Brain charts for the human lifespan.

Over the past few decades, neuroimaging has become a ubiquitous tool in basic research and clinical studies of the human brain. However, no reference standards currently exist to quantify individual differences in neuroimaging metrics over time, in contrast to growth charts for anthropometric traits such as height and weight1. Here we assemble an interactive open resource to benchmark brain morphology derived from any current or future sample of MRI data ( http://www.brainchart.io/ ). With the goal of basing these reference charts on the largest and most inclusive dataset available, acknowledging limitations due to known biases of MRI studies relative to the diversity of the global population, we aggregated 123,984 MRI scans, across more than 100 primary studies, from 101,457 human participants between 115 days post-conception to 100 years of age. MRI metrics were quantified by centile scores, relative to non-linear trajectories2 of brain structural changes, and rates of change, over the lifespan. Brain charts identified previously unreported neurodevelopmental milestones3, showed high stability of individuals across longitudinal assessments, and demonstrated robustness to technical and methodological differences between primary studies. Centile scores showed increased heritability compared with non-centiled MRI phenotypes, and provided a standardized measure of atypical brain structure that revealed patterns of neuroanatomical variation across neurological and psychiatric disorders. In summary, brain charts are an essential step towards robust quantification of individual variation benchmarked to normative trajectories in multiple, commonly used neuroimaging phenotypes.

Comment.

We congratulate the authors on their comments on innovative approaches to drug development that fall out of the traditional mold and may result in more quickly bringing safe and effective treatments to patients. Changes in the overall clinical develop approach are most relevant to "breakthrough" therapies, which have generally yielded exceptional efficacy data in early clinical studies, motivating exploration of accelerated development and regulatory approaches, as well as a potential ethical need for crossover upon progression in randomized controlled studies (Horning et al., 2015). As is clear from the manuscript, it will be important to develop an understanding of what works well and where the pitfalls in new approaches are. We comment briefly on the four topics mentioned by the authors, combining comments on items 2 and 3: 1) non-proportional hazards, 2) interpretability of extended Phase I trials, 3) single-arm trials as a basis for approval, and 4) recent innovations in trial design.

Clustering of clinical and environmental Escherichia coli O104 isolates using the DiversiLab™ repetitive sequence-based PCR system.

The DiversiLab™ rep-PCR system was used to amplify DNA regions of 28 well-characterized Escherichia coli O104 strains to generate a digital DNA fingerprint profile for strain differentiation. E. coli O104 strains from human stools and other sources were examined. The results indicate that this system can cluster similar O104 strains rapidly.

Conducting systematic reviews of intervention questions I: Writing the review protocol, formulating the question and searching the literature.

This article is the fourth of six articles addressing systematic reviews in animal agriculture and veterinary medicine. Previous articles in the series have introduced systematic reviews, discussed study designs and hierarchies of evidence, and provided details on conducting randomized controlled trials, a common design for use in systematic reviews. This article describes development of a review protocol and the first two steps in a systematic review: formulating a review question, and searching the literature for relevant research. The emphasis is on systematic reviews of questions related to interventions. The review protocol is developed prior to conducting the review and specifies the plan for the conduct of the review, identifies the roles and responsibilities of the review team and provides structured definitions related to the review question. For intervention questions, the review question should be defined by the PICO components: population, intervention, comparison and outcome(s). The literature search is designed to identify all potentially relevant original research that may address the question. Search terms related to some or all of the PICO components are entered into literature databases, and searches for unpublished literature also are conducted. All steps of the literature search are documented to provide transparent reporting of the process.

Normoxic or hypoxic CD44/CD41 a2 B1 integrin-positive prostate PC3 cell side fractions and cancer stem cells.

A CD44/a 2 B 1- (CD41 integrin) and B-catenin-labeled fraction of PC3 prostate cancer cells is able to reconstitute cells representative of the original tumor in immuno-deficient mice (Li et al. in Cancer Res 68:1820-1825, 2008). After 48 h of culture under nitrogen with a resulting medium pH of 7.8, sorted hypoxic PC3 cells yielded a higher percentage and concentration/10(5) of cells in a doubly labeled (DL) CD44(+)/CD41(+) side fraction compared with control cells cultured under normoxia (5 % CO2 in the ambient atmosphere at 37 °C). When the rise in pH was prevented (95 % N2, 5 % CO2), the difference in sorted hypoxic cell numbers remained. Sorted N and H DL cells and CD44(+)/CD41(-) cells were cultured under standard conditions. After 1-2 weeks, the number of attached colonies from formerly hypoxic cells, whether previously cultured with N2 or 95 % N2 + 5 % CO2, exceeded the number of doubly labeled normoxic cells, consistent with their greater initial concentration. Cultured sorted N or H CD44(+)/CD41(-) cells resulted in monolayers containing a small percentage of DL cells. Recovery of greater percentage and numbers of putative cancer stem cells, confirmed by quantitative cell sorting after culture under hypoxic conditions, is consistent with their greater relative numbers present in hypoxic niches. In addition, the report that neither CD44(+) nor CD41(+) epitopes were preferentially associated with FAM65B(high)/MF12(low)/LEF1(low) PC3 cells able to reconstitute tumors in immuno-deficient mice (Zhang and Waxman in Mol Cancer 9:319-330, 2010) suggests an in vitro mimic of tumor cell heterogeneity observed in epithelial cancers.

