Miracle-dz/UniFAR-FTUs huggingface dataset:
Receptor-specific Ubiquitination of β-Arrestin Directs Assembly and Targeting of Seven-transmembrane Receptor Signalosomes*

Angiotensin II type 1a (AT1a), vasopressin V2, and neurokinin 1 (NK1) receptors are seven-transmembrane receptors (7TMRs) that bind and co-internalize with the multifunctional adaptor protein, β-arrestin. These receptors also lead to robust and persistent activation of extracellular-signal regulated kinase 1/2 (ERK1/2) localized on endosomes. Recently, the co-trafficking of receptor-β-arrestin complexes to endosomes was demonstrated to require stable β-arrestin ubiquitination (Shenoy, S. K., and Lefkowitz, R. J. (2003) J. Biol. Chem. 278, 14498–14506). We now report that lysines at positions 11 and 12 in β-arrestin2 are specific and required sites for its AngII-mediated sustained ubiquitination. Thus, upon AngII stimulation the mutant β-arrestin2K11,12R is only transiently ubiquitinated, does not form stable endocytic complexes with the AT1aR, and is impaired in scaffolding-activated ERK1/2. Fusion of a ubiquitin moiety in-frame to β-arrestin2K11,12R restores AngII-mediated trafficking and signaling. Wild type β-arrestin2 and β-arrestin2K11R,K12R-Ub, but not β-arrestin2K11R,K12R, prevent nuclear translocation of pERK. These findings imply that sustained β-arrestin ubiquitination not only directs co-trafficking of receptor-β-arrestin complexes but also orchestrates the targeting of “7TMR signalosomes” to microcompartments within the cell. Surprisingly, binding of β-arrestin2K11R,K12R to V2R and NK1R is indistinguishable from that of wild type β-arrestin2. Moreover, ubiquitination patterns and ERK scaffolding of β-arrestin2K11,12R are unimpaired with respect to V2R stimulation. In contrast, a quintuple lysine mutant (β-arrestin2K18R,K107R,K108R,K207R,K296R) is impaired in endosomal trafficking in response to V2R but not AT1aR stimulation. Our findings delineate a novel regulatory mechanism for 7TMR signaling, dictated by the ubiquitination of β-arrestin on specific lysines that become accessible for modification due to the specific receptor-bound conformational states of β-arrestin2.
Associations between TNFAIP3 gene polymorphisms and systemic lupus erythematosus risk: an updated meta‐analysis

In order to determine whether tumor necrosis factor alpha inducible protein 3 (TNFAIP3) gene polymorphisms confers susceptibility to systemic lupus erythematosus (SLE) in ethnically different populations. A meta‐analysis was conducted to examine the association between TNFAIP3 polymorphisms and susceptibility to SLE. A systematic literature search was conducted to identify all relevant studies. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were used to estimate the strength of the association. A total of 23 studies from 20 articles, involving 18,501 patients with SLE and 30,435 healthy controls were included in this meta‐analysis. Overall, we found significant association between SLE and the TNFAIP3 rs2230926, rs5029937, rs5029939, and rs3757173 polymorphisms (all P < 0.001). Stratification by ethnicity indicated that rs5029939 polymorphism was associated with SLE in Europeans, while rs2230926, rs5029937, and rs3757173 polymorphisms were associated with SLE both in Europeans and Asians (all P < 0.001). The results of our meta‐analysis suggest that TNFAIP3 (rs2230926, rs5029937, rs5029939, and rs3757173) polymorphisms are associated with susceptibility to SLE.
KRYPTON GAS CYLINDERS AS A SOURCE OF RADIATION

