A new in vitro assay to test UVR protection of dermal extracellular matrix components by a flat spectrum sunscreen.

The efficacy of topical sunscreens is currently assessed by crude, costly and time consuming in vivo assays. We have previously demonstrated that components of the dermal extracellular matrix (ECM), rich in UV-absorbing amino acids, are susceptible to damage by solar simulated radiation (SSR) in vitro. Here we developed an in vitro method to test the ability of sunscreens to protect fibrillin-rich microfibrils (FRM) and fibronectin, key components of the dermal ECM from UV-induced damage. Solutions of FRM or fibronectin were irradiated without protection, in the presence of a vehicle or a commercially-available flat-spectrum sunscreen. The effect of SSR on molecular structure was determined by atomic force microscopy (FRM) and SDS-PAGE (fibronectin). Following irradiation, FRM periodicity became bi-modally distributed (peaks: 40nm & 59nm) compared to the unimodal distribution in unexposed controls (peak: 50nm). Irradiation in the presence of flat-spectrum sunscreen protected against this change, maintaining the unimodal distribution. SSR induced significant aggregation of fibronectin (p=0.005), which was abrogated by sunscreen. These results demonstrate that this in vitro assay system is sufficiently sensitive to act as an initial/additional screen of sunscreen efficacy. We conclude that sunscreen can reduce UV-mediated damage of key dermal ECM in vitro and thereby prevent remodelling associated with photoageing.

DNA methylation of Th2 lineage determination genes at birth is associated with allergic outcomes in childhood.

BACKGROUND: There is now increasing evidence that asthma and atopy originate in part in utero, with disease risk being associated with the altered epigenetic regulation of genes. OBJECTIVE AND METHODS: To determine the relationship between variations in DNA methylation at birth and the development of allergic disease, we examined the methylation status of CpG loci within the promoter regions of Th1/2 lineage commitment genes (GATA3, IL-4, IL-4R, STAT4 and TBET) in umbilical cord DNA at birth in a cohort of infants from the Southampton Women's Survey (n = 696) who were later assessed for asthma, atopic eczema and atopy. RESULTS: We found that higher methylation of GATA3 CpGs -2211/-2209 at birth was associated with a reduced risk of asthma at ages 3 (median ratio [median methylation in asthma group/median methylation in non-asthma group] = 0.74, P = .006) and 6-7 (median ratio 0.90, P = .048) years. Furthermore, we demonstrated that the GATA3 CpG loci associated with later risk of asthma lie within a NF-κB binding site and that methylation here blocks transcription factor binding to the GATA3 promoter in the human Jurkat T-cell line. Associations between umbilical cord methylation of CpG loci within IL-4R with atopic eczema at 12 months (median ratio 1.02, P = .028), and TBET with atopy (median ratio 0.98, P = .017) at 6-7 years of age were also observed. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings provide further evidence of a developmental contribution to the risk of later allergic disorders and suggest that involvement of epigenetic mechanisms in childhood asthma is already demonstrable at birth.

The developmental environment, epigenetic biomarkers and long-term health.

Evidence from both human and animal studies has shown that the prenatal and early postnatal environments influence susceptibility to chronic disease in later life and suggests that epigenetic processes are an important mechanism by which the environment alters long-term disease risk. Epigenetic processes, including DNA methylation, histone modification and non-coding RNAs, play a central role in regulating gene expression. The epigenome is highly sensitive to environmental factors in early life, such as nutrition, stress, endocrine disruption and pollution, and changes in the epigenome can induce long-term changes in gene expression and phenotype. In this review we focus on how the early life nutritional environment can alter the epigenome leading to an altered susceptibility to disease in later life.

Synthesis of fruity ethyl esters by acyl coenzyme A: alcohol acyltransferase and reverse esterase activities in Oenococcus oeni and Lactobacillus plantarum.

