NT‐3 at the MF‐CA3 pathway were blocked by the presence of a Trk receptor inhibitor (K252a). These findings support the idea that NT‐3 actions modify subsequent synaptic plasticity, a homeostatic mechanism thought to be essential for maintaining synapses in the adult mammalian brain.
Suppression of inhibitory GABAergic transmission by cAMP signaling pathway: alterations in learning and memory mutants
Authors: Ganguly Archan , Lee Daewoo
Published: February 2013

The cAMP signaling pathway mediates synaptic plasticity and is essential for memory formation in both vertebrates and invertebrates. In the fruit fly Drosophila melanogaster, mutations in the cAMP pathway lead to impaired olfactory learning. These mutant genes are preferentially expressed in the mushroom body (MB), an anatomical structure essential for learning. While cAMP‐mediated synaptic plasticity is known to be involved in facilitation at the excitatory synapses, little is known about its function in GABAergic synaptic plasticity and learning. In this study, using whole‐cell patch‐clamp techniques on Drosophila primary neuronal cultures, we demonstrate that focal application of an adenylate cyclase activator forskolin (FSK) suppressed inhibitory GABAergic postsynaptic currents (IPSCs). We observed a dual regulatory role of FSK on GABAergic transmission, where it increases overall excitability at GABAergic synapses, while simultaneously acting on postsynaptic GABA receptors to suppress GABAergic IPSCs. Further, we show that cAMP decreased GABAergic IPSCs in a PKA‐dependent manner through a postsynaptic mechanism. PKA acts through the modulation of ionotropic GABA receptor sensitivity to the neurotransmitter GABA. This regulation of GABAergic IPSCs is altered in the cAMP pathway and short‐term memory mutants dunce and rutabaga, with both showing altered GABA receptor sensitivity. Interestingly, this effect is also conserved in the MB neurons of both these mutants. Thus, our study suggests that alterations in cAMP‐mediated GABAergic plasticity, particularly in the MB neurons of cAMP mutants, account for their defects in olfactory learning.
Repair or replace? Exploiting novel gene and cell therapy strategies for muscular dystrophies
Authors: Benedetti Sara , Hoshiya Hidetoshi , Tedesco Francesco Saverio
Published: February 2013

Muscular dystrophies are genetic disorders characterized by skeletal muscle wasting and weakness. Although there is no effective therapy, a number of experimental strategies have been developed over recent years and some of them are undergoing clinical investigation. In this review, we highlight recent developments and key challenges for strategies based upon gene replacement and gene/expression repair, including exon‐skipping, vector‐mediated gene therapy and cell therapy. Therapeutic strategies for different forms of muscular dystrophy are discussed, with an emphasis on Duchenne muscular dystrophy, given the severity and the relatively advanced status of clinical studies for this disease.