A standard 40 foot shipping container with a cargo of pressurized krypton gas in 159 steel cylinders, which had triggered a radiation alarm, was investigated to address radiation safety and illicit nuclear trafficking concerns. The investigation included contamination and dose rate measurements as well as in situ high resolution gamma spectroscopy. The dose rate measurements gave a maximum value of 0.07 &mgr;Sv h−1 above background (0.08 to 0.11 &mgr;Sv h−1) on the cylinder surface and no detectable increase above background at distances of 1 m and higher. Contamination monitor readings showed a similar relative increase (plus 8 cpm) above background (about 12 cpm) to the dose rate readings. Quantitative gamma spectroscopy revealed a contamination of the gas with 85Kr at a level of 3.5 × 105 Bq kg−1. This value was found to be consistent with analytical and numerical estimates based on current data for atmospheric 85Kr, which is captured from ambient air together with stable krypton during the production process. This incident demonstrates an apparent lack of radiation-related knowledge by those who handle krypton gas, as well as by border control personnel and emergency responders. We therefore propose to improve labeling and documentation standards for such shipments. This effort may be facilitated by introducing the new category of “technically enhanced artificial radioactive material,” or “TEARM” (similar to the existing “naturally occurring radioactive material” or “NORM” and “technically enhanced naturally occurring radioactive material” or “TENORM” categories).
Cell Surface Heparan Sulfate Proteoglycans Participate in Factor VIII Catabolism Mediated by Low Density Lipoprotein Receptor-related Protein*

We have demonstrated previously that catabolism of a coagulation factor VIII (fVIII) from its complex with von Willebrand factor (vWf) is mediated by low density lipoprotein receptor-related protein (LRP) (Saenko, E. L., Yakhyaev, A. V., Mikhailenko, I., Strickland, D. K., and Sarafanov, A. G. (1999) J. Biol. Chem. 274, 37685–37692). In the present study, we found that this process is facilitated by cell surface heparan sulfate proteoglycans (HSPGs). This was demonstrated by simultaneous blocking of LRP and HSPGs in model cells, which completely prevented fVIII internalization and degradation from its complex with vWf. In contrast, the selective blocking of either receptor had a lesser effect. In vivo studies of clearance of125I-fVIII-vWf complex in mice also demonstrated that the simultaneous blocking of HSPGs and LRP led to a more significant prolongation of fVIII half-life (5.5-fold) than blocking of LRP alone (3.5-fold). The cell culture and in vivo experiments revealed that HSPGs are also involved in another, LRP-independent pathway of fVIII catabolism. In both pathways, HSPGs act as receptors providing the initial binding of fVIII-vWf complex to cells. We demonstrated that this binding occurs via the A2 domain of fVIII, since A2, but not other portions of fVIII or isolated vWf, strongly inhibited cell surface binding of fVIII-vWf complex, and the affinities of A2 and fVIII-vWf complex for the cells were similar. The A2 site involved in binding to heparin was localized to the region 558–565, based on the ability of the corresponding synthetic peptide to inhibit A2 binding to heparin, used as a model for HSPGs.
Diversity of Bacterial Symbionts in Populations of Sitobion miscanthi (Hemiptera: Aphididae) in China

ABSTRACT Aphids are a group of insects frequently associated with bacterial symbionts. Although Chinese aphids harbor a high level of species diversity, the associations between Chinese aphids and bacterial symbionts are less known. In this study, we uncovered the diversity of bacterial symbionts in a Chinese widespread aphid, Sitobion miscanthi (Takahashi). In this study, we detected the aphid obligate symbiont Buchnera aphidicola, and two secondary symbionts, Hamiltonella defensa and Regiella insecticola, with the diagnostic polymerase chain reaction method in S. miscanthi samples. In addition, symbiotic species of Acinetobacter, Aeromonas, Enterobacter, Pantoea, and Pseudomonas, and the family Enterobacteriaceae were also found. Geographically, sporadic occurrences were detected for H. defensa and R. insecticola. Moreover, the infection rates of them vary widely among the infected populations: H. defensa (5.26–95.2%) and R. insecticola (5.26–46.7%). Phylogenetic analyses indicated that the strain of B. aphidicola mirrored the history and divergence of S. miscanthi; however, the H. defensa and R. insecticola strains were probably experienced horizontal transmission among S. miscanthi and its distantly related species.
On the elucidation of a tumour suppressor role of 3p in lung cancer

Bij longkanker is het nog onduidelijk welke genen of regulerende stukjes DNA (‘regelsequenties’) verantwoordelijk zijn voor het ontstaan van de ziekte. Arja ter Elst richtte zich op de analyse van een gedeelte van chromosoom 3 dat vaak al verdwenen is in het beginstadium van longkankertumoren. Het vinden van genen of regelsequenties is van belang om de ziekte in een vroeg stadium te kunnen opsporen. De promovendus toont aan dat een gedeelte van het chromosoomgebied dat verdwenen is bij longtumoren, juist in staat is om tumoren te onderdrukken. Daarbij wist zij het gebied dat mogelijk tumor-onderdrukkende capaciteiten heeft, tot een kwart in te perken. Tot slot vond Ter Elst aanwijzingen dat genen in het betreffende gebied op een gelijke manier worden gereguleerd.
The clinical significance of cytomegaloviral inclusions in the allograft kidney.

