KLHL12 promotes non-lysine ubiquitination of the dopamine receptors D-4.2 and D-4.4, but not of the ADHD-associated D-4.7 variant

Dopamine D-4 Receptor Polymorphism : The dopamine D-4 receptor has an important polymorphism in its third intracellular loop that is intensively studied and has been associated with several abnormal conditions, among others, attention deficit hyperactivity disorder. KLHL12 Promotes Ubiquitination of the Dopamine D-4 Receptor on Non-Lysine Residues : In previous studies we have shown that KLHL12, a BTB-Kelch protein, specifically interacts with the polymorphic repeats of the dopamine D-4 receptor and enhances its ubiquitination, which, however, has no influence on receptor degradation. In this study we provide evidence that KLHL12 promotes ubiquitination of the dopamine D-4 receptor on non-lysine residues. By using lysine-deficient receptor mutants and chemical approaches we concluded that ubiquitination on cysteine, serine and/or threonine is possible. Differential Ubiquitination of the Dopamine D-4 Receptor Polymorphic Variants : Additionally, we show that the dopamine D-4.7 receptor variant, which is associated with a predisposition to develop attention deficient hyperactivity disorder, is differentially ubiquitinated compared to the other common receptor variants D-4.2 and D-4.4. Together, our study suggests that GPCR ubiquitination is a complex and variable process

Distinguished Brief

The Government of the Republic of Bretoria and the Kingdom of Pagonia have agreed to submit by Special Agreement the present controversy for final solution to the International Court of Justice pursuant to Article 36, paragraph 1 of the Statute of the International Court of Justice, in relation to Article 40, paragraph 1, of the Statute of the Court

Novel strategy for rapid functional in vivo validation of oncogenic drivers in haematological malignancies

In cancer research, it remains challenging to functionally validate putative novel oncogenic drivers and to establish relevant preclinical models for evaluation of novel therapeutic strategies. Here, we describe an optimized and efficient pipeline for the generation of novel conditional overexpression mouse models in which putative oncogenes, along with an eGFP/Luciferase dual reporter, are expressed from the endogenous ROSA26 (R26) promoter. The efficiency of this approach was demonstrated by the generation and validation of novel R26 knock-in (KI) mice that allow conditional overexpression of Jarid2, Runx2, MN1 and a dominant negative allele of ETV6. As proof of concept, we confirm that MN1 overexpression in the hematopoietic lineage is sufficient to drive myeloid leukemia. In addition, we show that T-cell specific activation of MN1 in combination with loss of Pten increases tumour penetrance and stimulates the formation of Lyl1(+) murine T-cell lymphoblastic leukemias or lymphomas (T-ALL/T-LBL). Finally, we demonstrate that these luciferase-positive murine AML and T-ALL/T-LBL cells are transplantable into immunocompromised mice allowing preclinical evaluation of novel antileukemic drugs in vivo

The effectiveness of non-native fish removal techniques in freshwater ecosystems: a systematic review

In aquatic systems, biological invasions can result in adverse ecological effects. Man-agement techniques available for non-native fish removal programs (including eradica-tion and population size control) vary widely, but include chemicals, harvest regimes, physical removal, or biological control. For management agencies, deciding on what non-native fish removal program to use has been challenging because there is little reliable information about the relative effectiveness of these measures in controlling or eradicat-ing non-native fish. We conducted a systematic review, including a critical appraisal of study validity, to assess the effectiveness of different non-native fish removal methods, and to identify the factors that influence the overall success rate of each type of method. We found 95 relevant studies, generating 158 data sets. The evidence base was dominated by poorly documented studies with inadequate experimental designs (76% of removal projects). When the management goal was non-native fish eradication, chemical treat-ments were relatively successful (antimycin 89%; rotenone 75%) compared to other in-terventions. Electrofishing and passive removal measure studies indicated successful eradication was possible (58% each respectively) but required intensive effort and multi-ple treatments over a number of years. Of these studies with sufficient information, elec-trofishing had the highest success for population size control (56% of data sets). Overall, inadequate data quality and completeness severely limited our ability to make strong con-clusions about the relationships between non-native fish abundance and different methods of eradication and population control, and the factors influencing the overall success rate of each method. Our review highlights that there is considerable scope for improving our evaluations of non-native fish removal methods. It is recommended that programs should have explicitly stated objectives, better data reporting, and study designs that (when pos-sible and appropriate) incorporate replicated and controlled investigations with rigorous, long-term quantitative monitoring. Future research on the effectiveness of non-native fish removal methods should focus on: (1) the efficacy of existing or potentially new removal measures in larger, more complex environments; (2) a broader range of removal measures in general, and (3) phenotypic characteristics of individual fish within a population that fail to be eradicated or controlled

CHARACTERIZATION OF F107 FIMBRIAE OF ESCHERICHIA-COLI 107/86, WHICH CAUSES EDEMA DISEASE IN PIGS, AND NUCLEOTIDE-SEQUENCE OF THE F107 MAJOR FIMBRIAL SUBUNIT GENE, FEDA

