bluuebunny/crossref_metadata_2025 huggingface dataset:
Mouse model recapitulating human Fcγ receptor structural and functional diversity
Authors: Smith Patrick , DiLillo David J. , Bournazos Stylianos , Li Fubin , Ravetch Jeffrey V.
Published: April 2012

The in vivo biological activities of IgG antibodies result from their bifunctional nature, in which antigen recognition by the Fab is coupled to the effector and immunomodulatory diversity found in the Fc domain. This diversity, resulting from both amino acid and glycan heterogeneity, is translated into cellular responses through Fcγ receptors (FcγRs), a structurally and functionally diverse family of cell surface receptors found throughout the immune system. Although many of the overall features of this system are maintained throughout mammalian evolution, species diversity has precluded direct analysis of human antibodies in animal species, and, thus, detailed investigations into the unique features of the human IgG antibodies and their FcγRs have been limited. We now report the development of a mouse model in which all murine FcγRs have been deleted and human FcγRs, encoded as transgenes, have been inserted into the mouse genome resulting in recapitulation of the unique profile of human FcγR expression. These human FcγRs are shown to function to mediate the immunomodulatory, inflammatory, and cytotoxic activities of human IgG antibodies and Fc engineered variants and provide a platform for the detailed mechanistic analysis of therapeutic and pathogenic IgG antibodies.
ATF3 protects against atherosclerosis by suppressing 25-hydroxycholesterol–induced lipid body formation
Authors: Gold Elizabeth S. , Ramsey Stephen A. , Sartain Mark J. , Selinummi Jyrki , Podolsky Irina , Rodriguez David J. , Moritz Robert L. , Aderem Alan
Published: April 2012

Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipid-loaded macrophages in the arterial wall. We demonstrate that macrophage lipid body formation can be induced by modified lipoproteins or by inflammatory Toll-like receptor agonists. We used an unbiased approach to study the overlap in these pathways to identify regulators that control foam cell formation and atherogenesis. An analysis method integrating epigenomic and transcriptomic datasets with a transcription factor (TF) binding site prediction algorithm suggested that the TF ATF3 may regulate macrophage foam cell formation. Indeed, we found that deletion of this TF results in increased lipid body accumulation, and that ATF3 directly regulates transcription of the gene encoding cholesterol 25-hydroxylase. We further showed that production of 25-hydroxycholesterol (25-HC) promotes macrophage foam cell formation. Finally, deletion of ATF3 in Apoe−/− mice led to in vivo increases in foam cell formation, aortic 25-HC levels, and disease progression. These results define a previously unknown role for ATF3 in controlling macrophage lipid metabolism and demonstrate that ATF3 is a key intersection point for lipid metabolic and inflammatory pathways in these cells.
Le ‘Romain’ au théâtre des XVII
e
et XVIII
e
siècles: naissance d'un type comique
Authors: Chaouche Sabine
Published: April 2012

Cet article examine un sujet qui a été peu étudié par la critique: la représentation des ‘Grands Comédiens’ de la fin du dix-septième siècle à la première moitié du dix-huitième siècle. La mise en scène des pratiques scéniques en usage chez les ‘Comédiens ordinaires du Roy’ et l'imitation de leur style de jeu ont eu, dès la fin du dix-septième siècle, un succès incontestable auprès d'une certaine frange du public parisien. Ce sont en effet les italiens, puis les forains, ennemis de la Comédie-Française, qui créèrent un personnage nouveau dans le paysage théâtral français: le tragédien caricatural ou ‘Romain’. Nous nous proposons d'analyser les différentes formes de cette figure ridicule afin de mieux en saisir la portée, notamment son influence sur le développement du théâtre français. Privilégiant les textes sources, nous abordons des thèmes tels que l'art de l'acteur et la production des spectacles sous l'Ancien Régime (comme, par exemple, le costume). Tout en donnant une vue générale du contexte culturel et théâtral de l'époque, nous nous attachons plus particulièrement aux rivalités entre troupes parisiennes afin de montrer comment le théâtre des foires a inventé une nouvelle forme de
mimèsis
à travers l'art du pastiche.
Panton–Valentine leukocidin gene sequence variation and phage in methicillin‐resistant and methicillin‐susceptible Staphylococcus aureus from children in mainland China
Authors: Li Xiangmei , Sun Jing , Wu Dejing , Wang Lijuan , Yang Yonghong , Wang Chuanqing , Liu Lan , Zhao Changan , Du Pengcheng , Yu Sangjie , Shen Xuzhuang
Published: April 2012

To determine the variation in the Panton–Valentine leukocidin (PVL) gene sequences and different PVL‐encoding phages of Staphylococcus aureus strains collected from children in mainland China, fifty‐eight strains with PVL collected from 2007 to 2009 were used. Their molecular characteristics were examined. Primers were designed to sequence the PVL genes. Six PVL‐encoding phages (ϕPVL, ϕ108PVL, ϕSLT, ϕSa2MW, ϕSa2958, and ϕSa2USA) were identified by PCR. Eleven sequence types (ST) were detected with ST59 (39.7%, 23/58) the most frequent ST, followed by 910 (22.4%, 13/58), and 338 (12.1%, 7/58). Single nucleotide polymorphisms (SNP) were identified at 11 locations in the PVL genes. SNP (nucleotide 1396, A→G) and SNP (nucleotide 1546, A→G) were observed in >10 sequences. Four additional SNP were non‐synonymous. Both SNP (nucleotide 16, C→A) and SNP (nucleotide 62, C→T) were present in the same ST59 strain. SNP (nucleotide 527, A→G) was present in five strains belonging to ST30, 121, 1, and 93. SNP (nucleotide 1436, A→C) was present in one ST30 strain. Fifteen strains belonging to ST910, ST217, and ST30 carried a PVL phage that had an icosahedral head morphology. Nine ST59 strains carried ϕ108PVL. Three ST88 strains carried a PVL phage that had an elongated head morphology. Twenty‐seven strains, including 60.9% (14/23) of ST59 and all ST338 strains, had no detectable phage. In conclusion, sequence variation in PVL genes and PVL‐encoding phages was generally related to the lineage. ST59 strains may indeed carry novel PVL phages.
Prokaryotic expression and immunogenicity of 56‐kDa protein of Orientia tsutsugamushi strain Karp
Authors: Wang Yuan‐Yuan , Chen Qiang , Yu Qiang , Zhang Li‐Juan
Published: April 2012

