Novel paradigm enables accurate monthly gestational screening to prevent congenital toxoplasmosis and more.

Background: Congenital toxoplasmosis is a treatable, preventable disease, but untreated causes death, prematurity, loss of sight, cognition and motor function, and substantial costs worldwide. Methods/Findings: In our ongoing USA feasibility/efficacy clinical trial, data collated with other ongoing and earlier published results proved high performance of an Immunochromatographic-test(ICT) that enables accurate, rapid diagnosis/treatment, establishing new paradigms for care. Overall results from patient blood and/or serum samples tested with ICT compared with gold-standard-predicate-test results found ICT performance for 4606 sera/1876 blood, 99.3%/97.5% sensitive and 98.9%/99.7% specific. However, in the clinical trial the FDA-cleared-predicate test initially caused practical, costly problems due to false-positive-IgM results. For 58 persons, 3/43 seronegative and 2/15 chronically infected persons had false positive IgM predicate tests. This caused substantial anxiety, concerns, and required costly, delayed confirmation in reference centers. Absence of false positive ICT results contributes to solutions: Lyon and Paris France and USA Reference laboratories frequently receive sera with erroneously positive local laboratory IgM results impeding patient care. Therefore, thirty-two such sera referred to Lyon's Reference laboratory were ICT-tested. We collated these with other earlier/ongoing results: 132 of 137 USA or French persons had false positive local laboratory IgM results identified correctly as negative by ICT. Five false positive ICT results in Tunisia and Marseille, France, emphasize need to confirm positive ICT results with Sabin-Feldman-Dye-test or western blot. Separate studies demonstrated high performance in detecting acute infections, meeting FDA, CLIA, WHO ASSURED, CEMark criteria and patient and physician satisfaction with monthly-gestational-ICT-screening. Conclusions/Significance: This novel paradigm using ICT identifies likely false positives or raises suspicion that a result is truly positive, rapidly needing prompt follow up and treatment. Thus, ICT enables well-accepted gestational screening programs that facilitate rapid treatment saving lives, sight, cognition and motor function. This reduces anxiety, delays, work, and cost at point-of-care and clinical laboratories. Author's Summary: Toxoplasmosis is a major health burden for developed and developing countries, causing damage to eyes and brain, loss of life and substantial societal costs. Prompt diagnosis in gestational screening programs enables treatment, thereby relieving suffering, and leading to > 14-fold cost savings for care. Herein, we demonstrate that using an ICT that meets WHO ASSURED-criteria identifying persons with/without antibody to Toxoplasma gondii in sera and whole blood with high sensitivity and specificity, is feasible to use in USA clinical practice. We find this new approach can help to obviate the problem of detection of false positive anti- T.gondii IgM results for those without IgG antibodies to T.gondii when this occurs in present, standard of care, predicate USA FDA cleared available assays. Thus, this accurate test facilitates gestational screening programs and a global initiative to diagnose and thereby prevent and treat T.gondii infection. This minimizes likelihood of false positives (IgG and/or IgM) while maintaining maximum sensitivity. When isolated IgM antibodies are detected, it is necessary to confirm and when indicated continue follow up testing in ∼2 weeks to establish seroconversion. Presence of a positive ICT makes it likely that IgM is truly positive and a negative ICT makes it likely that IgM will be a false positive without infection. These results create a new, enthusiastically-accepted, precise paradigm for rapid diagnosis and validation of results with a second-line test. This helps eliminate alarm and anxiety about false-positive results, while expediting needed treatment for true positive results and providing back up distinguishing false positive tests.

Netting and pan traps fail to identify the pollinator guild of an agricultural crop.

