We study temporal correlations and multifractal properties of long river discharge records from 41 hydrological stations around the globe. To detect long-term correlations and multifractal behaviour in the presence of trends, we apply several recently developed methods [detrended fluctuation analysis (DFA), wavelet analysis, and multifractal DFA] that can systematically detect and overcome non-stationarities in the data at all time scales. We find that above some crossover time that usually is several weeks, the daily runoffs are long-term correlated, being characterized by a correlation function C(s) that decays as C(s)ws Kg . The exponent g varies from river to river in a wide range between 0.1 and 0.9. The power-law decay of C(s) corresponds to a power-law increase of the related fluctuation function F 2 (s)ws H where HZ1Kg/2. We also find that in most records, for large times, weak multifractality occurs. The Renyi exponent t(q) for q between K10 and C10 can be fitted to the remarkably simple form tðqÞZKlnða q C b q Þ=ln2, with solely two parameters a and b between 0 and 1 with aCbR1. This type of multifractality is obtained from a generalization of the multiplicative cascade model. q

We study the multifractal temporal scaling properties of river discharge and precipitation records. We compare the results for the multifractal detrended fluctuation analysis method with the results for the wavelet transform modulus maxima technique and obtain agreement within the error margins. In contrast to previous studies, we find non-universal behaviour: On long time scales, above a crossover time scale of several months, the runoff records are described by fluctuation exponents varying from river to river in a wide range. Similar variations are observed for the precipitation records which exhibit weaker, but still significant multifractality. For all runoff records the type of multifractality is consistent with a modified version of the binomial multifractal model, while several precipitation records seem to require different models.The analysis of river flows has a long history. Already more than half a century ago the engineer H. E. Hurst found that runoff records from various rivers exhibit 'long-range statistical dependencies' [1]. Later, such long-term correlated fluctuation behaviour has also been reported for many other geophysical records including precipitation data [2,3], see also [4]. These original approaches exclusively focused on the absolute values or the variances of the full distribution of the fluctuations, which can be regarded as the first moment F 1 (s) [1][2][3] and the second moment F 2 (s) [5], respectively. In the last

We have developed a resource-based relative-value scale as an alternative to the system of payment based on charges for physicians' services. Resource inputs by physicians include (1) total work input performed by the physician for each service; (2) practice costs, including malpractice premiums; and (3) the cost of specialty training. These factors were combined to produce a relative-value scale denominated in nonmonetary units. We describe here the process by which the physician's work was defined and estimated. The study asked two questions: What is the physician's work for each service performed? and Can work be estimated reliably and validly? We concluded that a physician's work has four major dimensions: time, mental effort and judgment, technical skill and physical effort, and psychological stress. We found that physicians can rate the relative amount of work of the services within their specialty directly, taking into account all the dimensions of work. Moreover, these ratings are highly reproducible, consistent, and therefore probably valid.

Chlamydial infection of the host cell induces Gamma interferon (IFNγ), a central immunoprotector for humans and mice. The primary defense against Chlamydia infection in the mouse involves the IFNγ-inducible family of IRG proteins; however, the precise mechanisms mediating the pathogen's elimination are unknown. In this study, we identify Irga6 as an important resistance factor against C. trachomatis, but not C. muridarum, infection in IFNγ-stimulated mouse embryonic fibroblasts (MEFs). We show that Irga6, Irgd, Irgm2 and Irgm3 accumulate at bacterial inclusions in MEFs upon stimulation with IFNγ, whereas Irgb6 colocalized in the presence or absence of the cytokine. This accumulation triggers a rerouting of bacterial inclusions to autophagosomes that subsequently fuse to lysosomes for elimination. Autophagy-deficient Atg5−/− MEFs and lysosomal acidification impaired cells surrender to infection. Irgm2, Irgm3 and Irgd still localize to inclusions in IFNγ-induced Atg5−/− cells, but Irga6 localization is disrupted indicating its pivotal role in pathogen resistance. Irga6-deficient (Irga6−/−) MEFs, in which chlamydial growth is enhanced, do not respond to IFNγ even though Irgb6, Irgd, Irgm2 and Irgm3 still localize to inclusions. Taken together, we identify Irga6 as a necessary factor in conferring host resistance by remodelling a classically nonfusogenic intracellular pathogen to stimulate fusion with autophagosomes, thereby rerouting the intruder to the lysosomal compartment for destruction.