To express the 56‐kDa protein of O. tsutsugamushi strain Karp, this protein gene was cloned into pET30a(+) before transforming into host bacteria, E. coli Rossetta. Specificity of the recombinant protein was assessed by ELISA using rabbit sera against common members of the order Rickettsiae and 10 other pathogenic bacteria. After IPTG induction, SDS‐PAGE analysis of isolated protein demonstrated a band at approximately 46‐kDa. Western blot and mass spectrometry analysis proved that the recombinant protein was expressed successfully. Specificity analysis demonstrated that all sera were negative, except sera against O. tsutsugamushi strains TA763, TH1817 and Kato, B. quintana, A. phagocytophilum, E. chaffeensis and B. bacilliformis. The purified protein was used to immunize BALB/c mice and polyclonal antisera were harvested. By examination of IFA and ELISA, the highest titer of the polyclonal antibodies reaches 1:1600. The recombinant 56‐kDa protein in the study is valuable for developing a simple and rapid diagnostic test and vaccine for O. tsutsugamushi.
CTHRC1 is upregulated by promoter demethylation and transforming growth factor‐β1 and may be associated with metastasis in human gastric cancer
Authors: Wang Ping , Wang Ying‐Chao , Chen Xiao‐Yu , Shen Zhi‐Yong , Cao Hui , Zhang Yan‐Jie , Yu Jian , Zhu Jing‐De , Lu You‐Yong , Fang Jing‐Yuan
Published: April 2012

The gene, collagen triple helix repeat containing 1 (CTHRC1), has been reported to increase in several kinds of human solid cancers and is associated with tumor invasion and metastasis. To date, the expression and function of CTHRC1 in gastric cancer (GC) have not been reported. The aim of this study was to investigate the expression levels and regulatory transcription mechanisms of CTHRC1 in GC. Immunohistochemical analysis revealed that CTHRC1 expression was markedly increased in carcinoma compared with normal gastric mucosa, chronic atrophic gastritis, and intestinal metaplasia (P < 0.05 for all), and this overexpression in tumor was related to depth of tumor invasion. Moreover, RNA interference‐mediated knockdown and ectopic expression of CTHRC1 showed that CTHRC1 promoted tumor cell invasion in vitro. We then investigated the mechanisms underlying the aberrant expression of CTHRC1 in GC and found that CTHRC1 expression was restored after GC cell lines were treated with the demethylating agent, 5‐aza‐2'‐deoxycytidine. Transforming growth factor‐β1 led to an increase in levels of CTHRC1 mRNA and protein. Overall, our data revealed that the upregulated expression of CTHRC1 in gastric carcinogenesis contributes to tumor cell invasion and metastasis, and promoter demethylation and transforming growth factor‐β1 may co‐regulate the expression of CTHRC1. (Cancer Sci 2012; 103: 1327–1333)
Modelling the potential distribution of endangered Prunus africana (Hook.f.) Kalkm. in East Africa
Authors: Mbatudde Maria , Mwanjololo Majaliwa , Kakudidi Esezah Kyomugisha , Dalitz Helmut
Published: April 2012

Continued harvesting and climate change are affecting the distributions of many plant species and may lead to numerous extinctions over the next century. Endangered species are likely to be a special concern, but the extent to which they are sensitive to climate is currently unclear. Species distribution modelling, if carefully implemented, can be used to assess climate sensitivity and potential climate change impacts, of tree species. We used MaxEnt algorithm for species distribution modelling to assess the potential distribution and climate change risks for a threatened Prunus africana, in East Africa. Data from different herbaria on its distribution were linked to data on climate to test hypotheses on the factors determining its distribution. Predictive models were developed and projected onto a climate scenario for 2050 to assess climate change risks. Precipitation of driest quarter and annual precipitation appeared to be the main factors influencing its distribution. Climate change was predicted to result in reductions of the species' habitats (e.g. Erasmus et al., Glob. Change Biol. 2002; 8: 679). Prunus africana distribution is thus highly vulnerable to a warming climate and highlights the fact that both in‐situ and ex‐situ conservation will be a solution to global warming.
Intrasite variation in the ability to detect tropical forest mammals
Authors: Breuer Thomas , Breuer‐Ndoundou Hockemba Mireille
Published: April 2012