AIMS: To assess the abilities of commercial wine lactic acid bacteria (LAB) to synthesize potentially flavour active fatty acid ethyl esters and determine mechanisms involved in their production. METHODS AND RESULTS: Oenococcus oeni AWRI B551 produced significant levels of ethyl hexanoate and ethyl octanoate following growth in an ethanolic test medium, and ester formation generally increased with increasing pH (4.5 > 3.5), anaerobiosis and precursor supplementation. Cell-free extracts of commercial O. oeni strains and Lactobacillus plantarum AWRI B740 were also tested for ester-synthesizing capabilities in a phosphate buffer via: (i) acyl coenzyme A: alcohol acyltransferase (AcoAAAT) activity and (ii) reverse esterase activity. For both ester-synthesizing activities, strain-dependent variation was observed, with AcoAAAT activity generally greater than reverse esterase. Reverse esterase in O. oeni AWRI B551 also esterified 1-propanol to produce propyl octanoate, and deuterated substrates ([(2)H(6)]ethanol and [(2)H(15)]octanoic acid) to produce the fully deuterated ester, [(2)H(5)]ethyl [(2)H(15)]octanoate. CONCLUSIONS: Wine LAB exhibit ethyl ester-synthesizing capability and possess two different ester-synthesizing activities, one of which is associated with an acyl coenzyme A: alcohol acyltransferase. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that wine LAB exhibit enzyme activities that can augment the ethyl ester content of wine. This knowledge will facilitate greater control over the impacts of malolactic fermentation on the fruity sensory properties and quality of wine.

Prenatal famine exposure, health in later life and promoter methylation of four candidate genes.

Poor nutrition during fetal development can permanently alter growth, cardiovascular physiology and metabolic function. Animal studies have shown that prenatal undernutrition followed by balanced postnatal nutrition alters deoxyribonucleic acid (DNA) methylation of gene promoter regions of candidate metabolic control genes in the liver. The aim of this study was to investigate whether methylation status of the proximal promoter regions of four candidate genes differed between individuals exposed to the Dutch famine in utero. In addition, we determined whether methylation status of these genes was associated with markers of metabolic and cardiovascular disease and adult lifestyle. Methylation status of the GR1-C (glucocorticoid receptor), PPARγ (peroxisome proliferator-activated receptor gamma), lipoprotein lipase and phosphatidylinositol 3 kinase p85 proximal promoters was investigated in DNA isolated from peripheral blood samples of 759 58-year-old subjects born around the time of the 1944-45 Dutch famine. We observed no differences in methylation levels of the promoters between exposed and unexposed men and women. Methylation status of PPARγ was associated with levels of high-density lipoprotein cholesterol and triglycerides as well as with exercise and smoking. Hypomethylation of the GR promoter was associated with adverse adult lifestyle factors, including higher body mass index, less exercise and more smoking. The previously reported increased risk of cardiovascular and metabolic disease after prenatal famine exposure was not associated with differences in methylation status across the promoter regions of these candidate genes measured in peripheral blood. The adult environment seems to affect GR and PPARγ promoter methylation.

[Coronary CT angiography using prospective ECG triggering: high diagnostic accuracy with low radiation dose]. / CT-Angiographie der Koronarien mit prospektivem EKG-Triggering: Hohe diagnostische Genauigkeit bei niedriger Strahlendosis.

BACKGROUND: The purpose of this study was to evaluate the diagnostic performance of coronary CT angiography (coronary CTA) using prospective ECG triggering (PT) for the detection of significant coronary artery stenosis compared to invasive coronary angiography (ICA). METHODS: A total of 20 patients underwent coronary CTA with PT using a 128-slice CT scanner (Definition AS+, Siemens) and ICA. All coronary CTA studies were evaluated for significant coronary artery stenoses (>or=50% luminal narrowing) by 2 observers in consensus using the AHA-15-segment model. Findings in CTA were compared to those in ICA. RESULTS: Coronary CTA using PT had 88% sensitivity in comparison to 100% with ICA, 95% to 88% specificity, 80% to 92% positive predictive value and 97% to 100% negative predictive value for diagnosing significant coronary artery stenosis on per segment per patient analysis, respectively. Mean effective radiation dose-equivalent of CTA was 2.6+/-1 mSv. CONCLUSION: Coronary CTA using PT enables non-invasive diagnosis of significant coronary artery stenosis with high diagnostic accuracy in comparison to ICA and is associated with comparably low radiation exposure.

Iodinated contrast media: effect of osmolarity and injection temperature on erythrocyte morphology in vitro.

