Miracle-dz/UniFAR-FTUs huggingface dataset:
Cu(I)-dependent Biogenesis of the Galactose Oxidase Redox Cofactor*

Galactose oxidase is a copper metalloenzyme containing a novel protein-derived redox cofactor in its active site, formed by cross-linking two residues, Cys228 and Tyr272. Previous studies have shown that formation of the tyrosyl-cysteine (Tyr-Cys) cofactor is a self-processing step requiring only copper and dioxygen. We have investigated the biogenesis of cofactor-containing galactose oxidase from pregalactose oxidase lacking the Tyr-Cys cross-link but having a fully processed N-terminal sequence, using both Cu(I) and Cu(II). Mature galactose oxidase forms rapidly following exposure of a pregalactose oxidase-Cu(I) complex to dioxygen (t½ = 3.9satpH7). In contrast, when Cu(II) is used in place of Cu(I) the maturation process requires several hours (t½ = 5.1 h). EDTA prevents reaction of pregalactose oxidase with Cu(II) but does not interfere with the Cu(I)-dependent biogenesis reaction. The yield of cross-link corresponds to the amount of copper added, although a fraction of the pregalactose oxidase protein is unable to undergo this cross-linking reaction. The latter component, which may have an altered conformation, does not interfere with analysis of cofactor biogenesis at low copper loading. The biogenesis product has been quantitatively characterized, and mechanistic studies have been developed for the Cu(I)-dependent reaction, which forms oxidized, mature galactose oxidase and requires two molecules of O2. Transient kinetics studies of the biogenesis reaction have revealed a pH sensitivity that appears to reflect ionization of a protein group (pKa = 7.3) at intermediate pH resulting in a rate acceleration and protonation of an early oxygenated intermediate at lower pH competing with commitment to cofactor formation. These spectroscopic, kinetic, and biochemical results lead to new insights into the biogenesis mechanism.
Differentiation of myeloperoxidase and glandular peroxidase in biological fluids: Application to human saliva

A modification of the assay system for peroxidase activity in human mixed saliva using 2,2′‐azinobis‐(‐ethylbenzthiazoline‐6‐sulphonic acid) (ABTS) as the substrate is reported. The modifications permit differentiation between glandular peroxidase (SPX) and myeloperoxidase (MPX) activity. In addition, factors endogenous to saliva such as thiocyanate did not adversely affect the modified assay system. The basic peroxidase assay utilized 1.0 mM ABTS and 0.1 mM H2O2 in 0.1 M sodium acetate, pH 4.7. The addition of 100 mM NaCl to the assay inhibited 90% of the activity of purified MPX but only 40% of the SPX activity in parotid saliva presumed to be free of MPX. Addition of Cl− to the assay had varied effects on the mixed saliva SPX activity (44–68% inhibition), suggesting that MPX is present in varying amounts in mixed saliva. Assay of mixed saliva from patients with different states of periodontal health indicated that the amount of Cl− inhibition is dependent on amount of inflammation and thus amount of MPX present. The results suggest that the modified assay yields a reliable estimate of relative SPX activity in mixed saliva.
Mutational Analysis of the Hydrolytic Activity of Yeast RNA Polymerase III*

For 25 mutant alleles of ret1, encoding the second largest subunit of yeast RNA polymerase III, we have studied the polymerase III nuclease activity, measuring both the total yield and dinucleotide product composition. Mutations affecting amino acids 309–325 gave slightly elevated nuclease activity. In region 367–376, two mutations gave 12–15-fold increased nuclease activity. Our results do not support the catalytic role in nuclease activity proposed for the conserved DDRD motif in this region (Shirai, T., and Go, M. (1991) Proc. Natl. Acad. Sci. U. S. A. 88, 9056–9060). Mutations centered on a basic region from amino acids 480 to 490, which aligns with Escherichia coli β-subunit sequences between Rifr clusters I and II, produce changes in the relative yields of A- and G-containing dinucleotides. Four such mutant polymerases pause during elongation at GPy sequences and, in addition, have a reduced frequency of termination at T5 terminator sequences. We propose that the side chains of these mutationally altered amino acids are in direct contact with bases in the RNA-DNA hybrid very near the growing 3′-end. Two mutations in domain I near the C terminus produced very large increases in exonuclease activity and strongly increased termination, suggesting that this region also contacts the nascent RNA in the hybrid region.
Understanding Household Garbage Reduction Behavior: A Test of an Integrated Model