Monitoring programs are essential for an adequate management of large mammal populations. Within Central Africa data on abundance of important bushmeat species, such as duikers and primates are often lacking. Relative densities [(encounter rates (ERs)] are often the only available data, but there is little knowledge on the factors affectingERs and detection of these animals. Here we investigate how different environmental factors and observers influence large mammalERs in a pristine rainforest in theNouabalé‐NdokiNationalPark,Republic ofCongo. Our results show large daily and monthly variation inERs of duikers and primates.ERs varied between habitat types, seasons and observers′ ability to detect animals. Season and rainfall had a strong effect on understory visibility and duiker detection, respectively. Detection of duikers often happened by sound instead of visual observations. Our study shows that the use ofERis a poor monitoring approach and more robust methods need to be used to evaluate rates of population change reliably.
The effects of a novel metabotropic glutamate receptor 5 antagonist (AZD2066) on transient lower oesophageal sphincter relaxations and reflux episodes in healthy volunteers
Authors: Rohof W. O. , Lei A. , Hirsch D. P. , Ny L. , Astrand M. , Hansen M. B. , Boeckxstaens G. E.
Published: April 2012

Background

Selective metabotropic glutamate receptor 5 (mGluR5) antagonists inhibit transient lower oesophageal sphincter relaxations (TLESRs) in animals and acid reflux in humans.

Aim

To assess the effect of single doses of the mGluR5 antagonist AZD2066 on TLESRs and reflux in humans.

Methods

Healthy male volunteers received AZD2066 13 mg and placebo (part A), or AZD2066 2 mg and AZD2066 6 mg and placebo (part B), in a randomised crossover study. Postprandial manometry/pH‐impedance measurements were taken after each dose.

Results

A total of 13 individuals completed part A of the study and 19 individuals completed part B. There was a significant reduction in the geometric mean number of TLESRs (27%; P = 0.02) and the geometric mean number of reflux episodes (51%; P = 0.01) in subjects receiving AZD2066 13 mg compared with placebo. Adverse events in participants receiving AZD2066 13 mg were mostly related to the nervous system [dizziness (3/13); disturbance in attention (3/13)]. Adverse events were reversible and of mild intensity. There were no serious adverse events. The effects of AZD2066 appeared dose‐dependent, with smaller reductions in TLESRs and reflux episodes (relative to placebo) and fewer adverse events observed for AZD2066 2 mg and AZD2066 6 mg compared with AZD2066 13 mg.

Conclusion

The mGluR5‐mediated inhibition of TLESRs may be a useful approach for inhibiting gastro‐oesophageal reflux.
Trends in oral drug bioavailability following bariatric surgery: examining the variable extent of impact on exposure of different drug classes
Authors: Darwich Adam S. , Henderson Kathryn , Burgin Angela , Ward Nicola , Whittam Janet , Ammori Basil J. , Ashcroft Darren M. , Rostami‐Hodjegan Amin
Published: April 2012

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Changes to oral drug bioavailability have been observed post bariatric surgery. However, the magnitude and the direction of changes have not been assessed systematically to provide insights into the parameters governing the observed trends. Understanding these can help with dose adjustments.

WHAT THIS STUDY ADDS

• Analysis of drug characteristics based on a biopharmaceutical classification system is not adequate to explain observed trends in altered oral drug bioavailability following bariatric surgery, although the findings suggest solubility to play an important role.

AIMS

To identify the most commonly prescribed drugs in a bariatric surgery population and to assess existing evidence regarding trends in oral drug bioavailability post bariatric surgery.

METHODS

A retrospective audit was undertaken to document commonly prescribed drugs amongst patients undergoing bariatric surgery in an NHS hospital in the UK and to assess practice for drug administration following bariatric surgery. The available literature was examined for trends relating to drug permeability and solubility with regards to the Biopharmaceutics Classification System (BCS) and main route of elimination.

RESULTS

No significant difference in the ‘post/pre surgery oral drug exposure ratio’ (ppR) was apparent between BCS class I to IV drugs, with regards to dose number (Do) or main route of elimination. Drugs classified as ‘solubility limited’ displayed an overall reduction as compared with ‘freely soluble’ compounds, as well as an unaltered and increased ppR.

CONCLUSION

Clinical studies establishing guidelines for commonly prescribed drugs, and the monitoring of drugs exhibiting a narrow therapeutic window or without a readily assessed clinical endpoint, are warranted. Using mechanistically based pharmacokinetic modelling for simulating the multivariate nature of changes in drug exposure may serve as a useful tool in the further understanding of postoperative trends in oral drug exposure and in developing practical clinical guidance.
Evaluation of phototoxic potential of aerial components of the fig tree against human melanoma
Authors: Conforti F. , Menichini G. , Zanfini L. , Tundis R. , Statti G. A. , Provenzano E. , Menichini F. , Somma F. , Alfano C.
Published: April 2012

Objectives

To date, Ficus carica L. cultivar Dottato (F. carica) has not been studied from a phototoxic point of view. In the present work, aerial components of F. carica from Italy, were examined to assess their antioxidant and phototoxic activity on human melanoma cells. A relationship between antioxidant, phototoxic activities and chemical composition has also been investigated.

Materials and methods

Coumarin and fatty acid content in F. carica leaves, bark and woody parts were examined and compared by capillary GC and GC/MS. Polyphenolic content was also determined. Linoleic acid peroxidation and DPPH test were used to assess antioxidant activities, and MTT assay was used to evaluate anti‐proliferative activity, on C32 human melanoma cells, after irradiation with a UVA dose of 1.08 J/cm2.

Results

Leaves demonstrated the best antioxidant and anti‐proliferative activity in comparison to bark and wood. In particular, leaves were shown to possess the highest anti‐radical activity and inhibition of peroxidation, with IC50 values of 64 and 1.48 μg/ml respectively. The leaves had highest anti‐proliferative activity with IC50 value of 3.92 μg/ml. The phytochemical investigation revealed different composition between the coumarins, psoralen and bergapten, fatty acids, polyphenols and flavonoid content among plant parts.

