Achaete-scute homolog-1 linked to remodeling and preneoplasia of pulmonary epithelium

The basic helix-loop-helix protein achaete-scute homolog-1 (ASH1) is involved in lung neuroendocrine (NE) differentiation and tumor promotion in SV40 transgenic mice. Constitutive expression of human ASH-1 (hASH1) in mouse lung results in hyperplasia and remodeling that mimics bronchiolization of alveoli (BOA), a potentially premalignant lesion of human lung carcinomas. We now show that this is due to sustained cellular proliferation in terminal bronchioles and resistance to apoptosis. Throughout the airway epithelium the expression of anti-apoptotic Bcl-2 and c-Myb was increased and Akt/mTOR pathway activated. Moreover, the expression of matrix metalloproteases (MMPs) including MMP7 was specifically enhanced at the bronchiolo-alveolar duct junction and BOA suggesting that MMPs play a key role in this microenvironment during remodeling. We also detected MMP7 in 70% of human BOA lesions. Knockdown of hASH1 gene in human lung cancer cells in vitro suppressed growth by increasing apoptosis. We also show that forced expression of hASH1 in immortalized human bronchial epithelial cells decreases apoptosis. We conclude that the impact of hASH1 is not limited to cells with NE phenotype. Rather, constitutive expression of hASH1 in lung epithelium promotes remodeling through multiple pathways that are commonly activated during lung carcinogenesis. The collective results suggest a novel model of BOA formation via hASH1-induced suppression of the apoptotic pathway. Our study yields a promising new preclinical tool for chemoprevention of peripheral lung carcinomas. © 2007 USCAP, Inc All rights reserved.

RAGE Regulates Immune Cell Infiltration and Angiogenesis in Choroidal Neovascularization

Purpose: RAGE regulates pro-inflammatory responses in diverse cells and tissues. This study has investigated if RAGE plays a role in immune cell mobilization and choroidal neovascular pathology that is associated with the neovascular form of age-related macular degeneration (nvAMD).

Methods: RAGE null (RAGE−/−) mice and age-matched wild type (WT) control mice underwent laser photocoagulation to generate choroidal neovascularization (CNV) lesions which were then analyzed for morphology, S100B immunoreactivity and inflammatory cell infiltration. The chemotactic ability of bone marrow derived macrophages (BMDMs) towards S100B was investigated.

Results: RAGE expression was significantly increased in the retina during CNV of WT mice (p<0.001). RAGE−/− mice exhibited significantly reduced CNV lesion size when compared to WT controls (p<0.05). S100B mRNA was upregulated in the lasered WT retina but not RAGE−/− retina and S100B immunoreactivity was present within CNV lesions although levels were less when RAGE−/− mice were compared to WT controls. Activated microglia in lesions were considerably less abundant in RAGE−/− mice when compared to WT counterparts (p<0.001). A dose dependent chemotactic migration was observed in BMDMs from WT mice (p<0.05–0.01) but this was not apparent in cells isolated from RAGE−/− mice.

Conclusions: RAGE-S100B interactions appear to play an important role in CNV lesion formation by regulating pro-inflammatory and angiogenic responses. This study highlights the role of RAGE in inflammation-mediated outer retinal pathology.

PET-CT as a predictor of outcome in resectable colorectal liver metastases

Introduction
PET-computed tomography (PET-CT) is a useful staging imaging modality in colorectal liver metastases (CRLM). This study aimed to determine whether PET-CT parameters, standardized uptake value (SUV) and reconstructed tumour volume (RTV), are predictors of prognosis and survival.

Methods
A study of all resectable CRLM patients in the regional HPB unit from 2007–2009 was performed. Preoperative PET-CT scans were retrospectively reviewed; SUV, diameter and RTV for each lesion was recorded. Correlation analysis was performed with other pathological and biochemical parameters, by Pearson’s correlation analysis. Survival analysis was performed using Cox regression hazard model. A P value of less than 0.05 was considered statistically significant.

