Miracle-dz/UniFAR-FTUs huggingface dataset:
In vivo regulatory phosphorylation of the phosphoenolpyruvate carboxylase AtPPC1 in phosphate-starved Arabidopsis thaliana

PEPC [PEP(phosphoenolpyruvate) carboxylase] is a tightly controlled cytosolic enzyme situated at a major branchpoint in plant metabolism. Accumulating evidence indicates important functions for PEPC and PPCK (PEPC kinase) in plant acclimation to nutritional Pi deprivation. However, little is known about the genetic origin or phosphorylation status of native PEPCs from −Pi (Pi-deficient) plants. The transfer of Arabidopsis suspension cells or seedlings to −Pi growth media resulted in: (i) the marked transcriptional upregulation of genes encoding the PEPC isoenzyme AtPPC1 (Arabidopsis thaliana PEPC1), and PPCK isoenzymes AtPPCK1 and AtPPCK2; (ii) >2-fold increases in PEPC specific activity and in the amount of an immunoreactive 107-kDa PEPC polypeptide (p107); and (iii) In vivo p107 phosphorylation as revealed by immunoblotting of clarified extracts with phosphosite-specific antibodies to Ser-11 (which could be reversed following Pi resupply). Approx. 1.3 mg of PEPC was purified 660-fold from −Pi suspension cells to apparent homogeneity with a specific activity of 22.3 units · mg−1 of protein. Gel filtration, SDS/PAGE and immunoblotting demonstrated that purified PEPC exists as a 440-kDa homotetramer composed of identical p107 subunits. Sequencing of p107 tryptic and Asp-N peptides by tandem MS established that this PEPC is encoded by AtPPC1. Pi-affinity PAGE coupled with immunoblotting indicated stoichiometric phosphorylation of the p107 subunits of AtPPC1 at its conserved Ser-11 phosphorylation site. Phosphorylation activated AtPPC1 at pH 7.3 by lowering its Km(PEP) and its sensitivity to inhibition by L-malate and L-aspartate, while enhancing activation by glucose 6-phosphate. Our results indicate that the simultaneous induction and In vivo phosphorylation activation of AtPPC1 contribute to the metabolic adaptations of −Pi Arabidopsis.
Identification of Differentiation-Associated Brain-Specific Phosphate Transporter as a Second Vesicular Glutamate Transporter (VGLUT2)

Glutamate is the major excitatory neurotransmitter in mammalian CNS. In the presynaptic nerve terminal, glutamate is stored in synaptic vesicles and released by exocytosis. Previously, it has been shown that a transport protein originally identified as a brain-specific Na(+)-dependent inorganic phosphate transporter I (BNPI) functions as vesicular glutamate transporter and thus has been renamed VGLUT1. Recently, a protein highly homologous to VGLUT1, "differentiation-associated BNPI" (DNPI), has been discovered. Northern blot and in situ hybridization analyses indicate that DNPI mRNA is expressed in some brain regions in which VGLUT1 mRNA is not expressed. We now show that DNPI functions as vesicular glutamate transporter with properties very similar to VGLUT1 and propose to rename the protein VGLUT2. VGLUT2 is highly enriched in synaptic vesicles. Furthermore, VGLUT2 resides on a vesicle population that is distinct from vesicles containing the vesicular GABA transporter or VGLUT1, showing that the expression of VGLUT1 and VGLUT2 do not overlap. When VGLUT2 was expressed in BON cells, membrane fractions displayed ATP-dependent, carbonyl cyanide p-trifluoromethoxyphenylhydrazone-sensitive glutamate uptake. Overexpression of VGLUT2 in cultured autaptic GABAergic neurons yielded postsynaptic currents that were insensitive to the GABA(A) receptor antagonist bicuculline but blocked by the AMPA-receptor antagonist 2,3-dihydroxy-6-nitro-7-sulfonyl-benzo[F]quinoxaline. Thus, expression of VGLUT2 suffices to cause GABAergic neurons to release glutamate in addition to GABA in a manner very similar to that reported previously for VGLUT1.
C‐terminal splice variants of the μ‐opioid receptor: existence, distribution and functional characteristics

