Evolutionary dynamics of residual disease in human glioblastoma.

BACKGROUND: Glioblastoma is the most common and aggressive adult brain malignancy against which conventional surgery and chemoradiation provide limited benefit. Even when a good treatment response is obtained, recurrence inevitably occurs either locally (∼80%) or distally (∼20%), driven by cancer clones that are often genomically distinct from those in the primary tumour. Glioblastoma cells display a characteristic infiltrative phenotype, invading the surrounding tissue and often spreading across the whole brain. Cancer cells responsible for relapse can reside in two compartments of residual disease that are left behind after treatment: the infiltrated normal brain parenchyma and the sub-ventricular zone. However, these two sources of residual disease in glioblastoma are understudied because of the difficulty in sampling these regions during surgery. PATIENT AND METHODS: Here, we present the results of whole-exome sequencing of 69 multi-region samples collected using fluorescence-guided resection from 11 patients, including the infiltrating tumour margin and the sub-ventricular zone for each patient, as well as matched blood. We used a phylogenomic approach to dissect the spatio-temporal evolution of each tumour and unveil the relation between residual disease and the main tumour mass. We also analysed two patients with paired primary-recurrence samples with matched residual disease. RESULTS: Our results suggest that infiltrative subclones can arise early during tumour growth in a subset of patients. After treatment, the infiltrative subclones may seed the growth of a recurrent tumour, thus representing the 'missing link' between the primary tumour and recurrent disease. CONCLUSIONS: These results are consistent with recognised clinical phenotypic behaviour and suggest that more specific therapeutic targeting of cells in the infiltrated brain parenchyma may improve patient's outcome.

In utero betamethasone affects 3ß-hydroxysteroid dehydrogenase and inhibin-α immunoexpression during testis development.

Prenatal glucocorticoids, commonly used in women at risk of preterm delivery, can predispose the newborn to disease in later life. Since male reproductive function is likely to reflect testis development during fetal life, we studied the effects of prenatal glucocorticoids on two key intra-testicular factors that play roles in cellular proliferation and differentiation, 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and inhibin-α. Pregnant sheep (n=42) were treated with betamethasone (0.5 mg/kg) or saline (control) at 104, 111 and 118 days of gestation (DG). Testicular tissue was sampled from fetuses at 121 and 132DG, and from lambs at 45 and 90 postnatal days (PD). Within the betamethasone treated group, 3ß-HSD immunostaining area was greater at 121DG than at 90PD (P=0.04), but the intensity of immunostaining was higher at 90PD than at 121DG (P=0.04), 132DG (P=0.04) and 45PD (P=0.03). Control animals showed no changes in 3ß-HSD area or intensity of immunostaining. No significant differences were observed between treated and control animals in immunostaining area, but immunostaining was more intense in the treated group than in the control group at 90PD (P=0.03). For inhibin-α, the proportion of immunostaining area declined in treated offspring from 121DG to 45PD, in contrast to control values, but recovered fully by 90PD, concomitantly with the onset of spermatogenesis. In conclusion, prenatal betamethasone increased the postnatal testicular expression of inhibin-α but reduced the expression of 3ß-HSD. These effects could compromise androgen-mediated testicular development and therefore adult capacity for spermatogenesis.

Improving Fiber Alignment in HARDI by Combining Contextual PDE Flow with Constrained Spherical Deconvolution.

We propose two strategies to improve the quality of tractography results computed from diffusion weighted magnetic resonance imaging (DW-MRI) data. Both methods are based on the same PDE framework, defined in the coupled space of positions and orientations, associated with a stochastic process describing the enhancement of elongated structures while preserving crossing structures. In the first method we use the enhancement PDE for contextual regularization of a fiber orientation distribution (FOD) that is obtained on individual voxels from high angular resolution diffusion imaging (HARDI) data via constrained spherical deconvolution (CSD). Thereby we improve the FOD as input for subsequent tractography. Secondly, we introduce the fiber to bundle coherence (FBC), a measure for quantification of fiber alignment. The FBC is computed from a tractography result using the same PDE framework and provides a criterion for removing the spurious fibers. We validate the proposed combination of CSD and enhancement on phantom data and on human data, acquired with different scanning protocols. On the phantom data we find that PDE enhancements improve both local metrics and global metrics of tractography results, compared to CSD without enhancements. On the human data we show that the enhancements allow for a better reconstruction of crossing fiber bundles and they reduce the variability of the tractography output with respect to the acquisition parameters. Finally, we show that both the enhancement of the FODs and the use of the FBC measure on the tractography improve the stability with respect to different stochastic realizations of probabilistic tractography. This is shown in a clinical application: the reconstruction of the optic radiation for epilepsy surgery planning.