Conclusions

Data obtained indicate that this type of fig tree may constitute an excellent source of bioactive compounds, such as phenolics, coumarins and fatty acids. This study offers a new perspective in developing others formulations potentially useful in photodynamic therapy for treatment of non‐melanoma skin cancers.
Fathers' experiences with child welfare services
Authors: Coady Nick , Hoy Sandra L. , Cameron Gary
Published: April 2012

The lack of engagement of fathers by child welfare services is well‐documented in the literature as a serious problem. Towards addressing this problem, this paper reports the findings of interviews with 18 fathers about their involvement with child welfare services in Ontario, Canada. Qualitative analysis of the interviews yielded themes about what men saw as the positive and negative aspects of their involvement with child welfare. Positive aspects of service involvement for fathers included understanding and supportive workers, useful assistance from workers, being connected to useful resources and being given a ‘wake‐up call’. Negative aspects of service involvement included uncaring, unhelpful and unprofessional workers; prejudice against fathers; and experiencing the child welfare system as unresponsive, uncaring and rigid. Implications for practice are discussed with a view to improving the engagement of men in, and their experiences with, child welfare services.
Lung effects of inhaled corticosteroids in a rhesus monkey model of childhood asthma
Authors: Plopper C. G. , Joad J. P. , Miller L. A. , Schelegle E. S. , Fanucchi M. V. , Van Winkle L. S. , Tyler N. K. , Avdalovic M. V. , Evans M. J. , Lasley W. L. , Buckpitt A. R. , Pinkerton K. E. , Tarkington B. K. , Davis S. , Nishio S. J. , Gershwin L. J. , Wu R. , Hyde D. M.
Published: April 2012

Background

The risks for infants and young children receiving inhaled corticosteroid (ICS) therapy are largely unknown. Recent clinical studies indicate that ICS therapy in pre‐school children with symptoms of asthma result in decreased symptoms without influencing the clinical disease course, but potentially affect postnatal growth and development. The current study employs a primate experimental model to identify the risks posed by ICS therapy.

Objective

To (1) establish whether ICS therapy in developing primate lungs reverses pulmonary pathobiology associated with allergic airway disease (AAD) and (2) define the impact of ICS on postnatal lung growth and development in primates.

Methods

Infant rhesus monkeys were exposed, from 1 through 6 months, to filtered air (FA) with house dust mite allergen and ozone using a protocol that produces AAD (AAD monkeys), or to FA alone (Control monkeys). From three through 6 months, the monkeys were treated daily with ICS (budesonide) or saline.

Results

Several AAD manifestations (airflow restrictions, lavage eosinophilia, basement membrane zone thickening, epithelial mucin composition) were reduced with ICS treatment, without adverse effects on body growth or adrenal function; however, airway branching abnormalities and intraepithelial innervation were not reduced. In addition, several indicators of postnatal lung growth and differentiation: vital capacity, inspiratory capacity, compliance, non‐parenchymal lung volume and alveolarization, were increased in both AAD and Control monkeys that received ICS treatment.

Conclusions and Clinical Relevance

Incomplete prevention of pathobiological changes in the airways and disruption of postnatal growth and differentiation of airways and lung parenchyma in response to ICS pose risks for developing primate lungs. These responses also represent two mechanisms that could compromise ICS therapy's ability to alter clinical disease course in young children.
Experimental estimation of velocities and anisotropy of a series of Swedish crystalline rocks and ores
Authors: Malehmir Alireza , Andersson Magnus , Lebedev Maxim , Urosevic Milovan , Mikhaltsevitch Vassili
Published: April 2012

To provide a guide for future deep (<1.5 km) seismic mineral exploration and to better understand the nature of reflections imaged by surface reflection seismic data in two mining camps and a carbonatite complex of Sweden, more than 50 rock and ore samples were collected and measured for their seismic velocities. The samples are geographically from the northern and central parts of Sweden, ranging from metallic ore deposits, meta‐volcanic and meta‐intrusive rocks to deformed and metamorphosed rocks. First, ultrasonic measurements of P‐ and S‐wave velocities at both atmospheric and elevated pressures, using 0.5 MHz P‐ and S‐wave transducers were conducted. The ultrasonic measurements suggest that most of the measured velocities show positive correlation with the density of the samples with an exception of a massive sulphide ore sample that shows significant low P‐ and S‐wave velocities. The low P‐ and S‐wave velocities are attributed to the mineral texture of the sample and partly lower pyrite content in comparison with a similar type sample obtained from Norway, which shows significantly higher P‐ and S‐wave velocities. Later, an iron ore sample from the central part of Sweden was measured using a low‐frequency (0.1–50 Hz) apparatus to provide comparison with the ultrasonic velocity measurements. The low‐frequency measurements indicate that the iron ore sample has minimal dispersion and attenuation. The iron ore sample shows the highest acoustic impedance among our samples suggesting that these deposits are favourable targets for seismic methods. This is further demonstrated by a real seismic section acquired over an iron ore mine in the central part of Sweden. Finally, a laser‐interferometer device was used to analyse elastic anisotropy of five rock samples taken from a major deformation zone in order to provide insights into the nature of reflections observed from the deformation zone. Up to 10% velocity‐anisotropy is estimated and demonstrated to be present for the samples taken from the deformation zone using the laser‐interferometery measurements. However, the origin of the reflections from the major deformation zone is attributed to a combination of anisotropy and amphibolite lenses within the deformation zone.
Influence of climate oscillations on dentinal deposition in teeth of Commerson's dolphin
Authors: Dellabianca Natalia A. , Hohn Aleta A. , Goodall R. Natalie P. , Pousa Jorge L. , MacLeod Colin D. , Lima Mauricio
Published: April 2012