The distribution of the mRNA of different C‐terminal splice variants of the μ‐opioid receptor in rat CNS was assessed by RT‐PCR. The mRNA species for MOR1, MOR1A and MOR1B were readily detectable and distributed widely throughout the rat CNS, with levels of MOR1 and MOR1A mRNA being overall greater than for MOR1B. We did not find convincing evidence that significant levels of MOR1C, MOR1C1, MOR1C2 and MOR1D are present in rat CNS. To examine possible differences in the agonist‐induced regulation of MOR1, MOR1A and MOR1B, we expressed these constructs in HEK293 cells along with G‐protein‐coupled inwardly rectifying K+ channel subunits and measured the rate and extent of desensitisation of (d‐Ala2,N‐Me‐Phe4,glycinol5)‐enkephalin (DAMGO)‐ and morphine‐induced G‐protein‐coupled inwardly rectifying K+ currents. Morphine‐induced desensitisation was rapid for all three splice variants (t½: 1.2–1.7 min) but DAMGO‐induced desensitisation was significantly slower for MOR1B (t½ 4.2 min). Inhibition of endocytosis by expression of a dynamin‐dominant negative mutant increased the rate of DAMGO‐induced desensitisation of MOR1B. These data show that some splice variants of μ‐opioid receptor are widely expressed in rat CNS but question the existence of others that have been reported in the literature. In addition, whereas the rate of desensitisation of MOR1 and MOR1A is agonist‐independent, that for MOR1B is agonist‐dependent.
Heterogeneity of competition at decameter scale: patches of high canopy leaf area in a shade-intolerant larch stand transpire less yet are more sensitive to drought.

Small differences in the sensitivity of stomatal conductance to light intensity on leaf surfaces may lead to large differences in total canopy transpiration (EC) with increasing canopy leaf area (L). Typically, the increase of L would more than compensate for the decrease of transpiration per unit of leaf area (EL), resulting in concurrent increase of EC. However, highly shade-intolerant species, such as Larix principis-rupprechtii Mayr. , may be so sensitive to increased shading that such compensation is not complete. We hypothesized that in such a stand, windfall-induced spatial variation at a decameter scale would result in greatly reduced EL in patches of high L leading to lower EC than low competition patches of sparse canopy. We further hypothesized that quicker extraction of soil moisture in patches of lower competition will result in earlier onset of drought symptoms in these patches. Thus, patches of low L will transition from light to soil moisture as the factor dominating EL. This process should progressively homogenize EC in the stand even as the variation of soil moisture is increasing. We tested the hypotheses utilizing sap flux of nine trees, and associated environmental and stand variables. The results were consistent with only some of the expectations. Under non-limiting soil moisture, EL was very sensitive to the spatial variation of L, decreasing sharply with increasing L and associated decrease of mean light intensity on leaf surfaces. Thus, under the conditions of ample soil moisture maximum EC decreased with increasing patch-scale L. Annual EC and biomass production also decreased with L, albeit more weakly. Furthermore, variation of EC among patches decreased as average stand soil moisture declined between rain events. However, contrary to expectation, high L plots which transpired less showed a greater EL sensitivity to decreasing stand-scale soil moisture, suggesting a different mechanism than simple control by decreasing soil moisture. We offer potential explanations to the observed phenomenon. Our results demonstrate that spatial variation of L at decameter scale, even within relatively homogeneous, single-species, even-aged stands, can produce large variation of transpiration, soil moisture and biomass production and should be considered in 1-D soil-plant-atmosphere models.
The Characteristics of Factor Investing

In this article, the authors dissect the realized performance of factor-based equity portfolios using a characteristics-based multifactor return model. They show that generic single-factor portfolios, which invest in stocks with high scores on one particular factor, are suboptimal because they ignore the possibility that these stocks may be unattractive from the perspective of other factors. The authors also show that differences in performance between (1) integrated and mixed-sleeve multifactor portfolios, (2) small-cap and large-cap factor portfolios, and (3) equal and value-weighted factor portfolios can be fully attributed to the differences in their factor characteristics. They conclude that efficient factor investing requires an understanding of how factor characteristics drive portfolio returns.
How does mild hypothermia affect monoclonal antibody glycosylation?