Progeroid syndromes: models for stem cell aging?

Stem cells are responsible for tissue repair and maintenance and it is assumed that changes observed in the stem cell compartment with age underlie the concomitant decline in tissue function. Studies in murine models have highlighted the importance of intrinsic changes occurring in stem cells with age. They have also drawn the attention to other factors, such as changes in the local or systemic environment as the primary cause of stem cell dysfunction. Whilst knowledge in murine models has been advancing rapidly there has been little translation of these data to human aging. This is most likely due to the difficulties of testing the regenerative capacity of human stem cells in vivo and to substantial differences in the aging phenotype within humans. Here we summarize evidence to show how progeroid syndromes, integrated with other models, can be valuable tools in addressing questions about the role of stem cell aging in human degenerative diseases of older age and the molecular pathways involved.

A systems biology approach to Down syndrome: identification of Notch/Wnt dysregulation in a model of stem cells aging.

Stem cells are central to the development and maintenance of many tissues. This is due to their capacity for extensive proliferation and differentiation into effector cells. More recently it has been shown that the proliferative and differentiative ability of stem cells decreases with age, suggesting that this may play a role in tissue aging. Down syndrome (DS), is associated with many of the signs of premature tissue aging including T-cell deficiency, increased incidence of early Alzheimer-type, Myelodysplastic-type disease and leukaemia. Previously we have shown that both hematopoietic (HSC) and neural stem cells (NSC) in patients affected by DS showed signs of accelerated aging. In this study we tested the hypothesis that changes in gene expression in HSC and NSC of patients affected by DS reflect changes occurring in stem cells with age. The profiles of genes expressed in HSC and NSC from DS patients highlight pathways associated with cellular aging including a downregulation of DNA repair genes and increases in proapoptotic genes, s-phase cell cycle genes, inflammation and angiogenesis genes. Interestingly, Notch signaling was identified as a potential hub, which when deregulated may drive stem cell aging. These data suggests that DS is a valuable model to study early events in stem cell aging.

A new heavy internal tamponade in vitreoretinal surgery: an in vitro study.

AimTo study the solubility of perfluorohexyloctane (F6H8) in silicone oil (polydimethylsiloxane (PDMS) 1000) and to measure the viscosity and the specific gravity of the mixture obtained (heavy silicone oil or HSO tamponade) to define the ideal ratio of these components.MethodsThe solubility diagram of the mixture was obtained with the turbidimetric method, indicating the miscibility of F6H8 and silicone oil 1000 at all the useful temperatures. The viscosity was measured in steady shear conditions by using a controlled stress rheometer (Haake RS150) and a double cone/plate (DC 60/4) system, both at 25 and 37 degrees C for different volume per cent compositions of the mixture. The specific gravity was measured at 37 degrees C using a digital densimeter.ResultsA mixture of F6H8 30 v% and PDMS 70 v% was found to be transparent and stable at all the useful temperatures. By combining these proportions of the two substances, a resultant density of 1.06 g/cm(3) was obtained. The viscosity of the 30% F6H8 mixture was 203 mPa.s at 25 degrees C and 163 mPa.s at 37 degrees C respectively.ConclusionsThe ideal F6H8 and silicone oil mixture can be obtained combining 30% of F6H8 with 70% of silicone oil 1000. This mixture seems to have rheological properties useful for its use as an alternative intraocular heavy tamponade.Eye (2008) 22, 1082-1088; doi:10.1038/eye.2008.144; published online 6 June 2008.

Viabilidad de huevos de helmintos en biosólidos

El trabajo consistió en determinar el tiempo de almacenamiento (deshidratación natural en zona de clima templado) necesario para que los lodos acumulados en las lagunas de sedimentación, pudiesen ser utilizados en la agricultura sin riesgo sanitario, para lo cual se estudio la viabilidad de los huevos de helmintos a través del tiempo. Dado que la concentración de huevos de helmintos parásitos en los lodos de la localidad de Alcorta, prov. de Santa Fe, fue baja (1-2 huevos/g MS) se decidio adicionar huevos de Ascaris artificialmente, para estudiar su viabilidad

Viabilidad de huevos de helmintos en biosólidos

El trabajo consistió en determinar el tiempo de almacenamiento (deshidratación natural en zona de clima templado) necesario para que los lodos acumulados en las lagunas de sedimentación, pudiesen ser utilizados en la agricultura sin riesgo sanitario, para lo cual se estudio la viabilidad de los huevos de helmintos a través del tiempo. Dado que la concentración de huevos de helmintos parásitos en los lodos de la localidad de Alcorta, prov. de Santa Fe, fue baja (1-2 huevos/g MS) se decidio adicionar huevos de Ascaris artificialmente, para estudiar su viabilidad

Intraocular lens decentration and posterior capsule opacification: anatomo-pathologic findings after implantation of AMOSI40 IOLS.