Melorheostosis and central giant cell granuloma of the mandible in a 15-year-old girl.

Melorheostosis is a nonhereditary bone dysplasia primarily affecting the appendicular skeleton. Because clinical and histologic features are often nonspecific, the diagnosis is often based on the radiographic presentation. Involvement of the craniofacial skeleton is rare. We describe a case of a 15-year-old girl with appendicular and craniofacial melorheostosis with adjacent central giant cell granuloma. We discuss the possible significance of this previously unreported finding.

Robotic-assisted surgery and the evolution of the radical prostatectomy.

The surgical treatment of prostate cancer has evolved considerably since it was first described in 1905. With the introduction of a robotic, surgical-assist device, minimally invasive techniques for prostate removal have been increasingly utilized throughout the world. Currently, there is a large body of literature suggesting that robotic-assisted laparoscopic prostatectomy is associated with certain improved perioperative and postoperative outcomes and similar cancer control rates compared to open radical prostatectomy. The goal of this review is to objectively evaluate and describe the current state-of-the-art in surgical technique, perioperative and long-term outcomes, complications and the future of robotic-assisted laparoscopic radical prostatectomy.

Using sex pheromone trapping to explore threats to wheat from Hessian fly (Diptera: Cecidomyiidae) in the Upper Great Plains.

Before embarking on the 5-10 yr effort it can take to transfer plant resistance (R) genes to adapted crop cultivars, a question must be asked: is the pest a sufficient threat to warrant this effort? We used the recently discovered female-produced sex pheromone of the Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae),to explore this question for populations in the Upper Great Plains. Methods for pheromone trapping were established and trapping data were used to explore geographic distribution, phenology, and density. The pheromone lure remained attractive for up to 10 d and only attracted male Hessian flies. Traps placed within the crop canopy caught flies but traps placed above the crop canopy did not. Hessian flies were trapped throughout North Dakota starting in the spring and continuing through the summer and autumn. Densities were low in the spring but increased greatly during the early part of the summer, with peak adult emergence taking place at a time (July/August) when spring wheat was being harvested and winter wheat had not yet been planted. In the autumn, adults were found at a time when winter wheat seedlings are growing. The discovery of flies on Conservation Reserve Program land supports the idea that pasture grasses serve as alternate hosts. We conclude that the Hessian fly is a risk to wheat in the Upper Great Plains and predict that global warming and the increasing cultivation of winter wheat will add to this risk.

Cost-effectiveness analysis of targeted and sequential screening strategies for latent tuberculosis.

SETTING: No cost-effectiveness studies of testing for latent tuberculosis infection have incorporated both targeted testing and the use of interferon-gamma release assays (IGRAs) in heterogeneous populations. OBJECTIVE: To examine the cost-effectiveness of universal vs. targeted and sequential testing strategies and the use of tuberculin skin testing (TST) vs. IGRAs. DESIGN: Using a decision-analytic model, incremental cost-effectiveness ratios were calculated in 2009 among nine potential strategies for screening recruits. A societal perspective was taken over a 20-year analytic horizon, discounting future costs at 3% annually. Sensitivity analyses were conducted to determine how changes in assumptions affected the estimates. RESULTS: Targeted strategies cost over US$250 000 per case prevented, whereas universal testing strategies cost over US$700 000 per incremental case prevented in base case and most sensitivity analyses. CONCLUSION: Targeted testing offered the best value in this population, although it was still relatively expensive compared to no testing. Sequential testing with both TST and IGRAs provided a poor incremental value compared to targeted and universal testing strategies. Targeted testing using TST was slightly more cost-effective than targeted testing using either QuantiFERON®-TB Gold In-Tube or T-SPOT®.TB, but these estimates were very sensitive to changes in model assumptions.

A reproductive fitness cost associated with Hessian fly (Diptera: Cecidomyiidae) virulence to wheat's H gene-mediated resistance.