Effects of climatic oscillations on populations in marine environments are produced mainly through the lower trophic levels. Marine mammals, as top predators, are affected indirectly by these fluctuations, showing variations in their survival, growth and reproduction. The main objective of this study was to determine whether we could detect the effect of climatic oscillations on Commerson's dolphins (Cephalorhynchus c. commersonii) off Tierra del Fuego, Argentina, by examining the presence and proportion of anomalous dentinal growth layer groups (GLGs). The relative deposition of anomalous GLGs was determined using calendar years from 1960 to 2005, and related to the Southern Annular Mode (SAM) and El Niño Southern Oscillation (ENSO) events, and to mean annual sea surface temperatures (SST). Then, growth parameters between animals that did and did not show anomalous dentinal growth layer patterns in one or more of their GLGs were compared. The presence of anomalous GLGs was related to the SAM, increasing in frequency with negative values of the SAM. No relationship was found among anomalous GLGs, ENSO, and SST. There were no significant differences in growth parameters between animals with and without anomalous GLGs. Using recording structures such as teeth provided a unique way of reconstructing time series to evaluate the effects of these climatic patterns on a top predator species in an area where no similar studies have been conducted, primarily due to a lack of suitable long‐term data sets.
Angiotropism is an independent predictor of microscopic satellites in primary cutaneous melanoma
Authors: Wilmott James , Haydu Lauren , Bagot Martine , Zhang Yuxiao , Jakrot Valerie , McCarthy Stanley , Lugassy Claire , Thompson John , Scolyer Richard , Barnhill Raymond
Published: April 2012

Wilmott J, Haydu L, Bagot M, Zhang Y, Jakrot V, McCarthy S, Lugassy C, Thompson J, Scolyer R & Barnhill R  (2012) Histopathology 61, 889–898

Angiotropism is an independent predictor of microscopic satellites in primary cutaneous melanoma

Aims:  To establish whether microscopic angiotropism of melanoma cells correlates with microscopic satellite (MS) formation in cutaneous melanomas and thus is likely to explain the development of MS.

Materials and results:  Patients with MS and controls without MS from 1996 to 2009 were evaluated for the presence or absence of angiotropism. MS was defined as a dermal/subcutaneous tumour nodule >0.05 mm, separated from the primary tumour by at least 0.3 mm. Forty four cases and controls were matched for tumour thickness, mitotic rate, ulceration, age, gender and primary site. Angiotropism (23 of 44, 52%) and absent regression (19 of 44, 43%) were significantly more frequent in melanomas with MS than in those without MS (controls) (12 of 44, 27%) (P = 0.017) and (32 of 44, 73%) (P = 0.005), respectively. Factors correlating with angiotropism included increased Clark level (P = 0.046), regression absence (P = 0.02) and MS (P = 0.017). On multivariable analysis, MS formation was predicted by angiotropism (P = 0.026), Clark level V (P = 0.01), absent regression (P = 0.009) and acral site (P = 0.02).

Conclusions:  Angiotropism predicts MS development. These data provide additional evidence for the importance of angiotropism as a means of melanoma metastasis.
Microdensitometry of osteopontin as an immunohistochemical prognostic biomarker in colorectal carcinoma tissue microarrays: potential and limitations of the method in ‘biomarker pathology’
Authors: Prall Friedrich , Maletzki Claudia , Linnebacher Michael
Published: April 2012

Prall F, Maletzki C & Linnebacher M  (2012) Histopathology 61, 823–832

Microdensitometry of osteopontin as an immunohistochemical prognostic biomarker in colorectal carcinoma tissue microarrays: potential and limitations of the method in ‘biomarker pathology’

Aims:  To explore the potential and limitations of ‘biomarker pathology’ with quantitative immunohistochemistry on tissue microarrays, taking osteopontin and colorectal carcinoma as a model system.

Methods and results:  Microdensitometry for quantitative evaluation of osteopontin immunohistochemistry (clone OP3N) on digital microphotographs using the public domain software ImageJ was observed to be straightforward to perform and reliable. However, using colorectal carcinoma cell lines (n = 11) the correlation between densitometric evaluations of Western blots and microdensitometry of immunocytochemistry of slide cultures was only moderate. A virtual resampling method to simulate tissue microarrays showed that, due to heterogeneity of immunostaining, tumours were misclassified in nearly 20% of the arrays, even if four punches were used. With regard to prognosis, microdensitometric evaluation of a tissue microarray made of a clinicopathologically well‐characterized series of colorectal carcinomas with long‐term follow‐up (222 cases evaluable in the tissue microarray, UICC Stages I–III/R0) showed a moderate survival advantage of patients with high osteopontin expression by microdensitometry.