BACKGROUND: Some side effects of intravenously injected iodinated contrast media are thought to be linked to the biological properties of the various agents and their effect on blood components. PURPOSE: To assess the effect of osmolarity and injection temperature of iodinated contrast media on erythrocyte (RBC) morphology in vitro. MATERIAL AND METHODS: Blood from 20 volunteers was incubated with three different contrast media (320 mg I/ml iso-osmolar iodixanol, 300 mg I/ml low-osmolar iopromide, 300 mg I/ml low-osmolar iopamidol) injected at 37 degrees C, 43 degrees C, and 48 degrees C, and in two different volumes corresponding to the estimated concentration at the site of venous injection and after systemic distribution. After 10 min incubation, aliquots were removed for complete blood count analysis and blood smears. Two hematologists blindedly and independently reviewed all smears, and determined the grade of morphological RBC changes compared to a blank sample. RESULTS: There was excellent (kappa = 0.98) inter-reader correlation for grading RBC changes. At systemic concentration at 37 degrees C, the grade of RBC changes was significantly (P<0.05) less in blood samples exposed to iso-osmolar iodixanol (mean 0.21) as compared to low-osmolar iopromide (mean 0.26) and low-osmolar iopamidol (mean 0.58). These differences became more significant at higher volumes, corresponding to concentrations at the site of injection and higher injection temperatures. CONCLUSION: In vitro, RBC morphology is less affected by iso-osmolar as compared to low-osmolar contrast media. These differences become more significant at higher injection temperatures that are proposed to improve flow dynamics for high-speed injection.

[CT coronary angiography: indications, image acquisition, and interpretation]. / Angiografía por tomografía computarizada coronaria: indicaciones, adquisición de imágenes e interpretación.

Intense scientific and clinical evaluation have brought about great improvements in cardiac CT. This is no longer merely an experimental technique, rather it has become a clinical application that is ready to fulfill its promise of replacing invasive cardiac catheterization in certain patient populations. Among the proven indications is the evaluation of patients with atypical chest pain, the morphological evaluation of the coronary arteries in cases of suspected congenital anomalies, and before surgical intervention, as well as the evaluation of coronary revascularizations. The use of CT angiography for the exhaustive evaluation of cardiac and non-cardiac pathology in patients with acute chest pain in the emergency department is currently being investigated. Because the heart is continuously moving, CT coronary angiography represents a greater technical challenge than other applications of CT. On the other hand, rapid technical development requires acquisition protocols to be adjusted constantly. However, users that know the general techniques of computed tomography can overcome these challenges. The aim of this article is to provide those interested and involved in CT angiography with a manual to enable them to follow our method step by step. We include considerations regarding the correct selection of patients, patient medication, radiological protection, contrast enhancement, acquisition and reconstruction parameters, image display, image analysis techniques, and the radiological report. Our recommendations are based on our experience, which runs from the evolution of multiple-row detector CT scanners for cardiac applications from its beginnings to the most modern presentations of advanced acquisition modalities, including dual-source CT, which we consider to be the precursor of this test in routine clinical practice.

Angiografía por tomografía computarizada coronaria: indicaciones, adquisición de imágenes e interpretación / CT coronary angiography: indications, image acquisition, and interpretation

La tomografía computarizada (TC) cardíaca, debido al perfeccionamiento técnico y a la intensa evaluación científica y clínica en curso, ha abandonado el campo de la investigación y se ha convertido en una aplicación clínica a punto de cumplir su promesa de sustituir a la cateterización cardíaca invasiva en determinadas situaciones clínicas. Entre las indicaciones comprobadas de la angiografía por tomografía computarizada (angio-TC) coronaria se incluye la evaluación de pacientes con dolor torácico atípico, la evaluación morfológica de las arterias coronarias en caso de sospecha de anomalías congénitas y antes de una intervención quirúrgica, así como la evaluación del by-pass coronario. Actualmente se está investigando el uso de la angio-TC para la evaluación de la enfermedad cardíaca y no cardíaca en pacientes con dolor torácico agudo en Urgencias. Dada la naturaleza de su objetivo, el corazón en continuo movimiento, la angio-TC coronaria supone un desafío técnico mayor que el de otras aplicaciones de la TC. Por otra parte, el rápido desarrollo técnico requiere una adaptación constante de protocolos de adquisición. Sin embargo, estos retos no son, en modo alguno, insuperables para los usuarios que conozcan la técnica general de la TC. La finalidad de este trabajo es facilitar a quienes estén interesados e involucrados en la angio-TC coronaria un manual que presenta paso a paso nuestro método para realizarla. Se incluyen consideraciones referentes a la selección correcta de pacientes, la medicación del paciente, la protección radiológica, el uso de medios de contraste, los parámetros de adquisición y reconstrucción, la visualización y las técnicas de análisis, así como el informe radiológico. Nuestras recomendaciones se basan en una experiencia que abarca la evolución de la TC multidetector para aplicaciones cardíacas, desde sus comienzos hasta las presentaciones más actuales de modalidades de adquisición avanzadas, incluida la TC de doble fuente, que muy probablemente asentará la entrada de la angio-TC coronaria en la práctica clínica diaria