The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. In this study, we investigated the impact of mild hypothermia on protein glycosylation. Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. Experimental results showed higher cell viability with decreased metabolic rates. The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. Expression of the glycosyltransferases that are responsible for N‐linked glycan branching and elongation were significantly lower at 32°C. As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. Biotechnol. Bioeng. 2015;112: 1165–1176. © 2014 Wiley Periodicals, Inc.
The Caspase-8 Dimerization/Dissociation Balance Is a Highly Potent Regulator of Caspase-8, -3, -6 Signaling*

Apoptosis is driven by positive feedback activation between aspartate-specific cysteinyl proteases (caspases). These feedback loops ensure the swift and efficient elimination of cells upon initiation of apoptosis execution. At the same time, the signaling network must be insensitive to erroneous, mild caspase activation to avoid unwanted, excessive cell death. Sublethal caspase activation in fact was shown to be a requirement for the differentiation of multiple cell types but might also occur accidentally during short, transient cellular stress conditions. Here we carried out an in silico comparison of the molecular mechanisms that so far have been identified to impair the amplification of caspase activities via the caspase-8, -3, -6 loop. In a systems model resembling HeLa cervical cancer cells, the dimerization/dissociation balance of caspase-8 potently suppressed the amplification of caspase responses, surprisingly outperforming or matching known caspase-8 and -3 inhibitors such as bifunctional apoptosis repressor or x-linked inhibitor of apoptosis protein. These findings were further substantiated in global sensitivity analyses based on combinations of protein concentrations from the sub- to superphysiological range to screen the full spectrum of biological variability that can be expected within cell populations and between distinct cell types. Additional modeling showed that the combined effects of x-linked inhibitor of apoptosis protein and caspase-8 dimerization/dissociation processes can also provide resistance to larger inputs of active caspases. Our study therefore highlights a central and so far underappreciated role of caspase-8 dimerization/dissociation in avoiding unwanted cell death by lethal amplification of caspase responses via the caspase-8, -3, -6 loop.
Residents’ Experience of Screening, Brief Intervention, and Referral to Treatment (Sbirt) as A Clinical Tool following Practical Application: A Mixed-Methods Study

Background Screening, brief intervention, and referral to treatment (SBIRT), an evidence-based validated system for providing early detection and brief treatment of substance use disorders, has been widely used in the training of medical residents across specialties at a number of sites. This article investigates the effectiveness of SBIRT training during short-term follow-up at Albany Medical Center, one of the initial Substance Abuse and Mental Health Services Administration (SAMHSA) grantees. Methods Training outcomes were measured by training satisfaction following opportunities to apply SBIRT skills in clinical work, the rate at which these techniques were applied in clinical work, and the degree to which residents felt that the SBIRT training provided skills that were applicable to their practice. We examined differences in learning experience by postgraduate year and by program, and conducted a qualitative analysis in a convergent parallel mixed-methods design to elucidate barriers encountered by residents upon using SBIRT techniques in clinical practice. Results Residents remained highly satisfied with the training at 4-month follow-up, with 80.1% reporting that they had used SBIRT skills in their clinical work. Use of SBIRT techniques was high at 6-month follow-up as well, with 85.9% of residents reporting that they regularly screened their patients for substance use, 74.4% reporting that they had applied brief intervention techniques, and 78.2% indicating that SBIRT training had made them overall more effective in helping patients with substance use issues. Differences in application rates and satisfaction were found by specialty. Qualitative analyses indicated that residents encountered patient readiness and specific contextual factors, such as time constraints, externally imposed values, and clinical norms, as barriers to implementation. Conclusions Despite encountering obstacles such as time constraints and patient readiness, residents utilized many of the skills they had learned during SBIRT training in clinical practice and reported finding these skills useful in their management of patients with substance use issues.
The potential for urban surveillance to help support people who are homeless: Evidence from Cairns, Australia