Conclusions:  These results challenge the basic assumption that microdensitometry is a precise technique for the quantification of proteins detected by immunohistochemistry and delineate drawbacks encountered when working with tissue microarrays in clinicopathological studies.
Interleukin‐28B (IL‐28B) single‐nucleotide polymorphisms and interferon plus ribavirin treatment outcome in Italian chronically HCV‐infected patients
Authors: Riva E. , Scagnolari C. , Monteleone K. , Selvaggi C. , Picardi A. , Mazzarelli C. , Pizzigallo E. , Vincenzi B. , Carducci A. , Antonaci S. , Giannelli G. , Antonelli G.
Published: April 2012

Summary.  To determine the single or combined effect of both rs12979860 and rs8099917 SNPs on HCV treatment response, these variants were genotyped in samples from a cohort of 170 patients infected with different HCV genotypes (HCVGT). The favourable rs12979860 CC genotype was found only in patients with sustained or rapid virological responses (SVR/RVR) and at significantly high proportions in HCVGT1/4 SVR patients. A significant association was also found between the rs8099917 TT genotype and SVR in both HCVGT1/4 and HCVGT2/3 groups of patients. In contrast, we found that there was significantly more of the rs8099917 GG genotype in nonresponders (NR) than in SVR patients which suggests a good association of the minor homozygote GG with the lack of treatment response. The combination of rs12979860/rs8099917 CC/TT favourable genotypes was found only in SVR patients and matched the frequency observed for their rs12979860 CC genotypes alone. By contrast, the inverse unfavourable correlate rs12979860/rs8099917 TT/GG genotype was seen more in NR than in SVR patients as observed for the single GG genotype. This study confirms the impact of both rs12979860 and/or rs8099917 IL‐28B SNPs on treatment‐induced clearance of HCV‐RNA and demonstrates that the rs12979860 CC genotype is stronger than rs8099917 TT genotype in predicting a positive treatment response in HCVGT1/4 patients. The unfavourable rs8099917 GG genotype seems to be more important in predicting the failure of treatment response independently from HCV genotype.
The scaffolding and signalling functions of a localization factor impact polar development
Authors: Curtis Patrick D. , Quardokus Ellen M. , Lawler Melanie L. , Guo Xiaoyun , Klein David , Chen Joseph C. , Arnold Randy J. , Brun Yves V.
Published: April 2012

In the differentiating alphaproteobacterium Caulobacter crescentus, organelle synthesis at cell poles is critical to forming different progeny after cell division. Co‐ordination of polar organelle synthesis, including pili and holdfast, and flagellum ejection, is mediated in part by the scaffolding protein PodJ. At the time of cell division, PodJ undergoes regulated processing to a short form that persists at the flagellar pole of swarmer cells. This study analyses how PodJ's role in structural and signalling protein localization impacts organelle synthesis. A PodJ mutant with an internal deletion exhibits reduced sensitivity to pili‐tropic phage ΦCbK, resulting from reduced pilA gene expression, which can be linked to altered signalling protein localization. The phage sensitivity defect of a ΔpodJ mutant can be partially suppressed by ectopic pilA expression. Induction of PodJ processing, by manipulation of podJ itself or controlled perP expression, resulted in decreased pilus biogenesis and, when coupled with a podJ mutation that reduced pilA expression, led to complete loss of phage sensitivity. As a whole, the results show that PodJ's scaffolding role for structural and signalling proteins both contribute to flagellar pole organelle development.
The histone chaperone ASF1 is essential for sexual development in the filamentous fungus Sordaria macrospora
Authors: Gesing Stefan , Schindler Daniel , Fränzel Benjamin , Wolters Dirk , Nowrousian Minou
Published: April 2012

Ascomycetes develop four major types of fruiting bodies that share a common ancestor, and a set of common core genes most likely controls this process. One way to identify such genes is to search for conserved expression patterns. We analysed microarray data of Fusarium graminearum and Sordaria macrospora, identifying 78 genes with similar expression patterns during fruiting body development. One of these genes was asf1 (anti‐silencing function 1), encoding a predicted histone chaperone. asf1 expression is also upregulated during development in the distantly related ascomycete Pyronema confluens. To test whether asf1 plays a role in fungal development, we generated an S. macrospora asf1 deletion mutant. The mutant is sterile and can be complemented to fertility by transformation with the wild‐type asf1 and its P. confluens homologue. An ASF1–EGFP fusion protein localizes to the nucleus. By tandem‐affinity purification/mass spectrometry as well as yeast two‐hybrid analysis, we identified histones H3 and H4 as ASF1 interaction partners. Several developmental genes are dependent on asf1 for correct transcriptional expression. Deletion of the histone chaperone genes rtt106 and cac2 did not cause any developmental phenotypes. These data indicate that asf1 of S. macrospora encodes a conserved histone chaperone that is required for fruiting body development.
Enhanced formation of aerenchyma and induction of a barrier to radial oxygen loss in adventitious roots of Zea nicaraguensis contribute to its waterlogging tolerance as compared with maize (Zea mays ssp. mays)
Authors: ABIKO TOMOMI , KOTULA LUKASZ , SHIONO KATSUHIRO , MALIK AL IMRAN , COLMER TIMOTHY DAVID , NAKAZONO MIKIO
Published: April 2012