Pollinator decline is expected to cause significant reductions in food production and plant reproduction. Quantifying the impact of pollinator decline on food production requires survey methods that can identify insect and bee species responsible for pollination of specific crops. To address this issue, we compared the effectiveness of two survey methods, netting and pan traps, at capturing the pollinators of alfalfa, Medicago sativa. Alfalfa is a major component of forage for cows and an important ingredient in chicken feed. We also examined bee species richness and diversity with these two survey methods, and compared these measures among three different colors of pan traps. Netting was more effective at capturing known pollinators of alfalfa, especially those belonging to the Bombus and Apis genera. Pan traps captured a higher bee diversity relative to netting and, like previous studies, each survey method and each trap color was more efficient at capturing certain bee genera. However, without a priori knowledge of pollinators, neither survey method could identify which of the bee species captured could pollinate alfalfa. We therefore recommend direct observations when the goal of a study is to identify pollinators or link pollinator decline to food production.

Hot Rocks and Not-So-Hot Rocks on the Seashore: Patterns and Body-Size Dependent Consequences of Microclimatic Variation in Intertidal Zone Boulder Habitat.

Microclimatic variation has emerged as an important driver of many ecological and evolutionary processes. Nonetheless, fine-scale temperature data are still rare in most habitats, limiting our ability to understand the consequences of microclimatic variation under current and future conditions. We measured fine-scale thermal variation in a common, species-rich, but rarely studied habitat with respect to temperature: the airspaces under rocks on intertidal zone boulder shores. The effects of thermal variation were investigated using physiological, behavioral, and demographic responses of the porcelain crab Petrolisthes cinctipes. Habitat temperatures were measured at fine spatial and temporal resolution over 18 months, producing 424,426 temperature records. Microclimatic variation increased with increasing intertidal elevation, particularly with respect to heat extremes. However, mean temperatures were similar across the entire intertidal zone. Overheating risk for P. cinctipes increases with intertidal elevation but is size dependent, as large animals are more heat sensitive than small animals. Still, microclimatic variation high in the intertidal zone provided thermal refugia even under the warmest conditions. Size-dependent thermal responses predicted that large crabs should be rare high in the intertidal zone, which was supported by demographic data. Furthermore, simulations parameterized by our microclimate and organismal data recapitulated demographic patterns. Therefore, interactions between microclimatic variation and size-dependent thermal responses may have significant ecological repercussions that warrant greater attention.

Rocas calientes y rocas no tan calientes en la orilla del mar: patrones y consecuencias dependientes del tamaño del cuerpo de la variación microclimática en el hábitat de rocas intermareales (Hot rocks and not-so-hot rocks on the seashore: patterns and body-size dependent consequences of microclimatic variation in intertidal zone boulder habitat) La variación microclimática se ha convertido en un importante impulsor de muchos procesos ecológicos y evolutivos. No obstante, los datos de temperatura a pequeña escala aún son raros en la mayoría de los hábitats, lo que limita nuestra capacidad de comprender las consecuencias de la variación microclimática en las condiciones actuales y futuras. Medimos la variación térmica a pequeña escala en un hábitat común, rico en especies, pero rara vez estudiado con respecto a la temperatura: los espacios aéreos debajo de las rocas en las costas de rocas de zonas intermareales. Los efectos de la variación térmica se investigaron utilizando respuestas fisiológicas, conductuales y demográficas de los cangrejos de porcelana Petrolisthes cinctipes. Las temperaturas del hábitat se midieron con una resolución espacial y temporal fina durante 18 meses, produciendo 424.426 registros de temperatura. La variación microclimática aumentó con el aumento de la elevación intermareal, particularmente con respecto a los extremos de calor. Sin embargo, las temperaturas medias fueron similares en toda la zona intermareal. El riesgo de sobrecalentamiento de P. cinctipes aumenta con la elevación intermareal, pero depende del tamaño, ya que los animales grandes son más sensibles al calor que los animales pequeños. Aún así, la variación microclimática alta en la zona intermareal proporcionó refugios térmicos incluso en las condiciones más cálidas. Las respuestas térmicas dependientes del tamaño predijeron que los cangrejos grandes deberían ser raramente altos en la zona intermareal, lo que fue respaldado por datos demográficos. Además, las simulaciones parametrizadas por nuestros microclimas y datos organismales recapitularon patrones demográficos. Por lo tanto, las interacciones entre la variación microclimática y las respuestas térmicas dependientes del tamaño pueden tener importantes repercusiones ecológicas que justifican una mayor atención. Translated to Spanish by YE Jimenez (yordano_jimenez@brown.edu).

