A novel method for assessment of fragmentation and beam-material interactions in helium ion radiotherapy with a miniaturized setup.

Radiotherapy with protons and carbon ions enables to deliver dose distributions of high conformation to the target. Treatment with helium ions has been suggested due to their physical and biological advantages. A reliable benchmarking of the employed physics models with experimental data is required for treatment planning. However, experimental data for helium interactions is limited, in part due to the complexity and large size of conventional experimental setups. We present a novel method for the investigation of helium interactions with matter using miniaturized instrumentation based on highly integrated pixel detectors. The versatile setup consisted of a monitoring detector in front of the PMMA phantom of varying thickness and a detector stack for investigation of outgoing particles. The ion type downstream from the phantom was determined by high-resolution pattern recognition analysis of the single particle signals in the pixelated detectors. The fractions of helium and hydrogen ions behind the used targets were determined. As expected for the stable helium nucleus, only a minor decrease of the primary ion fluence along the target depth was found. E.g. the detected fraction of hydrogen ions on axis of a 220MeV/u 4He beam was below 6% behind 24.5cm of PMMA. Monte-Carlo simulations using Geant4 reproduce the experimental data on helium attenuation and yield of helium fragments qualitatively, but significant deviations were found for some combinations of target thickness and beam energy. The presented method is promising to contribute to the reduction of the uncertainty of treatment planning for helium ion radiotherapy.

Locomotor training through a novel robotic platform for gait rehabilitation in pediatric population: short report.

BACKGROUND: Cerebral Palsy (CP) is a disorder of posture and movement due to a defect in the immature brain. The use of robotic devices as alternative treatment to improve the gait function in patients with CP has increased. Nevertheless, current gait trainers are focused on controlling complete joint trajectories, avoiding postural control and the adaptation of the therapy to a specific patient. This paper presents the applicability of a new robotic platform called CPWalker in children with spastic diplegia. FINDINGS: CPWalker consists of a smart walker with body weight and autonomous locomotion support and an exoskeleton for joint motion support. Likewise, CPWalker enables strategies to improve postural control during walking. The integrated robotic platform provides means for testing novel gait rehabilitation therapies in subjects with CP and similar motor disorders. Patient-tailored therapies were programmed in the device for its evaluation in three children with spastic diplegia for 5 weeks. After ten sessions of personalized training with CPWalker, the children improved the mean velocity (51.94 ± 41.97 %), cadence (29.19 ± 33.36 %) and step length (26.49 ± 19.58 %) in each leg. Post-3D gait assessments provided kinematic outcomes closer to normal values than Pre-3D assessments. CONCLUSIONS: The results show the potential of the novel robotic platform to serve as a rehabilitation tool. The autonomous locomotion and impedance control enhanced the children's participation during therapies. Moreover, participants' postural control was substantially improved, which indicates the usefulness of the approach based on promoting the patient's trunk control while the locomotion therapy is executed. Although results are promising, further studies with bigger sample size are required.

An anthropomorphic multimodality (CT/MRI) head phantom prototype for end-to-end tests in ion radiotherapy.

With the increasing complexity of external beam therapy "end-to-end" tests are intended to cover every step from therapy planning through to follow-up in order to fulfill the higher demands on quality assurance. As magnetic resonance imaging (MRI) has become an important part of the treatment process, established phantoms such as the Alderson head cannot fully be used for those tests and novel phantoms have to be developed. Here, we present a feasibility study of a customizable multimodality head phantom. It is initially intended for ion radiotherapy but may also be used in photon therapy. As basis for the anthropomorphic head shape we have used a set of patient computed tomography (CT) images. The phantom recipient consisting of epoxy resin was produced by using a 3D printer. It includes a nasal air cavity, a cranial bone surrogate (based on dipotassium phosphate), a brain surrogate (based on agarose gel), and a surrogate for cerebrospinal fluid (based on distilled water). Furthermore, a volume filled with normoxic dosimetric gel mimicked a tumor. The entire workflow of a proton therapy could be successfully applied to the phantom. CT measurements revealed CT numbers agreeing with reference values for all surrogates in the range from 2 HU to 978 HU (120 kV). MRI showed the desired contrasts between the different phantom materials especially in T2-weighted images (except for the bone surrogate). T2-weighted readout of the polymerization gel dosimeter allowed approximate range verification.

Developing a competence-based addiction medicine curriculum in Indonesia: the training needs assessment.