Enhancement of oxygen transport from shoot to root tip by the formation of aerenchyma and also a barrier to radial oxygen loss (ROL) in roots is common in waterlogging‐tolerant plants. Zea nicaraguensis (teosinte), a wild relative of maize (Zea mays ssp. mays), grows in waterlogged soils. We investigated the formation of aerenchyma and ROL barrier induction in roots of Z. nicaraguensis, in comparison with roots of maize (inbred line Mi29), in a pot soil system and in hydroponics. Furthermore, depositions of suberin in the exodermis/hypodermis and lignin in the epidermis of adventitious roots of Z. nicaraguensis and maize grown in aerated or stagnant deoxygenated nutrient solution were studied. Growth of maize was more adversely affected by low oxygen in the root zone (waterlogged soil or stagnant deoxygenated nutrient solution) compared with Z. nicaraguensis. In stagnant deoxygenated solution, Z. nicaraguensis was superior to maize in transporting oxygen from shoot base to root tip due to formation of larger aerenchyma and a stronger barrier to ROL in adventitious roots. The relationships between the ROL barrier formation and suberin and lignin depositions in roots are discussed. The ROL barrier, in addition to aerenchyma, would contribute to the waterlogging tolerance of Z. nicaraguensis.
Barriers to conducting effective obstetric audit in Ifakara: a qualitative assessment in an under‐resourced setting in Tanzania
Authors: van Hamersveld Koen T. , den Bakker Emil , Nyamtema Angelo S. , van den Akker Thomas , Mfinanga Elirehema H. , van Elteren Marianne , van Roosmalen Jos
Published: April 2012

Objective  To explore barriers to and solutions for effective implementation of obstetric audit at Saint Francis Designated District Hospital in Ifakara, Tanzania, where audit results have been disappointing 2 years after its introduction.

Methods  Qualitative study involving participative observation of audit sessions, followed by 23 in‐depth interviews with health workers and managers. Knowledge and perceptions of audit were assessed and suggestions for improvement of the audit process explored.

Results  During the observational period, audit sessions were held irregularly and only when the head of department of obstetrics and gynaecology was available. Cases with evident substandard care factors were audited. In‐depth interviews revealed inadequate knowledge of the purpose of audit, despite the fact that participants regarded obstetric audit as a potentially useful tool. Insufficient staff commitment, managerial support and human and material resources were mentioned as reasons for weak involvement of health workers and poor implementation of recommendations resulting from audit. Suggestions for improvement included enhancing feedback to all staff and managers to attend sessions and assist with the effectuation of audit recommendations.

Conclusion  Obstetric staff in Ifakara see audit as an important tool for quality improvement. They recognise, however, that in their own situation, insufficient staff commitment and poor managerial support are barriers to successful implementation. They suggested training in concept and principles of audit as well as strengthening feedback of audit outcomes, to achieve structural health care improvements through audit.
Evolutionary stasis in Euphorbiaceae pollen: selection and constraints
Authors: MATAMORO‐VIDAL A. , FURNESS C. A. , GOUYON P.‐H. , WURDACK K. J. , ALBERT B.
Published: April 2012

Although much attention has been paid to the role of stabilizing selection, empirical analyses testing the role of developmental constraints in evolutionary stasis remain rare, particularly for plants. This topic is studied here with a focus on the evolution of a pollen ontogenetic feature, the last points of callose deposition (LPCD) pattern, involved in the determination of an adaptive morphological pollen character (aperture pattern). The LPCD pattern exhibits a low level of evolution in eudicots, as compared to the evolution observed in monocots. Stasis in this pattern might be explained by developmental constraints expressed during male meiosis (microsporogenesis) or by selective pressures expressed through the adaptive role of the aperture pattern. Here, we demonstrate that the LPCD pattern is conserved in Euphorbiaceae s.s. and that this conservatism is primarily due to selective pressures. A phylogenetic association was found between the putative removal of selective pressures on pollen morphology after the origin of inaperturate pollen, and the appearance of variation in microsporogenesis and in the resulting LPCD pattern, suggesting that stasis was due to these selective pressures. However, even in a neutral context, variation in microsporogenesis was biased. This should therefore favour the appearance of some developmental and morphological phenotypes rather than others.
Proteins within the intracellular calcium store determine cardiac RyR channel activity and cardiac output
Authors: Dulhunty Angela F , Wium Elize , Li Linwei , Hanna Amy D , Mirza Shamaruh , Talukder Sadik , Ghazali Nuur AA , Beard Nicole A
Published: April 2012

The contractile function of the heart requires the release of Ca2+ from intracellular Ca2+ stores in the sarcoplasmic reticulum (SR) of cardiac muscle cells. The efficacy of Ca2+ release depends on the amount of Ca2+ loaded into the Ca2+ store and the way in which this ‘Ca2+ load’ influences the activity of the cardiac ryanodine receptor Ca2+ release channel (RyR2).

The effects of the Ca2+ load on Ca2+ release through RyR2 are facilitated by: (i) the sensitivity of RyR2 itself to luminal Ca2+ concentrations; and (ii) interactions between the cardiac Ca2+‐binding protein calsequestrin (CSQ) 2 and RyR2, transmitted through the ‘anchoring’ proteins junctin and/or triadin.

Mutations in RyR2 are linked to catecholaminergic polymorphic ventricular tachycardia (CPVT) and sudden cardiac death. The tachycardia is associated with changes in the sensitivity of RyR2 to luminal Ca2+. Triadin‐, junctin‐ or CSQ‐null animals survive, but their longevity and ability to tolerate stress is compromised. These studies reveal the importance of the proteins in normal muscle function, but do not reveal the molecular nature of their functional interactions, which must be defined before changes in the proteins leading to CPVT and heart disease can be understood.