BACKGROUND Renal allografts are a frequent site of subclinical cytomegalovirus (CMV) infection diagnosed by culture, but histologic inclusions occur in less than 1% of biopsies. The natural history of this subgroup of patients has been reported only occasionally, mostly before the availability of ganciclovir therapy. METHODS We analyzed the clinical parameters and pathologic findings in 10 patients with CMV inclusions at allograft biopsy. RESULTS The patients were 29-72 years old, and 9 of 10 (90%) had previous episodes of acute rejection, 3 of whom needed OKT3 administration. Histopathologic examination of the allografts showed interstitial inflammation with tubulitis in 7 of 10 (70%) patients; in 3 of 10 (30%) patients, viral inclusions were present in the glomerular capillary endothelia without any associated inflammatory response. Morphologic criteria for acute transplant glomerulopathy or proliferative glomerulonephritis were not satisfied. Extrarenal viral inclusions were documented in the gastrointestinal tracts of 2 of 10 (20%) patients. The patients were treated with reduced immunosuppression and ganciclovir. Five patients lost their grafts 56.6+/-86.6 days (range, 4-210 days; median, 21 days) after initial diagnosis. The serum creatinine in the remaining five patients was 3.3+/-2.0 mg/dl (range, 1.2-6.5 mg/dl; median, 2.5 mg/dl) 77+/-16 days (range, 56-101 days; median, 77 days) after transplantation. Histopathologic examination showed no residual viral inclusions in 5 of 7 (71.4%) follow-up specimens available for examination. CONCLUSIONS CMV inclusions in renal allograft biopsies typically occur after treatment for rejection. Ganciclovir eradicates replicative virus, but graft outcome is determined by coexisting acute rejection and chronic allograft nephropathy. Graft loss primarily attributable to CMV was not observed.
Mutants of Mucor hiemalis Endo-β-N-acetylglucosaminidase Show Enhanced Transglycosylation and Glycosynthase-like Activities*

Endo-β-N-acetylglucosaminidase from Mucor hiemalis (Endo-M), a family 85 glycoside hydrolase, acts on the β1,4 linkage of N,N′-diacetylchitobiose moiety in the N-linked glycans of glycoproteins and catalyzes not only the hydrolysis reaction but also the transglycosylation reaction that transfers the releasing sugar chain to an acceptor other than water to form a new glycosidic linkage. The transglycosylation activity of Endo-M holds a great promise for the chemo-enzymatic synthesis and glyco-engineering of glycoproteins, but the inherent hydrolytic activity for product hydrolysis and low transglycosylation have hampered its broad applications. This paper describes the site-directed mutagenesis on residues in the putative catalytic region of Endo-M to generate mutants with superior transglycosylation activity. Two interesting mutants were discovered. The Y217F mutant was found to possess much enhanced transglycosylation activity and yet much diminished hydrolytic activity in comparison with the wild-type Endo-M. Kinetic analyses revealed that the Km value of Y217F for an acceptor substrate 4-methylumbelliferyl-β-l-N-acetylglucosaminide was only one-tenth of that of the wild-type, implicating a much higher affinity of Y217F for the acceptor substrate than the wild-type. The other mutant, N175A, acts like a glycosynthase. It was found that mutation at Asn175“knocked out” the hydrolytic activity, but the mutant was able to take the highly active sugar oxazolines (the transition state mimics) as donor substrates for transglycosylation. This is the first glycosynthase derived from endo-β-N-acetylglucosaminidases that proceed via a substrate-assisted mechanism. Our findings provide further insights on the substrate-assisted mechanism of GH85. The usefulness of the novel glycosynthase was exemplified by the efficient synthesis of a human immunodeficiency virus, type 1 (HIV-1) glycopeptide with potent anti-HIV activity.
Diet during pregnancy and levels of maternal pregnancy hormones in relation to the risk of breast cancer in the offspring