Numerous studies have documented how surveillance practices, such as CCTV, are deployed to support ‘revanchist’ responses to homelessness wherein punitive policing and urban design practices are used to exclude people who are homeless from prime urban areas. However, few studies have considered the capacity of surveillance to facilitate supportive responses to homelessness. In this paper, we explore this supportive capacity through an ethnographic case study of responses to homelessness in the regional Australian city of Cairns. We demonstrate that, whilst surveillance is deployed to police the homeless in Cairns, it is also used to facilitate social services to access and engage with them, for example by using CCTV as a means to coordinate supportive street outreach activities. We conclude from this that there is no necessary relationship between surveillance and punitive/revanchist responses to homelessness, therefore efforts should be made to document and promote its positive uses alongside critiquing its punitive ones.
Reduced utilization and cost of primary care clinic visits resulting from self-care education for patients with osteoarthritis of the knee.

OBJECTIVE To determine the extent to which the cost of an effective self-care intervention for primary care patients with knee osteoarthritis (OA) was offset by savings resulting from reduced utilization of ambulatory medical services. METHODS In an attention-controlled clinical trial, 211 patients with knee OA from the general medicine clinic of a municipal hospital were assigned arbitrarily to conditions of self-care education (group E) or attention control (group AC). Group E (n = 105) received individualized instruction and followup emphasizing nonpharmacologic management of joint pain. Group AC (n = 106) received a standard public education presentation and attention-controlling followup. A comprehensive clinical database provided data concerning utilization and cost of health services during the following year. RESULTS Only 25 subjects (12%) were lost to followup. The 94 subjects remaining in group E made 528 primary care visits during the year following intervention, compared with 616 visits by the 92 patients remaining in group AC (median visits 5 versus 6, respectively; P < 0.05). Fewer visits translated directly into reduced clinic costs in group E, relative to controls (median costs [1996 dollars] $229 versus $305, respectively; P < 0.05). However, self-care education had no significant effects on utilization and costs of outpatient pharmacy, laboratory, or radiology services over the ensuing year. The cost per patient to deliver the self-care intervention was estimated to be $58.70. CONCLUSION Eighty percent of the cost of delivering effective self-care education to the knee OA patients in this study was offset within 1 year by the reduced frequency and costs of primary care visits. For >50% of patients receiving the intervention, the savings associated with fewer primary care visits exceeded the cost of self-care education.
Expression dynamics of the pea rbcS multigene family and organ distribution of the transcripts

We have determined the nucleotide sequence of two members (rbcS‐3A and ‐3C) of the pea nuclear gene family encoding the small subunit (rbcS) of ribulose‐1,5‐bisphosphate carboxylase. Both rbcS‐3A and ‐3C are interrupted by two introns located at the same positions as those of the other three pea rbcS genes. Compared with the other pea rbcS genes the rbcS‐3C gene has the most divergent 5′‐ and 3′‐flanking sequences while the rbcS‐3A gene has a larger and highly divergent intron 1. All five pea rbcS genes are conserved in their coding regions but show considerable sequence differences in their 3′‐untranslated portion. The 3′ sequence divergence of the rbcS genes has allowed us to use S1 nuclease mapping procedures to compare their expression levels in different organs and during light induction. All the rbcS genes are differentially expressed in various organs of the pea plants; moreover, specific rbcS transcripts are under‐represented in seeds and petals. In leaves there is a 10‐fold difference between the highest and lowest specific rbcS transcript levels. By quantitating the distribution of rbcS transcripts during light, phytochrome and blue light induction of immature (etiolated), and mature (green), pea leaves, we show that the genes are differentially activated during leaf development.
An Alternative Way to Make Knowledge Sharing Work in Online Communities? The Effects of Hidden Knowledge Facilitators