Chikungunya virus (CHIKV) is a globally spreading alphavirus against which there is no commercially available vaccine or therapy. Here we use a genome-wide siRNA screen to identify 156 proviral and 41 antiviral host factors affecting CHIKV replication. We analyse the cellular pathways in which human proviral genes are involved and identify druggable targets. Twenty-one small-molecule inhibitors, some of which are FDA approved, targeting six proviral factors or pathways, have high antiviral activity in vitro, with low toxicity. Three identified inhibitors have prophylactic antiviral effects in mouse models of chikungunya infection. Two of them, the calmodulin inhibitor pimozide and the fatty acid synthesis inhibitor TOFA, have a therapeutic effect in vivo when combined. These results demonstrate the value of loss-of-function screening and pathway analysis for the rational identification of small molecules with therapeutic potential and pave the way for the development of new, host-directed, antiviral agents.

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Single-cell PCR and sequencing of full-length Ig heavy (Igh) and Igk and Igl light chain genes is a powerful tool to measure the diversity of antibody repertoires and allows the functional assessment of B-cell responses through direct Ig gene cloning and the generation of recombinant mAbs. However, the current methodology is not high-throughput compatible. Here we developed a two-dimensional bar-coded primer matrix to combineKeywords: Antibody repertoire r B cell r Index cell sorting Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionAntibodies are B-cell antigen receptors encoded by two independent Ig heavy (Igh) and light (Igk or Igl) chain genes. Therefore, the accurate assessment of antibody repertoires and clonal relationships between individual B cells requires the combined analysis Correspondence: Hedda Wardemann e-mail: wardemann@mpiib-berlin.mpg.de of Igh and Igk/Igl genes from the same cell. Conventional singlecell (sc) based antibody repertoire studies use Sanger sequencing of Igh and corresponding Igk/Igl PCR products to assess V(D)J gene usage, levels of somatic hypermutations, and B-cell clonality [1][2][3][4][5]. This process is highly efficient, unbiased, and has been widely used to assess antibody repertoires in health and disease in mice and humans. Available single-cell PCR (scPCR) protocols provide full-length gene information and are compatible with the direct cloning of Igh and corresponding Igk/Igl genes [3,4]. This facilitates the production of recombinant mAbs, which have beenwww.eji-journal.eu 598Christian E. Busse et al. Eur. J. Immunol. 2014. 44: 597-603 used to study B-cell repertoires based on Ig reactivity and function. Here we combined the scPCR-based amplification of Igh and Igk/Igl transcripts and NGS into a unified pipeline that allows the high-throughput antibody repertoire analysis of thousands of single isolated B cells. The platform is scalable and provides highquality sequence information by consensus building of multiple reads originating from the same cell. It integrates sequencing and flow cytometry data obtained by indexed cell sorting into a single data set and thereby allows for the retrospective phenotypic characterization of B cells according to their antibody genotype. The strategy provides full-length Ig gene sequences of corresponding Igh and Igk/Igl genes and is therefore ideally suited for phylogenetic B-cell repertoire analyses and fully compatible with established procedures for antibody cloning and expression [3]. Importantly, the tag matrix can be used universally to barcode primer sets for the single-cell-based amplification of Ig genes from any species including humans [4]. Results Primer matrix designThe aim of our experimental strategy was to combine the advantages of scPCR and NGS to facilitate the massive parallel analysis of B-cell antibody repertoires (Fig. 1). To perform high-throughput parallel NGS of pooled PCR products without loss of Igh and Igk/Igl linkage, we desig...

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