Rochas quentes e rochas não-tão-quentes à beira-mar: padrões e conseqüências dependentes do tamanho coporal para a variação microclimática no habitat de rochas da zona entremarés (Hot rocks and not-so-hot rocks on the seashore: patterns and body-size dependent consequences of microclimatic variation in intertidal zone boulder habitat) A variação microclimática emergiu como um importante causador de muitos processos ecológicos e evolutivos. No entanto, dados de temperatura em escala fina ainda são raros para a maioria dos habitats, limitando nossa capacidade de compreender as conseqüências da variação microclimática sob condições atuais e futuras. Nós medimos a variação térmica numa escala fina em um habitat comum, rico em espécies, mas raramente estudado em relação à temperatura: os espaços aéreos sob as rochas nas margens dos blocos de pedra da zona entremarés. Os efeitos da variação térmica foram investigados usando respostas fisiológicas, comportamentais e demográficas do caranguejo Petrolisthes cinctipes. As temperaturas do habitat foram medidas em resolução espacial e temporal ao longo de 18 meses, produzindo 424.426 registros de temperatura. A variação microclimática aumentou com o aumento da elevação intertidal, particularmente com relação aos extremos de calor. No entanto, as temperaturas médias foram semelhantes em toda a zona entremarés. O risco de superaquecimento de P. cinctipes aumenta com a elevação intertidal, mas é dependente do seu tamanho, já que os animais grandes são mais sensíveis ao calor do que os pequenos. Ainda assim, a variação microclimática alta na zona entremarés forneceu refúgios térmicos mesmo sob as condições mais quentes. Respostas térmicas dependentes do tamanho previam que os grandes caranguejos deveriam ser raros na zona entremarés, o que era apoiado por dados demográficos. Além disso, as simulações parametrizadas pelos nossos dados de microclima e organismal recapitularam padrões demográficos. Portanto, as interações entre a variação microclimática e as respostas térmicas dependentes do tamanho corporal podem ter repercussões ecológicas significativas que merecem maior atenção. Translated to Portuguese by G Sobral (gabisobral@gmail.com).

Adaptation of genetically monomorphic bacteria: evolution of copper resistance through multiple horizontal gene transfers of complex and versatile mobile genetic elements.

Copper-based antimicrobial compounds are widely used to control plant bacterial pathogens. Pathogens have adapted in response to this selective pressure. Xanthomonas citri pv. citri, a major citrus pathogen causing Asiatic citrus canker, was first reported to carry plasmid-encoded copper resistance in Argentina. This phenotype was conferred by the copLAB gene system. The emergence of resistant strains has since been reported in Réunion and Martinique. Using microsatellite-based genotyping and copLAB PCR, we demonstrated that the genetic structure of the copper-resistant strains from these three regions was made up of two distant clusters and varied for the detection of copLAB amplicons. In order to investigate this pattern more closely, we sequenced six copper-resistant X. citri pv. citri strains from Argentina, Martinique and Réunion, together with reference copper-resistant Xanthomonas and Stenotrophomonas strains using long-read sequencing technology. Genes involved in copper resistance were found to be strain dependent with the novel identification in X. citri pv. citri of copABCD and a cus heavy metal efflux resistance-nodulation-division system. The genes providing the adaptive trait were part of a mobile genetic element similar to Tn3-like transposons and included in a conjugative plasmid. This indicates the system's great versatility. The mining of all available bacterial genomes suggested that, within the bacterial community, the spread of copper resistance associated with mobile elements and their plasmid environments was primarily restricted to the Xanthomonadaceae family.

Natural Infection of Cashew (Anacardium occidentale) by Xanthomonas citri pv. mangiferaeindicae in Burkina Faso.

