Redox-Linked Domain Movements in the Catalytic Cycle of Cytochrome P450 Reductase

SummaryNADPH-cytochrome P450 reductase is a key component of the P450 mono-oxygenase drug-metabolizing system. There is evidence for a conformational equilibrium involving large-scale domain motions in this enzyme. We now show, using small-angle X-ray scattering (SAXS) and small-angle neutron scattering, that delivery of two electrons to cytochrome P450 reductase leads to a shift in this equilibrium from a compact form, similar to the crystal structure, toward an extended form, while coenzyme binding favors the compact form. We present a model for the extended form of the enzyme based on nuclear magnetic resonance and SAXS data. Using the effects of changes in solution conditions and of site-directed mutagenesis, we demonstrate that the conversion to the extended form leads to an enhanced ability to transfer electrons to cytochrome c. This structural evidence shows that domain motion is linked closely to the individual steps of the catalytic cycle of cytochrome P450 reductase, and we propose a mechanism for this

Repeated translocation of a gene cassette drives sex-chromosome turnover in strawberries

Turnovers of sex-determining systems represent important diversifying forces across eukary- otes. Shifts in sex chromosomes—but conservation of the master sex-determining genes— characterize distantly related animal lineages. Yet in plants, in which separate sexes have evolved repeatedly and sex chromosomes are typically homomorphic, we do not know whether such translocations drive sex-chromosome turnovers within closely related taxo- nomic groups. This phenomenon can only be demonstrated by identifying sex-associated nucleotide sequences, still largely unknown in plants. The wild North American octoploid strawberries (Fragaria) exhibit separate sexes (dioecy) with homomorphic, female heteroga- metic (ZW) inheritance, yet sex maps to three different chromosomes in different taxa. To characterize these turnovers, we identified sequences unique to females and assembled their reads into contigs. For most octoploid Fragaria taxa, a short (13 kb) sequence was observed in all females and never in males, implicating it as the sex-determining region (SDR). This female-specific “SDR cassette” contains both a gene with a known role in fruit and pollen production and a novel retrogene absent on Z and autosomal chromosomes. Phy- logenetic comparison of SDR cassettes revealed three clades and a history of repeated translocation. Remarkably, the translocations can be ordered temporally due to the capture of adjacent sequence with each successive move. The accumulation of the “souvenir” sequence—and the resultant expansion of the hemizygous SDR over time—could have been adaptive by locking genes into linkage with sex. Terminal inverted repeats at the inser- tion borders suggest a means of movement. To our knowledge, this is the first plant SDR shown to be translocated, and it suggests a new mechanism (“move-lock-grow”) for expan- sion and diversification of incipient sex chromosomes

Writing Difficulties Faced by Politeknik Kuching Sarawak Commerce Diploma Students in Doing their Assignments

Writing is one of the four main skills in learning and it is important especially in the academic context where students have to write all sorts of texts such as assignments, essays and reports. Often, second language students find that it is difficult to express their thoughts in writing, especially when they have to write in English. Therefore, the purpose of this study was to investigate the writing difficulties faced by the Politeknik Kuching Sarawak Diploma students when writing their assignments. The respondents of this study comprised 132 students who had enrolled in the A4004 English for Commercial Purposes course. Questionnaires were used to collect data from the respondents. The information gathered from the questionnaires was then analyzed using descriptive statistics (frequency count and percentage). The findings showed that the students faced major difficulties in completing writing tasks that required creativity and/or critical thinking. They also faced problems in proofreading and in writing grammatical sentences. It was also found that students often thought in their first language and translated their thoughts into English when writing assignments. Other difficulties encountered by the students included not being able to decide on the correct tenses, and differentiating between British English and American English. Some also encountered difficulties in finding suitable words to say what they meant

Disruption of Smad4 impairs TGF-β/Smad3 and Smad7 transcriptional regulation during renal inflammation and fibrosis in vivo and in vitro

The mechanism by which TGF-β regulates renal inflammation and fibrosis is largely unclear; however, it is well accepted that its biological effects are mediated through Smad2 and Smad3 phosphorylation. Following activation, these Smads form heteromeric complex with Smad4 and translocate into the nucleus to bind and regulate the expression of target genes. Here we studied the roles of Smad4 to regulate TGF-β signaling in a mouse model of unilateral ureteral obstruction using conditional Smad4 knockout mice and in isolated Smad4 mutant macrophages and fibroblasts. Disruption of Smad4 significantly enhanced renal inflammation as evidenced by a greater CD45+ leukocyte and F4/80+ macrophage infiltration and upregulation of IL-1β, TNF-α, MCP-1, and ICAM-1 in the obstructed kidney and in IL-1β-stimulated macrophages. In contrast, deletion of Smad4 inhibited renal fibrosis and TGF-β1-induced collagen I expression by fibroblasts. Further studies showed that the loss of Smad4 repressed Smad7 transcription, leading to a loss of functional protein. This, in turn, inhibited IκBα expression but enhanced NF-κB activation, thereby promoting renal inflammation. Interestingly, deletion of Smad4 influenced Smad3-mediated promoter activities and the binding of Smad3 to the COL1A2 promoter, but not Smad3 phosphorylation and nuclear translocation, thereby inhibiting the fibrotic response. Thus, Smad4 may be a key regulator for the diverse roles of TGF-β1 in inflammation and fibrogenesis by interacting with Smad7 and Smad3 to influence their transcriptional activities in renal inflammation and fibrosis

