Nine microsatellite loci were identified in the western rock lobster (Panulirus cygnus) using two different methods. The first method involved the screening of a small, fragment, partial genomic library with a radioactive (CA) 6 probe. The second method, was based upon an enrichment method and used biotinylated, tetranucleotide microsatellite oligonucleotide capture probes. The nine loci described are all very polymorphic, with 11 to 34 alleles observed for each locus and heterozygosities ranging from 0.58 to 0.86. These microsatellite loci will be useful in analysing both the population structure and the mating systems used by this species and will add important information for the management of the wild stocks of this economically important species.Keywords Panulirus cygnus Á Microsatellite Á SSR The western rock lobster (Panulirus cygnus) is Western Australia's most important single species fishery with annual harvests over the last decade ranging between approximately 9 and 14.5 thousand tonnes per annum. The elucidation and understanding of population and mating structures for this species will allow the information obtained to be incorporated into an improved management strategy for the brood stock. In order to understand the population structure and mating systems of the western rock lobster, polymorphic, microsatellite genetic markers have been isolated and characterised. Two strategies were used to develop these microsatellites, the first consisting of the construction of a Hae III small fragment library which was screened with a (CA)n probe and the second library was made using an enrichment technique followed by screening with tetranucleotide microsatellite motifs. Nine microsatellite loci have been fully characterised and these are described in this report.A DNA genomic library was made using Qiagen Tissue Kit (Qiagen) to isolate DNA extracted from tail muscle tissue. The DNA was digested with HaeIII restriction endonuclease (Promega, CA, USA) followed by excision of the 100-500 bp region after agarose (1%) gel electrophoresis. Fragments were ligated into Sma I (Promega) digested, dephosphorylased pUC18 plasmid vector and transformed into competent cells (DH5a Invitrogen). Colonies were transferred to Hybond N? membranes and screened with radiolabelled (CA)10 probe. Thirty clones were identified after the first round of screening. Following successive screening rounds to further isolate the clones, twenty of these were sequenced. Plasmid DNA was isolated from the positive clones and sequenced using cyclesequencing combined with IRD 800 labelled primers on a Li-Cor gene sequencer. Primers were designed using Mac Vector (Eastman-Kodak, USA) or PRIMER 3 software (Rozen and Skaletsky 2000). Many of the loci identified contained insufficient flanking DNA sequences or produced microsatellites containing few repeat units. Three loci (WRL 1, WRL 2 and WRL 3) identified by using this method were deemed to be useful.A second library enriched for tetranucleotide microsatellites was prepared using genomic DNA. Br...

The olfactory system combines input from multiple receptor types to represent odor information, but there are few explicit examples relating olfactory receptor (OR) activity patterns to odor perception. To uncover these relationships, we performed genome-wide scans on odor-perception phenotypes for ten odors in 1003 Han Chinese and validated results for six of these odors in an ethnically diverse population (n=364). In both populations, we replicated three previously reported associations (β-ionone/OR5A, androstenone/OR7D4, cis-3-hexen-1-ol/OR2J3 LD-band), suggesting that olfactory phenotype/genotype studies are robust across populations. Two novel associations between an OR and odor perception contribute to our understanding of olfactory coding. First, we found a SNP in OR51B2 that associated with trans-3-methyl-2-hexenoic acid, a key component of human underarm odor. Second, we found two linked SNPs associated with the musk Galaxolide in a novel musk receptor, OR4D6, which is also the first OR shown to drive specific anosmia to a musk compound. We also found that the derived alleles of the SNPs reportedly associated with odor perception tend to reduce odor intensity, supporting the hypothesis that the primate olfactory gene repertoire has degenerated over time. This study provides information about coding for human body odor, and gives us insight into broader mechanisms of olfactory coding, such as how differential OR activation can converge on a similar percept.

Introduction Standard FCR therapy in untreated CLL patients (F-25 mg/m2 d1–3 q 4wk; C-250 mg/m2 d 1–3 q 4wk; R-500 mg/m2 d1 q 4wk for 6 cycles) was recently reported to have complete remission (CR) of 70% and overall responses (OR) of 95% (J Clin Oncol2005;23:4079). The major toxicity was grade 3/4 neutropenia during 52% of courses. One approach to decrease neutropenia without compromising efficacy could be by reducing the doses of F and C and increasing the dose of R. Methods We conducted a phase II study for previously untreated advanced CLL patients treated with FCR-Lite (F-20mg/m2 d1–3 q 4 wk; C-150 mg/m2 d1–3 q 4 wk; R-500mg/m2 d1 and d14 q 4wks; maintenance R-500 mg/m2 q 3 months until progression). A Simon two-stage design was used where 15 patients were accrued in the first stage and because of acceptable toxicity and response rate in stage I an additional 35 patients will be treated. The primary endpoint was response rate. Results Twenty eight patients (19 male, 9 female), age 36–85 years (median 58) were treated with a total of 154 FCR-Lite courses. All 28 patients were evaluable for toxicity. Grade 3/4 neutropenia occurred during 12 (8%) courses. There was one episode of neutropenic fever. Grade 3/4 thrombocytopenia occurred during 4 (3%) courses. Grade III anemia occurred during 2 (1%) courses. One patient had pneumonia. Among the 21 evaluable patients for response, the CR rate was 86%, PR rate was 14% with an OR rate of 100%. Seventeen of the CR patients were tested by flow cytometry and had no evidence of CD5+/CD19+ cells in their bone marrow after therapy. Preliminary data from pharmacokinetics performed on 16 patients showed time dependent increase in trough and peak antibody concentrations throughout treatment (Table 1). Conclusions Our results in the first 28 patients suggest FCR-Lite is highly effective with considerably less grade 3/4 neutropenia than standard FCR. Complete responders had no detectable CD5+/CD19+ cells in their bone marrow following FCR-Lite. Table 1. Serum Rituximab Concentrations Before and After Infusion (N=16 patients) Infusion No. Concentration (μg/ml) (Mean) C1,d1, before 0 C1,d1, after 116.1 C1,d14, before 10.0 C1,d14, after 207.8 C2,d1, before 33.2 C2,d1, after 231.4 C3,d14, before 125.3 C3,d14, after 345.5 C4,d1, before 173.2 C4,d1, after 390.8

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Congenital diaphragmatic hernia (CDH) is a severe congenital anomaly that is often accompanied by other anomalies. Although the role of genetics in the pathogenesis of CDH has been established, only a small number of disease genes have been identified. To further investigate the genetics of CDH, we analyzed de novo coding variants in 827 proband-parent trios and confirmed an overall significant enrichment of damaging de novo variants, especially in constrained genes. We identified LONP1 (Lon Peptidase 1, Mitochondrial) and ALYREF (Aly/REF Export Factor) as novel candidate CDH genes based on de novo variants at a false discovery rate below 0.05. We also performed ultra-rare variant association analyses in 748 cases and 11,220 ancestry-matched population controls and identified LONP1 as a risk gene contributing to CDH through both de novo and ultra-rare inherited largely heterozygous variants clustered in the core of the domains and segregating with CDH in familial cases. Approximately 3% of our CDH cohort was heterozygous with ultra-rare predicted damaging variants in LONP1 who have a range of clinical phenotypes including other anomalies in some individuals and higher mortality and requirement for extracorporeal membrane oxygenation. Mice with lung epithelium specific deletion of Lonp1 die immediately after birth and have reduced lung growth and branching that may at least partially explain the high mortality in humans. Our findings of both de novo and inherited rare variants in the same gene may have implications in the design and analysis for other genetic studies of congenital anomalies.