Xanthomonas citri pv. mangiferaeindicae is the causal agent of bacterial canker of mango (Mangifera indica, Anacardiaceae), a disease of international importance. Since the original description of the bacterium in the 1940s, the status of cashew (Anacardium occidentale, Anacardiaceae) as a host species has been unclear. Here, we report the first outbreak of a cashew bacterial disease in Burkina Faso (Western Africa) where X. citri pv. mangiferaeindicae recently emerged on mango. A comprehensive molecular characterization, based on multilocus sequence analysis, supplemented with pathogenicity assays of isolates obtained during the outbreak, indicated that the causal agent on cashew in Burkina Faso is X. citri pv. mangiferaeindicae and not X. citri pv. anacardii, which was previously reported as the causal agent of a cashew bacterial leaf spot in Brazil. Pathogenicity data supported by population biology in Burkina Faso suggest a lack of host specialization. Therefore, the inoculum from each crop is potentially harmful to both host species. Symptoms induced on cashew leaves and fruit by X. citri pv. mangiferaeindicae and nonpigmented strains of X. citri pv. anacardii are similar, although the causative bacteria are genetically different. Thus, xanthomonads pathogenic on cashew may represent a new example of pathological convergence in this bacterial genus.

Bridgehead invasion of a monomorphic plant pathogenic bacterium: Xanthomonas citri pv. citri, an emerging citrus pathogen in Mali and Burkina Faso.

Molecular epidemiology studies further our understanding of migrations of phytopathogenic bacteria, the major determining factor in their emergence. Asiatic citrus canker, caused by Xanthomonas citri pv. citri, was recently reported in Mali and Burkina Faso, a region remote from other contaminated areas. To identify the origin and pathways of these emergences, we used two sets of markers, minisatellites and microsatellites, for investigating different evolutionary scales. Minisatellite typing suggested the introduction of two groups of strains in Mali (DAPC 1 and DAPC 2), consistent with microsatellite typing. DAPC 2 was restricted to Bamako district, whereas DAPC 1 strains were found much more invasive. The latter strains formed a major clonal complex based on microsatellite data with the primary and secondary founders detected in commercial citrus nurseries and orchards. This suggests that human activities played a major role in the spread of DAPC 1 strains via the movement of contaminated propagative material, further supported by the frequent lack of differentiation between populations from geographically distant nurseries and orchards. Approximate Bayesian Computation analyses supported the hypothesis that strains from Burkina Faso resulted from a bridgehead invasion from Mali. Multi-locus variable number of tandem repeat analysis and Approximate Bayesian Computation are useful for understanding invasion routes and pathways of monomorphic bacterial pathogens.

The regional sensitivity of chondrocyte gene expression to coactive mechanical load and exogenous TNF-α stimuli.

Both mechanical load and elevated levels of proinflammatory cytokines have been associated with the risk for developing osteoarthritis (OA), yet the potential interaction of these mechanical and biological factors is not well understood. The purpose of this study was to evaluate the response of chondrocytes to the effects of dynamic unconfined compression, TNF-α, and the simultaneous effects of dynamic unconfined compression and TNF-α. The response to these three treatments was markedly different and, taken together, the response in the gene expression of chondrocytes to the different treatment conditions suggest a complex interaction between structure, biology, and mechanical loading.

First Report of Xanthomonas citri pv. mangiferaeindicae Causing Mango Bacterial Canker on Mangifera indica in Ivory Coast.