PURPOSE: To evaluate the incidence of intraocular lens (IOL) decentration and posterior capsule opacification (PCO) after implantation of a three-piece posterior chamber silicone IOL in a series of eyes examined postmortem. METHODS: Twenty-three pseudophakic enucleated human cadaver eyes, implanted with AMO SI40NB IOLs after phacoemulsification, were analyzed. Eyes obtained postmortem were sectioned at the equatorial plane and the anterior segment photographed from a posterior view. Location of IOL optic and haptics, type of fixation, and centration of IOL was evaluated. PCO was graded and the presence of Nd:YAG laser posterior capsulotomy was noted. RESULTS: Mean age at the time of surgery was 77.83 years, mean time since implantation was 18.26 months. In all the eyes examined, IOL haptics were positioned in the capsular bag. Mean decentration was 0.20+/-0.16 mm. No correlation was found between IOL decentration and time since implantation. The degree of peripheral PCO ranged from none (13.0%) to mild (39.1%) to moderate (26.1%) to severe (21.7%). The degree of central PCO ranged from none (52.2%) to mild (30.4%) to moderate (4.3%). Three patients (13.0%) underwent Nd:YAG laser posterior capsulotomy. CONCLUSIONS: A very good centration can be obtained when silicone AMOSI40NB IOLs are correctly implanted with the haptics inside the capsular bag. About half of the implants showed no central PCO while Nd:YAG laser posterior capsulotomy rates documented a relatively low PCO 18 months after surgery. A careful in the bag haptics placement is needed in order to reduce the IOL decentration and to prevent central PCO.

Investigating helminth eggs and Salmonella sp. in stabilization ponds treating septage.

Sludge management arises as a relevant problem after being accumulated in primary ponds of septage treatment plants. One of the most attractive options for sludge disposal is its use in agriculture and then specific guidelines regarding hygienic quality must be fulfilled. This study aimed at evaluating the storage time needed to inactivate Ascaris eggs and Salmonella in sludge accumulated in a primary pond treating septage. Raw septage exhibited very low concentrations of viable Ascaris eggs, thus experiments with Ascaris suum eggs spiking were conducted. The concentration of Ascaris eggs in the solids accumulated at the bottom of the pond was 20 eggs/g of total solids (g TS) at the time of pond closure. Although it decreased, some eggs remained viable (0.59 mean viable eggs/g TS) up to 20 months of in-pond storage of the biosolids. Salmonella survival was studied after developing an analytical method that inhibited the native flora. Sludge was seeded with Salmonella enteritidis. An equation adequately describing Salmonella die-off in biosolids subjected to 115 days of in-pond storage/dewatering, was found to be represented by the regression: y = log MPN Salmonella/g TS = 6.67 x t(-0.086), with t = storage time elapsed in days. The initial concentration was 7.0 x 10(6) MPN/g TS and the removal efficiency was 99%.

Isolation of Salmonella sp. in sludge from septage treatment plant.

Waste stabilization ponds (WSP) are an often-used option to treat faecal sludges collected from on-site sanitation systems. Since agricultural use is one of the most attractive options for sludge disposal, specific guidelines on the hygienic sludge quality must be fulfilled, such as for viable helminth eggs and Salmonella sp. Although Salmonella isolation methods are well known for other types of samples, they are not suitable for faecal sludge. The reason can be attributed to the co-existence of a native bacterial sludge flora masking Salmonella development, especially if this bacteria is present at low concentrations. In order to select the best methodology for Salmonella recovery from septage sludge, different culture media were assayed at different incubation periods and temperatures. The proposed methodology for Salmonella recovery from sludge can be summarised as follows: (1) enrichment in Rappaport-Vassiliadis broth at 43 degrees C, 48 hours, and (2) isolation in XLD agar at 40 degrees C, 24 hours. Identification of suspected colonies by biochemical tests: TSI, LIA, urease and serological confirmation with Group O Antigen.