Intense scientific and clinical evaluation have brought about great improvements in cardiac CT. This is no longer merely an experimental technique, rather it has become a clinical application that is ready to fulfill its promise of replacing invasive cardiac catheterization in certain patient populations. Among the proven indications is the evaluation of patients with atypical chest pain, the morphological evaluation of the coronary arteries in cases of suspected congenital anomalies, and before surgical intervention, as well as the evaluation of coronary revascularizations. The use of CT angiography for the exhaustive evaluation of cardiac and non-cardiac pathology in patients with acute chest pain in the emergency department is currently being investigated. Because the heart is continuously moving, CT coronary angiography represents a greater technical challenge than other applications of CT. On the other hand, rapid technical development requires acquisition protocols to be adjusted constantly. However, users that know the general techniques of computed tomography can overcome these challenges. The aim of this article is to provide those interested and involved in CT angiography with a manual to enable them to follow our method step by step. We include considerations regarding the correct selection of patients, patient medication, radiological protection, contrast enhancement, acquisition and reconstruction parameters, image display, image analysis techniques, and the radiological report. Our recommendations are based on our experience, which runs from the evolution of multiple-row detector CT scanners for cardiac applications from its beginnings to the most modern presentations of advanced acquisition modalities, including dual-source CT, which we consider to be the precursor of this test in routine clinical practice

MR-guided interventions of the prostate gland.

In recent years MR imaging has played an increasingly important role in the diagnosis and treatment of prostate cancer. MR imaging of the prostate allows a clear delineation of the anatomic structures and prostate tumors when performing interventions such as biopsies, brachytherapy or thermal therapy of the prostate gland. MRI robotic assistance will improve the accuracy of the interventions. Due to the advantages of MR imaging MR-guided prostate interventions will play an increasing role in future.

64 slice cardiovascular CT in the emergency department: concepts and first experiences.

PURPOSE: The purpose of this study was to evaluate contrast-enhanced electrocardiogram (ECG)-gated 64-slice computed tomography (CT) angiography of the thorax as a triage tool in patients with acute equivocal chest pain. MATERIAL AND METHODS: Technical principles and diagnostic algorithms for using a single ECG-gated 64-slice CT scan for triple rule-out of acute pulmonary embolism, aortic dissection, acute coronary syndromes and other diseases of the chest are introduced. Our experience using this test in 23 patients are reviewed and exemplary cases are illustrated. The total length of hospitalisation and charges for Emergency Department care at the time of discharge were compared with a matched control population that underwent catheter angiography for emergent cardiac workup. Statistical analyses were performed with an independent Student's t test. Mann-Whitney rank sum test was also used on variables that did not have equal variance. RESULTS: Of the 23 patients, 11 presented without pathological findings, two with extensive pulmonary embolism, two with definite coronary artery disease (CAD) but stenosis <50% and eight with significant CAD (>50% stenosis). Catheter angiography was performed in the latter group, confirming the CT findings in all cases. Nine patients without CT findings were discharged on the same day. In comparison with the control group, length of hospitalisation (p=0.009) and total hospital charges (p<0.001) were significantly reduced. CONCLUSIONS: Our initial experience shows that ECG-gated 64- slice CT angiography of the entire thorax is technically feasible and enables rapid triage of patients to determine underlying cardiac and noncardiac reasons for chest pain. This test may thus help to significantly reduce costs and length of hospitalisation. Prospective studies involving larger groups of patients are required to confirm these findings.