Xanthomonas citri pv. mangiferaeindicae causing bacterial canker (or black spot) is a major mango (Mangifera indica L.) pathogen in tropical and subtropical areas (3). The bacterium infects a wide range of mango cultivars, and induces raised, angular, black leaf lesions, sometimes with a yellow chlorotic halo. Fruit symptoms first appear as small water-soaked spots on the lenticels turning into star-shaped, erumpent lesions, which exude an infectious gum, yielding tear-stain patterns. Severe infections cause severe defoliation and/or premature fruit drop. Twig cankers are potential sources of inoculum and weaken branch resistance to winds. Drastic yield losses have been reported at grove scale for susceptible cultivars (3). Mango leaves showing typical angular, black, raised leaf lesions were first observed and collected in April 2014 from trees cv. Kent in five localities of the Korhogo province of Ivory Coast (i.e., the major commercial mango-growing area in this country). Non-pigmented Xanthomonas-like colonies were isolated on KC semi-selective medium (4). Five strains (LL60-1, LL61-1, LL62-1, LL63-1, and LL64-1), one from each locality, were compared by multilocus sequence analysis (MLSA) to the type strain of X. citri and the pathotype strain of several X. citri pathovars, including pvs. anacardii and mangiferaeindicae. This assay targeted the atpD, dnaK, efp, and gyrB genes, as described previously (2). Nucleotide sequences were 100% identical to those of the pathotype strain of X. citri pv. mangiferaeindicae whatever the gene assayed, but differed from any other assayed X. citri pathovar. Leaves of mango cv. Maison Rouge from the youngest vegetative flush were infiltrated (10 inoculation sites/leaf for three replicate leaves on different plants/bacterial strain) as detailed previously (1) with the same five strains. Bacterial suspensions (~1 × 105 cfu/ml) were prepared in 10 mM Tris buffer (pH 7.2) from 16-h-old cultures on YPGA (7 g yeast, 7 g peptone, 7 g glucose, and 18 g agar/liter, pH 7.2). The negative control treatment consisted of three leaves infiltrated with sterile Tris buffer (10 sites/leaf). Plants were incubated in a growth chamber at 30 ± 1°C by day and 26 ± 1°C by night (12-h day/night cycle) at 80 ± 5% RH. All leaves inoculated with the strains from Ivory Coast showed typical symptoms of bacterial canker a week after inoculation. No lesions were recorded from the negative controls. The pathogen was recovered at high population densities (>1 × 106 cfu/lesion) from leaf lesions, typical of a compatible interaction (1) and isolated colonies were identified as the target by atpD sequencing (2). Koch's postulates have therefore been fully verified. This is the first report of the disease in Ivory Coast, a country which has been an internationally significant mango exporter (up to 15,000 tons per year) over the last two decades. A high disease incidence and severity were observed, outlining the need for implementing integrated pest management in mango groves and the production of disease-free nursery stock. This report further expands the distribution of the pathogen in West Africa after its first description from Ghana in 2011 (5) and subsequently in other neighboring countries. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) L. Gagnevin and O. Pruvost. Plant Dis. 85:928, 2001. (4) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (5) O. Pruvost et al. Plant Dis. 95:774, 2011.

First Report of Xanthomonas citri pv. citri Pathotype A Causing Asiatic Citrus Canker in Grande Comore and Anjouan.