Indonesia has one of the fastest growing, injecting drugs user-driven, human immunodeficiency virus (HIV) epidemics in Asia. Coverage of needle and syringe programs (NSPs), opioid substitution therapy (OST), and antiretroviral treatment (ART) is increasing, but is still low, whereas professional training in addiction medicine is not yet established. Urgent development and scaling-up of professional capacity in comprehensive, evidence-based addiction medicine is needed. In this article the results of the first step is presented, being the training needs assessment (TNA) and the process of further developing a national evidence- and competence-based addiction medicine curriculum in Indonesia.

Characterizing head motor disorders to create novel interfaces for people with cerebral palsy: creating an alternative communication channel by head motion.

This paper aims to validate a head mounted inertial interface to characterize disorder movements in people with cerebral palsy (CP). The kinematic patterns extracted from this study will be used to design an alternative communication channel (using head motion) adapted to user's capabilities and limitations. Four people with CP participated (GMFCS level V) and three healthy subjects as reference group. The main outcome measures were divided into 1) Time-domain, 2) Frequency-domain and 3) Spatial domain. Results showed that the inertial interface succeeds assessing the pathological motion. Firstly, the system differentiates between voluntary and involuntary motion in terms of motor control, frequency and range of motion. Secondly some motion disorders such as hypertonia, hypotonia can be identified. These results suggest that people with motor disorders could benefit from the developed inertial system in three fields: 1) diagnosis of motor disorder by means of an objective quantification, 2) physical and cognitive rehabilitation by means of proprioceptive enhancement through visual-motor feedback and 3) functional compensation by means of an inertial person-machine interface for controlling computer and assistive devices (e.g. wheelchairs or walkers).

Sensitivity of capsular-producing Streptococcus thermophilus strains to bacteriophage adsorption.

AIMS: To determine whether the presence and type of exopolysaccharides (EPS), slime-EPS or capsular, and the structural characteristics of the polymers produced by Streptococcus thermophilus strains could interfere with or be involved in phage adsorption. METHODS AND RESULTS: Phage-host interactions between eight EPS-producing Strep. thermophilus strains (CRL419, 638, 804, 810, 815, 817, 821, 1190) and five streptococcus specific phages (phiYsca, phi3, phi5, phi6, phi8) isolated from Argentinean faulty fermentation failed yoghurts were evaluated. No relationship was found between the EPS chemical composition and the phage sensitivity/resistance phenotype. In general, the capsular-producing strains were more sensitive to phage attacks than the noncapsular-producing strains. Streptococcus thermophilus CRL1190 (capsular-producing) was the only strain sensitive to all bacteriophages and showed the highest efficiency of plating. Phage adsorption to a capsular-negative, EPS low-producing mutant of strain CRL1190 was reduced, especially for phiYcsa and phi8. CONCLUSIONS: The presence of capsular polysaccharide surrounding the cells of Strep. thermophilus strains could play a role in the adsorption of specific phages to the cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Capsular-producing Strep. thermophilus strains should be evaluated for their bacteriophage sensitivity if they are included in starter cultures for the fermented food industry.

Prevalence of Escherichia coli O157 and O157:H7-infecting bacteriophages in feedlot cattle feces.

AIM: To estimate the distribution and prevalence of both Escherichia coli O157 and O157:H7-infecting bacteriophages within a 50,000 head commercial beef feedlot. METHODS AND RESULTS: Escherichia coli O157 was detected in approximately 27% of the individual samples, distributed across seven of the 10 pens screened. In a simple initial screen to detect O157:H7-infecting phages, none were detected in any pen or individual sample. In contrast, after a series of enrichment procedures O157:H7-infecting phages were detected in every pen and in the majority of the samples from most pens; virulent bacteriophages active against E. coli O157:H7 were detected post-enrichment from 39/60 (65%) of the feedlot samples, and 58/60 (approximately 97%) contained phage that infected E. coli B or O157:H7. CONCLUSIONS: The data we present here indicates that we may be grossly underestimating the prevalence of O157:H7-infecting phages in livestock if we simply screen samples and that enrichment screening is required to truly determine the presence of phages in these ecosystems. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data suggest that O157:H7-infecting phages may play a role in the ecology and transient colonization of cattle by E. coli O157:H7. Further, this and previous data suggest that before starting in vivo pathogen eradication studies using phage or any other regime, test animals should be enrichment screened for phage to avoid erroneous results.

