Zymography for Picogram Detection of Lipase and Esterase Activities

Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.     

Non-Toxic Dimeric Peptides Derived from the Bothropstoxin-I Are Potent SARS-CoV-2 and Papain-like Protease Inhibitors

The COVID-19 outbreak has rapidly spread on a global scale, affecting the economy and public health systems throughout the world. In recent years, peptide-based therapeutics have been widely studied and developed to treat infectious diseases, including viral infections. Herein, the antiviral effects of the lysine linked dimer des-Cys¹¹, Lys¹²,Lys¹³-(pBthTX-I)₂K ((pBthTX-I)₂K)) and derivatives against SARS-CoV-2 are reported. The lead peptide (pBthTX-I)₂K and derivatives showed attractive inhibitory activities against SARS-CoV-2 (EC₅₀ = 28–65 µM) and mostly low cytotoxic effect (CC₅₀ > 100 µM). To shed light on the mechanism of action underlying the peptides’ antiviral activity, the Main Protease (M^pro) and Papain-Like protease (PL^pro) inhibitory activities of the peptides were assessed. The synthetic peptides showed PL^pro inhibition potencies (IC₅₀s = 1.0–3.5 µM) and binding affinities (K_d = 0.9–7 µM) at the low micromolar range but poor inhibitory activity against M^pro (IC₅₀ > 10 µM). The modeled binding mode of a representative peptide of the series indicated that the compound blocked the entry of the PL^pro substrate toward the protease catalytic cleft. Our findings indicated that non-toxic dimeric peptides derived from the Bothropstoxin-I have attractive cellular and enzymatic inhibitory activities, thereby suggesting that they are promising prototypes for the discovery and development of new drugs against SARS-CoV-2 infection.     

Labdane Diterpenoids from Salvia tingitana Etl. Synergize with Clindamycin against Methicillin-Resistant Staphylococcus aureus

Quorum-sensing (QS) is a regulatory mechanism in bacterial communication, important for pathogenesis control. The search for small molecules active as quorum-sensing inhibitors (QSI) that can synergize with antibiotics is considered a good strategy to counteract the problem of antibiotic resistance. Here the antimicrobial labdane diterpenoids sclareol (1) and manool (2) extracted from Salvia tingitana were considered as potential QSI against methicillin-resistant Staphylococcus aureus. Only sclareol showed synergistic activity with clindamycin. The quantification of these compounds by LC–MS analysis in the organs and in the calli of S. tingitana showed that sclareol is most abundant in the flower spikes and is produced by calli, while manool is the major labdane of the roots, and is abundant also in the leaves. Other metabolites of the roots were abietane diterpenoids, common in Salvia species, and pentacyclic triterpenoids, bearing a γ-lactone moiety, previously undescribed in Salvia. Docking simulations suggested that 1 and 2 bind to key residues, involved in direct interactions with DNA. They may prevent accessory gene regulator A (AgrA) binding to DNA or AgrA activation upon phosphorylation, to suppress virulence factor expression. The antimicrobial activity of these two compounds probably achieves preventing upregulation of the accessory gene regulator (agr)-regulated genes.     

Understanding the Chloride Affinity of Barbiturates for Anion Receptor Design

Due to their potential binding sites, barbituric acid (BA) and its derivatives have been used in metal coordination chemistry. Yet their abilities to recognize anions remain unexplored. In this work, we were able to identify four structural features of barbiturates that are responsible for a certain anion affinity. The set of coordination interactions can be finely tuned with covalent decorations at the methylene group. DFT-D computations at the BLYP-D3(BJ)/aug-cc-pVDZ level of theory show that the C−H bond is as effective as the N−H bond to coordinate chloride. An analysis of the electron charge density at the C−H⋅⋅⋅Cl⁻ and N−H⋅⋅⋅Cl⁻ bond critical points elucidates their similarities in covalent character. Our results reveal that the special acidity of the C−H bond shows up when the methylene group moves out of the ring plane and it is mainly governed by the orbital interaction energy. The amide and carboxyl groups are the best choices to coordinate the ion when they act together with the C−H bond. We finally show how can we use this information to rationally improve the recognition capability of a small cage-like complex that is able to coordinate NaCl.     