The causal agent of Asiatic citrus canker, Xanthomonas citri pv. citri, is a bacterium of major economic importance in tropical and subtropical citrus-producing areas. X. citri pv. citri pathotype A can cause severe infection in a wide range of citrus species and induces erumpent, callus-like lesions with water-soaked margins evolving to corky cankers and leading to premature fruit, leaf drop, and twig dieback on susceptible cultivars. This quarantine organism can strongly impact citrus markets so it has consequently been subjected to eradication efforts and international quarantine regulations. Asiatic citrus canker occurs on most islands in the Southwest Indian Ocean region including the Mascarene and Seychelles archipelagos. In the Comoros archipelago, the disease was observed for the first time in Mohéli island in 1966 (2), but had not yet been reported in neighboring islands, Grande Comore and Anjouan. In September 2013, leaves of key lime (Citrus aurantifolia) and sweet orange (C. sinensis) showing symptoms of citrus canker were collected from Anjouan, Grande Comore, and Mohéli. Nine Xanthomonas-like strains (three from each of the three islands) were isolated using KC semi-selective medium (5) from diseased samples (LK126-3, LK127-7, LK128-2, LK131-10, LK137-1, LK141-3, LK144-5, LK145-5, LK146-2). Based on a specific PCR assay with 4/7 primers (4), all Xanthomonas-like strains were tentatively identified as X. citri pv. citri. All strains produced a 468-bp amplicon similar to X. citri pv. citri strain IAPAR 306 used as a positive control. Negative control reactions with sterile tris buffer did not produce amplicons. Multilocus sequence analysis (MLSA) targeting six housekeeping genes (atpD, dnaK, efp, gltA, gyrB, and lepA) (1,3) fully identified all strains from the Comoros as X. citri pv. citri. More specifically, eight strains were identified as sequence type ST2 composed of pathotype A strains of X. citri pv. citri (3) (including all strains from the Southwest Indian Ocean region) while one of them (LK141-3 from Mohéli) was identified as a new sequence type based on a non-synonymous single nucleotide polymorphism in gyrB (accession KJ941208). All strains were inoculated by a detached leaf assay (3) onto Mexican lime SRA 140 (C. aurantifolia), Tahiti lime SRA 58 (C. latifolia), sweet orange New Hall Navel SRA 343 (C. sinensis), grapefruit Henderson SRA 336 (C. paradisi), and Ortanique tangor SRA 110 (C. reticulata × C. sinensis). All citrus species inoculated produced typical erumpent, callus-like tissue at wound sites. Xanthomonas-like yellow colonies were re-isolated from lesions produced on Mexican lime. Boiled bacterial suspensions were assayed by PCR with 4/7 primers (4) and produced the expected amplicon, fulfilling Koch's postulates. No lesions developed on the negative control consisting of inoculations with sterile tris buffer. This is the first report of X. citri pv. citri-A causing Asiatic citrus canker in Grande Comore and Anjouan islands confirming the wide distribution of the pathogen in Southwest Indian Ocean islands. Canker-free nurseries and grove sanitation should be implemented to decrease the prevalence of Asiatic canker in the Comoros. References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (2) J. Brun. Fruits 26:533, 1971. (3) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (4) J. S. Hartung et al. Phytopathology 86:95, 1996. (5) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005.

Application of principal component analysis in clinical gait research: identification of systematic differences between healthy and medial knee-osteoarthritic gait.

For a successful completion of a movement task the motor control system has to observe a multitude of internal constraints that govern the coordination of its segments. The purpose of this study was to apply principal component (PC) analysis to detect differences in the segmental coordination between healthy subjects and patients with medial knee osteoarthritis (OA). It was hypothesized that (1) systematic differences in patterns of whole body movement would be identifiable with this method even in small sample sized groups and that (2) these differences will include compensatory movements in the OA patients in both the lower and upper body segments. Marker positions and ground reaction forces of three gait trials of 5 healthy and 5 OA participants with full body marker sets were analyzed using a principal component analysis. Group differences in the PC-scores were determined for the first 10 PC-vectors and a linear combination of those PC-vectors where differences were found defined a discriminant vector. Projecting the original trials onto this discriminant vector yielded significant group differences (t(d=8)=3.011; p=0.017) with greater upper body movement in patients with knee OA that was correlated with the medial-lateral ground reaction force. These results help to characterize the adaptation of whole-body gait patterns to knee OA in a relatively small population and may provide an improved basis for the development of interventions to modify knee load. The PC-based motion analysis offered a highly sensitive approach to identify characteristic whole body patterns of movement associated with pathological gait.

[Parkes-Weber syndrome and pregnancy: anaesthetic implications]. / Syndrome de Parkes-Weber et grossesse : implications anesthésiques.

We report two deliveries in a patient with a Parkes-Weber syndrome. This parturient had a complex angiodysplasia including a soft tissue hypertrophy of a lower limb, a cutaneous angioma and arteriovenous malformations. The risk of perimedullar arteriovenous malformations was ruled out by angiographic magnetic resonance imaging of the spinal cord. We also describe other aspects of the management, including prepartum cardiovascular assessment, mode of delivery, the use of epidural analgesia and the prevention of haemorrhagia and thromboembolism.

Kinematic adaptations to a variable stiffness shoe: mechanisms for reducing joint loading.

