The Urea Carboxylase and Allophanate Hydrolase Activities of Urea Amidolyase Are Functionally Independent

Urea amidolyase (UAL) is a multifunctional biotin-dependent enzyme that contributes to both bacterial and fungal pathogenicity by catalyzing the ATP-dependent cleavage of urea into ammonia and CO2. UAL is comprised of two enzymatic components: urea carboxylase (UC) and allophanate hydrolase (AH). These enzyme activities are encoded on separate but proximally related genes in prokaryotes while, in most fungi, they are encoded by a single gene that produces a fusion enzyme on a single polypeptide chain. It is unclear whether the UC and AH activities are connected through substrate channeling or other forms of direct communication. Here, we use multiple biochemical approaches to demonstrate that there is no substrate channeling or interdomain/intersubunit communication between UC and AH. Neither stable nor transient interactions can be detected between prokaryotic UC and AH and the catalytic efficiencies of UC and AH are independent of one another. Furthermore, an artificial fusion of UC and AH does not significantly alter the AH enzyme activity or catalytic efficiency. These results support the surprising functional independence of AH from UC in both the prokaryotic and fungal UAL enzymes and serve as an important reminder that the evolution of multifunctional enzymes through gene fusion events does not always correlate with enhanced catalytic function

Evidence for Quantum Interference in SAMs of Arylethynylene Thiolates in Tunneling Junctions with Eutectic Ga-In (EGaIn) Top-Contacts

This paper compares the current density (J) versus applied bias (V) of self-assembled monolayers (SAMs) of three different ethynylthiophenol-functionalized anthracene derivatives of approximately the same thickness with linear-conjugation (AC), cross-conjugation (AQ), and broken-conjugation (AH) using liquid eutectic Ga-In (EGaIn) supporting a native skin (~1 nm thick) of Ga2O3 as a nondamaging, conformal top-contact. This skin imparts non-Newtonian rheological properties that distinguish EGaIn from other top-contacts; however, it may also have limited the maximum values of J observed for AC. The measured values of J for AH and AQ are not significantly different (J ≈ 10-1 A/cm2 at V = 0.4 V). For AC, however, J is 1 (using log averages) or 2 (using Gaussian fits) orders of magnitude higher than for AH and AQ. These values are in good qualitative agreement with gDFTB calculations on single AC, AQ, and AH molecules chemisorbed between Au contacts that predict currents, I, that are 2 orders of magnitude higher for AC than for AH at 0 < |V| < 0.4 V. The calculations predict a higher value of I for AQ than for AH; however, the magnitude is highly dependent on the position of the Fermi energy, which cannot be calculated precisely. In this sense, the theoretical predictions and experimental conclusions agree that linearly conjugated AC is significantly more conductive than either cross-conjugated AQ or broken conjugate AH and that AQ and AH cannot necessarily be easily differentiated from each other. These observations are ascribed to quantum interference effects. The agreement between the theoretical predictions on single molecules and the measurements on SAMs suggest that molecule-molecule interactions do not play a significant role in the transport properties of AC, AQ, and AH.

Alcohol Abstinence Does Not Fully Reverse Abnormalities of Mucosal-Associated Invariant T Cells in the Blood of Patients With Alcoholic Hepatitis

OBJECTIVES: Alcoholic hepatitis (AH) develops in approximately 30% of chronic heavy drinkers. The immune system of patients with AH is hyperactivated, yet ineffective against infectious diseases. Mucosal-associated invariant T (MAIT) cells are innate-like lymphocytes that are highly enriched in liver, mucosa, and peripheral blood and contribute to antimicrobial immunity. We aimed to determine whether MAIT cells were dysregulated in heavy drinkers with and without AH and the effects of alcohol abstinence on MAIT cell recovery. METHODS: MR1 tetramers loaded with a potent MAIT cell ligand 5-(2-oxopropylideneamino)-6-d-ribitylaminouracil were used in multiparameter flow cytometry to analyze peripheral blood MAIT cells in 59 healthy controls (HC), 56 patients with AH, and 45 heavy drinkers without overt liver disease (HDC) at baseline and 6- and 12-month follow-ups. Multiplex immunoassays were used to quantify plasma levels of cytokines related to MAIT cell activation. Kinetic Turbidimetric Limulus Amebocyte Lysate Assay and ELISA were performed to measure circulating levels of 2 surrogate markers for bacterial translocation (lipopolysaccharide and CD14), respectively. RESULTS: At baseline, patients with AH had a significantly lower frequency of MAIT cells than HDC and HC. HDC also had less MAIT cells than HC (median 0.16% in AH, 0.56% in HDC, and 1.25% in HC). Further, the residual MAIT cells in patients with AH expressed higher levels of activation markers (CD69, CD38, and human leukocyte antigen [HLA]-DR), the effector molecule granzyme B, and the immune exhaustion molecule PD-1. Plasma levels of lipopolysaccharide and CD14 and several cytokines related to MAIT cell activation were elevated in patients with AH (interferon [IFN]-α, interleukin [IL]-7, IL-15, IL-17, IL-18, IL-23, IFN-γ, and tumor necrosis factor α). Decreased MAIT cell frequency and upregulated CD38, CD69, and HLA-DR correlated negatively and positively, respectively, with aspartate aminotransferase level. MAIT cell frequency negatively correlated with IL-18. HLA-DR and CD38 levels correlated with several cytokines. At follow-ups, abstinent patients with AH had increased MAIT cell frequency and decreased MAIT cell activation. However, MAIT cell frequency was not fully normalized in patients with AH (median 0.31%). DISCUSSION: We showed that HDC had a reduction of blood MAIT cells despite showing little evidence of immune activation, whereas patients with AH had a severe depletion of blood MAIT cells and the residual cells were highly activated. Alcohol abstinence partially reversed those abnormalities