Integrin-linked kinase, a promising cancer therapeutic target: biochemical and biological properties.

Integrin-linked kinase (ILK) is an ankyrin repeat-containing Ser/Thr kinase that interacts with the cytoplasmic domains of beta(1) and beta(3) integrins. ILK is widely expressed in tissues throughout the body, and, as might be expected, appears to mediate a diversity of functions relating to its role in coupling integrins and growth factor receptors to downstream signaling pathways. Through its downstream targets protein kinase B/Akt and glycogen synthase kinase-3beta, ILK appears to be involved in several oncogenesis-related events, including suppression of apoptosis and promotion of cell survival, as well as cell migration and invasion. Over-expression of ILK in epithelial cells results in anchorage-independent cell growth with increased cell cycle progression. Inoculation of nude mice with ILK over-expressing cells leads to tumor formation. Furthermore, increased ILK expression and activity have been correlated with malignancy in several human tumor types, including breast, prostate, brain, and colon carcinomas. Based on these findings, ILK represents an excellent therapeutic target for the prevention of tumor progression. Here, we provide an overview of the physical and biochemical properties of ILK, and present data describing the impact of small-molecule ILK inhibitors on several ILK-mediated cellular functions.

Inhibition of beta 2 integrin receptor and Syk kinase signaling in monocytes by the Src family kinase Fgr.

While beta 2 integrin ligand-receptor recognition interactions are well characterized, less is known about how these events trigger signal transduction cascades to regulate the transition from tethering to firm adhesion, spreading, and transendothelial migration. We have identified critical positive and negative regulatory components of this cascade in monocytes. Whereas the Syk tyrosine kinase is essential for beta 2 integrin signaling and cell spreading, the Src family kinase Fgr is a negative regulator of this pathway. Fgr selectively inhibits beta 2 but not beta 1 integrin signaling and Syk kinase function via a direct association between the Fgr SH2 domain and Syk tyrosine Y342. The inhibitory effects of Fgr are independent of its kinase activity, are dose dependent, and can be overcome by chemokines and inflammatory mediators.

Disease mechanisms in psoriasis and psoriatic arthritis.

Any hypothesis that tries to explain disease mechanisms in psoriasis and psoriatic arthritis (PsA) must take into account the containment of the inflammatory process to three specific sites: the skin, synovium, and enthesis. This article reviews the recent literature that advances our understanding of disease mechanisms at these specific sites. Additional progress will be achieved by research that focuses on common pathogenic pathways at these sites, in particular when searching for foreign candidates or auto-antigens triggering the T-cell-mediated immune response.

The radiology of pulmonary cryptococcosis in a tertiary medical center.

Pulmonary cryptococcal infections occur in both immunocompetent and immunocompromised individuals, with a reported increased incidence of diffuse pulmonary disease in acquired immune deficiency syndrome (AIDS) patients. The authors observed no differences in the radiographic appearances of pulmonary cryptococcal disease between human immunodeficiency virus (HIV) patients and other immunocompromised individuals. Chest computed tomography (CT) contributes to a more comprehensive understanding of pulmonary cryptococcal infections.

Acquired esotropia: subjective and objective outcomes.

PURPOSE: The measurement of outcomes of pediatric therapy will be increasingly important to third-party payers as they allocate health care resources. We undertook this study to assess the effectiveness of treatment of acquired esotropia, as measured subjectively by parents and objectively by examination. METHODS: A chart review was used to gather objective clinical data from all patients presenting between 1994 and 1995 with esotropia after their first birthday. Parents' impressions of the quality and impact of treatment were tabulated by using a telephone questionnaire. RESULTS: Of 48 patients studied (mean age, 3.8 years; 44 months' follow-up), the 26 with amblyopia were effectively treated in 81% of cases. Mean esotropic angles decreased, with glasses and/or surgery, from 33.1 to 4.9 PD at distance and from 40.0 to 11.4 PD at near. The average estimated cost of care was 547 dollars per patient per year. Parents considered treatment "extremely important" to their child's future happiness and success in 90% of cases. CONCLUSION: Our experience has shown that treatment of acquired esotropia in childhood, while relatively inexpensive, is highly effective, both objectively and subjectively.