El conocimiento del desarrollo embrionario del corazón como una herramienta para la terapia celular cardíaca / No disponible

La regeneración del corazón dañado ha sido objeto de intensa investigación durante la década pasada. Las diferentes estrategias que han sido desarrrolladas (como por ejemplo la terapia celular basada en células madre (ES)), esclarecen la información acerca de las moléculas y los factores de transcripción involucrados en las cardiomiogénesis. Sin embargo, todavía no es posible programar eficientemente las ES para que se desarrollen a cardiomiocitos. Los mecanismos celulares y moleculares inherentes en el desarrollo embrionario del corazón, así como las interconexiones entre ellos, pueden aportar datos acerca de las rutas bioquímicas necesarias para la diferenciación de las ES embrionarias a células cardíacas. Nosotros proponemos que un modelo cuantitativo que puede servir para descifrar las elaboradas rutas involucradas en la cardiomiogénesis. Esta aproximación podría revelar la etiología de los defectos cardíacos y permitiría producir cardiomiocitos con propósitos clínicos en la regeneracíon y la toxicología entre otros (AU)

No disponible

A functional genomics approach to the mode of action of apratoxin A.

The cyanobacterial metabolite apratoxin A (1) demonstrates potent cytotoxicity against tumor cell lines by a hitherto unknown mechanism. We have used functional genomics to elucidate the molecular basis for this activity. Gene expression profiling and DNA content analysis showed that apratoxin A induces G1-phase cell cycle arrest and apoptosis. Cell-based functional assays with a genome-wide collection of expression cDNAs showed that ectopic induction of fibroblast growth factor receptor (FGFR) signaling attenuates the apoptotic activity of apratoxin A. This natural product inhibited phosphorylation and activation of STAT3, a downstream effector of FGFR signaling. It also caused defects in FGF-dependent processes during zebrafish development, with concomitant reductions in expression levels of the FGF target gene mkp3. We conclude that apratoxin A mediates its antiproliferative activity through the induction of G1 cell cycle arrest and an apoptotic cascade, which is at least partially initiated through antagonism of FGF signaling via STAT3.

Retinoic acid signalling links left-right asymmetric patterning and bilaterally symmetric somitogenesis in the zebrafish embryo.

During embryogenesis, cells are spatially patterned as a result of highly coordinated and stereotyped morphogenetic events. In the vertebrate embryo, information on laterality is conveyed to the node, and subsequently to the lateral plate mesoderm, by a complex cascade of epigenetic and genetic events, eventually leading to a left-right asymmetric body plan. At the same time, the paraxial mesoderm is patterned along the anterior-posterior axis in metameric units, or somites, in a bilaterally symmetric fashion. Here we characterize a cascade of laterality information in the zebrafish embryo and show that blocking the early steps of this cascade (before it reaches the lateral plate mesoderm) results in random left-right asymmetric somitogenesis. We also uncover a mechanism mediated by retinoic acid signalling that is crucial in buffering the influence of the flow of laterality information on the left-right progression of somite formation, and thus in ensuring bilaterally symmetric somitogenesis.

Activation of Notch signaling pathway precedes heart regeneration in zebrafish.

Several vertebrates display the ability to regenerate parts of their body after amputation. During this process, differentiated cells reenter the cell cycle and proliferate to generate a mass of undifferentiated cells. Repatterning mechanisms act on these cells to eventually shape a regenerated tissue or organ that replaces the amputated one. Experiments with regenerating limbs/fins in newts and zebrafish have shown that members of the Msx family of homeodomain-containing transcription factors play key roles during blastema formation and patterning. Here we show that adult zebrafish have a remarkable capacity to regenerate the heart in a process that involves up-regulation of msxB and msxC genes. We present evidence indicating that heart regeneration involves the execution of a specific genetic program, rather than redeployment of a cardiac development program. Preceding Msx activation, there is a marked increase in the expression of notch1b and deltaC, which we show are also up-regulated during fin regeneration. These data suggest a role for the Notch pathway in the activation of the regenerative response. Taken together, our results underscore the use of zebrafish as a model for investigating the process of regeneration in particular and the biology of stem cells in general. Advances in these fields will undoubtedly aid in the implementation of strategies for regenerative medicine.

Notch activity induces Nodal expression and mediates the establishment of left-right asymmetry in vertebrate embryos.

Left-sided expression of Nodal in the lateral plate mesoderm is a conserved feature necessary for the establishment of normal left-right asymmetry during vertebrate embryogenesis. By using gain- and loss-of-function experiments in zebrafish and mouse, we show that the activity of the Notch pathway is necessary and sufficient for Nodal expression around the node, and for proper left-right determination. We identify Notch-responsive elements in the Nodal promoter, and unveil a direct relationship between Notch activity and Nodal expression around the node. Our findings provide evidence for a mechanism involving Notch activity that translates an initial symmetry-breaking event into asymmetric gene expression.