Birth weight is positively associated with the risk of breast cancer in the offspring and the underlying process is likely to involve the pregnancy endocrine milieu. We have examined the association of diet and related factors during pregnancy with the levels (at the 16th and 27th gestational week) of maternal pregnancy oestradiol, oestriol, sex hormone-binding globulin (SHBG), progesterone and prolactin, in a cohort of 270 Caucasian women who delivered in a major hospital in Boston, USA. Oestradiol and oestriol were not strongly associated with any of the diet-related variables, but SHBG was significantly and consistently related inversely to pre-pregnancy body mass index and weight gain during pregnancy, and positively to vegetable and pulses intake. Pregnancy progesterone was associated positively with alcohol and inversely with polyunsaturated lipid and vitamin B12 intake, whereas pregnancy prolactin was inversely associated with cereal consumption. If the pregnancy hormones studied are indeed involved in the intra-uterine origin of breast cancer, these findings, if confirmed, would focus dietary advice to pregnant women, with a view to reducing the risk of breast cancer in the offspring, towards avoidance of excess energy intake and an emphasis on plant foods. This advice does not contradict current dietary advice on prudent diet during pregnancy and throughout life.
Targeting Bacterial Cell Wall Peptidoglycan Synthesis by Inhibition of Glycosyltransferase Activity

Synthesis of bacterial cell wall peptidoglycan requires glycosyltransferase enzymes that transfer the disaccharide–peptide from lipid II onto the growing glycan chain. The polymerization of the glycan chain precedes cross‐linking by penicillin‐binding proteins and is essential for growth for key bacterial pathogens. As such, bacterial cell wall glycosyltransferases are an attractive target for antibiotic drug discovery. However, significant challenges to the development of inhibitors for these targets include the development of suitable assays and chemical matter that is suited to the nature of the binding site. We developed glycosyltransferase enzymatic activity and binding assays using the natural products moenomycin and vancomycin as model inhibitors. In addition, we designed a library of disaccharide compounds based on the minimum moenomycin fragment with peptidoglycan glycosyltransferase inhibitory activity and based on a more drug‐like and synthetically versatile disaccharide building block. A subset of these disaccharide compounds bound and inhibited the glycosyltransferase enzymes, and these compounds could serve as chemical entry points for antibiotic development.
Dihydrotestosterone administration does not increase intraprostatic androgen concentrations or alter prostate androgen action in healthy men: a randomized-controlled trial.

CONTEXT Concern exists that androgen treatment might adversely impact prostate health in older men. Dihydrotestosterone (DHT), derived from local conversion of testosterone to DHT by 5α-reductase enzymes, is the principal androgen within the prostate. Exogenous androgens raise serum DHT concentrations, but their effects on the prostate are not clear. OBJECTIVE To determine the impact of large increases in serum DHT concentrations on intraprostatic androgen concentrations and androgen action within the prostate. DESIGN Double-blind, randomized, placebo-controlled. SETTING Single academic medical center. PARTICIPANTS 31 healthy men ages 35-55. INTERVENTION Daily transdermal DHT or placebo gel. MAIN OUTCOME MEASURES Serum and prostate tissue androgen concentrations and prostate epithelial cell gene expression after 4 wk of treatment. RESULTS Twenty-seven men completed all study procedures. Serum DHT levels increased nearly sevenfold, while testosterone levels decreased in men treated with daily transdermal DHT gel but were unchanged in the placebo-treated group (P < 0.01 between groups). In contrast, intraprostatic DHT and testosterone concentrations on d 28 were not different between groups (DHT: placebo = 2.8 ± 0.2 vs. DHT gel = 3.1 ± 0.5 ng/g; T: placebo = 0.6 ± 0.2 vs. DHT gel = 0.4 ± 0.1, mean ± se). Similarly, prostate volume, prostate-specific antigen, epithelial cell proliferation, and androgen-regulated gene expression were not different between groups. CONCLUSIONS Robust supraphysiologic increases in serum DHT, associated with decreased serum T, do not significantly alter intraprostatic levels of DHT, testosterone, or prostate epithelial cell androgen-regulated gene expression in healthy men. Changes in circulating androgen concentrations are not necessarily mimicked within the prostate microenvironment, a finding with implications for understanding the impact of androgen therapies in men.
Id3 Controls Cell Death of 2B4+ Virus-Specific CD8+ T Cells in Chronic Viral Infection