The authors examine the antecedents of household garbage reduction behavior in the context of an integrated waste management behavior model. This model incorporates a wide variety of important factors from previous research on environmental behavior into a single theoretical framework provided by the Theory of Planned Behavior. The model was tested using data from a sample of over 1400 individual respondents, each of whom completed a survey and a two-week diary of his or her waste management behaviors. Overall, the results suggest that this model fits the data well and provides significant insight into the factors that influence waste reduction behavior. The authors draw public policy implications from the results.
Occurrence of a Trypanosoma cruzi-like organism in some mammals from southwestern Georgia and northwestern Florida.

Report of the occurrence of American trypanosomiasis (Chagas' disease) in the United States (Woody and Woody, 1955) has alerted workers in the field to the possibility that the disease may occur elsewhere in the southern United States. The etiologic agent of the disease, Trypanosoma cruzi, is known to have been present in this country since 1933 when it was isolated from Triatoma protracta by Kofoid and Donat (Wood and Wood, 1941). Since then, it has been found in other Triatoma spp. and in mammalian hosts as well. The following mammals have been reported as hosts of T. cruzi in the United States: armadillo, opossum, pallid bat, house mouse, 3 species of wood rats (Usinger, 1944), and the raccoon (Walton et al, 1956). In the present study, a T. cruzi-like organism was detected in wild mammals from southwestern Georgia and northwestern Florida while studying the epizootiology of leptospirosis in that area. This paper reports the isolation of the organism from 107 animals representing 4 species.
Changes in Body Composition with Conjugated Linoleic Acid

Conjugated linoleic acid has been shown to reduce body fat accumulation in several animal models. We have conducted several studies in AKR/J mice showing that CLA reduces body fat accumulation whether animals are fed a high-fat or low-fat diet, with no effect on food intake. One mechanism by which CLA reduces body fat is by increased energy expenditure, which is observed within one week of CLA feeding and is sustained for at least six weeks. The increased energy expenditure is sufficient to account for the decreased fat accumulation. Increased uncoupling protein gene expression does not appear to be involved in the increased energy expenditure. We have observed increased fat oxidation but no decrease in de novo fat biosynthesis with CLA feeding. We have also observed increased liver weights and plasma insulin levels with higher doses of CLA. In all of the studies we have conducted to date we have used a CLA preparation that contains several isomers, primarily c9,t11 and t10,c12. It was assumed that the active form was c9,t11, as CLA was identified as an anticarcinogenic compound from cooked beef, of which the c9,t11 form accounts for 60% to 80% of the CLA. Most of the studies conducted so far must be repeated using the purified isomers in order to determine which isomers are responsible for each of the identified actions of CLA.
Socioeconomic disadvantage and periodontal disease: the Dental Atherosclerosis Risk in Communities study.

OBJECTIVES We used data from the Dental Atherosclerosis Risk in Communities study to examine whether individual- and neighborhood-level socioeconomic characteristics were associated with periodontal disease. METHODS We assessed severe periodontitis with a combination of clinical attachment loss and pocket depth measures. Marginal logistic regression modeling was used to estimate the association between individual and neighborhood socioeconomic indicators and prevalence of severe periodontitis before and after control for selected covariates. Residual intraneighborhood correlations in outcomes were taken into account in the analyses. RESULTS Individual-level income and education were associated with severe periodontitis among Whites and African Americans, and these associations remained significant after adjustment for age, gender, recruitment center, and neighborhood socioeconomic score. Low-income Whites residing in disadvantaged neighborhoods had 1.8-fold (95% confidence interval=1.2, 2.7) higher odds of having severe periodontitis than high-income Whites residing in advantaged neighborhoods. CONCLUSIONS Individual income and education were associated with severe periodontitis independently of neighborhood socioeconomic circumstances. Although the association between neighborhood socioeconomic status and severe periodontitis was not statistically significant, poverty and residence in a disadvantaged neighborhood were associated with higher odds of severe periodontitis among Whites.
In Vitro Demonstration Using 19F Magnetic Resonance to Augment Molecular Imaging With Paramagnetic Perfluorocarbon Nanoparticles at 1.5 Tesla