Remoción biológica de hierro y manganeso, una tecnología apropiada

El estudio tuvo como objetivo el desarrollo de un proceso adecuado de remoción de hierro y manganeso en aguas subterráneas, para ser usado en el diseño de plantas para pequeñas y medianas poblaciones. El sistema estudiado comprende las siguientes etapas: aeración, prefiltración ascendente en mantos de grava, y filtración lenta en arena. Los ensayos en plantas piloto utilizaron dos calidades de aguas subterráneas, y en pruebas previas a los mismos se detectó la presencia de bacterias del hierro en agua cruda. Los ensayos demostraron que la prefiltración juega un rol preponderante en la línea de tratamiento

The challenge of faecal sludge management in urban areas--strategies, regulations and treatment options.

In urban centres of industrialising countries, the majority of houses are served by on-site sanitation systems such as septic tanks and unsewered toilets. The faecal sludges (FS) collected from these systems are usually discharged untreated into the urban and peri-urban environment, posing great risks to water resources and to public health. Contrary to wastewater management, the development of strategies to cope with faecal sludges, adapted to the conditions prevailing in developing countries, have long been neglected. The authors describe the current situation and discuss selected issues of FS management. A proposal is made for a rational setting of sludge quality or treatment standards in economically emerging countries. The authors stipulate that regulatory setting should take into account local economic, institutional and technical conditions. Defining suitable treatment options as critical control points in securing adequate sludge quality is better than setting and relying on numerical sludge quality standards. A separate section is devoted to the practice and to regulatory aspects of (faecal) sludge use in Argentina. An overview of treatment options, which may prove sustainable in less industrialized countries is provided. Planted sludge drying beds are one of these options. It has been piloted in Thailand for four years and details on its performance and operation are presented along with data on the hygienic quality of treated biosolids.

Cotreatment of sewage and septage in waste stabilization ponds.

A one year study was carried out in a waste stabilization ponds system where septage and sewage are cotreated. The system consists of two septage ponds which operate alternately followed by two ponds in series which receive the combined effluent. The septage ponds also act as evaporation ponds for the accumulated sludge. The monitoring program was divided in two phases. The results of the first phase indicate that the effluent of septage ponds has an adequate quality to be discharged into the waste stabilization ponds designed to treat sewage and that is possible to use the septage ponds to dry the accumulated sludge. Further investigation is needed to find suitable post-treatment of the sludge in order to use it in agriculture.

Comparative evaluation of corneal epithelial permeability after the use of diclofenac 0.1% and flurbiprofen 0.03% after phacoemulsification.

PURPOSE: To assess the corneal epithelial function after prolonged topical administration of diclofenac 0.1% and flurbiprofen 0.03% single-dose eyedrops. SETTING: University Eye Clinic of Trieste, Trieste, Italy. METHODS: This randomized prospective study comprised 24 patients scheduled for phacoemulsification. The patients were randomly assigned to receive diclofenac or flurbiprofen eyedrops for 2 months after surgery. Corneal epithelial permeability was determined by fluorophotometry 7, 37, and 67 days after surgery. RESULTS: An increase in corneal epithelial permeability was observed in the diclofenac group 37 and 67 days after surgery. No epithelial function alterations occurred in the flurbiprofen group. CONCLUSIONS: Subclinical impairment of the epithelial function was observed during topical treatment with diclofenac 0.1% single-dose eyedrops after phacoemulsification. The mechanism responsible for this effect remains unknown.

Failure of urinary trans,trans-muconic acid as a biomarker for indoor environmental benzene exposure at PPB levels.

Benzene is a widespread pollutant whose main source in the environment is automotive emission. There is increasing interest in the exposure of the population to this pollutant as benzene is present also in the indoor environment due to cigarette smoke, drinking water, and food. The aim of this study was to evaluate, in an adult nonsmoking population not occupationally exposed to benzene, whether it is possible to detect differences in the urinary concentration of trans,trans-muconic acid (t,t-MA) between low and high environmental exposure to benzene. A study sample of 31 employees working in pharmacies in a large town in Italy with low environmental exposure to benzene (4.8 microg/m3) was compared to a high (8.1 microg/m3) benzene exposure group. Analysis of urinary t,t-MA was carried out by high-performance liquid chromatography (HPLC; photodiode array detector); analysis of environmental benzene samples was by gas chromatography/mass spectroscopy (GC-MS). The statistical analysis revealed no significant differences in urinary levels of t,t-MA of subjects with high (mean concentration: 157.9 microg/g creatinine) versus low exposure (mean concentration: 114.2 microg/g creatinine). Data show that it is difficult to correlate urinary t,t-MA with benzene exposure at parts per billion levels.