Sustained Ag persistence in chronic infection results in a deregulated CD8+ T cell response that is characterized by T cell exhaustion and cell death of Ag-specific CD8+ T cells. Yet, the underlying transcriptional mechanisms regulating CD8+ T cell exhaustion and cell death are poorly defined. Using the experimental mouse model of lymphocytic choriomeningitis virus infection, we demonstrate that the transcriptional regulator Id3 controls cell death of virus-specific CD8+ T cells in chronic infection. By comparing acute and chronic infection, we showed that Id3− virus-specific CD8+ T cells were less abundant, whereas the absolute numbers of Id3+ virus-specific CD8+ T cells were equal in chronic and acute infection. Phenotypically, Id3− and Id3+ cells most prominently differed with regard to expression of the surface receptor 2B4; although Id3− cells were 2B4+, almost all Id3+ cells lacked expression of 2B4. Lineage-tracing experiments showed that cells initially expressing Id3 differentiated into Id3−2B4+ cells; in turn, these cells were terminally differentiated and highly susceptible to cell death under conditions of persisting Ag. Enforced Id3 expression specifically increased the persistence of 2B4+ virus-specific CD8+ T cells by decreasing susceptibility to Fas/Fas ligand–mediated cell death. Thus, our findings reveal that the transcriptional regulator Id3 promotes the survival of virus-specific CD8+ T cells in chronic infection and suggest that targeting Id3 might be beneficial for preventing cell death of CD8+ T cells in chronic infection or for promoting cell death of uncontrolled, hyperactive CD8+ T cells to prevent immunopathology.
Paracoccidioides brasiliensis: attenuation of yeast cells by gamma irradiation

Paracoccidioides brasiliensis is the agent of paracoccidioidomycosis, the most prevalent mycosis in Latin America, and currently there is no effective vaccine. The aim of this study was to attenuate the yeast form of P. brasiliensis by gamma irradiation for further studies on vaccine research. Paracoccidioides brasiliensis (strain Pb 18) cultures were irradiated at doses between 0.5 and 8.0 kGy. After each dose the viability, reproductive ability and protein metabolism were evaluated. The comparison between the antigenic profile of irradiated and control yeast was made by Western blot and the virulence evaluated by the inoculation in C57Bl/J6 mice. At 6.5 kGy the yeast lost its reproductive capacity. The viability and the incorporation of [L‐35S]‐methionine were the same in control and up to 6.5 kGy irradiated cells, but 6.5 kGy‐irradiated yeast secreted 40% less proteins. The Western blot profile was clearly similar in control and 6.5 kGy‐irradiated yeast. No colony‐forming unit (CFU) could be recovered from the tissues of the mice infected with the radioattenuated yeast. We concluded that for P. brasiliensis yeast it is possible to find a dose in which the pathogen loses its reproductive ability and virulence, while retaining its viability, metabolic activity and the antigenic profile.
Prefrontal D2‐receptor stimulation mediates flexible adaptation of economic preference hierarchies

Advantageous economic decision making requires flexible adaptation of gain‐based and loss‐based preference hierarchies. However, where the neuronal blueprints for economic preference hierarchies are kept and how they may be adapted remains largely unclear. Phasic cortical dopamine release likely mediates flexible adaptation of neuronal representations. In this PET study, cortical‐binding potential (BP) for the D2‐dopamine receptor ligand [11C]FLB 457 was examined in healthy participants during multiple sessions of a probabilistic four‐choice financial decision‐making task with two behavioral variants. In the changing‐gains/constant‐losses variant, the implicit gain‐based preference hierarchy was unceasingly changing, whereas the implicit loss‐based preference hierarchy was constant. In the constant‐gains/changing‐losses variant, it was the other way around. These variants served as paradigms, respectively, contrasting flexible adaptation versus maintenance of loss‐based and gain‐based preference hierarchies. We observed that in comparison with the constant‐gains/changing‐losses variant, the changing‐gains/constant‐losses variant was associated with a decreased D2‐dopamine receptor‐BP in the right lateral frontopolar cortex. In other words, lateral frontopolar D2‐dopamine receptor stimulation was specifically increased during continuous adaptation of mental representations of gain‐based preference hierarchies. This finding provides direct evidence for the existence of a neuronal blueprint of gain‐based decision‐making in the lateral frontopolar cortex and a crucial role of local dopamine in the flexible adaptation of mental concepts of future behavior. Hum Brain Mapp, 2013. © 2011 Wiley Periodicals, Inc.