ABSTRACT Some firms use hidden knowledge facilitators (HKFs) to facilitate knowledge sharing among employees within intrafirm online communities. These firms hope for enhanced knowledge sharing outcomes within their organizations without letting employees know that HKFs exist. Yet, the extent to which HKFs’ interventions are effective remains unknown to researchers and managers. Built on the knowledge sharing (KS) literature, this study explores the unique roles of HKFs as moderators between a company and its employees. We develop several hypotheses to test the impact of the quantity and quality of HKFs’ online interventions on several KS outcomes. By analyzing log data of a Chinese corporation's online R&D community, we find that (1) the quantity of HKFs’ intervention has a mostly positive impact on KS outcomes; (2) the quality of HKFs’ intervention has a mixed impact on several KS outcomes, depending on which aspect of quantity is considered; and (3) the quality of HKFs’ intervention also moderates the positive impact of the quantity of HKFs’ intervention in different ways on different intended KS outcomes. This study makes a clear contribution to the literature on knowledge sharing and knowledge facilitation by demonstrating the impact of HKFs on KS outcomes in a Chinese context.
Time-domain bie analysis of large electromagnetic scattering problems with Impedance Boundary Condition

Methods for solving the electromagnetic scattering of computationally large problems in a fast and efficient way are well established. However their application is limited to PEC scattering bodies, whilst when applied to lossy or conducting materials they become inefficient and less desirable. In this paper we extend the capabilities of these fast methods to solve for non-dispersive conductors by using the approximation of the Impedance Boundary Condition.
The stability of child abuse reports: a longitudinal study of the reporting behaviour of young adults

Background. The aims of this study were to use longitudinal report data on physical and sexual abuse to examine the stability and consistency of abuse reports. Methods. The study was based on the birth cohort of young people studied in the Christchurch Health and Development Study. At ages 18 and 21 years, these young people were questioned about their childhood exposure to physical punishment and sexual abuse. Concurrent with these assessments, sample members were also assessed on measures of psychiatric disorder and suicidal behaviour. Results. Reports of childhood sexual abuse and physical punishment were relatively unstable and the values of kappa for test–retests of abuse reporting were in the region of 0·45. Inconsistencies in reporting were unrelated to the subject's psychiatric state. Latent class analyses suggested that: (a) those not abused did not falsely report being abused; and (b) those who were abused provided unreliable reports in which the probability of a false negative response was in the region of 50%. Different approaches to classifying subjects as abused led to wide variations in the estimated prevalence of abuse but estimates of the relative risk of psychiatric adjustment problems conditional on abuse exposure remained relatively stable. Conclusions. There was substantial unreliability in the reporting of child abuse. This unreliability arose because those who were subject to abuse often provided false negative reports. The consequences of errors in reports appear to be: (a) that estimates of abuse prevalence based on a single report are likely to seriously underestimate the true prevalence of abuse; while (b) estimates of the relative risk of psychiatric adjustment problems conditional on abuse appear to be robust to the effects of reporting errors.
RETRACTED: Bromhexine-Selective PVC Membrane Electrode Based on Bromhexinium Tetraphenylborate

A bromhexine ion-selective PVC membrane electrode based on bromhexinium tetraphenylborate has been prepared with dioctyl phthalate as plasticizer. The electrode showed a linear response with a slope factor of 57.5 mV/concentration decade at 20°C over the concentration range from 4 x 10 -4 to 10 -1 M bromhexine. The effects of membrane composition, pH of the test solution, and the time of soaking on the electrode performance were studied. The electrode exhibited good selectivity for bromhexine with respect to a large number of inorganic cations and organic substances of biological importance. The standard addition method and potentiometric titrations were used to determine bromhexine concentrations in pure solutions and in a pharmaceutical preparation, with satisfactory results.