Results
A total of 79 patients were included. SUV moderately correlated with tumour diameter, both PET-CT (r=0.4927; P<0.0001) and histology (r=0.4513; P=0.0003); RTV (r=0.4489; P<0.001), preoperative carcinoembryonic antigen (CEA) (r=0.4977; P=0.0001), and postoperative CEA (r=0.3727; P=0.004). Multivariate analysis found that an independent predictor of SUVmax was preoperative CEA (P=0.03). RTV strongly correlated with preoperative CEA (r=0.9389; P<0.0001). SUV and RTV had a negative effect on survival.

Conclusion
PET-CT, in the setting of CRLM, may have a prognostic role in assessing survival. Although no definite conclusions can be drawn regarding the prognostic role of SUV and RTV, it acts to reinforce the need for further prospective studies to validate these findings.

Intravitreal Injection of Normal Saline Induces Retinal Degeneration in the C57BL/6J Mouse

Purpose: To investigate the adverse effect of intravitreal injection of normal saline (NS) and phosphate buffered saline (PBS) in mouse eyes.

Methods: NS or PBS was injected intravitreally into C57BL/6J mouse eyes. Retinal lesions were monitored by fundus imaging, spectral-domain optical coherence tomography (SD-OCT), and histological investigations. Retinal immune gene expression was determined by real-time polymerase chain reaction (PCR). The toxic effect of NS and PBS or retinal protein from NS- or PBS-injected eyes on retinal pigment epithelium (RPE) was tested in B6-RPE-07 mouse RPE cell cultures.

Results: Intravitreal injection of NS dose-dependently induced localized retinal lesion in mice. Histological investigations revealed multiple vacuoles in photoreceptor outer segments and RPE cells. The lesions recovered over time and by 3 weeks post injection the majority of lesions vanished in eyes receiving 1 μl NS. Inflammatory genes, including TNF-α, IL-1β, IL-6, iNOS, and VEGF were upregulated in NS injected eyes. Intravitreal injection of PBS did not cause any pathology. The treatment of B6-RPE07 cells with 30% PBS or 30% NS did not affect RPE viability. However, incubation of 1-μg/ml retinal protein from NS-injected eyes, but not PBS-injected eyes induced RPE cell death.

Conclusion: NS is toxic to the C57BL/6J mouse retina and should not be used as a vehicle for intraocular injection. PBS is not toxic to the retina and is a preferred vehicle.

Translational Relevance: NS is not a physiological solution for intraocular injection in the C57BL/6J mice and questions its suitability for intraocular injection in other species, including human.

Field-Isolated Genotypes of Mycobacterium bovis Vary in Virulence and Influence Case Pathology but Do Not Affect Outbreak Size

Strains of many infectious agents differ in fundamental epidemiological parameters including transmissibility, virulence and pathology. We investigated whether genotypes of Mycobacterium bovis (the causative agent of bovine tuberculosis, bTB) differ significantly in transmissibility and virulence, combining data from a nine-year survey of the genetic structure of the M. bovis population in Northern Ireland with detailed records of the cattle population during the same period. We used the size of herd breakdowns as a proxy measure of transmissibility and the proportion of skin test positive animals (reactors) that were visibly lesioned as a measure of virulence. Average breakdown size increased with herd size and varied depending on the manner of detection (routine herd testing or tracing of infectious contacts) but we found no significant variation among M. bovis genotypes in breakdown size once these factors had been accounted for. However breakdowns due to some genotypes had a greater proportion of lesioned reactors than others, indicating that there may be variation in virulence among genotypes. These findings indicate that the current bTB control programme may be detecting infected herds sufficiently quickly so that differences in virulence are not manifested in terms of outbreak sizes. We also investigated whether pathology of infected cattle varied according to M. bovis genotype, analysing the distribution of lesions recorded at post mortem inspection. We concentrated on the proportion of cases lesioned in the lower respiratory tract, which can indicate the relative importance of the respiratory and alimentary routes of infection. The distribution of lesions varied among genotypes and with cattle age and there were also subtle differences among breeds. Age and breed differences may be related to differences in susceptibility and husbandry, but reasons for variation in lesion distribution among genotypes require further investigation. © 2013 Wright et al.

The management of radial scars of the breast - does core biopsy help?

Purpose: To compare the effectiveness of fine needle aspiration cytology (FNAC) with core biopsy (CB) in the pre-operative diagnosis of radial scar (RS) of the breast.