Objectives:This study explored the use of 19F spectroscopy and imaging with targeted perfluorocarbon nanoparticles for the simultaneous identification of multiple biosignatures at 1.5 T. Materials and Methods:Two nanoparticle emulsions with perfluoro-15-crown-5-ether (CE) or perfluorooctylbromide (PFOB) cores were targeted in vitro to fibrin clot phantoms (n = 12) in 4 progressive ratios using biotin–avidin interactions. The CE nanoparticles incorporated gadolinium. Fluorine images were acquired using steady-state gradient-echo techniques; spectra using volume-selective and nonselective sampling. Results:On conventional T1-weighted imaging, clots with CE nanoparticles enhanced as expected, with intensity decreasing monotonically with CE concentration. All clots were visualized using wide bandwidth fluorine imaging, while restricted bandwidth excitation permitted independent imaging of CE or PFOB nanoparticles. Furthermore, 19F imaging and spectroscopy allowed visual and quantitative confirmation of relative perfluorocarbon nanoparticle distributions. Conclusions:19F MRI/S molecular imaging of perfluorocarbon nanoparticles in vitro suggests that noninvasive phenotypic characterization of pathologic biosignatures is feasible at clinical field strengths.
Ganciclovir Pharmacokinetic Parameters Do Not Change When Extending Valganciclovir Cytomegalovirus Prophylaxis From 100 to 200 Days

Background. A 3-month course of prophylaxis is usually recommended for cytomegalovirus (CMV) D+/R− renal transplant recipients. Based on recent data, up to 6 months of prophylaxis may be used. A subanalysis was performed to evaluate the pharmacokinetics of ganciclovir after valganciclovir administration and to perform an exploratory pharmacokinetic/pharmacodynamic analysis. Methods. In Improved Protection Against Cytomegalovirus in Transplant, a phase III, randomized, double blind, placebo-controlled, multicenter study, 318 CMV D+/R− kidney transplant recipients received valganciclovir prophylaxis (900 mg once daily) for 200 or 100 days. A population pharmacokinetic analysis was conducted on a subgroup of patients (n=120). The relationships between ganciclovir exposure (AUC0–24 hr) and clinical outcomes were explored. Results. The final population parameter estimates (95% confidence interval) were as follows: apparent clearance of ganciclovir, 12 L/hr (11.3–12.7 L/hr); volume of distribution, 18.5 L (14.4–22.6 L); and peripheral volume, 44.4 L (40.2–48.6 L). No differences were apparent between the two treatment groups and these estimates. These results are consistent with previously published pharmacokinetic models. There were no direct correlations between the likelihood of developing hematologic adverse events and ganciclovir exposure at the time of the event. The incidence of CMV disease was not correlated with ganciclovir exposure. Conclusion. The pharmacokinetics of ganciclovir were similar between the two dosing groups (100 vs. 200 days), with the majority of patients achieving an area under the concentration time curve in the target therapeutic range (40–60 &mgr;g hr/mL). The fact that the majority of patients were within the target therapeutic range and the absence of a control arm (no treatment) precluded any attempt to validate a correlation with clinical parameters (i.e., CMV disease).
Design and Simulation of a Novel Bipolar Digital Logic Technology for a Balanced Performance in 4H-SiC

High performance and stable operation are desired features of today's integrated circuits (ICs). Due to its inherent properties, devices based on silicon carbide (SiC) exhibit excellent temperature and radiation tolerance, with performance levels similar to or better than the ones in silicon (Si). Instead of specifically creating circuit topologies suited for SiC, current research efforts are focused on adapting existing Si-based technologies to SiC. In this letter, we report a novel bipolar logic technology designed to exploit the electrical characteristics of 4H-SiC and to outperform conventional technologies. It has fewer transistors, simpler topology, and features simple-to-design complex gates. The circuit shows simulated gate delays as low as 3.3 ns at 27 °C/4.2 ns at 500 °C and noise margins higher than 1 V at 27 °C-500 °C. The proposed technology performs remarkably well across all performance parameters, validating its potential for cost effective, smaller footprint, simple to design, and easily reconfigurable digital ICs in SiC for small-scale logic applications.
Psychiatric Disorders in Pregnancy