Apparent horizons in the quasi-spherical Szekeres models

The notion of an apparent horizon (AH) in a collapsing object can be carried over from the Lema\^{\i}tre -- Tolman (L--T) to the quasispherical Szekeres models in three ways: 1. Literally by the definition -- the AH is the boundary of the region, in which every bundle of null geodesics has negative expansion scalar. 2. As the locus, at which null lines that are as nearly radial as possible are turned toward decreasing areal radius $R$. These lines are in general nongeodesic. The name "absolute apparent horizon" (AAH) is proposed for this locus. 3. As the boundary of a region, where null \textit{geodesics} are turned toward decreasing $R$. The name "light collapse region" (LCR) is proposed for this region (which is 3-dimensional in every space of constant $t$); its boundary coincides with the AAH. The AH and AAH coincide in the L--T models. In the quasispherical Szekeres models, the AH is different from (but not disjoint with) the AAH. Properties of the AAH and LCR are investigated, and the relations between the AAH and the AH are illustrated with diagrams using an explicit example of a Szekeres metric. It turns out that an observer who is already within the AH is, for some time, not yet within the AAH. Nevertheless, no light signal can be sent through the AH from the inside. The analogue of the AAH for massive particles is also considered.Comment: 14 pages, 9 figures, includes little extensions and style corrections made after referee's comments, the text matches the published versio

Shari’ah Board, The Task of Fatwa, and Ijtihad in Islamic Economics, and Finance

The rulings of Mua‟malat in today‟s Islamic Economics, and Finance can be adapted through the process of Ijtihad. While the basic principles or doctrines of the Mua‟malat are given in Shari‟ah, the interpretation of these principles to suit circumstances in different times and places constitutes the Fiqh Mua‟malat. New rulings can be reached by understanding the effective cause (Illah) and rationale (hikmah) of the original ruling and the importance of Maslaha (benefit) under the changed circumstances (Usul Fiqh); which is normally evaluated by the Shari‟ah Board members of the concerned entity. Fatwa issuing via Ijtihad is used to derive laws from the basic principles of Shari‟ah to address the needs of people in different places and times. The important aspect of these new rules is that they may at times change depending on the context of application. Islamic Finance contemporary practices of Ijtihad through various bodies like Islamic Fiqh Academy, have resolved the practice of taqlid (limitation). The doctrine of maqasid al-Shari‟ah establishes Maslahah as an essential element of the ends of law, so that it becomes an important goal in framing new rules (Shari‟ah parameters and guidelines) through Ijtihad. Thus, both the principles set by Shari‟ah and use of Ijtihad to frame new rules has Maslahah or benefit of people as the underlying basis and goal. On the other hand; the standardization of Shari‟ah may become against the fundamental premise of Ijtihad which has existed for centuries and especially in today‟s finance. If rules become standards, and imposed by legal authorities, then Ijtihad cannot be applied towards a critical and dynamic industry like Islamic Finance today. This will eventually damage the very reason that we are able to apply Shari‟ah in all times and places, that is, Ijtihad is the main reason why Shari‟ah is dynamic and is able to be applied in different circumstances. In addition; to standardize Shari‟ah rulings may mean the precedence of one Islamic school of thought over the other, which cannot be universally acceptable. There is no doubt that the synchronization of these two views has to be done through mutual understanding and collaboration between Shari‟ah scholars and various Shari‟ah key board members, market leaders, and regulators. To be very clear and accurate, the question of whether Shari‟ah standards can be harmonized is a matter to be dealt with by Shari‟ah scholars and not market professionals or regulators. The simple reason for this is because Shari‟ah scholars are specialized in their field and whether a Fatwa can be standardized or not is a matter of religious reasoning and should be taken from Shari‟ah own instructions and judgments.Fatwa , fatawa , Ijtihad , Shari’ah Board, Islamic Economics Jurisprudence , GCC , Ijma,

Antimicrobials: a global alliance for optimizing their rational use in intra-abdominal infections (AGORA)

Intra-abdominal infections (IAI) are an important cause of morbidity and are frequently associated with poor prognosis, particularly in high-risk patients. The cornerstones in the management of complicated IAIs are timely effective source control with appropriate antimicrobial therapy. Empiric antimicrobial therapy is important in the management of intra-abdominal infections and must be broad enough to cover all likely organisms because inappropriate initial antimicrobial therapy is associated with poor patient outcomes and the development of bacterial resistance. The overuse of antimicrobials is widely accepted as a major driver of some emerging infections (such as C. difficile), the selection of resistant pathogens in individual patients, and for the continued development of antimicrobial resistance globally. The growing emergence of multi-drug resistant organisms and the limited development of new agents available to counteract them have caused an impending crisis with alarming implications, especially with regards to Gram-negative bacteria. An international task force from 79 different countries has joined this project by sharing a document on the rational use of antimicrobials for patients with IAIs. The project has been termed AGORA (Antimicrobials: A Global Alliance for Optimizing their Rational Use in Intra-Abdominal Infections). The authors hope that AGORA, involving many of the world's leading experts, can actively raise awareness in health workers and can improve prescribing behavior in treating IAIs