Level structure of 100Tc

Gamma and electron spectra following thermal neutron capture on ⁹⁹Tc have been studied with a bent-crystal spectrometer, Ge(Li) and Si(Li) detectors and a magnetic spectrometer. Prompt and delayed γ-γ coincidences with Ge(Li) detectors have been performed. A level scheme is proposed for ¹⁰⁰Tc comprising 21 excited states up to 640 keV. The binding energy of the last neutron in ¹⁰⁰Tc was deduced. For most levels, spin and parity values were assigned. Two isometric transitions of respective half-lives 10.2 and 4.6 μs have been identified using the ¹⁰⁰Mo(d, 2n)¹⁰⁰Tc reaction with a pulsed beam of deuterons. From the comparison of the present (n, γ) study and the collaborative study of the ⁹⁹Tc(d, p) reaction, several members of the multiplets $\text{π}\text{g}{}_{\mathrm{<mtext>9</mtext><mtext>2</mtext>}}\text{ν}\text{g}{}_{\mathrm{<mtext>7</mtext><mtext>2</mtext>}}$, $\text{π}\text{g}{}_{\mathrm{<mtext>9</mtext><mtext>2</mtext>}}\text{ν}\text{g}{}_{\mathrm{<mtext>5</mtext><mtext>2</mtext>}}$, $\text{π}\text{g}{}_{\mathrm{<mtext>9</mtext><mtext>2</mtext>}}\text{ν}\text{s}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ and $\text{π}\text{p}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{ν}\text{d}{}_{\mathrm{<mtext>5</mtext><mtext>2</mtext>}}$ have been identified.     

Crystalline thiophene. II: a comprehensive study of stable and metastable phases by means of heat capacity,thermally stimulated currents and raman spectroscopy measurements

This calorimetric study of crystalline thiophene between 77 and 250 K reveals the existence of four phase transitions at 174.50, 170.49, 136.8 and 111.26 K, respectively. The transition II-III (170.49 K) is easily avoided during cooling, leading to metastable phases, which, in turn, exhibit their own phase transitions.These findings explain previously confused observations. The existence of stable and metastable phases was confirmed by thermally stimulated current measurements. Intermolecular Raman spectra were observed between 10 and 300 K and assigned to the observed phases. Some predictions are made concerning the low-temperature behaviour of thiophene. Tentative rules are proposed to number metastable phases relative to stable ones.     

Weak measurements are universal

It is well known that any projective measurement can be decomposed into a sequence of weak measurements, which cause only small changes to the state. Similar constructions for generalized measurements, however, have relied on the use of an ancilla system. We show that any generalized measurement can be decomposed into a sequence of weak measurements without the use of an ancilla, and give an explicit construction for these weak measurements. The measurement procedure has the structure of a random walk along a curve in state space, with the measurement ending when one of the end points is reached. This shows that any measurement can be generated by weak measurements, and hence that weak measurements are universal. This may have important applications to the theory of entanglement.     

Halo structure of (14)Be

The two-neutron halo nucleus (14)Be has been investigated in a kinematically complete measurement of the fragments ((12)Be and neutrons) produced in dissociation at 35 MeV/nucleon on C and Pb targets. Two-neutron removal cross sections, neutron angular distributions, and invariant mass spectra were measured, and the contributions from electromagnetic dissociation (EMD) were deduced. Comparison with three-body model calculations suggests that the halo wave function contains a large nu(2s(1/2))(2) admixture. The EMD invariant mass spectrum exhibited enhanced strength near threshold consistent with a nonresonant soft-dipole excitation.     

High-resolution optical correlation imaging in a scattering medium

An optical correlation setup is used to image transparent objects through scattering media, and 10-mum longitudinal and 2.5-mum transverse resolution are achieved. Spectral-bandwidth sampling of the light source is made possible by a tunable dye laser and leads to signal enhancement as a result of sampling interferogram filtering. An optical system allows observation of sample slices without the need for a translation stage.