The care and treatment of women with psychiatric disorders in pregnancy is both challenging and complex. The decision to treat or not treat can pose a risk to the mother, the developing fetus or infant and the pregnancy outcome. Treatment options can be both pharmacologic and non-pharmacologic. The risk and benefit of treatment is a shared decision between the woman and her obstetrical and mental health providers. There is no definitive answer to the optimal treatment of psychiatric disorders in pregnancy. The ideal treatment would minimize the risk to the developing fetus and enable the mother to have few or no psychiatric symptoms during pregnancy.
Dividend catering, life-cycle, and policy: Evidence from Indonesia

Abstract This study examines the behavior of firms in Indonesia in relation to the life-cycle and catering theories under the assumption that investors expect optimum returns on stock investments through dividends, capital gains, or both. To this end, we examine 212 firms listed on the Indonesia Stock Exchange during 2010 to 2016 and investigate dividend policy, our dependent variable, in terms of: (1) dividend payers and non-payers and (2) higher, lower, and non-dividend payers. The independent variables in the basic model of this study are retained earnings-over-total-equity, return-on-assets, market-to-book value, firm size, and dividend premium, and the control variables are systematic and idiosyncratic risks. For hypothesis testing, this study conducts two analyses, namely logistic regression and its extension to multinomial regression. The findings confirm that pseudo R-squared and confidence improve under the dividend policy when controlling for risk and dividend payers. We find that mature Indonesian firms pay higher dividends as they are larger and more profitable, with more free cash and insignificant growth opportunities. Conversely, growing Indonesian firms with significant future opportunities pay lower dividends. The findings of this study imply that the dividend policy of mature Indonesian firms supports the life-cycle theory and is inconsistent with the catering theory.
Nanolocalized single cell membrane nanoelectroporation

Today single cell research is a great interest to analyze cell to cell or cell to environment behavior with their intracellular compounds, where bulk measurement can provide average value. To deliver biomolecules precise and localized way into single living cell with high transfection rate and high cell viability is a challenging and promisible task for biological and therapeutic research. In this report, we present a nano-localized single cell nano-electroporation technique, where electroporation take place in a very precise and localized area on a single cell membrane to achieve high efficient delivery with high cell viability. We fabricated 60nm gap with 40 nm triangular Indium Tin Oxide (ITO) based nano-eletcrode tip, which can intense electric field in a nano-localized area of a single cell to permeabilize cell membrane and deliver exogenous biomolecules from outside to inside of the cell. This device successfully deliver dyes, proteins into single cell with high cell viability (98%). The process not only control precise delivery mechanism into single cell with membrane reversibility, but also it can provide special, temporal and qualitative dosage control, which might be beneficial for therapeutic and biological cell studies.
Distinct roles for Rev1p and Rev7p during translesion synthesis in Saccharomyces cerevisiae

Translesion synthesis (TLS) in Saccharomyces cerevisiae requires at least Rev1p and polymerase ζ (Pol ζ), a complex of the Rev3 polymerase and its accessory factor Rev7p. Although their precise role(s) are poorly characterized, in vitro studies suggest that each protein contributes to TLS in a manner dependent on the particular lesion and surrounding DNA sequence. In the present study, strand segregation analysis is used to attempt to identify the role(s) of the Rev1 and Rev7 proteins during TLS. This assay uses double‐stranded plasmids containing a genetic marker opposite to a replication blocking lesion (N‐2‐acetylaminofluorene; AAF) to measure TLS quantitatively and qualitatively in vivo. The AAF adduct is localized within a repetitive sequence in a manner that allows the formation of misaligned primer–template replication intermediates. Elongation from a misaligned intermediate fixes a frameshift mutation (slipped TLS), while extension of the correctly aligned lesion terminus yields error‐free (non‐slipped) TLS. The results indicate that there is a strong requirement for Rev7p during Pol ζ‐mediated TLS measured in vivo. Furthermore, Rev1p is needed only for non‐slipped TLS; slipped TLS remains efficient in its absence, revealing a previously uncharacterized Rev1p activity similar to Escherichia coli UmuDC function. Specifically, this activity is required for elongation from a correctly aligned lesion terminus.
Phosphorylation of Spc110p by Cdc28p-Clb5p kinase contributes to correct spindle morphogenesis in S. cerevisiae