F107 fimbriae were isolated and purified from edema disease strain 107/86 of Escherichia coli. Plasmid pIH120 was constructed, which contains the gene cluster that codes for adhesive F107 fimbriae. The major fimbrial subunit gene, fedA, was sequenced. An open reading frame that codes for a protein with 170 amino acids, including a 21-amino-acid signal peptide, was found. The protein without the signal sequence has a calculated molecular mass of 15,099 Da. Construction of a nonsense mutation in the open reading frame of fedA abolished both fimbrial expression and the capacity to adhere to isolated porcine intestinal villi. In a screening of 28 reference edema disease strains and isolates from clinically ill piglets, fedA was detected in 24 cases (85.7%). In 20 (83.3%) of these 24 strains, fedA was found in association with Shiga-like toxin II variant genes, coding for the toxin that is characteristic for edema disease strains of E. coli. The fimbrial subunit gene was not detected in enterotoxigenic E. coli strains. Because of the capacity of E. coli HB101(pIH120) transformants to adhere to isolated porcine intestinal villi, the high prevalence of fedA in edema disease strains, and the high correlation with the Shiga-like toxin II variant toxin-encoding genes, we suggest that F107 fimbriae are an important virulence factor in edema disease strains of E. coli

The ecology of the two-spined blackfish Gadopsis Bispinosus (Pisces: Gadopsidae)

The Gadopsidae is an endemic mono-generic freshwater fish family of south-eastern Australia. The family was thought to be mono-specific, containing the River Blackfish Gadopsis marmoratus, until 1984 when a second species, the Two-spined Blackfish G. bispinosus was described (Sanger 1984). Due its relatively recent description, little information was available on the ecology or conservation status of G. bispinosus in NSW and the ACT. The distribution and abundance of G. bispinosus in southern NSW and the ACT was investigated with the species recorded at 16 of 119 sites surveyed. At most sites where they were recorded, G. bispinosus was abundant, and was invariably found in association with one of the trout species. G. bispinosus was found to be restricted to cool, clear upland streams with rocky, cobble bottoms and relatively intact forest vegetation. It was hypothesised that the presence of the species in the upper Murrumbidgee drainage was due to stream capture, in which the headwater streams from the Murray drainage have been captured by the Murrumbidgee drainage. Interpretation of historical reports of blackfish distribution in NSW indicated that G. bispinosus had suffered declines in some rivers with the species now apparently absent from the Yarrangobilly River and the Murrumbidgee, Naas, and Paddys rivers in the ACT. The decline is thought to have been due to habitat degradation, particularly sediment addition, which has reduced the cover available by filling the interstitial spaces in the cobble substrate favoured by G. bispinosus. The invariable association of G. bispinosus with introduced trout species and habitats with abundant cover, suggests that trout may have played some part in the current distribution of G bispinosus by excluding them from sub-optimal habitats. The movements of G. bispinosus was found to be very restricted with a home range of approximately 15 metres estimated for adult fish. Recapture rates were high, particularly in adult fish, in comparison with other studies of freshwater fish, indicating that adult G. bispinosus are particularly sedentary. The home ranges of G. bispinosus were found to be stable from year to year with fish able to maintain their position in the stream over the high flow periods of winter and spring. On the basis of aquarium observations of pugnacious and aggressive behaviour between adult G. bispinosus, it was considered that these home ranges may be considered territories under the definition of (Gerking 1953) who defined a territory as "any defended area". The diet of G. bispinosus was investigated with distinct seasonal and ontogenetic differences apparent in the diet. Juvenile fish consumed predominantly smaller items such as early instar ephemeropterans and chironomid larvae with some trichopterans present in the diet. The proportion of trichopterans in the diet increased with increasing fish size, with the importance of ephemeropterans and dipterans inversely related to fish size. terrestrial items were not present at all in the stomachs of juvenile fish, were of minor importance to immature fish, but were a major dietary item of adults. Terrestrial items were most abundant in the diet of both immature and adult fish in summer and autumn. The diet of Rainbow Trout 0. mykiss was also examined, with significant dietary overlap apparent between 0. mykiss and G. bispinosus. As with G. bispinosus, the diet of 0. mykiss was dominated by ephemeropterans, trichopterans and terrestrial items. Dietary overlap was greatest between similar size classes of both species, with some seasonal pattern evident. Significant overlap occurred between the non-adult G. bispinosus and juvenile 0. mykiss all seasons except autumn. Overlap was greatest between mature blackfish and non-juvenile trout with significant dietary overlap recorded in all seasons. It was considered that the consistent significant overlap values indicate the dietary competition is likely between these two fish species. The reproductive ecology of G. bispinosus was found to be similar to that recorded for G. marmoratus by Jackson (1978a). Both blackfish species deposit large, yolky, adhesive, demersal eggs in late spring/early summer when water temperatures exceed 16-17 °C. Fecundity in G. bispinosus is low with less than 300 eggs carried by most females. The natural spawning site was not located but is thought to be in the interstitial spaces between cobble and boulders on the river bed. Artificial P.V.C. spawning tubes proved successful with a total of 15 egg masses deposited in them over the course of the study. The numbers of eggs in each egg mass were within the fecundity estimates of individual fish, and all eggs within a mass were at the same stage of development , indicating that a single fish is probably responsible for each egg mass. A large adult male was present with each egg mass. Eggs hatched after approximately 15-17 days, with the embryo emerging from the chorion but the yolk sac remaining inside, effectively tethering the young to the spawning substrate. Parental care by the male continued for approximately 3-4 weeks after hatching by which time the yolk sac was almost fully utilised and young blackfish could swim well