Patients and methods: A retrospective analysis was made of all radial scars diagnosed on surgical histology over an 8-year period. Comparison was made between the results of different preoperative needle biopsy techniques and surgical histology findings.

Results: Forty of 47 patients with a preoperative radiological diagnosis of radial scar were included in this analysis. Thirty-eight patients had impalpable lesions diagnosed on mammography and two presented with a palpable lump. FNAC (n=17) was inadequate in 47% of patients, missed two co-existing carcinomas found in this group, and gave a false positive or suspicious result for malignancy in 4 patients. CB (n=23) suggested a RS in 15 patients, but only diagnosed 4 out of 7 co-existing carcinomas found in this group.

Conclusion: CB is more accurate than FNAC in the diagnosis of RS. However, these data demonstrate that CB may offer little to assist in the management of patients with RS. In summary, this paper advocates the use of CB in any lesion with a radiological suspicion of carcinoma and diagnostic excision of all lesions thought to be typical of RS on mammography.

Genome-wide association study identifies novel loci associated with resistance to bovine tuberculosis

Tuberculosis (TB) caused by Mycobacterium bovis is a re-emerging disease of livestock that is of major economic importance worldwide, as well as being a zoonotic risk there is significant heritability for host resistance to bovine TB (bTB) in dairy cattle. To identify resistance loci for bTB, we undertook a genome-wide association study in female Holstein-Friesian cattle with 592 cases and 559 age-matched controls from case herds. Cases and controls were categorised into distinct phenotypes: skin test and lesion positive vs skin test negative on multiple occasions, respectively these animals were genotyped with the Illumina BovineHD 700K BeadChip. Genome-wide rapid association using linear and logistic mixed models and regression (GRAMMAR), regional heritability mapping (RHM) and haplotype-sharing analysis identified two novel resistance loci that attained chromosome-wise significance, protein tyrosine phosphatase receptor T (PTPRT; P=4.8 × 10 -7) and myosin IIIB (MYO3B; P=5.4 × 10 -6). We estimated that 21% of the phenotypic variance in TB resistance could be explained by all of the informative single-nucleotide polymorphisms, of which the region encompassing the PTPRT gene accounted for 6.2% of the variance and a further 3.6% was associated with a putative copy number variant in MYO3B the results from this study add to our understanding of variation in host control of infection and suggest that genetic marker-based selection for resistance to bTB has the potential to make a significant contribution to bTB control.

Post-tensioning of glulam timber with steel tendons

This paper describes a series of four-point bending tests that were conducted, under service loads and to failure, on unreinforced, reinforced and post-tensioned glulam timber beams, where the reinforcing tendon used was 12 mm diameter toughened steel bar. The research was designed to evaluate the benefits offered by including an active reinforcement in contrast to the passive reinforcement typically used within timber strengthening works, in addition to establishing the effect that bonding the reinforcing tendon has on the materials performance.

The laboratory investigations established that the flexural strength and stiffness increased for both the reinforced and post-tensioned timbers compared to the unreinforced beams. The flexural strength of the reinforced timber increased by 29.4%, while the stiffness increased by 28.1%. Timber that was post-tensioned with an unbonded steel tendon showed a flexural strength increase of 17.6% and an increase in stiffness of 8.1%. Post-tensioned beams with a bonded steel tendon showed increases in flexural strength and stiffness of 40.1% and 30% respectively.

Post-tensioning glulam timber beams with basalt FRP tendons

Improvements in the structural performance of glulam timber beams by the inclusion of reinforcing materials can increase both the service performance and ultimate capacity. This paper describes a series of four-point bending tests conducted, under service loads and to failure, on unreinforced, reinforced and post-tensioned glulam timber beams, where the reinforcing tendon used is 12 mm dia. basalt fibre-reinforced polymer. The research is designed to evaluate the benefits offered by including an active reinforcement in contrast to the passive reinforcement typically used within timber strengthening works, in addition to establishing the effect that bonding the reinforcing tendon has on the material's performance. Further experimental tests have also been developed to investigate the long-term implications of this research, with emphasis placed upon creep and loss of post-tensioning; however, this is ongoing and is not presented in this paper. The laboratory investigations establish that the flexural strength and stiffness increase for both the unbonded and bonded post-tensioned timbers compared to the unreinforced and reinforced beams. Timber that is post-tensioned with an unbonded basalt fibre-reinforced polymer tendon shows a flexural strength increase of 2ṡ8% and an increase in stiffness of 8ṡ7%. Post-tensioned beams with a bonded basalt fibre-reinforced polymer tendon show increases in flexural strength and stiffness of 15ṡ4% and 11ṡ5% respectively.