[Variables related to the first presbyopia correction]. / Edad de la primera compensación de la presbicia, factores asociados.

PURPOSE: We carried out a prospective study based on variables related to the age of appearance of presbyopia (AAP): refraction defect, sex, iris color, profession and pupil diameter. METHOD: Our sample population was made up of 179 persons who present consecutively for the first optical correction of presbyopia as sole reason, having a refractive defect equal to 1 diopter or less. RESULTS: In our series the AAP was not influenced by the small refraction defect, neither by sex or iris color. Different professions showed a significant AAP difference later in life (farmers, x = 51.76 D.E. 5.47 years) or earlier (services workers, x = 46.15 D.E. 3.55 years) (p<0.000003, ANOVA), than other professions such as housewife (48.42 D.E. 4.71 years) and construction workers (47.54 D.E. 3.93 years). We found significant differences in AAP among subjects with intense miosis (<2,5 mm, x = 51 D.E. 5.39 years) and without miosis (3 to 7 mm, x = 47.47 D.E. 4.71 years) (p<0.02 t Student). CONCLUSIONS: The variables which determine a significant delay in the AAP are: profession (farmer and housewife) and miosis, a 1 mm pupilar diameter reduction, in near vision, could contribute to a delay of 1.53 years in AAP.

Edad de la primera compensación de la presbicia, factores asociados / Variables related to the first presbyopia correction

Objetivo: Hemos realizado un estudio prospectivo sobre aquellas variables relacionadas con la edad de aparición de la presbicia (AAP): defecto de refracción, sexo, color del iris, profesión y diámetro pupilar. Métodos: Nuestra muestra poblacional consta de 179 personas que acudían consecutivamente demandando su primera corrección óptica de presbicia, como único motivo, presentando un defecto de refracción igual o inferior a 1 dioptría. Resultados: En nuestra serie la AAP no estaba influenciada por el pequeño defecto de refracción, ni por el sexo, ni por el color del iris. Las diferentes profesiones demostraban una diferencia significativa en la media de AAP, más tardía (agricultores, x= 51,76 D.E. 5,47 años), o más temprana (trabajadores del sector Servicios, x= 46,15 D.E. 3,55 años) (p<0,000003, ANOVA), que otras profesiones tales como amas de casa (48,42 D.E. 4,71 años) y trabajadores de la construcción (47,54 D.E. 3,93 años).Nosotros hemos encontrado diferencias significativas en la AAP entre personas con intensa miosis (<2,5 mm, x=51 D.E. 5,39 años) y pacientes sin miosis (3 a 7 mm, x= 47,47 D.E. 4,71 años) (p<0,02, t a Student 0,05).Conclusiones: Las variables que determinan un retraso significativo en la AAP son: la profesión (agricultores y amas de casa) y la miosis, 1 mm de reducción del diámetro pupilar, en la visión de cerca, podía contribuir en un retraso en la AAP de 1,53 años (AU)

Selection of bacteriocin producer strains of lactic acid bacteria from a dairy environment.

Two strains showing bacteriocin production were selected from a total of 206 lactic acid bacteria isolated from samples of milk, milk serum, whey and homemade cheeses in Southern Cordoba, Argentina. This property was detected by means of well diffusion assays. The strains were identified as Enterococcus hirae and Enterococcus durans. The protein nature of those substances was proved by showing their sensitivity to type IV and XXV proteases, papaine, trypsin, pepsin and K proteinase. The bacteriocins inhibited the growth of Listeria monocytogenes, Bacillus cereus, Clostridium perfringes and two strains of Staphylococcus aureus, an A-enterotoxin and a B-enterotoxin producers. All of these bacteria are common pathogens usually associated with food borne diseases (ETA). These lactic acid bacteria or their bacteriocins could be suitable candidates for food preservation and specially useful in the our regional dairy industry.

Lactic acid bacteria and their effect on the immune system.

Lactic acid bacteria (LAB) are present in the intestine of most animals. The beneficial role played by these microorganisms in the humans and other animals, including the effect on the immune system, has been extensively reported. They are present in many foods and are frequently used as probiotics to improve some biological functions in the host. The activation of the systemic and secretory immune response by LAB requires many complex interactions among the different constituents of the intestinal ecosystem (microflora, epithelial cells and immune cells). Through different mechanisms they send signals to activate immune cells. Thus the knowledge of the normal intestinal microflora, the contribution of LAB and their role in the numerous functions in the digestive tract as well as the functioning of the mucosal immune system form the basis for the study and selection of a probiotic strain with immunostimulatory properties. In the selection of LAB by their immunostimulatory capacity it helps to know not only the effect which they have on the mucosal immune system, but the specific use to which these oral vaccine vectors are being put. Although there are reports of the protection of animals and humans against diseases such as microbial infections and cancer, more work remains to be done on the factors affecting the design of oral vaccine vectors and the use of LAB for therapeutic purposes. The basic knowledge of LAB immunostimulation and the criteria for selection of LAB by their immunostimulatory capacity, will be extensively discussed and appraised in this review.