We studied whether adaptation of the Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), to plant resistance incurs fitness costs. In this gene-for-gene interaction, adaptation to a single H resistance gene occurs via loss of a single effector encoded by an Avirulence gene. By losing the effector, the adapted larva now survives on the H gene plant, presumably because it evades the plant's H gene-mediated surveillance system. The problem is the Hessian fly larva needs its effectors for colonization. Thus, for adapted individuals, there may be a cost for losing the effector, with this then creating a trade-off between surviving on H-resistant plants and growing on plants that lack H genes. In two different tests, we used wheat lacking H genes to compare the survival and growth of a nonadapted strain to two H-adapted strains. The two adapted strains differed in that one had been selected for adaptation to H9, whereas the other strain had been selected for adaptation to H13. Tests showed that two H-adapted strains were similar to the nonadapted strain in egg-to-adult survival but that they differed in producing adults with smaller wings. By using known relationships between wing length and reproductive potential, we found that losses in wing length underestimate losses in reproductive potential. For example, H9- and H13-adapted females had 9 and 3% wing losses, respectively, but they were estimated to have 32 and 12% losses in egg production. Fitness costs of adaptation will be investigated further via selection experiments comparing Avirulence allele frequencies for Hessian fly populations exposed or not exposed to H genes.

The effect of normoxia and hypoxia on a prostate (PC-3) CD44/CD41 cell side fraction.

It has been reported that human prostate-derived PC-3 cells that are CD44- and CD 41 (a2 B1)-positive are enriched in cancer stem cells. This study compared the effect of PC-3 cell proliferation under normoxia or hypoxia on the initial and subsequent expression of this doubly-labeled side-fraction. Despite the numerical advantage of attached normoxic cells, 48 h of culture under nitrogen, an environment containing minimal oxygen and CO(2) resulting in an elevated pH of the medium, was associated with a higher percentage, absolute and relative number of doubly-labeled (DL) hypoxic compared to normoxic cells. At 24 h, the reverse was found. When the pH was controlled with the use of 95% nitrogen and 5% carbon dioxide, the percentage and number of normoxic DL cells exceeded hypoxic ones at both 24 and 48 h. At 24 h, 2-deoxy-L-glucose or sodium arsenate reduced normoxic DL cell numbers more than hypoxic ones. The interplay between hypoxia, increased medium pH and the effect of inhibitors as they might influence therapy are considered.

H-gene-mediated resistance to Hessian fly exhibits features of penetration resistance to fungi.

Features shared by host-specific phytophagous insects and biotrophic plant pathogens include gene-for-gene interactions and the ability to induce susceptibility in plants. The Hessian fly shows both. To protect against Hessian fly, grasses have H genes. Avirulent larvae die on H-gene-containing resistant plants but the cause of death is not known. Imaging techniques were used to examine epidermal cells at larval attack sites, comparing four resistant wheat genotypes (H6, H9, H13, and H26) to a susceptible genotype. Present in both resistant and susceptible plants attacked by larvae were small holes in the tangential cell wall, with the size of the holes (0.1 microm in diameter) matching that of the larval mandible. Absent from attacked resistant plants were signs of induced susceptibility, including nutritive tissue and ruptured cell walls. Present in attacked resistant plants were signs of induced resistance, including cell death and fortification of the cell wall. Both presumably limit larval access to food, because the larva feeds on the leaf surface by sucking up liquids released from ruptured cells. Resistance was associated with several subcellular responses, including elaboration of the endoplasmic reticulum-Golgi complex and associated vesicles. Similar responses are observed in plant resistance to fungi, suggesting that "vesicle-associated penetration resistance" also functions against insects.

Are cancer stem cells concentrated in more alkaline hypoxic regions of tumors?

We wonder if the most viable hypoxic cancer stem cells concentrate in more alkaline regions of tumors, favoring their survival and evolution. Alternately, or in addition, do some cancer stem cells themselves maintain a more alkaline internal environment, achieving the same result. Based upon the response of cultured cells, including stem cells, to a certain degree of hypoxia and of most if not all proliferating cells to a somewhat more alkaline ambient and especially endogenous pH, their survival and proliferation should be favored. The broad outline of the argument, abstracted from a number of the available examples is developed: that the survival of cancer stem cells is favored by these conditions, contributing to their limited response to various therapies and their subsequent development of more malignant properties.

In vitro effects of dichloroacetate and CO2 on hypoxic HeLa cells.