Charmless Three-Body Baryonic B Decays

Motivated by recent data on B-> p pbar K decay, we study various charmless three-body baryonic B decay modes, including Lambda pbar pi, Sigma0 pbar pi, p pbar pi, p pbar Kbar0, in a factorization approach. These modes have rates of order 10^{-6}. There are two mechanisms for the baryon pair production, current-produced and transition. The behavior of decay spectra from these baryon production mechanisms can be understood by using QCD counting rules. Predictions on rates and decay spectra can be checked in the near future.Comment: 26 pages, 9 figures; version to appear in Phys. Rev.

Association between statins and the risk of suicide attempt, depression, anxiety, and seizure: A population-based, self-controlled case series study

Background: Risk of suicide attempt, depression, anxiety and seizure and the association with statins is an ongoing debate. We aim to investigate the association between statins and the above neuropsychological outcomes, in specific pre- and post-exposure time windows./ Methods: We identified patients aged 40–75 years old who were dispensed a statin between January 1, 2003 and December 31, 2012 from the Hong Kong Clinical Data Analysis & Reporting System (CDARS), an electronic medical records database. Patients with new onset of suicide attempt, depression, anxiety and seizure were derived from the original dataset separately, in a self-controlled case series study design. A non-parametric spline-based self-controlled case series model was built to measure continuous changes of risk./ Results: We identified 396,614 statin users. The risk of each outcome was elevated prior to statin initiation with incidence rate ratios of 1.38 (95 % CI, 1.09–1.74) for suicide attempt, 1.29 (95 % CI, 1.15–1.45) for depression, 1.35 (95 % CI, 1.19–1.53) for anxiety, and 1.45 (95 % CI, 1.21–1.73) for seizure. The incidence rate ratios remained elevated after the initiation of statins during the first 90 and 91–365 days after statin prescription and decreased to the baseline level after 1 year of continuous prescription./ Limitations: CDARS includes prescription data but not adherence data, which could lead to misclassification of exposure periods./ Conclusions: Our study does not support a direct association between statin use and suicide attempt, depression, anxiety and seizure, whose risks could be explained by cardiovascular events, for which statins were prescribed

Alterations in the processing of non-drug-related affective stimuli in abstinent heroin addicts.

Long-term exposure to drug may alter the neural system associated with affective processing, as evidenced by both clinical observations and behavioral data documenting dysfunctions in emotional experiences and processing in drug addicts. Although many imaging studies examined neural responses to drug or drug-related cues in addicts, there have been few studies explicitly designed to reveal their neural abnormalities in processing non-drug-related natural affective materials. The present study asked abstinent heroin addicts and normal controls to passively view standardized affective pictures of positive, negative, or neutral valence and compared their brain activities with functional MRI. Compared to normal controls, addicts showed reduced activation in right amygdala in response to the affective pictures, consistent with previous reports of blunted subjective experience for affective stimuli in addicts. Furthermore, in two visual cortical areas BA 19 and 37, while the controls showed greater responses to positive pictures than to negative ones replicating literature findings, the addicts showed the opposite pattern. The results reveal a complex pattern of altered processing of non-drug-related affective materials in addicts showing both heightened and blunted neural responses in different brain regions and for different stimulus valence. The present study highlights the importance of brain imaging research on drug addicts' processing of affective stimuli in understanding disruptions in their emotion circuitry

Radiative Corrections to Neutrino Mixing and CP Violation in the Minimal Seesaw Model with Leptogenesis

Radiative corrections to neutrino mixing and CP violation are analyzed in the minimal seesaw model with two heavy right-handed neutrinos. We find that textures of the effective Majorana neutrino mass matrix are essentially stable against renormalization effects. Taking account of the Frampton-Glashow-Yanagida ansatz for the Dirac neutrino Yukawa coupling matrix, we calculate the running effects of light neutrino masses, lepton flavor mixing angles and CP-violating phases for both $m_1 =0$ (normal mass hierarchy) and $m_3 =0$ (inverted mass hierarchy) cases in the standard model and in its minimal supersymmetric extension. Very instructive predictions for the cosmological baryon number asymmetry via thermal leptogenesis are also given with the help of low-energy neutrino mixing quantities.Comment: 21 pages, 6 figures; more references adde