POST-TENSIONING OF TIMBER BEAMS WITH BASALT FIBRE REINFORCED POLYMER

Improvements in the structural performance of glulam timber beams by the inclusion of reinforcing materials can improve both the service performance and ultimate capacity. In recent years research focusing on the addition of fibre reinforced polymers to strengthen members has yielded positive results. However, the FRP material is still a relatively expensive material and its full potential has not been realised in combination with structural timber. This paper describes a series of four-point bending tests that were conducted, under service and ultimate loads, on post-tensioned glulam timber beams where the reinforcing tendon used was 12 mm diameter Basalt Fibre Reinforced Polymer (BFRP). The research was designed to evaluate the additional benefits of including an active type of reinforcement, by post-tensioning the BFRP tendon, as opposed to the passive approach of simply reinforcing the timber beam.
From the laboratory investigations, it was established that there was a 16% increase in load carrying capacity, in addition to a 14% reduction in deflection under service loads when members containing the post-tensioned BFRP composite are compared with control timber specimens. Additionally a more favourable ductile failure mode was witnessed compared to the brittle failure of an unreinforced timber beam. The results support the assumption that by initially stressing the embedded FRP tendon the structural benefits experienced by the timber member increase in a number of ways, indicating that there is significant scope for this approach in practical applications.

Development of Novel Post-Tensioned Glulam Timber Composites

Improvements in the structural performance of glulam timber beams by the inclusion of reinforcing materials can increase both the service performance and ultimate capacity. In recent years research focusing on the addition of fibre reinforced polymers (FRP) to strengthen members has yielded positive results. However, the FRP material is still relatively expensive and its full potential in combination with structural timber has not been realised. This paper describes a series of four-point bending tests that were conducted, under service loads and to failure, on unreinforced, reinforced and post-tensioned glulam timber beams, where the reinforcing tendon used was 12mm diameter basalt fibre reinforced polymer (BFRP). The research was designed to evaluate the benefits offered by including an active reinforcement in contrast to the passive reinforcement typically used within timber strengthening works, in addition to establishing the affect that bonding the reinforcing tendon has on the material’s performance. Further experimental tests have been developed to investigate the long-term implications of this research, with emphasis placed upon creep and loss of post-tensioning.
The laboratory investigations established that the flexural strength and stiffness increased for both the unbonded and bonded post-tensioned timbers compared to the unreinforced beams. Timber that was post-tensioned with an unbonded BFRP tendon showed a flexural strength increase of 2.8% and an increase in stiffness of 8.7%. Post-tensioned beams with a bonded BFRP tendon showed increases in flexural strength and stiffness of 16.6% and 11.5% respectively.

BENIGN URETHRAL POLYPS

Ten male patients presented between 1985 and 1991 with benign urethral polyps. This lesion is believed to represent a developmental error in the invagination process of the submucous glandular material of the inner zone of the prostate. The clinicopathological features of the condition are discussed and the literature is reviewed.

Inhibition of Rho-kinase protects cerebral barrier from ischaemia-evoked injury through modulations of endothelial cell oxidative stress and tight junctions