HeLa and PANC-1 cells were exposed to conflicting signals promoting anaerobic or aerobic energy-generating processes and their viability, cell numbers and the ability of HeLa cells to form colonies were assessed. Under conventional aerobic cell culture with 5% CO(2), dichloroacetate (DCA), an inhibitor of the enzyme pyruvate dehydrogense kinase with subsequent stimulation of pyruvate dehydrogenase that redirects energy metabolism toward the Kreb cycle, reduced HeLa and PANC-1 cellular proliferation and viability. With nitrogen-induced hypoxia, the number of control cells and cells cultured with 12.5 mM DCA paradoxically was greater than that of normoxic controls under similar conditions. A higher medium pH of cells cultured under nitrogen contributed to these differences. In 96-well experiments, 95% nitrogen with 5% CO(2) reduced the numbers of hypoxic cells and medium pH toward that of the aerobic controls, with retention of the DCA-induced hypoxic compared to normoxic cell numbers. The media of these cells cultured with DCA still exhibited an increased pH. Increased hypoxia-inducible factor 1, alpha subunit (HIF1A) mRNA expression in hypoxic HeLa cells and their greater reliance on D-glucose for metabolic energy confirmed the reliability of the incubation conditions. Compared with normoxic cells, hypoxic cells initially increased their synthesis of ATP, but once proliferation ceased, this no longer closely correlated with cell numbers. Type 1 apoptosis, which was somewhat greater in hypoxic than normoxic cells, contributed to hypoxia and DCA-induced cell death. Colony counts of hypoxic, DCA-inhibited cells subsequently switched to normoxia exceeded those of similarly treated normoxic DCA cells. Despite inhibition in certain hypoxic environments of pyruvate dehydrogenase kinase by DCA and its contribution to increased cellular apoptosis and necrosis, hypoxic cells generally outnumbered normoxic control cells, as did hypoxic DCA-treated cells compared with comparable DCA-treated normoxic cells. Since in vivo hypoxic cells are considered a major factor contributing to therapeutic failure, and as DCA redirects energy metabolism toward the more energy efficient Kreb citric acid cycle, associated with increased medium (and inferred cellular) pH, similar circumstances in vivo could promote proliferation and survival of hypoxic cell clones with the potential for developing unwanted properties.

Multinucleate cell angiohistiocytoma: an uncommon mucosal tumour.

Multinucleate cell angiohistiocytoma (MCAH) is an uncommon benign soft-tissue lesion with characteristic histological and immunohistochemical features. It is unclear if it represents a benign neoplasm or a reactive/inflammatory process. The overwhelming majority of these tumors have been described on cutaneous surfaces. One case has been reported on the skin of the lip. This is the first documentation of MCAH within the oral cavity. The clinical presentation, histopathological features and immunohistochemical reactivity pattern are described. Because of the benign nature of this lesion, a conservative approach is recommended in its management. Surgical excision appears to be curative.

Do intra-tumor alkaline micro-regions represent additional therapeutically privileged sites?

We briefly review some implications for therapeutic resistance in solid cancers that could be associated with more alkaline intra-tumor micro-regions reported to exist. Regions of increased alkalinity may provide a proliferative advantage for cancer "stem" or other cells, as more alkaline intra- and extra-cellular environments often are associated with increased cellular proliferation. If increased alkalinity is present in aerobic, but perhaps more especially in hypoxic micro-environments, proliferation of cells less susceptible to programmed cell death, with reduced expression of multi-drug resistance membrane proteins and altered efficacy of some therapeutic drugs should develop. Such cells are also more likely to generate aberrant clones resistant to additional therapy, particularly those dependent upon mitochondrial oxidative processes with greater generation of free radicals, compared to cells reliant on anaerobic glycolysis for metabolic energy. The interplay between alkalinity and normoxia, hypoxia or anoxia may differentially advantage some cancer "stem" cells.

Does homeobox-related "positional" genomic information contribute to implantation of metastatic cancer cells at non-random sites?

Reasons for the lodgment of metastases from several types of solid cancer at apparently non-random sites have not been established. Recently, a group of genes expressed in human fibroblasts obtained from different anatomic locations was implicated in "positional" genomic information. Essentially, a Cartesian coordinate system identifying fibroblasts originally resident at anterior or more posterior, proximal or distal and dermal or non-dermal (heart, lung, etc.) locations was proposed. The determinants used for these identifications included HOX genes, central to embryonic segmental development, some of which are expressed in differentiated, post-embryonic cells. To the extent that HOX or other homeobox genes are expressed in ectodermal, mesodermal or endodermally-derived, malignantly transformed cells, they might contribute "positional" information to nidation of specific malignant clones at non-random sites. As understood in the past, interdiction of HOX or homeobox-related gene expression might reduce the probability of cancer cell implantation or alter their destinations in complex ways. Ideally, by interfering with HOX or other homeobox gene-related expression of antigenic determinants potentially contributing to their "homing" and nidation, reduced implantation of circulating cancer cells could render them more susceptible to systemic chemotherapy or immunotherapy, as demonstrated in mice. Furthermore, HOX or other homeobox genes or their products could provide novel intra- or extracellular targets for therapy.