Herein, we discuss known interactions between the RyR, triadin, junctin and CSQ with emphasis on the cardiac isoforms of the proteins. Where there is little known about the cardiac isoforms, we discuss evidence from skeletal isoforms.
Pharmacokinetics and pharmacodynamics of insulin aspart in patients with Type 2 diabetes: Assessment using a meal tolerance test under clinical conditions
Authors: Ando Hitoshi , Kurita Seiichiro , Shimizu Akiko , Kato Ken‐ichiro , Ishikura Kazuhide , Taji Koumei , Uno Masafumi , Takeshita Yumie , Misu Hirofumi , Fujimura Akio , Kaneko Shuichi , Takamura Toshinari
Published: April 2012

Few studies have evaluated the pharmacokinetics of rapid‐acting insulin analogues in patients with Type 2 diabetes, especially under clinical conditions. The aim of the present study was to assess both the pharmacokinetics and pharmacodynamics of insulin aspart in Type 2 diabetic patients who were being treated with the analogue alone.

Meal tolerance tests with and without self‐injection of a customary dose of insulin aspart (0.05–0.22 U/kg) were conducted in 20 patients in a randomized cross‐over study.

The dose of insulin aspart (per bodyweight) was significantly correlated with both the maximum concentration (r2 = 0.59; P < 0.01) and area under the concentration–time curve for insulin aspart (r2 = 0.53; P < 0.01). However, the time to maximum concentration (Tmax), which varied widely from < 60 to ≥ 120 min, was not associated with either dosage (r2 = 0.02; P = 0.51) or body mass index (r2 = 0.02; P = 0.57). Injection of insulin aspart exacerbated delayed hyperinsulinaemia after meal loading, mainly in patients with Tmax ≥ 120 min. With regard to pharmacodynamics, insulin aspart had favourable effects on postprandial hyperglycaemia, hyperglucagonaemia and hyperlipidaemia.

The Tmax for this insulin analogue differed greatly between individuals and delayed hyperinsulinaemia was particularly exacerbated in patients with higher Tmax values. Identification of the factors contributing to interindividual variation in the absorption lag time is essential for improving the efficacy and safety of insulin aspart.
Protective role of chrysin against oxidative stress in d‐galactose‐induced aging in an experimental rat model
Authors: Anand Kalaiselvi Velayutham , Mohamed Jaabir Mohamed Sultan , Thomas Philip A , Geraldine Pitchairaj
Published: April 2012

Aim:  To evaluate the putative protective role of chrysin, an isoflavone, on d‐galactose‐induced aging in an experimental rat model.

Methods:  Rats were divided into five groups of five each. Group I received 0.9% saline only. Groups II, III and IV received d‐galactose (50 mg/kg bodyweight) intraperitoneally, additionally group III and group IV received chrysin (20 mg/kg bodyweight) and α‐tocopherol acetate (200 mg/kg bodyweight), respectively. Group V received chrysin alone. The experiment period was for a period of 8 weeks. After the rats were killed, the tissue samples were analyzed for mean activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione‐S‐transferase, glutathione reductase, glucose‐6‐phosphate dehydrogenase, reduced glutathione, vitamin C, vitamin E, malondialdehyde and protein carbonyl. Histopathological studies were also carried out for morphological conformation.

Results:  Tissue samples from d‐galactose‐exposed untreated rats showed significantly (P < 0.05) lower levels of enzymatic and non‐enzymatic anti‐oxidants, and significantly (P < 0.05) higher levels of malondialdehyde and protein carbonyl when compared with group I and group III rats. Oral administration of chrysin for a period of 8 weeks, concomitant with the exposure to d‐galactose, appeared to protect against oxidative damage and maintained all parameters at near normal levels. Histopathological studies confirmed the oxidative damage caused by d‐galactose alone in tissues and also showed the tissue protective role of chrysin in rats receiving d‐galactose and chrysin.

Conclusion:  These results suggest that chrysin protects against oxidative stress‐induced tissue damage in d‐galactose‐induced aging in an experimental rat model. Geriatr Gerontol Int 2012; ••: ••–••.
Predicting the in‐patient outcomes of acute medical admissions from the nursing home: The experience of St James's Hospital, Dublin, 2002–2010
Authors: Romero‐Ortuno Roman , O'Shea Diarmuid , Silke Bernard
Published: April 2012

Aim:  To identify predictors of negative in‐patient outcomes (prolonged hospital stay and death) in nursing home (NH) residents admitted to the hospital as medical emergencies.

Methods:  This was a retrospective patient series set at St James's Hospital (Dublin, Ireland). The participants were all NH patients requiring acute medical admission under the on‐call medical team between 1 January 2002 and 31 December 2010. Patient characteristics on admission, such as demographics, comorbidity level, major diagnostic categories, vital signs and laboratory profile, were measured. The outcomes of the study were prolonged hospital stay (≥30 days) and in‐hospital mortality. The characteristics of NH patients were compared with those of non‐NH patients aged ≥65 years. Multivariate analyses were based on generalized estimating equations and classification trees.

Results:  There were 55 763 acute medical admissions over the period, of which 1938 (3.5%) were from NH. As compared with non‐NH patients aged ≥65 years, NH patients had greater acute illness severity. NH patients had a median length of stay of 7 days, and 17% had a prolonged admission. Their overall mortality rate was 23%. However, the classification analysis showed substantial patient heterogeneity; the subgroup with the highest mortality (54%, n = 100) had positive serum troponin and a respiratory major diagnosis. The lowest mortality rate (4%) was seen in those without positive troponin, urea of 12 mmol/L or less, and albumin of more than 37 mg/L (n = 226).

Conclusions:  Simple serum markers, such as troponin, urea and albumin, might predict mortality in medically admitted NH patients. This might help health‐care practitioners to anticipate their clinical course at an early stage. Geriatr Gerontol Int 2012; ••: ••–••.