A recently described variable-stiffness shoe has been shown to reduce the adduction moment and pain in patients with medial-compartment knee osteoarthritis. The mechanism associated with how this device modifies overall gait patterns to reduce the adduction moment is not well understood. Yet this information is important for applying load modifying intervention for the treatment of knee osteoarthritis. A principal component analysis (PCA) was used to test the hypothesis that there are differences in the frontal plane kinematics that are correlated with differences in the ground reaction forces (GRFs) and center of pressure (COP) for a variable-stiffness compared to a constant-stiffness control shoe. Eleven healthy adults were tested in a constant-stiffness control shoe and a variable-stiffness shoe while walking at self-selected speeds. The PCA was performed on trial vectors consisting of all kinematic, GRF and COP data. The projection of trial vectors onto the linear combination of four PCs showed there were significant differences between shoes. The interpretation of the PCs indicated an increase in the ankle eversion, knee abduction and adduction, decreases in the hip adduction and pelvic obliquity angles and reduced excursion of both the COP and peak medial-lateral GRFs for the variable-stiffness compared to the control shoe. The variable-stiffness shoe produced a unique dynamic change in the frontal plane motion of the ankle, hip and pelvis that contributed to changes in the GRF and COP and thus reduced the adduction moment at a critical instant during gait suggesting a different mechanism that was seen with fixed interventions (e.g. wedges).

Predictive-focus illumination for reducing photodamage in live-cell microscopy.

Due to photobleaching and phototoxicity induced by high-intensity excitation light, the number of fluorescence images that can be obtained in live cells is always limited. This limitation becomes particularly prominent in multidimensional recordings when multiple Z-planes are captured at every time point. Here we present a simple technique, termed predictive-focus illumination (PFI), which helps to minimize cells' exposure to light by decreasing the number of Z-planes that need to be captured in live-cell 3D time-lapse recordings. PFI utilizes computer tracking to predict positions of objects of interest (OOIs) and restricts image acquisition to small dynamic Z-regions centred on each OOI. Importantly, PFI does not require hardware modifications and it can be easily implemented on standard wide-field and spinning-disc confocal microscopes.

Evidence for associations between the purinergic receptor P2X(7) (P2RX7) and toxoplasmosis.

Congenital Toxoplasma gondii infection can result in intracranial calcification, hydrocephalus and retinochoroiditis. Acquired infection is commonly associated with ocular disease. Pathology is characterized by strong proinflammatory responses. Ligation of ATP by purinergic receptor P2X(7), encoded by P2RX7, stimulates proinflammatory cytokines and can lead directly to killing of intracellular pathogens. To determine whether P2X(7) has a role in susceptibility to congenital toxoplasmosis, we examined polymorphisms at P2RX7 in 149 child/parent trios from North America. We found association (FBAT Z-scores +/-2.429; P=0.015) between the derived C(+)G(-) allele (f=0.68; OR=2.06; 95% CI: 1.14-3.75) at single-nucleotide polymorphism (SNP) rs1718119 (1068T>C; Thr-348-Ala), and a second synonymous variant rs1621388 in linkage disequilibrium with it, and clinical signs of disease per se. Analysis of clinical subgroups showed no association with hydrocephalus, with effect sizes for associations with retinal disease and brain calcifications enhanced (OR=3.0-4.25; 0.004

Computer-aided evaluation of neuroblastoma on whole-slide histology images: Classifying grade of neuroblastic differentiation.

Neuroblastoma (NB) is one of the most frequently occurring cancerous tumors in children. The current grading evaluations for patients with this disease require pathologists to identify certain morphological characteristics with microscopic examinations of tumor tissues. Thanks to the advent of modern digital scanners, it is now feasible to scan cross-section tissue specimens and acquire whole-slide digital images. As a result, computerized analysis of these images can generate key quantifiable parameters and assist pathologists with grading evaluations. In this study, image analysis techniques are applied to histological images of haematoxylin and eosin (H&E) stained slides for identifying image regions associated with different pathological components. Texture features derived from segmented components of tissues are extracted and processed by an automated classifier group trained with sample images with different grades of neuroblastic differentiation in a multi-resolution framework. The trained classification system is tested on 33 whole-slide tumor images. The resulting whole-slide classification accuracy produced by the computerized system is 87.88%. Therefore, the developed system is a promising tool to facilitate grading whole-slide images of NB biopsies with high throughput.