Isolation and characterization of a slowly milk-coagulating variant of Lactobacillus helveticus deficient in purine biosynthesis.

A slowly milk-coagulating variant (Fmc(-)) of Lactobacillus helveticus CRL 1062, designated S1, was isolated and characterized. Strain S1 possessed all the known essential components required to utilize casein as a nitrogen source, which include functional proteinase and peptidase activities as well as functional amino acid, di- and tripeptide, and oligopeptide transport systems. The amino acid requirements of strain S1 were similar to those of the parental strain. However, on a purine-free, chemically defined medium, the growth rate of the Fmc(-) strain was threefold lower than that of the wild-type strain. L. helveticus S1 was found to be defective in IMP dehydrogenase activity and therefore was deficient in the ability to synthesize XMP and GMP. This conclusion was further supported by the observation that the addition of guanine or xanthine to milk, a substrate poor in purine compounds, restored the Fmc(+) phenotype of L. helveticus S1.

Nutritional requirements and nitrogen-dependent regulation of proteinase activity of Lactobacillus helveticus CRL 1062.

The nutritional requirements of Lactobacillus helveticus CRL 1062 were determined with a simplified chemically defined medium (SCDM) and compared with those of L. helveticus CRL 974 (ATCC 15009). Both strains were found to be prototrophic for alanine, glycine, asparagine, glutamine, and cysteine. In addition, CRL 1062 also showed prototrophy for lysine and serine. The microorganisms also required riboflavin, calcium pantothenate, pyridoxal, nicotinic acid, and uracil for growth in liquid SCDM. The growth rate and the synthesis of their cell membrane-bound serine proteinases, but not of their intracellular leucyl-aminopeptidases, were influenced by the peptide content of the medium. The highest proteinase levels were found during cell growth in basal SCDM, while the synthesis of this enzyme was inhibited in SCDM supplemented with Casitone, Casamino Acids, or beta-casein. Low-molecular-mass peptides (<3,000 Da), extracted from Casitone, and the dipeptide leucylproline (final concentration, 5 mM) play important roles in the medium-dependent regulation of proteinase activity. The addition of the dipeptide leucylproline (5 mM) to SCDM reduced proteinase activity by 25%.

Characterization of natural isolates of Lactobacillus strains to be used as starter cultures in dairy fermentation.

The technological relevant characteristics of five homofermentative lactobacilli strains, isolated from natural fermented hard cheeses, were studied. Isolates CRL 581 and CRL 654, from Argentinian artesanal hard cheeses, and isolates CRL 1177, CRL 1178, and CRL 1179, from Italian Grana cheeses, were identified as Lactobacillus delbrueckii subsp. lactis and Lactobacillus helveticus, respectively, by physiological and biochemical tests, SDS-PAGE of whole-cell proteins and sequencing of the variable (V1) region of the 16S ribosomal DNA. All strains showed high levels of beta-galactosidase activity. However, proteolytic activity varied widely among isolates. Strains CRL 581, CRL 654, and CRL 1177 hydrolyzed alpha- and beta-caseins and were able to coagulate reconstituted skim milk in less than 16 h at 42 degrees C. According to the substrate specificity, these proteinases have a caseinolytic activity comparable to that of the P(III)-type of lactococcal proteinases. No strains produced inhibitor substances (bacteriocin) and all were insensitive to attack by 14 L. helveticus- and L. delbrueckii subsp. lactis-specific bacteriophages.

Deoxyribonuclease activities in Lactobacillus delbrueckii.

DNase activity was examined in the extracellular and subcellular fractions of six non-transformable strains belonging to Lactobacillus delbrueckii subsp. lactis (L. lactis) and Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) and compared with the activity present in Lactobacillus johnsonii NCK 65, a transformable strain of Lactobacillus. In the extracellular fraction of the L. delbrueckii strains, a common protein band of 36 kDa was detected, while a band of 29 kDa was found in the same fraction of L. johnsonii. No nuclease activity was detected in the cytoplasmic fraction of this strain, indicating that the localization of the DNase activity could be a key factor in the uptake of foreign DNA.