Spindle morphogenesis is regulated by cyclin-dependent kinases and monitored by checkpoint pathways to accurately coordinate chromosomal segregation with other events in the cell cycle. We have previously dissected the contribution of individual B-type cyclins to spindle morphogenesis in Saccharomyces cerevisiae. We showed that the S-phase cyclin Clb5p is required for coupling spindle assembly and orientation. Loss of Clb5p-dependent kinase abolishes intrinsic asymmetry between the spindle poles resulting in lethal translocation of the spindle into the bud with high penetrance in diploid cells. This phenotype was exploited in a screen for high dosage suppressors that yielded spc110Δ13, encoding a truncation of the spindle pole body component Spc110p (the intranuclear receptor for the γ-tubulin complex). We found that Clb5p-GFP was localised to the spindle poles and intranuclear microtubules and that Clb5p-dependent kinase promoted cell cycle dependent phosphorylation of Spc110p contributing to spindle integrity. Two cyclin-dependent kinase consensus sites were required for this phosphorylation and were critical for the activity of spc110Δ13 as a suppressor. Together, our results point to the function of cyclin-dependent kinase phosphorylation of Spc110p and provide, in addition, support to a model for Clb5p control of spindle polarity at the level of astral microtubule organisation.
Distant HNF1 Site as a Master Control for the Human Class I Alcohol Dehydrogenase Gene Expression*

Gene duplication and divergence have contributed to the biochemical diversity of the alcohol dehydrogenase (ADH) family. Class I ADH is the major enzyme that catalyzes alcohol to acetaldehyde in the liver. To investigate the mechanism(s) controlling tissue-specific and temporal regulation of the three human class I ADH genes (ADH1A, ADH1B, and ADH1C), we compared genomic sequences for the human and mouse ADH loci and analyzed human ADH gene expression in BAC transgenic mice carrying different lengths of the upstream sequences of the class I ADH. A conserved noncoding sequence, located between the class I and class IV ADH (ADH7) genes, was found to be essential for directing class I ADH gene expression in fetal and adult livers. Within this region, a 275-bp fragment displaying liver-specific DNase I hypersensitivity was bound by HNF1. The HNF1-containing upstream sequence enhanced all three class I ADH promoters in an orientation-dependent manner, and the transcriptional activation depended on binding to the HNF1 site. Deletion of the conserved HNF1 site in the BAC led to the shutdown of human class I ADH gene expression in the transgenic livers, leaving ADH1C gene expression in the stomach unchanged. Moreover, interaction between the upstream element and the class I ADH gene promoters was demonstrated by chromosome conformation capture, suggesting a DNA looping mechanism is involved in gene activation. Taken together, our data indicate that HNF1 binding, at ∼51 kb upstream, plays a master role in controlling human class I ADH gene expression and may govern alcohol metabolism in the liver.
Biomimetic Design of Artificial Micro-vasculatures for Tissue Engineering

Over the last decade, highly innovative micro-fabrication techniques have been developed that are set to revolutionise the biomedical industry. Fabrication processes, such as photolithography, wet and dry etching, moulding, embossing and lamination, have been developed for a range of biocompatible and biodegradable polymeric materials. One area where these fabrication techniques could play a significant role is in the development of artificial micro-vasculatures for the creation of tissue samples for drug screening and clinical applications. Despite the enormous technological advances in the field of tissue engineering, one of the major challenges is the creation of miniaturised fluid distribution networks to transport nutrients and waste products, in order to sustain the viability of the culture. In recent years, there has been considerable interest in the development of microfluidic manifolds that mimic the hierarchical vascular and parenchymal networks found in nature. This article provides an overview of microfluidic tissue constructs, and also reviews the hydrodynamic scaling laws that underpin the fluid mechanics of vascular systems. It shows how Murray's law, which governs the optimum ratio between the diameters of the parent and daughter branches in biological networks, can be used to design the microfluidic channels in artificial vasculatures. It is shown that it is possible to introduce precise control over the shear stress or residence time in a hierarchical network, in order to aid cell adhesion and enhance the diffusion of nutrients and waste products. Finally, the paper describes the hydrodynamic extensions that are necessary in order to apply Murray's law to the rectangular channels that are often employed in artificial micro-vasculatures.