Long-term proton pump inhibitor use in children: a retrospective review of safety.

The objective of this work was to assess the efficacy of continuous proton pump inhibitor (PPI) therapy in children and to evaluate changes in biochemical, endoscopic, and histologic parameters during such treatment. A retrospective review of children receiving PPI therapy continuously for 1 year or more with baseline and follow-up esophageal and gastric biopsies on treatment was conducted to assess type, frequency, and duration of PPI dosing, symptom relief, gastrin levels, histologic findings, and adverse events. A total of 113 children (59 male, median age 4.5 years) were identified. Of these, 31% (35/113) were neurologically impaired. The median treatment duration was 35.2 months. Elevated serum gastrin levels occurred in 73% of children with no statistically significant differences in gastrin level by PPI type, dose, and dosing frequency or treatment duration. Adverse events were reported by 12% of children: diarrhea (5%) and constipation (4%) were the most frequent. Long-term PPI therapy appears to be effective, safe and well tolerated in children despite some biochemical, endoscopic, and histologic changes.

Modeling of clusters in a strong 248-nm laser field by a three-dimensional relativistic molecular dynamic model.

A relativistic time-dependent three-dimensional particle simulation model has been developed to study the interaction of intense ultrashort KrF (248 nm) laser pulses with small Xe clusters. The trajectories of the electrons and ions are treated classically according to the relativistic equation of motion. The model has been applied to a different regime of ultrahigh intensities extending to 10(21) W/ cm(2). In particular, the behavior of the interaction with the clusters from intensities of approximately 10(15) W/cm(2) to intensities sufficient for a transition to the so-called "collective oscillation model" has been explored. At peak intensities below 10(20) W/cm(2), all electrons are removed from the cluster and form a plasma. It is found that the "collective oscillation model" commences at intensities in excess of 10(20) W/cm(2), the range that can be reached in stable relativistic channels. At these high intensities, the magnetic field has a profound effect on the shape and trajectory of the electron cloud. Specifically, the electrons are accelerated to relativistic velocities with energies exceeding 1 MeV in the direction of laser propagation and the magnetic field distorts the shape of the electron cloud to give the form of a pancake.

Retinal thickness measurements from optical coherence tomography using a Markov boundary model.

We present a system for detecting retinal boundaries in optical coherence tomography (OCT) B-scans. OCT is a relatively new imaging modality giving cross-sectional images that are qualitatively similar to ultrasound. However, the axial resolution with OCT is much higher, on the order of 10 microm. Objective, quantitative measures of retinal thickness may be made from OCT images. Knowledge of retinal thickness is important in the evaluation and treatment of many ocular diseases. The boundary-detection system presented here uses a one-dimensional edge-detection kernel to yield edge primitives. These edge primitives are rated, selected, and organized to form a coherent boundary structure by use of a Markov model of retinal boundaries as detected by OCT. Qualitatively, the boundaries detected by the automated system generally agreed extremely well with the true retinal structure for the vast majority of OCT images. Only one of the 1450 evaluation images caused the algorithm to fail. A quantitative evaluation of the retinal boundaries was performed as well, using the clinical application of automatic retinal thickness determination. Retinal thickness measurements derived from the algorithm's results were compared with thickness measurements from manually corrected boundaries for 1450 test images. The algorithm's thickness measurements over a 1-mm region near the fovea differed from the corrected thickness measurements by less than 10 microm for 74% of the images and by less than 25 microm (10% of normal retinal thickness) for 98.4% of the images. These errors are near the machine's resolution limit and still well below clinical significance. Current, standard clinical practice involves a qualitative, visual assessment of retinal thickness. A robust, quantitatively accurate system such as ours can be expected to improve patient care.