Ischaemic strokes evoke blood-brain barrier (BBB) disruption and oedema formation through a series of mechanisms involving Rho-kinase activation. Using an animal model of human focal cerebral ischaemia, this study assessed and confirmed the therapeutic potential of Rho-kinase inhibition during the acute phase of stroke by displaying significantly improved functional outcome and reduced cerebral lesion and oedema volumes in fasudil- versus vehicle-treated animals. Analyses of ipsilateral and contralateral brain samples obtained from mice treated with vehicle or fasudil at the onset of reperfusion plus 4 h post-ischaemia or 4 h post-ischaemia alone revealed these benefits to be independent of changes in the activity and expressions of oxidative stress- and tight junction-related parameters. However, closer scrutiny of the same parameters in brain microvascular endothelial cells subjected to oxygen-glucose deprivation ± reperfusion revealed marked increases in prooxidant NADPH oxidase enzyme activity, superoxide anion release and in expressions of antioxidant enzyme catalase and tight junction protein claudin-5. Cotreatment of cells with Y-27632 prevented all of these changes and protected in vitro barrier integrity and function. These findings suggest that inhibition of Rho-kinase after acute ischaemic attacks improves cerebral integrity and function through regulation of endothelial cell oxidative stress and reorganization of intercellular junctions. Inhibition of Rho-kinase (ROCK) activity in a mouse model of human ischaemic stroke significantly improved functional outcome while reducing cerebral lesion and oedema volumes compared to vehicle-treated counterparts. Studies conducted with brain microvascular endothelial cells exposed to OGD ± R in the presence of Y-27632 revealed restoration of intercellular junctions and suppression of prooxidant NADPH oxidase activity as important factors in ROCK inhibition-mediated BBB protection.

Discovering subgroups of patients from DNA copy number data using NMF on compacted matrices

In the study of complex genetic diseases, the identification of subgroups of patients sharing similar genetic characteristics represents a challenging task, for example, to improve treatment decision. One type of genetic lesion, frequently investigated in such disorders, is the change of the DNA copy number (CN) at specific genomic traits. Non-negative Matrix Factorization (NMF) is a standard technique to reduce the dimensionality of a data set and to cluster data samples, while keeping its most relevant information in meaningful components. Thus, it can be used to discover subgroups of patients from CN profiles. It is however computationally impractical for very high dimensional data, such as CN microarray data. Deciding the most suitable number of subgroups is also a challenging problem. The aim of this work is to derive a procedure to compact high dimensional data, in order to improve NMF applicability without compromising the quality of the clustering. This is particularly important for analyzing high-resolution microarray data. Many commonly used quality measures, as well as our own measures, are employed to decide the number of subgroups and to assess the quality of the results. Our measures are based on the idea of identifying robust subgroups, inspired by biologically/clinically relevance instead of simply aiming at well-separated clusters. We evaluate our procedure using four real independent data sets. In these data sets, our method was able to find accurate subgroups with individual molecular and clinical features and outperformed the standard NMF in terms of accuracy in the factorization fitness function. Hence, it can be useful for the discovery of subgroups of patients with similar CN profiles in the study of heterogeneous diseases.

Docking the value of pigmeat? Prevalence and financial implications of welfare lesions in Irish slaughter pigs.

Expansion of the meat inspection process to incorporate animal-based welfare measurements could contribute towards significant improvements in pig (Sus scrofa domesticus) welfare and farm profitability. This study aimed to determine the prevalence of different welfare-related lesions on the carcase and their relationship with carcase condemnations (CC) and carcase weight (CW). The financial implications of losses associated with CC and CW reductions related to the welfare lesions were also estimated. Data on tail lesions, loin bruising and bursitis, CW and condemnation/trimming outcome (and associated weights) were collected for 3,537slaughter pigs (mean [± SEM] carcase weight: 79.2 [± 8.82] kg). Overall, 72.5% of pigs had detectable tail lesions, whilst 16.0 and 44.0% were affected by severe loin bruising and hind limb bursitis, respectively. There were 2.5% of study carcases condemned and a further 3.3% were trimmed. The primary cause of CC was abscessation. While tail lesion severity did not increase the risk of abscessation, it was significantly associated with CC. Male pigs had a higher risk of tail lesions and of CC. The financial loss to producers associated with CC and trimmings was estimated at €1.10 per study pig. CW was reduced by up to 12 kg in cases of severe tail lesions. However, even mild lesions were associated with a significant reduction in CW of 1.2 kg. The value of the loss in potential CW associated with tail lesions was €0.59 per study pig. Combined with losses attributable to CC and trimmings this represented a loss of 43% of the profit margin per pig, at the time of the study, attributable to tail biting. These findings illustrate the magnitude of the impact of tail biting on pig welfare and on profitability of the pig industry. They also emphasise the potential contribution that the inclusion of welfare parameters at meat inspection could make to pig producers in informing herd health and welfare management plans.